Volleyball Hires Allison Palmi as Assistant Coach2/25/2025 5:00:00 PM  | Volleyball
<a href="/articles/s41598-024-77236-6/metrics" data-track="click" data-track-action="view metrics" data-track-label="link" rel="nofollow">Metrics details</a>
Insecticidal potential of seven commonly available alkaloids against melon thrips (Thrips palmi Karny) was investigated through in vivo experiments and the bioactivity was explained via in silico approaches
In vivo screening showed highest mortality of T
closely followed by tropinone (41%) after 24 h of incubation
tropinone surpassed reserpine with 83% mortality
indicating its prolonged insecticidal activity
A detailed bioassay of tropinone revealed its LC50 values as 1187.9 and 686.9 µg mL−1 after 24 and 48 h
While studying the molecular interactions between the alkaloids and four physiologically important target proteins of T
tropinone demonstrated the highest ligand efficiency and lowest predicted inhibitory constant
particularly when forming complexes with CathB protein
binding energy calculations of the docked complexes showed most favorable binding of reserpine with CathB
considering both binding energy and ligand efficiency as the evaluation parameters
a molecular dynamics study was carried out
which predicted higher stability of CathB-tropinone complex than CathB-reserpine complex in terms of the total energy of the system
These in silico findings aligned well with the in vivo results
confirming tropinone as a promising candidate for effective thrips management programs
Though the alkaloids selected for the present study can be naturally obtained from different plant species (e.g
atropine and tropinone from Atropa belladonna or deadly nightshade; cinchonine from Cinchona officinalis; noscapine from Papaver somniferum or opium poppy; piperine from Piper nigrum or black pepper; reserpine from Rauvolfia serpentina or Indian snakeroot; strychnine from Strychnos nux-vomica etc.)
their synthetic routes of preparation are well known
the results of the in vivo studies were corroborated with the findings of the computer-assisted in silico simulation techniques to predict the probable site of action of the alkaloids
As the biological activity of a molecule depends on its binding affinity with target proteins
the investigation of ligand-target interaction through in silico docking studies helps to predict the potential target sites of activities without carrying out the in vitro experiments
the interaction of the selected alkaloids with four target proteins specific to thrips physiology was assessed through molecular simulation to explain their differential biological activity
this is the first systematic study of the anti-thrips properties of commonly available alkaloids where the in vivo efficacy has been explained at the molecular level with the help of computational simulations
and tropinone (99%) were purchased from Sigma-Aldrich Chemicals Pvt
and their route of chemical synthesis are well known
Polyethylene glycol (average molecular weight 400 Da) was supplied by SD Fine-Chem Ltd.
Acetamiprid 20% SP formulation was gifted by Insecticide (India) Ltd.
Double distilled water was used for carrying out the studies
Stock solutions of the test alkaloids were prepared by weighing the required quantity of the alkaloids (~ 0.1 g) individually in volumetric flasks of 10 mL capacity
The compounds were dissolved in a PEG 400-water (1:4
v/v) solvent system to get a stock solution (~ 1% strength) of individual alkaloids
A working standard of 1000 µg mL−1 concentration was prepared from the stock solution by dilution with the same solvent system for every alkaloid
where C is the leaf area fed in control and T is the leaf area fed in treatment
A high AFI value indicates the effectiveness of the test alkaloid to inhibit feeding
The most active alkaloid from this screening study was selected for subsequent screening at various concentrations to determine the LC50 values
Tropinone was found to be most effective in the screening study and was chosen further for detailed bioassay. As tropinone is relatively water-soluble (<a href="https://foodb.ca/compounds/FDB031231">https://foodb.ca/compounds/FDB031231</a>)
a 0.5% stock solution of the compound was prepared by dissolving accurately 125 mg of tropinone directly in double distilled water followed by a volume makeup to 25 mL in a volumetric flask of the same capacity
Standard solutions of tropinone of different concentrations (500–2000 µg mL−1) were prepared from the stock solution by serial dilution with double distilled water and used for the bioassay experiment
In silico molecular docking and dynamics simulation studies were carried out to understand the rationale behind the in vivo screening results of the alkaloids against T
The idea of the study was to establish a molecular basis for the results of the bioassay experiments through a virtual platform
Structure-based virtual screening of selected alkaloids was performed using a variety of algorithms
The Research Collaboratory for Structural Bioinformatics Protein Data Bank (RCSB PDB) and the PubChem database were employed as online tools for data retrieval
The amino acid sequences and biological functions of the selected protein targets have been represented in Supplementary Information S1
Every residue in the ensemble had its phi-psi torsion angles displayed on the Ramachandran map
The 3D structures of the selected alkaloids were obtained in SDF (structured data file) format from the PubChem database
Employing Open Babel GUI (graphical user interface)
an open-source chemical toolkit for the interconversion of chemical structures
all atomic coordinates were modified to a .mol2 setup
Universal Force Field was used for the minimization of the internal energy of the ligands
The 3D structures of the seven alkaloids were then organized into a study table in .sdf format for conformation generation in MolSoft
palmi target proteins were imported into MolSoft ICM Pro v 3.8
The atomic coordinates of the target proteins were transformed into MolSoft format by converting them to ICM objects
The grid box was centered on the protein structures for molecular docking
and all other parameters were kept at their default values
After screening the docking results for the docking score
all potential docked conformations for the alkaloids were generated
Only conformations that specifically interact with the targeted proteins’ active-site residues were chosen after being evaluated with Discovery Studio and PyMOL
Discovery Studio was used to investigate detailed non-bonding interactions and their types
and van der Waals contacts produced between the alkaloids and the target proteins
and Kipred is the predicted inhibitory constant
Ligand efficiency (LE) is a commonly applied parameter for selecting favorable ligands by comparing the values of average binding energy per atom
The following formula was applied to calculate LE:
where LE is the ligand efficiency (kcal mol−1 non-H atom−1)
∆G is the binding affinity (kcal mol−1) and N is the number of non-hydrogen atoms in the ligand
an explicit solvation model with pre-equilibrated TIP3P (transferable intermolecular potential with 3 points) water molecules was used
filling a truncated octahedral box with a minimum distance of 10 Å between the box faces and any atom of the protein: ligand complex
and 3 Cl– ions (counter ion concentration of 0.145 M) other than the apo-protein/complex
The complex geometry was minimized in two stages: protein/protein-ligands were first kept fixed and only the position of the water molecules was minimized; afterward
the first step was heating for 100 picoseconds (ps) [time step of 2 femtoseconds (fs)
followed by equilibration at constant volume and temperature (considering periodic boundaries conditions)
and finally a production run by 50 nanoseconds (ns) of MD simulation with NPT (isothermal-isobaric) ensemble
in which Langevin dynamics was used (collision frequency of 1.0 ps−1) to control the temperature at 302.3 K
These simulations were carried out using the Standard Dynamics Cascade module
the bond lengths involving hydrogen atoms were constrained using the SHAKE algorithm
and the equations of motion were integrated with a 2 fs time step using the Verlet leapfrog algorithm
The Particle-Mesh Ewald (PME) method was used to include the long-range interactions
and the non-bonded interactions were truncated with a 10 Å cutoff
The MD trajectory was saved every 2 ps and the MD results (300 conformations) were analyzed with the Analyze Trajectory module of Discovery Studio 4.1
The Solvation Energy protocol was used to calculate the electrostatic contribution to the solvation energy using the DelPhi program in a two-step protocol
The solvation energy was calculated as the difference of the total grid energies between the molecule embedded in the solvent (solvated state) and the molecule surrounded by a reference solvent
A completely randomized design (CRD) was followed for conducting the in vivo experiments with at least three replicates for each treatment
Data were expressed as mean ± SD (standard deviation) values
Statistical analysis was performed using JASP (version 0.17.2.1)
The significance of the differences between variables was tested by one-way ANOVA (analysis of variance)
The p-values less than 0.05 were considered statistically significant and the Tukey’s HSD comparison test was used to compare the mean values
</a>Mortality (A) and average area of leaf fed (B) by T
palmi on cucumber leaf treated with 1000 µg mL−1 concentration of the selected alkaloids and 50 µg mL−1 concentration of acetamiprid [Tukey’s HSD test was performed to flag significant differences among the treatment means of an experiment
Different alphabets above the bars suggested significant differences at 95% confidence limit within a group.]
strychnine showed maximum antifeedancy (89.99 ± 3.97%)
The lowest AFI was observed in the case of reserpine (50.65 ± 3.25%) and tropinone (50.97 ± 4.47%)
the scenario was almost similar with the highest AFI for strychnine (70.48 ± 0.70%)
while those of reserpine and tropinone were lowest (47.25 ± 8.37 and 46.67 ± 4.07%
and piperine exhibited almost statistically similar trends irrespective of duration
feeding was found to be increased with incubation time
The results represent an interesting trend between mortality and antifeedancy for the alkaloids
an inverse relationship between mortality and AFI was observed
palmi mortality was found to be highest and vice versa
AFI values were quite high for acetamiprid (positive control)
which can be correlated to the very high toxicity of the insecticide and a few bites were able to kill the test insects
The data fit well within the 95% confidence limit
the LC50 values for acetamiprid were recorded as 18.9 and 9.6 µg mL−1 after 24 and 48 h
When the tropinone-treated cucumber leaves were carefully observed
it was found that with the increase in tropinone concentration from 500 to 2000 µg mL−1
palmi larvae was gradually decreased (Supplementary Information S9)
As compared to blank (no treatment) and negative control (treated with water only)
the silvery scars created by the feeding of the larvae on tropinone-treated leaves were significantly less
On the leaves treated with the highest concentration of tropinone (2000 µg mL−1)
with the extension of exposure time from 24 h to 48 h
the fed area was found to be increased in all the treatments
</a>Antifeedant index (%) of Thrips palmi at different concentration of tropinone
Seven alkaloids with different chemical structures were screened against four representative receptor proteins of T
and CysP to understand their inhibition potential of the target sites
The dock scores, binding affinities (∆G), ligand efficiencies (LE) and predicted inhibitory constants (pKi) of the screened alkaloid-target complexes are reported in Table <a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/s41598-024-77236-6#Tab2">2</a>
In silico binding affinities (∆G) in the order of favorability were as follows:
strychnine &gt; noscapine &gt; reserpine &gt; piperine &gt; cinchonine &gt; atropine &gt; tropinone (for HSP70); strychnine &gt; piperine &gt; reserpine &gt; cinchonine &gt; noscapine &gt; atropine &gt; tropinone (for EnoPh); strychnine &gt; noscapine &gt; reserpine &gt; piperine &gt; cinchonine &gt; atropine &gt; tropinone (for CathB); strychnine &gt; reserpine &gt; cinchonine &gt; noscapine &gt; piperine &gt; atropine &gt; tropinone (for CysP)
tropinone &gt; piperine &gt; strychnine &gt; cinchonine &gt; atropine &gt; noscapine &gt; reserpine (for HSP70); tropinone &gt; piperine &gt; strychnine &gt; cinchonine &gt; atropine &gt; noscapine &gt; reserpine (for EnoPh); tropinone &gt; piperine &gt; strychnine &gt; atropine &gt; cinchonine &gt; noscapine &gt; reserpine (for CathB); tropinone &gt; strychnine &gt; cinchonine = piperine &gt; atropine &gt; noscapine &gt; reserpine (for CysP)
interesting trends (in the order of decreasing favorability) were observed for the pKi values of the alkaloids-receptor targets complexes:
tropinone &gt; atropine &gt; cinchonine &gt; piperine &gt; reserpine &gt; noscapine &gt; strychnine (for HSP70); tropinone &gt; atropine &gt; noscapine &gt; cinchonine &gt; reserpine &gt; piperine &gt; strychnine (for EnoPh); tropinone &gt; atropine &gt; cinchonine &gt; piperine &gt; reserpine &gt; noscapine &gt; strychnine (for CathB); tropinone &gt; atropine &gt; piperine &gt; noscapine &gt; cinchonine &gt; reserpine &gt; strychnine (for CysP)
On careful perusal of the molecular docking results
it was found that the values of Hbond were the lowest (most favorable) for atropine in the case of HSP70 and CathB complexes
the values were most favorable for noscapine and cinchonine in the case of EnoPh and CysP complexes
the lowest values of Hphob and VwInt were obtained for all the reserpine-protein complexes
but Eintl was lowest for all the tropinone-protein complexes
and acceptors were lowest in the case of all the tropinone-protein complexes
The in silico complexional interactions for the four protein targets
in the order of their quanta/abundance were as follows:
</a>2D representation of binding sites of HSP70 with (a) Atropine
</a>2D representation of binding sites of EnoPh with (a) Atropine
</a>2D representation of binding sites of CathB with (a) Atropine
</a>2D representation of binding sites of CysP with (a) Atropine
the tropinone-protein complexes showed the highest LE and lowest pKi values
the minimum of which was its binding against CathB
The depiction of the hydrophobic surface of the receptor pockets for all four tropinone-protein complexes is shown in Supplementary Information S14
The increasing order of the ease of binding of the ligands to the CathB protein was found to be as: tropinone &gt; strychnine &gt; atropine &gt; cinchonine &gt; noscapine &gt; piperine &gt; reserpine
electrostatic energy (b) and van der Waals energy (c) through molecular dynamic simulation of CathB apoprotein
CathB-reserpine complex and CathB-tropinone complex
Further experiments with tropinone established its potential against melon thrips
The LC50 of 686.9 µg mL−1 at 48 h suggested its activity even at low concentrations
palmi larvae fed more on treated leaves to acquire the toxic amount of the alkaloid
in the leaves treated with higher concentrations of tropinone
just a few bites were enough to get a sufficient amount of toxic compound
there was not much difference between the AFI at 24 h and 48 h at higher concentration as a major percentage of the larvae were killed within 24 h of exposure
the commercial insecticide acetamiprid showed a much higher toxicity than tropinone
no previous study reported the effect of tropane alkaloids
The present study is the first report on the insecticidal properties of tropinone against T
which further established the fact that tropinone might easily be touted as the best putative inhibitor of the studied target proteins
These results endorsed the highest mortality of T
palmi by tropinone in the in vivo screening experiment
Though the favorable binding affinity of strychnine with the target proteins hinted at its potential insecticidal effect
the feeding deterrence perspective made it much less effective in vivo
preferred hydrophobic and van der Waals interaction of the reserpine-protein complexes over the others might be responsible for the insecticidal activity of reserpine despite showing the least LE values
which is just the reverse of the increasing order for the predicted inhibitory constant (pKi)
This clearly showed that the effectiveness of tropinone as a thrips inhibitor lies in the overall greater hydrophilic distribution of the ligand-receptor interface
The present study further emphasized the importance of those parameters while carrying out the in silico studies as they could satisfactorily substantiate the in vivo results
Binding energy calculations of the protein-ligand complexes through MD simulation established those compounds as better agonists than the commercial ones
The present study explored the potential of some commonly available alkaloids against T
palmi through in vivo experiments which was further attempted to rationalize via in silico approaches
The in vivo experiments established tropinone as the best candidate among the tested alkaloids to show insecticidal activity against T
Though its efficacy is not comparable with the proven insecticidal activity of acetamiprid
considering the environmental risks associated with the commercial insecticide
tropinone can be explored further as a safer alternative for pest management
The docking studies of the alkaloids against the selected four target proteins gave a molecular basis for their differential insecticidal activities against T
It was also established in this study that instead of a simple docking score
and binding energy can help in better understanding of the ligand-protein interactions in finding out potential molecules against specific targets
Not only the high average binding energy per atom for the ligand-protein complexes but also the stability of such complexes calculated through the molecular dynamics simulation studies are important in selecting the best putative inhibitor of the target proteins
Though all the alkaloids studied in the experiments can be obtained from natural origin
the synthetic routes of their preparation are well known
The most active compound found in this study i.e
a stable intermediate in the biosynthesis pathway of tropane alkaloids can be synthesized by the famous Robinson’s reaction
in the pure form will not be a limiting factor for developing alkaloid-based pest control products
though the delivery of such compounds safely and effectively to the target is a researchable issue for the future
potential side-effects to non-target organisms
and ecosystem should be studied in detail to establish itself as a safer insecticidal agent
Supervised field trials are also necessary to evaluate the practical effectiveness of the compound in real-time pest management scenarios
The present study is a novel attempt to explore the insecticidal potential of some commonly available alkaloids against Thrips palmi and to justify the bioactivity through in silico studies
Such kind of studies with other biomolecules against different pest species can help to identify potential candidates for effective pest management strategies
More advanced modelling techniques such as flexible docking considering protein dynamics
and biological factors can further improve the predictions of ligand-protein interactions
it is envisaged to open up an avenue to test the potential biomolecules as alternatives to environmentally hazardous synthetic pesticides to carry out pest management activities in a rational manner
The datasets used and/or analyzed during the current study are available from the corresponding authors on reasonable request
A review of the pest status and natural enemies of Thrips palmi
276 (Springer Science &amp; Business Media
&amp; Basavaraj &amp; Genetics of Thrips palmi (Thysanoptera: Thripidae)
Progression of watermelon bud necrosis virus infection in its vector
Thrips: pests of concern to China and the United States
A review of the pest status and control options for Thrips palmi
A decade of a thrips invasion in China: lessons learned
Insecticide resistance management strategies against the western flower thrips
is possibly related to pyrethroid-resistant in Thrips palmi
Monitoring on insecticide resistance of major insect pests in plastic house
Occurrence of spinosad and chlorfenapyr resistance population of Thrips palmi Karny
is conferred by G275E mutation in α6 subunit of nicotinic acetylcholine receptor and cytochrome P450 detoxification
Establishment of discriminating concentration based assessment for insecticide resistance monitoring of palm thrips
Residual contact vial method for the rapid on-site detection of insecticide resistance in Thrips palmi
Botanical insecticides inspired by plant-herbivore chemical interactions
A review of bioinsecticidal activity of Solanaceae alkaloids
Plant-derived alkaloids acting on dengue virus and their vectors: from chemistry to pharmacology
Recent advances for alkaloids as botanical pesticides for use in organic agriculture
Molecular mapping of resistance to thrips in potato
NMR metabolomics of thrips (Frankliniella occidentalis) resistance in Senecio hybrids
The relationship between structurally different pyrrolizidine alkaloids and western flower thrips resistance in F2 hybrids of Jacobaea vulgaris and Jacobaea aquatica
Alkaloid metabolism in thrips-Papaveraceae interaction: recognition and mutual response
Insecticidal tests with oils and alkaloids of larkspur (Delphinium consolida) and stavesacre (Delphinium Staphisagria)
Toxicity of anabasine to the citrus thrips
Differences in effects of pyrrolizidine alkaloids on five generalist insect herbivore species
The effect of structurally related metabolites on insect herbivores: a case study on pyrrolizidine alkaloids and western flower thrips
Phytochemical background mediates effects of pyrrolizidine alkaloids on western flower thrips
Insecticidal effects of matrine against flower thrips
Thrips hawaiiensis Morgan on ‘Cavendish’ banana
Antifeedant discrimination thresholds for two populations of western corn rootworm
Insecticide activity of piperine: toxicity to eggs of Spodoptera frugiperda (Lepidoptera: Noctuidae) and Diatraea Saccharalis (Lepidoptera: Pyralidae) and phytotoxicity on several vegetables
effects of reserpine on reproduction and serotonin immunoreactivity in the stable fly Stomoxys calcitrans (L)
Larvicidal activity of synthetic tropane alkaloids against Ascia monuste orseis (Lepidoptera: Pieridae)
Synthesis of novel 9R/S-acyloxy derivatives of cinchonidine and cinchonine as insecticidal agents
Species of the genus Thrips from India (Thysanoptera)
A photo-based key of thrips (Thysanoptera) associated with horticultural crops in Florida
Preformulation studies on vasicinone – a bronchodilatory alkaloid (study of some physico-chemical aspects)
Evaluation of three bioassay techniques for citrus thrips resistance and correlation of the leaf dip method to field mortality
A method of computing the effectiveness of an insecticide
Antifeedant effects of azadirachtin and structurally related compounds on lepidopterous larvae
genomic organization and expression profiles of four heat shock protein genes in the western flower thrips
Chromosome-level assembly of the melon thrips genome yields insights into evolution of a sap‐sucking lifestyle and pesticide resistance
Effects of cysteine protease inhibitors on oviposition rate of the western flower thrips
Specific cysteine protease inhibitors act as deterrents of western flower thrips
Silicosilico analyses of molecular interactions between groundnut bud necrosis virus and its vector
Recognition of errors in three-dimensional structures of proteins
ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins
More and better reference data for improved all-atom structure validation
&amp; InstaDock A single-click graphical user interface for molecular docking-based virtual high-throughput screening
Molecular docking analysis of rutin reveals possible inhibition of SARS-CoV-2 vital proteins
Ligands binding and molecular simulation: the potential investigation of a biosensor based on an insect odorant binding protein
Molecular docking of nimbolide extracted from leaves of Azadirachta indica with protein targets to confirm the antifungal
γ-Cyclodextrin as inhibitor of the precipitation reaction between berberine and glycyrrhizin in decoctions of natural medicines: interaction studies of cyclodextrins with glycyrrhizin and glycyrrhetic acid by 1H‐NMR spectroscopy and molecular‐dynamics calculation
Influence of dietary alkaloids on survival and growth of Spodoptera Littoralis
The effect of varying alkaloid concentrations on the feeding behavior of gypsy moth larvae
Lymantria dispar (L.) (Lepidoptera: Lymantriidae)
Assessment of insecticidal activity of benzylisoquinoline alkaloids from Chilean Rhamnaceae plants against fruit-fly Drosophila melanogaster and the lepidopteran crop pest Cydia Pomonella
A feeding deterrent for Thrips palmi (Thysanoptera: Thripidae) found in tomato leaves: isolation and identification
Methyl jasmonate increases the tropane alkaloid scopolamine and reduces natural herbivory in Brugmansia suaveolens: is scopolamine responsible for plant resistance
Selection on tropane alkaloids in native and non‐native populations of Datura stramonium
Tropane alkaloids and terpenes synthase genes of Datura stramonium (Solanaceae)
Insecticidal activity of Hyoscyamus niger L
The role of molecular size in ligand efficiency
The role of ligand efficiency metrics in drug discovery
In silico molecular docking of niloticin with acetylcholinesterase 1 (AChE1) of Aedes aegypti L
(Diptera: Culicidae): a promising molecular target
Toxicity of bioactive molecule andrographolide against Spodoptera litura Fab and its binding potential with detoxifying enzyme cytochrome P450
Molecular modeling studies of some phytoligands from Ficus sycomorus fraction as potential inhibitors of cytochrome CYP6P3 enzyme of Anopheles coluzzii
New ecdysone receptor agonists: a computational approach for rational discovery of insecticides for crop protection
<a data-track="click" data-track-action="download citation references" data-track-label="link" rel="nofollow" href="https://citation-needed.springer.com/v2/references/10.1038/s41598-024-77236-6?format=refman&amp;flavour=references">Download references</a>
This work was carried out under the in-house project of the ICAR-Indian Agricultural Research Institute (CRSCIARISIL2014035267)
The first author has performed the work as a part of his post-graduate research program
ICAR-Indian Agricultural Research Institute
ICAR-Indian Institute of Horticultural Research
ICAR-National Institute of Secondary Agriculture
The authors declare no competing interests
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations
Below is the link to the electronic supplementary material
<a data-test="citation-link" data-track="click" data-track-action="download article citation" data-track-label="link" data-track-external="" rel="nofollow" href="https://citation-needed.springer.com/v2/references/10.1038/s41598-024-77236-6?format=refman&amp;flavour=citation">Download citation</a>
DOI: https://doi.org/10.1038/s41598-024-77236-6
Anyone you share the following link with will be able to read this content:
a shareable link is not currently available for this article
Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology
Get the best experience and stay connected to your community with our Spectrum News app. <a href="https://spectrumlocalnews.com/app">Learn More</a>
—&nbsp;When the prep volleyball season begins in the fall
Osceola High School in Kissimmee will return with one of the nation’s top players in rising junior
Her passion for the sport is something that runs in the family
“The game is always something I love to do,” said Palmi
“I’m very excited for the high school season
I think it will be one of our better years yet.”
Palmi comes from a very athletic background
with both parents being college athletes at Jacksonville University
and all my brothers played baseball and all the girls played volleyball in my family,” she said
“We are all very athletic and very competitive.”
That competitive drive helps her on and off the court
“Definitely getting into the gym everyday is one of my main goals and I have been doing that,” said Palmi
“It’s been definitely helping to improve from the 4 day tournaments
One of those 4-day tournaments was held at the Orange County Convention Center in June
Her team placed third in the open division and she was named All-American.&nbsp;
“I was very proud of my team to get to that moment
because I wouldn’t have done it without them,” said Palmi
There is a sense of pride to see the Palmi’s name on the record board
<a href="https://mynews13.com/fl/orlando/sports/2019/09/01/osceola-s-allison-palmi-shatters-county-record-for-volleyball-kills--named-athlete-of-the-week" target="_blank">Her sister, Allison</a>
went on to play at the University of the Incarnate Word in San Antonio
But she isn’t the only family member connected to the program: her mom is the head coach — something she doesn’t take for granted
“It’s something I wouldn’t want to trade ever
it’s definitely something very special,” she said
Palmi is hoping to play at the next level when she is done with her high school career
“I definitely can’t see myself walk away from it all these years
It’s just something I love to do,” she said
Osceola will begin their 2024 season at home against Celebration on Aug
<a href="https://www.youtube.com/channel/UCHJXaNHan3UWwwUKozSB2BQ" target="_blank" rel="noopener">Check out more of CinemaDaily US’ video interviews on our YouTube channel</a>
Icelandic filmmaker Baltasar Kormákur has been making movies both in his home country and in Hollywood for decades
is something that hits a lot closer to home
It’s not just because it begins in Iceland and is based on an Icelandic novel (by Olaf Olafsson)
but also because it’s more like the Icelandic films he made before teaming with Mark Wahlberg for Contraband in 2012
which took his career on a whole different path
We watch this relationship unfold via flashback as the older Kristófer tries to find answers in the present day
Cinema Daily US had a chance to speak with Baltasar and Pálmi over Zoom a few weeks back to learn what got the director interested in doing something different from his American films
and what it was like working with Kôki and some of the other Japanese actors
look for Cinema Daily US’s interview with Kôki very soon.)
Director: <a href="https://www.imdb.com/name/nm0466349/">Baltasar Kormákur</a>
Rating: R (Sexuality|Language|A Scene of Violence)
<a href="https://cinemadailyus.com/author/edward-douglasgmail-com/" target="_blank" rel="noopener">Check out more of Edward Douglas’ articles</a>
The tragic death of actor Christopher Reeve in 2004 was devastating for those who knew and loved him
both in real life and his millions of fans from playing the pivotal role as Superman in four movies from 1978 to 1987
His death was made worse by the fact that nine years earlier
Check out more of CinemaDaily US’ video interviews on our YouTube channel In 2021
Michael Sarnoski directed the Nicolas Cage thriller
and he’s back now with the sci-fi thriller
a prequel to the blockbuster John Krasinski movies from 2018 and 2021
A Quiet Place: Day One stars Oscar winner&#8230;
Ox-Head Village is a 2022 Japan-Japanese language movie
directed by Takashi Shimizu and written by Daisuke Hosaka
Synopsis : In a so-called &#8220;spirit video&#8221;
Kanon (Kôki) is astonished to see a high-school girl who looks exactly like her
Exclusive Interview with Director Takashi Shimizu Q: What kind of place is Tsubono Kosen (Mineral&#8230;
©Courtesy of Sundance Film Festival Nobuhiro Hosoki grew up watching American films since he was a kid; he decided to go to the United States thanks to seeing the artistry of Stanley Kubrick&#8217;s &#8220;A Clockwork Orange.” After graduating from film school
he worked as an assistant director on TV Tokyo’s program called &#8220;Morning Satellite&#8221; at&#8230;
and website in this browser for the next time I comment
&#916;document.getElementById( "ak_js_1" ).setAttribute( "value"
Courtesy of Paramount Pictures Get a behind-the-scenes glimpse into how Mission: Impossible &#8211; The Final Reckoning&#8216;s underwater scenes were filmed by Tom Cruise and film crew members
The upcoming film stars Tom Cruise as Ethan Hunt
©Courtesy of Rolling Pictures Fighting against any obstacle in order to report the news without fear of death or reprisal is an admirable sign of the world&#8217;s most courageous journalists
is a prime example of such an innovative writer
Her story is chronicled in the new biographical movie,&#8230;
©Courtesy of Saban Films and Roadside Attractions Saban Films and Roadside Attractions have released a new trailer for “The Unholy Trinity” which is starring Samuel L
Jackson has returned to the Western arena a decade after starring in Quentin Tarantino&#8217;s &#8220;The Hateful Eight.&#8221; This time
Photo by Nobuhiro Hosoki Nobuhiro Hosoki grew up watching American films since he was a kid; he decided to go to the United States thanks to seeing the artistry of Stanley Kubrick&#8217;s &#8220;A Clockwork Orange.” After graduating from film school
he worked as an assistant director on TV Tokyo’s program called &#8220;Morning Satellite&#8221; at the&#8230;
Y"},"category":false,"taxonomy":{"active":false,"name":"category"}},"markup":{"custom_html":true,"wpp-start":"","wpp-end":"<\/ul>","title-start":"","title-end":"<\/h2>","post-html":"{thumb} {title} {stats}<\/span><\/li>"},"theme":{"name":""}}
If you like our contents, please subscribe our news letter!
Volume 9 - 2022 | <a class="ArticleLayoutHeader__info__doi" href="https://doi.org/10.3389/fmolb.2022.853339">https://doi.org/10.3389/fmolb.2022.853339</a>
This article is part of the Research TopicMethods and Applications in Molecular Diagnostics<a class="Link Link--linkType Link--maincolor Link--medium Link--icon Link--chevronRight Link--right" aria-label="View all 9 articles" href="https://www.frontiersin.org/research-topics/27235/methods-and-applications-in-molecular-diagnostics/magazine" target="_self" data-event="customLink-linkType-a_viewAll9Articles">View all 9 articles</a>
Thrips cause considerable economic losses to a wide range of food
it is often difficult to distinguish thrips species in crops and large consignments by conventional methods
Thysanoptera: Thripidae) is an invasive insect pest of vegetables
and ornamentals besides being vector to several viruses
It poses a threat to domestic and international plant biosecurity and can invade and establish in new areas
we report a polymerase spiral reaction (PSR)-based isothermal assay for rapid
this is the first application of PSR in the identification of any insect species
A primer pair designed based on 3&#x02032;-polymorphism of mtCOIII region can specifically identify T
palmi without any cross-reactivity with predominant thrips species
The assay uses crude lysate of a single thrips saving time and reagents involved in nucleic acid extraction
palmi is visualized by the appearance of bright fluorescence under ultraviolet light or a change in reaction color thus avoiding gel electrophoresis steps
The entire process can be completed in 70&#x000a0;min on-site using only an ordinary water bath
The assay is sensitive to detecting as little as 50 attograms of T
The assay was validated with known thrips specimens and found to be efficient in diagnosing T
The described method will be useful for non-expert personnel to detect an early infestation
restrict the spread of diseases and formulate appropriate management strategies
This is the first experimental demonstration where PSR has been utilized for the detection of insect species
The assay can be executed using only a water bath and the results can be seen with the naked eye
The assay reported here is relatively simple
and easy to use and can be optimized for other invasive insects
The outcome of the study aims to minimize crop losses by early detection
and adopting adequate pest management strategies
palmi were collected from the stock population using a fine Camel hairbrush (Kokuyo Camlin Ltd.
The assay was initially performed using purified DNA and later optimized with crude lysate of thrips
Total genomic DNA was isolated from a single thrips adult using DNeasy Blood and Tissue Kit (Qiagen
the insect was crushed in 180&#x000a0;&#x000b5;l of ATL buffer with a sterile micro-pestle (Dewsil Scientific Pvt
India) and the lysate was incubated at 56&#x000b0;C for 1&#x000a0;h
The incubation was followed by the addition of 200&#x000a0;&#x000b5;l of AL buffer and 200&#x000a0;&#x000b5;l of molecular grade ethanol (Merck
This was followed by washing with 500&#x000a0;&#x000b5;l wash buffers AW1 and AW2
the DNA was eluted in 20&#x000a0;&#x000b5;l of sterile water (Puregene
India) and stored at &#x02212;20&#x000b0;C until further use
The length of the primers was kept between 20&#x02013;23 bp excluding the adaptor sequence
There was no mismatch at the 3&#x02032;-end of the primers and &#x02264; 3 mismatches in the entire primer sequence
The specificity of the primer pairs was confirmed using Primer-BLAST
PSR primer pairs were first validated in a gradient PCR
The 25 &#x003bc;l PCR reaction comprised of 50&#x000a0;ng DNA template
1X DreamTaq buffer (Thermo Fisher Scientific
0.4&#x000a0;&#x000b5;M each forward and reverse primer (Integrated DNA Technologies
260&#x000a0;&#x000b5;M dNTP mix (Thermo Fisher Scientific)
and 2 U DreamTaq DNA Polymerase (Thermo Fisher Scientific)
PCR was carried out in a T100 Thermal Cycler (Bio-Rad
United States) at the following reaction conditions: 94&#x000b0;C for 3&#x000a0;min
35 cycles of 94&#x000b0;C for 30&#x000a0;s
annealing at 60&#x02013;65&#x000b0;C depending upon the primer pairs for 50&#x000a0;s
and a final extension at 72&#x000b0;C for 10&#x000a0;min
PCR products were resolved on 2% agarose gel (Lonza
United States) stained with GoodView (BR Biochem
India) and visualized in a gel documentation system (MaestroGen Inc
Taiwan) with a 1&#x000a0;kb plus DNA ladder (Thermo Fisher Scientific)
The PSR assay was done using a temperature gradient of 60&#x02013;69&#x000b0;C and reaction time of 60&#x02013;90&#x000a0;min
The primer concentration was also optimized
The reaction mixture comprised of 2.5&#x000a0;&#x000b5;l of 10X Thermopol reaction buffer (New England Biolabs
United States) containing 20&#x000a0;mM Tris-HCl
10&#x02013;40&#x000a0;&#x000b5;M each forward and reverse primer
1.4&#x000a0;mM dNTP mix (Thermo Fisher Scientific)
8&#x02013;16 U of Bst DNA polymerase large fragment (New England Biolabs)
and the final volume was adjusted to 25&#x000a0;&#x000b5;l with sterile distilled water
No-template water control (NTC) was used with each run
Restriction digestion of PSR products was done using 5&#x000a0;&#x000b5;l of PSR product
2&#x000a0;&#x000b5;l of NcoI FastDigest enzyme (Thermo Fisher Scientific)
2&#x000a0;&#x000b5;l 10X FastDigest Green Buffer (Thermo Fisher Scientific)
in a final volume of 20&#x000a0;&#x000b5;l for 1&#x000a0;h at 37&#x000b0;C
The digested products were resolved in 2% agarose gel electrophoresis as described above
primer pair AG339F-AG340R was further assessed for cross-reactivity and sensitivity
Potential cross-reactivity of the PSR primers was first assessed in conventional PCR and then in PSR
The primer pair AG339F-AG340R was assessed for cross-reactivity with the other congeneric and predominant thrips vectors viz
MN594552) maintained at Advanced Centre for Plant Virology
PCR was done in a 25&#x000a0;&#x000b5;l reaction mixture as described above with DNA templates from T
PCR amplicons were resolved on 2% agarose gel (Lonza) as described above
The specificity of the primer pairs was further confirmed in PSR
PSR assay was performed in a 25&#x000a0;&#x000b5;l reaction mixture with the DNA templates from T
The amplified products were digested by NcoI FastDigest enzyme
The amplified and digested PSR products were resolved on 2% agarose gel as described above
The sensitivity of the PSR assay using primer pair AG339F-AG340R was determined using a 10-fold serial dilution of template DNA
The initial DNA concentration of 50&#x000a0;ng/&#x003bc;l was serially diluted up to 5&#x000a0;ng/&#x003bc;l &#x000d7; 10&#x02212;8&#x000a0;ng/&#x003bc;l and was used in PSR assays as described above
The amplified PSR products were resolved on 2% agarose
the sensitivity of the PSR assay was compared with PCR using the same dilutions of template DNA
PCR was carried out in a 25&#x000a0;&#x000b5;l reaction mixture as above and products were resolved on 2% agarose gel
DNA extraction steps were eliminated by using a crude lysate of thrips
palmi was collected in a 1.5&#x000a0;ml microcentrifuge tube (Tarsons
India) and 20&#x000a0;&#x000b5;l of sterile water (Genetix Biotech Asia Pvt
The specimen was crushed within the tube with the help of a micro-pestle (Dewsil Scientific Pvt
The tube was placed in a water bath (Jaibro
The lysate so obtained was directly used for amplification in PSR as described above
To simplify the end-point detection and make the PSR assay portable
the gel electrophoresis step was eliminated by using DNA intercalating fluorescent and colorimetric dyes
1&#x000a0;&#x000b5;l of SYBR Green I (Thermo Fischer Scientific) was added to each reaction
To make the visual detection more cost-effective
SYBR Green I was replaced with GoodView (BR Biochem)
1&#x000a0;&#x000b5;l of GoodView was added to each tube after the completion of the reaction
The presence or absence of fluorescence was detected under ultraviolet (UV) light
End-point detection was further simplified using a colorimetric dye
Two micro liter of 3 mM hydroxy naphthol blue (HNB
India) was added at the start of the reaction and mixed well by pipetting
PSR was done by incubation at 65&#x000b0;C for 60&#x000a0;min in a water bath
A change in reaction color indicated the presence of T
The PSR assay was validated using known thrips species
palmi from randomly collected field specimens
Thrips specimens were collected from eggplant
and ridge gourd at experimental fields of IARI
Thrips were collected from both leaves and flowers
Specimens were packed in sealed sample bags and carried to the laboratory
Crude lysate was extracted from single thrips individuals and PSR was done as described above
palmi was confirmed by the GoodView fluorescence using a UV torch and/or change in color while using HNB
MtCOI region of representative field specimens was sequenced to substantiate the specificity of the PSR assay
A homogeneous population developed from a single adult female of T
The quadrangular head had three brick red ocelli in a triangular formation
A pair of setae were located outside this ocellar triangle
The females had sharp ovipositors at the apex of abdomen
whereas the apex of males was round and blunt
Males were slightly smaller than females and faster in their movement
PCR using T. palmi-specific primers for ITS2 region (<a href="#B32">Jangra et al., 2020a</a>) yielded the expected amplicon of 568 bp on an agarose gel (accession number MN194202). Further, the nucleotide sequence of a 660 bp PCR amplified mtCOI product using primer pair LCO1490 and HCO2198 (<a href="#B17">Folmer et al., 1994</a>) showed 100% sequence identity with T
The sequence can be retrieved from NCBI with accession number OK398218
Four pairs of primers viz. AG329F-AG330R, AG341F-AG342R, AG337F-AG338R and AG339F-AG340R were designed based on sequence polymorphism in mtCOI, mtCOIII, and ITS2 regions. The length and GC content of the primer pairs ranged from 43&#x02013;46 nt and 40&#x02013;68.4%, respectively (<a href="#T1">Table 1</a>)
There was no mismatch at the 3&#x02032;-end of the primers
AG329F-AG330R were highly conserved without any mismatch among 44 T
palmi ITS2 sequences used to design the primers
The rest of the primer sequences had &#x02264; 3 mismatches across the entire primer length
The melting temperature of all the primer pairs ranged between 63.6&#x02013;67&#x000b0;C
Primer-BLAST analysis showed a low probability for secondary structure formation
The primer pairs were found to be specific to T
and AG341F-AG342R predicated amplicon sizes of 179
respectively without any cross-reactivity to sequences of other thrips species
In gradient PCR at 60&#x02013;65&#x000b0;C
primer pair AG329F-AG330R yielded a &#x0223c;180 bp amplicon of ITS2 at annealing temperatures of 62&#x02013;65.3&#x000b0;C
Sharp DNA bands of &#x0223c;200 bp were observed in PCR with primer pairs AG337F-AG338R and AG339F-AG340R at all tested annealing temperatures
a distinct band of &#x0223c;175 bp was observed with primer pair
The amplified products were sequenced to confirm the specificity of the PCR reactions
The sequences can be retrieved with the GenBank accession no
provided in the data availability statement
Among the four pairs of primers tested in gradient PCR
the primer pair AG339F-AG340R could only amplify the DNA template of T
The PSR assay was optimized for primer concentration
Primer concentration of 40&#x000a0;&#x000b5;M produced the best amplification and hence was adopted throughout the assay
A reaction temperature range of 60&#x02013;69&#x000b0;C was assessed
the best amplification was observed at 65&#x000b0;C
the concentration of Bst DNA polymerase was standardized at 16 U per reaction
The optimized PSR reaction mixture was comprised of 2.5&#x000a0;&#x000b5;l of 10X Thermopol reaction buffer
40&#x000a0;&#x000b5;M each forward and reverse primer
The incubation time of PSR at 65&#x000b0;C was assessed for 60&#x02013;90&#x000a0;min
we did not observe any significant difference in amplification between 60 and 90&#x000a0;min
hence a reaction time of 60&#x000a0;min was adopted
FP and RP denote forward and reverse primers targeting the mtCOIII region of T
The 3&#x02032; sequence of forward primer is denoted as &#x0201c;F&#x0201d; and that of reverse primer is denoted as &#x0201c;R&#x0201d; and is complementary to the target mtCOIII sequence
An adapter sequence (A) was added at 5&#x02032; of revere primer
The adapter sequence of forward primer (Ar) is reverse to adapter sequence of reverse primer (A)
the double-stranded template DNA unfolds in presence of Betaine
F segment of the forward primer (FP) anneals to the complementary single-strand of DNA (step 1) and extends (step 2)
the R segment of the reverse primer (RP) binds to it (step 3) and extends (step 4)
both the strands melt and form a single chain (step 5)
As the sequences of A and Arc are reverse complementary to each other
The 3&#x02032; end continues to extend and gives spiral amplification (step 7)
mechanism of amplification happens for another single-stranded chain in the right panel
PSR-amplified products using primer pair AG339F-AG340R resolved on 2% agarose gel
lanes 5&#x02013;7: NcoI-digested PSR amplicons
Assessment of cross-reactivity of primer pair AG339F-AG340R in (A)
lanes 2&#x02013;6: PCR with DNA templates of T
Lanes 9&#x02013;13: PSR with DNA templates of T
Lanes 16&#x02013;19: NcoI-digested PSR amplicons from DNA templates of T
(A) Sensitivity of PSR assay using primer pair AG339F-AG340R
palmi DNA was used as a template and products resolved on 2% agarose gel
10&#x02013;15: PSR amplicons of serially-diluted T
palmi template of 5&#x000a0;ng &#x000d7; 10&#x000a0;ng (2)
5&#x000a0;ng &#x000d7; 10&#x02212;1&#x000a0;ng (4)
5&#x000a0;ng &#x000d7; 10&#x02212;2&#x000a0;ng (5)
5&#x000a0;ng &#x000d7; 10&#x02212;3&#x000a0;ng (6)
5&#x000a0;ng &#x000d7; 10&#x02212;4&#x000a0;ng (7)
5&#x000a0;ng &#x000d7; 10&#x02212;5&#x000a0;ng (8)
5&#x000a0;ng &#x000d7; 10&#x02212;6&#x000a0;ng (10)
5&#x000a0;ng &#x000d7; 10&#x02212;7&#x000a0;ng (11)
5&#x000a0;ng &#x000d7; 10&#x02212;8&#x000a0;ng (12)
5&#x000a0;ng &#x000d7; 10&#x02212;9&#x000a0;ng (13)
5&#x000a0;ng &#x000d7; 10&#x02212;10&#x000a0;ng (14)
5&#x000a0;ng &#x000d7; 10&#x02212;11&#x000a0;ng (15)
Sensitivity of PCR assessed using the same primer pair and template
lanes 18&#x02013;23: PCR amplicons of serially diluted T
palmi template of 5&#x000a0;ng &#x000d7; 10&#x000a0;ng (18)
5&#x000a0;ng &#x000d7; 10&#x02212;1&#x000a0;ng (20)
5&#x000a0;ng &#x000d7; 10&#x02212;2&#x000a0;ng (21)
5&#x000a0;ng &#x000d7; 10&#x02212;3&#x000a0;ng (22)
5&#x000a0;ng &#x000d7; 10&#x02212;4&#x000a0;ng (23)
The entire process could be completed in around 70&#x000a0;min
palmi as positive (&#x0002b;ve) and water as negative (-ve) control
Addition of 1&#x000a0;&#x000b5;l of SYBR Green I and GoodView after completion of PSR reaction showed fluorescence under UV light in positive samples
whereas no fluorescence was observed in negative samples
The PSR reaction was mixed with 2&#x000a0;&#x000b5;l of 3&#x000a0;mM HNB prior to amplification and showed a change in color from violet to sky blue in the case of T
while no corresponding color change was observed in negative samples
palmi as indicated by the PSR assay results
MtCOI sequences of random positive and negative specimens (14 samples) further confirmed the specificity of the assay
The sequences can be retrieved from NCBI using GenBank accession no
palmi collected from different crop plants using PSR
Crude lysate of thrips was used in PSR with primer pair AG339F-AG340R
18&#x02013;23: unknown thrips collected from chili (2)
The present study reports an alternative isothermal assay for on-site, rapid identification of thrips species. PSR is a unique amalgamation of isothermal LAMP and conventional PCR (<a href="#F1">Figure 1</a>)
Isothermal assay like LAMP uses more than four primers or a DNA helicase (HDA)
PSR assay described in this study uses only one pair of primers and follows isothermal amplification
Designing PSR primers is also simpler like PCR primers
Ar) of exogenous origin was added on 5&#x002b9; end of the primers (F
The melting temperature of the stuffer sequence (A and/or Ar) was set 5&#x000b0;C lower than the primer sequence (F
R) to ensure binding of primers to the target genes before the formation of &#x0201c;spiral structure&#x0201d; for amplification
the stuffer sequences were derived from an exogenous origin to avoid non-specific reactions
The PSR assay does not require a denaturation step
and the reaction starts as soon as the temperature reaches around 65&#x000b0;C
The sensitivity of the PSR assay is higher than previously reported real-time PCR and LAMP and comparable to that of RPA
The advantages of PSR over RPA are the easy availability of enzymes and reagents and cost-effectiveness
We found that the cost-effectiveness ratio of RPA
PSR utilizes the same Bst polymerase as LAMP but needs only one pair of primers with an exogenous adapter sequence for spiral isothermal amplification
palmi collected from a leaf using a Camel hairbrush and placed in a 1.5&#x000a0;mlmicrocentrifuge tube
Thrips crushed with the help of a micro-pestle in sterile distilled water
Micro-centrifuge tube placed in a floating rack and incubated at 100&#x000b0;C in a water bath for 2&#x000a0;min
PSR mixture prepared in 0.5&#x000a0;ml PCR tubes
PSR reaction incubated in a water bath at 65&#x000b0;C for 60&#x000a0;min
Visual detection of presence or absence of T
Fluorescence was observed in the positive samples while no fluorescence was detected in negative samples
Change in color from violet to sky blue indicates the presence of T
while no color change was observed in negative samples
The use of HNB for visual detection offers advantages over other diagnostics tools as the results can be viewed by the naked eye without opening the tubes
The assay was validated over a large number of known specimens and tested randomly on collected thrips populations from natural vegetation
The availability of several molecular methods will strengthen the on-site
This would be a better alternative to presently available molecular tools for on-site diagnosis
The detection assay can be performed by non-expert personnel at the field level without any sophisticated laboratory equipment
The assay will strengthen the biosecurity infrastructure and decision support system for insect pest management
Rapid and early detection will help prevent outbreaks of T
palmi in endemic areas and restrict its introduction or spread into new areas
Similar assays can also be developed for the diagnosis of other insect species of interest
The datasets generated and/or analyzed during the current study are available in the NCBI database and can be accessed using the accession numbers MN194202
and VB suggested improvements in the methods
All authors read and approved the manuscript
The study was supported by funding from DBT (BT/PR26136/AGIII/103/1005/2018)
The funding agency played no role in the design of the study
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations
Any product that may be evaluated in this article
or claim that may be made by its manufacturer
is not guaranteed or endorsed by the publisher
The authors acknowledge the support received from IARI
New Delhi and the Queensland Alliance for Agriculture and Food Innovation
Exploring the Use of High-Resolution Melting Analysis and Helicase-dependent Amplification for Discrimination of Bemisia Tabaci (Hemiptera: Aleyrodidae) Cryptic Species and Trialeurodes Vaporariorum
<a href="https://pubmed.ncbi.nlm.nih.gov/32841358/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1093/jee/toaa180">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Exploring+the+Use+of+High-Resolution+Melting+Analysis+and+Helicase-dependent+Amplification+for+Discrimination+of+Bemisia+Tabaci+(Hemiptera:+Aleyrodidae)+Cryptic+Species+and+Trialeurodes+Vaporariorum&amp;btnG=">Google Scholar</a>
Thrips on Vegetable Crops in the Philippines
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Thrips+on+Vegetable+Crops+in+the+Philippines&amp;btnG=">Google Scholar</a>
Species of the Genus Thrips from India (Thysanoptera)
<a href="https://doi.org/10.1111/j.1365-3113.1980.tb00404.x">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Species+of+the+Genus+Thrips+from+India+(Thysanoptera)&amp;btnG=">Google Scholar</a>
From Laboratory to point of Entry: Development and Implementation of a Loop-Mediated Isothermal Amplification (LAMP)-based Genetic Identification System to Prevent Introduction of Quarantine Insect Species
<a href="https://pubmed.ncbi.nlm.nih.gov/29363271/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1002/ps.4866">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=From+Laboratory+to+point+of+Entry:+Development+and+Implementation+of+a+Loop-Mediated+Isothermal+Amplification+(LAMP)-based+Genetic+Identification+System+to+Prevent+Introduction+of+Quarantine+Insect+Species&amp;btnG=">Google Scholar</a>
nouveaux d&#x000e9;pr&#x000e9;dateurs du cotonnier en C&#x000f4;te d&#x02019;Ivoire
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Deux+Thysanoptères,+nouveaux+déprédateurs+du+cotonnier+en+Côte+d’Ivoire&amp;btnG=">Google Scholar</a>
On the Geographical Distribution of Thrips Palmi Karny
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=On+the+Geographical+Distribution+of+Thrips+Palmi+Karny&amp;btnG=">Google Scholar</a>
Population Analysis Reveals Genetic Structure of an Invasive Agricultural Thrips Pest Related to Invasion of Greenhouses and Suitable Climatic Space
<a href="https://pubmed.ncbi.nlm.nih.gov/31700532/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1111/eva.12847">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Population+Analysis+Reveals+Genetic+Structure+of+an+Invasive+Agricultural+Thrips+Pest+Related+to+Invasion+of+Greenhouses+and+Suitable+Climatic+Space&amp;btnG=">Google Scholar</a>
A Photo-Based Key of Thrips (Thysanoptera) Associated with Horticultural Crops in Florida
<a href="https://doi.org/10.1653/024.100.0208">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=A+Photo-Based+Key+of+Thrips+(Thysanoptera)+Associated+with+Horticultural+Crops+in+Florida&amp;btnG=">Google Scholar</a>
Infestation of the Southern Yellow Thrips Thrips Palmi
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Infestation+of+the+Southern+Yellow+Thrips+Thrips+Palmi,+in+Vegetables&amp;btnG=">Google Scholar</a>
Phylogenetics of Social Behavior in Australian Gall-Forming Thrips: Evidence From Mitochondrial DNA Sequence
<a href="https://pubmed.ncbi.nlm.nih.gov/9479705/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1006/mpev.1997.0449">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Phylogenetics+of+Social+Behavior+in+Australian+Gall-Forming+Thrips:+Evidence+From+Mitochondrial+DNA+Sequence,+Adult+Morphology+and+Behavior,+and+Gall+Morphology.&amp;btnG=">Google Scholar</a>
and Vector Competence of Thrips Palmi (Thysanoptera: Thripidae)
<a href="https://doi.org/10.1007/s12600-021-00893-0">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Host+Plant+Influences+Life+Cycle,+Reproduction,+Feeding,+and+Vector+Competence+of+Thrips+Palmi+(Thysanoptera:+Thripidae),+a+Vector+of+Tospoviruses&amp;btnG=">Google Scholar</a>
<a name="B15" id="B15"></a>EPPO (2021). EPPO Global Database. AvaliableAt: <a href="https://www.cabi.org/isc/abstract/20210216039">https://www.cabi.org/isc/abstract/20210216039</a> (accessed on 11 23
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=EPPO+Global+Database&amp;btnG=">Google Scholar</a>
Genetic Variation of Thrips Populations Assessed by Amplified Fragment Length Polymorphism (Thysanoptera: Thripidae)
doi:10.1603/0013-8746(2005)098[0351:gvotpa]2.0.co;2
<a href="https://doi.org/10.1603/0013-8746(2005)098%5B0351:gvotpa%5D2.0.co;2">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Genetic+Variation+of+Thrips+Populations+Assessed+by+Amplified+Fragment+Length+Polymorphism+(Thysanoptera:+Thripidae)&amp;btnG=">Google Scholar</a>
DNA Primers for Amplification of Mitochondrial Cytochrome C Oxidase Subunit I from Diverse Metazoan Invertebrates
<a href="https://pubmed.ncbi.nlm.nih.gov/7881515/">PubMed Abstract</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=DNA+Primers+for+Amplification+of+Mitochondrial+Cytochrome+C+Oxidase+Subunit+I+from+Diverse+Metazoan+Invertebrates&amp;btnG=">Google Scholar</a>
&#x0201c;Taq DNA Polymerase,&#x0201d; in PCR Technology: Principles and Applications for DNA Amplifcation
<a href="https://doi.org/10.1007/978-1-349-20235-5_2">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Taq+DNA+Polymerase&amp;btnG=">Google Scholar</a>
Exposure to Watermelon Bud Necrosis Virus and Groundnut Bud Necrosis Virus Alters the Life History Traits of Their Vector
<a href="https://pubmed.ncbi.nlm.nih.gov/31440810/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1007/s00705-019-04381-z">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Exposure+to+Watermelon+Bud+Necrosis+Virus+and+Groundnut+Bud+Necrosis+Virus+Alters+the+Life+History+Traits+of+Their+Vector,+Thrips+Palmi+(Thysanoptera:+Thripidae)&amp;btnG=">Google Scholar</a>
&#x0201c;Thrips as the Vectors of Tospoviruses in Indian Agriculture,&#x0201d; in A century of Plant Virology in India
<a href="https://doi.org/10.1007/978-981-10-5672-7_24">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Thrips+as+the+Vectors+of+Tospoviruses+in+Indian+Agriculture&amp;btnG=">Google Scholar</a>
Genetics of Thrips Palmi (Thysanoptera: Thripidae)
<a href="https://doi.org/10.1007/s10340-019-01160-2">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Genetics+of+Thrips+Palmi+(Thysanoptera:+Thripidae)&amp;btnG=">Google Scholar</a>
Frontiers Approaches to the Diagnosis of Thrips (Thysanoptera): How Effective Are the Molecular and Electronic Detection Platforms
<a href="https://pubmed.ncbi.nlm.nih.gov/34680689/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.3390/insects12100920">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Frontiers+Approaches+to+the+Diagnosis+of+Thrips+(Thysanoptera):+How+Effective+Are+the+Molecular+and+Electronic+Detection+Platforms?&amp;btnG=">Google Scholar</a>
Progression of Watermelon Bud Necrosis Virus Infection in its Vector
<a href="https://pubmed.ncbi.nlm.nih.gov/33672941/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.3390/cells10020392">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Progression+of+Watermelon+Bud+Necrosis+Virus+Infection+in+its+Vector,+Thrips+Palmi&amp;btnG=">Google Scholar</a>
Assessment of Loci for DNA Barcoding in the Genus Thrips (Thysanoptera:Thripidae)
<a href="https://pubmed.ncbi.nlm.nih.gov/21564990/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1111/j.1755-0998.2009.02723.x">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Assessment+of+Loci+for+DNA+Barcoding+in+the+Genus+Thrips+(Thysanoptera:Thripidae)&amp;btnG=">Google Scholar</a>
Visual Isothermal Amplification Method for Detection of Canine Parvovirus 2 Genomic DNA
<a href="https://pubmed.ncbi.nlm.nih.gov/28349355/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1007/s00705-017-3321-5">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Polymerase+Spiral+Reaction+(PSR):+a+Novel,+Visual+Isothermal+Amplification+Method+for+Detection+of+Canine+Parvovirus+2+Genomic+DNA&amp;btnG=">Google Scholar</a>
Revue bibliographique et premi&#x000e8;res observations en Guadeloupe sur Thrips palmi Karny
<a href="https://doi.org/10.1051/agro:19880701">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Revue+bibliographique+et+premières+observations+en+Guadeloupe+sur+Thrips+palmi+Karny&amp;btnG=">Google Scholar</a>
&#x0201c;Pest Status and Biological Control of Thrips Palmi in Southeast Asia,&#x0201d; in Proceedings of a regional consultation workshop
Taiwan: Asian Vegetable Research and Development Center)
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Pest+Status+and+Biological+Control+of+Thrips+Palmi+in+Southeast+Asia&amp;btnG=">Google Scholar</a>
Risk Map for the Range Expansion of Thrips Palmi in Korea under Climate Change: Combining Species Distribution Models with Land-Use Change
<a href="https://doi.org/10.1016/j.aspen.2019.04.013">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Risk+Map+for+the+Range+Expansion+of+Thrips+Palmi+in+Korea+under+Climate+Change:+Combining+Species+Distribution+Models+with+Land-Use+Change&amp;btnG=">Google Scholar</a>
Successive Chelatometric Titration of Calcium and Magnesium Using Hydroxy Naphthol Blue (HNB) Indicator
<a href="https://doi.org/10.2116/bunsekikagaku.19.393">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Successive+Chelatometric+Titration+of+Calcium+and+Magnesium+Using+Hydroxy+Naphthol+Blue+(HNB)+Indicator&amp;btnG=">Google Scholar</a>
In Silico analyses of Molecular Interactions between Groundnut Bud Necrosis Virus and its Vector
<a href="https://pubmed.ncbi.nlm.nih.gov/31179363/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1007/s13337-019-00521-w">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=In+Silico+analyses+of+Molecular+Interactions+between+Groundnut+Bud+Necrosis+Virus+and+its+Vector,+Thrips+Palmi&amp;btnG=">Google Scholar</a>
Watermelon Bud Necrosis Tospovirus Is a Distinct Virus Species Belonging to Serogroup IV
<a href="https://pubmed.ncbi.nlm.nih.gov/9739341/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1007/s007050050405">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Watermelon+Bud+Necrosis+Tospovirus+Is+a+Distinct+Virus+Species+Belonging+to+Serogroup+IV&amp;btnG=">Google Scholar</a>
An Observation on the Embryonic Development in Thrips Palmi (Thysanoptera: Thripidae) Eggs Obtained by an Artificial Oviposition Setup
<a href="https://doi.org/10.1016/j.aspen.2020.03.012">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=An+Observation+on+the+Embryonic+Development+in+Thrips+Palmi+(Thysanoptera:+Thripidae)+Eggs+Obtained+by+an+Artificial+Oviposition+Setup&amp;btnG=">Google Scholar</a>
AMultiplex PCR Assay for Rapid Identification of Major Tospovirus Vectors Reported in India
<a href="https://pubmed.ncbi.nlm.nih.gov/32070289/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1186/s12864-020-6560-x">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=AMultiplex+PCR+Assay+for+Rapid+Identification+of+Major+Tospovirus+Vectors+Reported+in+India&amp;btnG=">Google Scholar</a>
Novel Polymerase Spiral Reaction Assay for the Visible Molecular Detection of Porcine Circovirus Type 3
<a href="https://pubmed.ncbi.nlm.nih.gov/31492192/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1186/s12917-019-2072-9">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Novel+Polymerase+Spiral+Reaction+Assay+for+the+Visible+Molecular+Detection+of+Porcine+Circovirus+Type+3&amp;btnG=">Google Scholar</a>
Population Trends of a Newly Introduced Species
on Commercial Watermelon Plantings in Hawaii
<a href="https://doi.org/10.1093/jee/79.3.718">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Population+Trends+of+a+Newly+Introduced+Species,+Thrips+Palmi+(Thysanoptera:+Thripidae),+on+Commercial+Watermelon+Plantings+in+Hawaii&amp;btnG=">Google Scholar</a>
Golden Jubilee Symposium on Horticultural Research: Changing Scenario
India: Indian Institute of Horticultural Research
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Golden+Jubilee+Symposium+on+Horticultural+Research:+Changing+Scenario&amp;btnG=">Google Scholar</a>
MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms
<a href="https://pubmed.ncbi.nlm.nih.gov/29722887/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1093/molbev/msy096">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=MEGA+X:+Molecular+Evolutionary+Genetics+Analysis+across+Computing+Platforms&amp;btnG=">Google Scholar</a>
Polymerase Spiral Reaction (PSR): A Novel Isothermal Nucleic Acid Amplification Method
<a href="https://pubmed.ncbi.nlm.nih.gov/26220251/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1038/srep12723">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Polymerase+Spiral+Reaction+(PSR):+A+Novel+Isothermal+Nucleic+Acid+Amplification+Method&amp;btnG=">Google Scholar</a>
An Assessment of the Potential Economic Impact of Thrips Palmi on Horticulture in England and the Significance of a Successful Eradication Campaign
<a href="https://doi.org/10.1016/j.cropro.2003.11.010">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=An+Assessment+of+the+Potential+Economic+Impact+of+Thrips+Palmi+on+Horticulture+in+England+and+the+Significance+of+a+Successful+Eradication+Campaign&amp;btnG=">Google Scholar</a>
Groundnut Bud Necrosis Virus Modulates the Expression of Innate Immune
and Cuticle Development-Associated Genes to Circulate and Propagate in its Vector
<a href="https://pubmed.ncbi.nlm.nih.gov/35369489/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.3389/fmicb.2022.773238">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Groundnut+Bud+Necrosis+Virus+Modulates+the+Expression+of+Innate+Immune,+Endocytosis,+and+Cuticle+Development-Associated+Genes+to+Circulate+and+Propagate+in+its+Vector,+Thrips+Palmi&amp;btnG=">Google Scholar</a>
Molecular Markers of Two Sympatric Species of the Genus Frankliniella (Thysanoptera: Thripidae)
<a href="https://doi.org/10.1016/j.aspen.2008.03.001">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Molecular+Markers+of+Two+Sympatric+Species+of+the+Genus+Frankliniella+(Thysanoptera:+Thripidae)&amp;btnG=">Google Scholar</a>
Sensitive and Visual Detection of SARS-CoV-2 Using Polymerase Spiral Reaction Assay
<a href="https://doi.org/10.1016/j.ijid.2021.12.102">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Sensitive+and+Visual+Detection+of+SARS-CoV-2+Using+Polymerase+Spiral+Reaction+Assay&amp;btnG=">Google Scholar</a>
Novel Polymerase Spiral Reaction (PSR) for Rapid Visual Detection of Bovine Herpesvirus 1 Genomic DNA from Aborted Bovine Fetus and Semen
<a href="https://pubmed.ncbi.nlm.nih.gov/29104164/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1016/j.gene.2017.11.004">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Novel+Polymerase+Spiral+Reaction+(PSR)+for+Rapid+Visual+Detection+of+Bovine+Herpesvirus+1+Genomic+DNA+from+Aborted+Bovine+Fetus+and+Semen&amp;btnG=">Google Scholar</a>
Effective Insecticide against Melon Thrips
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Effective+Insecticide+against+Melon+Thrips&amp;btnG=">Google Scholar</a>
Molecular Characterization of Tospovirus Transmitting Thrips Populations from India
<a href="https://doi.org/10.3844/ajbbsp.2005.167.172">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Molecular+Characterization+of+Tospovirus+Transmitting+Thrips+Populations+from+India&amp;btnG=">Google Scholar</a>
Development of a Novel Polymerase Spiral Reaction (PSR) Assay for Rapid and Visual Detection of Clostridium perfringens in Meat
<a href="https://pubmed.ncbi.nlm.nih.gov/33490689/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1016/j.heliyon.2021.e05941">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Development+of+a+Novel+Polymerase+Spiral+Reaction+(PSR)+Assay+for+Rapid+and+Visual+Detection+of+Clostridium+perfringens+in+Meat&amp;btnG=">Google Scholar</a>
Development of a Novel Polymerase Spiral Reaction (PSR) Assay for Rapid and Visual Detection of Staphylococcus aureus in Meat
<a href="https://doi.org/10.1016/j.lwt.2020.110507">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Development+of+a+Novel+Polymerase+Spiral+Reaction+(PSR)+Assay+for+Rapid+and+Visual+Detection+of+Staphylococcus+aureus+in+Meat&amp;btnG=">Google Scholar</a>
Notes on the Thrips (Thysanoptera) Occurring on the Soybean in Java
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Notes+on+the+Thrips+(Thysanoptera)+Occurring+on+the+Soybean+in+Java&amp;btnG=">Google Scholar</a>
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=New+State+Record:+Thrips+Palmi+Karny.+Hawaii+Pest+Report&amp;btnG=">Google Scholar</a>
Loop-mediated Isothermal Amplification of DNA
<a href="https://doi.org/10.1093/nar/28.12.e63">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Loop-mediated+Isothermal+Amplification+of+DNA&amp;btnG=">Google Scholar</a>
A Rapid Field-Based Assay Using Recombinase Polymerase Amplification for Identification of Thrips Palmi
<a href="https://doi.org/10.1007/s10340-020-01284-w">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=A+Rapid+Field-Based+Assay+Using+Recombinase+Polymerase+Amplification+for+Identification+of+Thrips+Palmi,+a+Vector+of+Tospoviruses&amp;btnG=">Google Scholar</a>
Real-time PCR Assay for Distinguishing Frankliniella Occidentalis and Thrips Palmi Arnika Przybylska
Aleksandra Obr&#x00119;palska-St&#x00119;plowska
<a href="https://pubmed.ncbi.nlm.nih.gov/28245887/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1017/s0007485317000177">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Real-time+PCR+Assay+for+Distinguishing+Frankliniella+Occidentalis+and+Thrips+Palmi+Arnika+Przybylska,+Żaneta+Fiedler,+Aleksandra+Obrępalska-Stęplowska&amp;btnG=">Google Scholar</a>
Detection of the Quarantine Species Thrips Palmi by Loop-Mediated Isothermal Amplification
<a href="https://pubmed.ncbi.nlm.nih.gov/25793743/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1371/journal.pone.0122033">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Detection+of+the+Quarantine+Species+Thrips+Palmi+by+Loop-Mediated+Isothermal+Amplification&amp;btnG=">Google Scholar</a>
PCR-RFLP Method to Distinguish Frankliniella Occidentalis
Frankliniella Pallida and Frankliniella Tenuicornis
<a href="https://doi.org/10.1515/jppr-2016-0009">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=PCR-RFLP+Method+to+Distinguish+Frankliniella+Occidentalis,+Frankliniella+Intonsa,+Frankliniella+Pallida+and+Frankliniella+Tenuicornis&amp;btnG=">Google Scholar</a>
<a href="https://pubmed.ncbi.nlm.nih.gov/26340723/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1016/j.coviro.2015.08.003">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Thrips+Transmission+of+Tospoviruses&amp;btnG=">Google Scholar</a>
Molecular Identification Key for Pest Species of Scirtothrips (Thysanoptera: Thripidae)
<a href="https://pubmed.ncbi.nlm.nih.gov/17066817/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1093/jee/99.5.1813">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Molecular+Identification+Key+for+Pest+Species+of+Scirtothrips+(Thysanoptera:+Thripidae)&amp;btnG=">Google Scholar</a>
Loop Mediated Isothermal Amplification: An Innovative Gene Amplification Technique for Animal Diseases
<a href="https://pubmed.ncbi.nlm.nih.gov/27284221/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.14202/vetworld.2016.465-469">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Loop+Mediated+Isothermal+Amplification:+An+Innovative+Gene+Amplification+Technique+for+Animal+Diseases&amp;btnG=">Google Scholar</a>
Abundance of AdventiveThrips palmi(Thysanoptera: Thripidae) Populations in Florida during the First Sixteen Years&#x000a7;
<a href="https://doi.org/10.1653/024.096.0312">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Abundance+of+AdventiveThrips+palmi(Thysanoptera:+Thripidae)+Populations+in+Florida+during+the+First+Sixteen+Years§&amp;btnG=">Google Scholar</a>
Thrips Palmi Karny (Thysanoptera: Thripidae) in South Florida
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Combating+Melon+Thrips,+Thrips+Palmi+Karny+(Thysanoptera:+Thripidae)+in+South+Florida&amp;btnG=">Google Scholar</a>
Polymerase Spiral Reaction (PSR) as a Novel Rapid Colorimetric Isothermal point of Care Assay for Detection of Trypanosoma Evansi Genomic DNA
<a href="https://pubmed.ncbi.nlm.nih.gov/35032829/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1016/j.vetpar.2021.109644">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Polymerase+Spiral+Reaction+(PSR)+as+a+Novel+Rapid+Colorimetric+Isothermal+point+of+Care+Assay+for+Detection+of+Trypanosoma+Evansi+Genomic+DNA&amp;btnG=">Google Scholar</a>
Development of Magnetic Bead Based Sample Extraction Coupled Polymerase Spiral Reaction for Rapid On-Site Detection of Chikungunya Virus
<a href="https://pubmed.ncbi.nlm.nih.gov/32669639/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1038/s41598-020-68469-2">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Development+of+Magnetic+Bead+Based+Sample+Extraction+Coupled+Polymerase+Spiral+Reaction+for+Rapid+On-Site+Detection+of+Chikungunya+Virus&amp;btnG=">Google Scholar</a>
&#x0201c;Status and Control Strategy of Peanut Bud Necrosis Disease in Uttar Pradesh,&#x0201d; in Recent Studies on Peanut Bud Necrosis Disease: Proceedings of a Meeting
1995 International Crop Research Institute for Semi-arid Tropics
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Status+and+Control+Strategy+of+Peanut+Bud+Necrosis+Disease+in+Uttar+Pradesh&amp;btnG=">Google Scholar</a>
Assessment of Yield Losses Due to Potato Stem Necrosis Disease
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Assessment+of+Yield+Losses+Due+to+Potato+Stem+Necrosis+Disease&amp;btnG=">Google Scholar</a>
Watermelon Bud Necrosis: A New Tospovirus Disease
<a href="https://doi.org/10.17660/actahortic.1996.431.6">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Watermelon+Bud+Necrosis:+A+New+Tospovirus+Disease&amp;btnG=">Google Scholar</a>
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Quarantine+Pests+for+Europe&amp;btnG=">Google Scholar</a>
Thrips: Pests of Concern to China and the United States
<a href="https://doi.org/10.1016/s1671-2927(11)60073-4">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Thrips:+Pests+of+Concern+to+China+and+the+United+States&amp;btnG=">Google Scholar</a>
&#x0201c;Thrips in Southeast Asia,&#x0201d; in Proceedings of a regional consultation workshop
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Thrips+in+Southeast+Asia&amp;btnG=">Google Scholar</a>
<a name="B67" id="B67"></a>ThripsWiki (2022). ThripsWiki - Providing Information on the World&#x00027;s Thrips. AvaliableAt: <a href="http://thrips.info/wiki/Main_Page">http://thrips.info/wiki/Main_Page</a>(Accessed on 03 19
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=ThripsWiki+-+Providing+Information+on+the+World's+Thrips&amp;btnG=">Google Scholar</a>
Polymerase Spiral Reaction Assay for Rapid and Real Time Detection of West Nile Virus from Clinical Samples
<a href="https://pubmed.ncbi.nlm.nih.gov/32984063/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.3389/fcimb.2020.00426">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Polymerase+Spiral+Reaction+Assay+for+Rapid+and+Real+Time+Detection+of+West+Nile+Virus+from+Clinical+Samples&amp;btnG=">Google Scholar</a>
Development of a Loop-Mediated Isothermal Amplification Method for the Rapid Detection of Phytopythium Vexans
<a href="https://pubmed.ncbi.nlm.nih.gov/34552572/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.3389/fmicb.2021.720485">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Development+of+a+Loop-Mediated+Isothermal+Amplification+Method+for+the+Rapid+Detection+of+Phytopythium+Vexans&amp;btnG=">Google Scholar</a>
SCAR Marker for Rapid Identification of the Western Flower Thrips
Frankliniella Occidentalis (Pergande) (Thysanoptera: Thripidae)
<a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=SCAR+Marker+for+Rapid+Identification+of+the+Western+Flower+Thrips,+Frankliniella+Occidentalis+(Pergande)+(Thysanoptera:+Thripidae)&amp;btnG=">Google Scholar</a>
Evaluation and Improvement of Isothermal Amplification Methods for point-of-need Plant Disease Diagnostics
<a href="https://pubmed.ncbi.nlm.nih.gov/32598374/">PubMed Abstract</a> &#x0007c; <a href="https://doi.org/10.1371/journal.pone.0235216">CrossRef Full Text</a> &#x0007c; <a href="https://scholar.google.com/scholar?hl=en&amp;as_sdt=0%2C5&amp;q=Evaluation+and+Improvement+of+Isothermal+Amplification+Methods+for+point-of-need+Plant+Disease+Diagnostics&amp;btnG=">Google Scholar</a>
Baranwal VK and Dietzgen RG (2022) Development of a Polymerase Spiral Reaction-Based Isothermal Assay for Rapid Identification of Thrips palmi
Received: 12 January 2022; Accepted: 23 March 2022;Published: 02 May 2022
Copyright &#x000a9; 2022 Jangra, Ghosh, Mukherjee, Baranwal and Dietzgen. This is an open-access article distributed under the terms of the <a rel="license" href="http://creativecommons.org/licenses/by/4.0/" target="_blank">Creative Commons Attribution License (CC BY).</a> The use
distribution or reproduction in other forums is permitted
provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited
in accordance with accepted academic practice
distribution or reproduction is permitted which does not comply with these terms
&#x0002a;Correspondence: Amalendu Ghosh, <a id="encmail">QW1hbGVuZHUuR2hvc2hAaWNhci5nb3YuaW4=</a>, <a id="encmail">b3JjaWQub3JnLzAwMDAtMDAwMS02NjM0LTU3NzE=</a>
Disclaimer:  All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations
Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher
94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish
 https://www.facebook.com/comunedipalmipaginaufficiale
On Sunday 9 March 2025, the streets of Palmi will come alive with the 41st edition of the Carnival of the Plain, an event that every year attracts masked groups, street artists and spectacular floats from all over Calabria.
The day will begin with a rally at 13:00, while the parade will start from Piazzale Concordato at 14:30, kicking off an explosion of music, colour and creativity. The best floats and masked groups will be awarded with trophies and plaques, in an exciting competition that will highlight the talent and imagination of the participants.&nbsp;
Street view</a>Interested<a href="https://calabriastraordinaria.it/en/events/carnival-celebration-in-santa-severina">Street PerformanceCarnival celebration in Santa SeverinaMar 2, 2025
</a><a href="https://calabriastraordinaria.it/en/places/francesco-cilea-and-antonino-manfroce-museum-palmi">MuseumPalmi
</a><a href="https://calabriastraordinaria.it/en/places/de-rosa-antiquarium-palmi">MuseumPalmi
</a><a href="https://calabriastraordinaria.it/en/places/raffaele-corso-museum-of-ethnography-and-folklore-palmi">MuseumPalmi
</a>Last update: Mar 9, 2025 6:01 AMContent owned by the Calabria Region | Tourism and Commerce Department issued under CC-BY License
</a><a href="https://www.youtube.com/channel/UCq-pw-Zs8qKid082U_weaUA" target="_blank" title="YouTube" class="cursor-pointer rounded-full p-2 hover:bg-secondary"></a><a href="https://instagram.com/calabria_straordinaria" target="_blank" title="Instagram" class="cursor-pointer rounded-full p-2 hover:bg-secondary"></a><a href="https://calabriastraordinaria.it/en/download" target="_self" class="group flex cursor-pointer items-center justify-start gap-2 rounded-full p-1">Download
OFFICIAL TOURIST INFORMATION SITE © 2022 Calabria Region Tourism and Commerce Department
Please enable JS and disable any ad blocker
<a href="https://helenafuneralhome.com/">Anderson Stevenson Wilke &amp; Retz Funeral Home</a>
Assisting the Montana communities of Helena
2021 from a battle with cancer since October 2019
She was raised in Minnesota with her brothers Ron and Roger Palmi
Carol went to nursing school in Minnesota and obtained her LPN license
Carol married Curtis Griffis in 1959 and moved to Montana where they raised their four children
The family loved to spend time outdoors camping and hiking
cross country skiing and spending time with family
Carol was famous for her delicious cooking
She also liked to quilt and be outdoors as much as possible
She was also a business owner operating the Wisdom Market in Wisdom
The family sends special thanks to the Whitehall Church
A Memorial Service for Carol will be held at a later date and will be announced when set
MT 59638 or visit below to share a memory of Carol
Services are pending at this time or no services will be held
Please visit the <a href="http://aswfuneralhome.com/floral-shops/" target="_blank">floral shop page</a> for assistance in choosing a local florist
No photos have been submitted yet. <a href="https://helenafuneralhome.com/add-your-photo/?obit_post_id=17106">Click here to share yours.</a>
Read the thoughts and memories, then feel free to <a href="#respond">add your own.</a>
<a class="comment-time-link" href="https://helenafuneralhome.com/obituaries/carol-h-focher-age-80-of-helena/#comment-460615" itemprop="url">February 15, 2021 at 1:45 pm</a>
<a class="comment-time-link" href="https://helenafuneralhome.com/obituaries/carol-h-focher-age-80-of-helena/#comment-460709" itemprop="url">February 20, 2021 at 11:25 pm</a>
&#x000A9;&nbsp;2025 · <a href="https://helenafuneralhome.com">Anderson Stevenson Wilke &amp; Retz Funeral Home</a> · Website by <a href="http://milescitywebsites.com/" target="_blank">Luci's Office</a>
sweet romance that takes place in two time periods: 50 years ago and during the COVID lockdown
There’s the Baltasar Kormákur who directs high octane thrillers like the killer lion movie “Beast” and the buddy cop movie “2 Guns,” and there’s a sweet
remarkably subtle Baltasar Kormákur who doesn’t get out as much these days
but occasionally reveals himself in the quieter moments of environmental adventures like “Everest” and “Adrift.”
“Touch,” has nothing explosive to distract us from the drama
It’s an understated and kind-hearted story about an old Icelandic widower named Kristofer (Egill Ólafsson
“Trapped”) who’s searching for the love of his life
Their relationship ended 50 years ago and now he’s a lonely man
But he just found out he probably doesn’t have long to live
so even though the COVID-19 pandemic just broke out — the film takes place in March of 2020 — he won’t let that stop him from traveling the world to see her one last time
Setting “Touch” at the beginning of the global pandemic adds surprisingly little tension to the story
Kristofer is frequently told he has to check out of his hotel because of lockdowns
but these were the early days and not everyone was taking the same preventative measures
There are scenes in “Touch” that take place in crowded restaurants where almost nobody is masking up
Not every country in the world took the same precautions but at least COVER YOUR NOSE
most of “Touch” actually takes place 50 years earlier
the director’s son) is a political activist going to school in London
His snotty friends tell him to join the proletariat if he really cares so much about worker’s rights so he impulsively quits school and walks into the first building he sees with a “Help Wanted” sign: a Japanese restaurant run by Takahashi (Masahiro Motoki
who quickly bonds with the young man and gives him a job washing dishes
Kristofer swiftly falls in love with Takahashi’s daughter
Baltasar Kormákur is in no rush whatsoever to get those two kooky kids together
Kristofer reveals himself to be mild of temperament when he’s separated from his confrontational school friends
who welcomes his understated affections despite her extroverted personality
Pálmi Kormákur and Kōki are exceptionally sweet performers
falling into natural rhythms and convincingly portraying moments of romantic intimacy
There’s an undercurrent of anxiety in the form of an untold story between Miko and her father that Kristofer doesn’t know at the time
and may never learn if he can’t find her in 50 years in the future
The way “Touch” resolves this storyline adds a sense of genuine significance to the story
but the film can’t quite help itself and eventually resorts to formulaic melodrama right at the end
It doesn’t ruin “Touch,” but it’s still a touch too much
Baltasar Kormákur directs “Touch” with tender confidence
knowing exactly how to absorb us in the scene with what looks like the minimum amount of effort — which usually means it took a spectacular amount of effort
Bergsteinn Björgúlfsson (“Wild Game”) photographs the present in dreary
muted tones and gradually warms up as Kristofer gets closer to his one true love
The past has a more tranquil palette; Takahashi’s restaurant is as inviting as any fictional restaurant can be
full of earthy comforts and little details
“Touch” can be distracting in its portrayal of the early pandemic
and it tries a little too hard to crowd-please in its final minutes
but what’s worth taking away from the film is its peacefulness
There are moments of friendship and family and workplace camaraderie that are real and charming
There’s a romance that plays out at its own pace
to the mild frustration of its own participants
who don’t want to push each other’s boundaries too fast
Baltasar Kormákur rejects the cinematic bombast of Baltasar Kormákur
&#8220;Touch&#8221; opens in theaters on Friday
Cate Palmi (left) takes a few minutes to visit with older sister Allison at the Junior Olympics National Volleyball Championship in Las Vegas PHOTO/PALMI FAMILY
Josh (center) and Nick (left) — are all involved in travel league baseball this summer
If you think juggling work and getting your grass cut in the same week is tough
try walking a few miles in the shoes of Carrie and Brett Palmi for a few weeks
Chances are you will never complain about time management again
who is starting her 17th season as the varsity volleyball coach at Osceola High School
who teaches history at Orlando Virtual School while also working for Southwest Airlines
are concluding what may just be the most hectic summer of any couple in Central Florida
That crazy schedule centers on the lives of six extremely active and athletic children
are currently playing travel league baseball — Nick for Power Baseball based in Orlando and Chris for Hardball Baseball in Gainesville
a rising sophomore will likely be the starting shortstop and a pitcher for the Bulldogs
This summer has been a whirlwind of travel for those two
as they have participated in four and five-day tournaments throughout Florida since school let out
the have played in several out-of-state tournaments in Alabama and Georgia
As if this wasn’t enough to keep the Palmi’s busy
plays club volleyball for nationally prominent Gamepoint
where she has participated in tournaments at the AAU and USA Volleyball levels in Dallas
Omaha and several Florida cities this summer
Gamepoint was one of three Florida teams that qualified for the National Junior Olympics Tournament
is also on a baseball travel ball roster for Team Florida and plays on the St
which finished third in the state this year and had to travel to district and regional tournaments
An older daughter Allison is finishing up her freshman year at the University of the Incarnate Word in San Antonio
Texas where she was on the volleyball team whose schedule was pushed to the spring because of COVID-19
That led to the family traveling to the school’s conference tournament in San Antonio this spring
a sixth child – six-year old Maddy – participates in summer volleyball camps
Carrie and Brett have made it a point that at least one of them and for the most part both of them would attend every tournament
difficult and exhausting but we manage to get it done,” Carrie said
“We have a great support system around us with friends and relatives that make it possible
It has taken a lot of planning and organization
but our family’s lives revolve around sports and frankly we love it.”
Still attending the games and tournaments can be challenging
Brett posted on his Facebook page a few weeks ago that he attended three different baseball games
in three different Florida cities — all in the same day
That family and relative cooperation is important when two or three events are going on in the same week
When Carrie and Brett were in Las Vegas watching Cate’s Junior Olympic Volleyball Tournament
Carrie has to rely on her father Chris Arago to drive Nick and Chris to a baseball tournament in Hoover
The Palmi’s then flew directly to Alabama from Las Vegas to attend that tournament
“Surprisingly both Brett and I have been able to get to almost all the events together as a family,” Carrie said
“Brett’s job with Southwest has made somewhat easier
we feel extremely blessed to be able to do what we do and to be able almost always do it as a family.”
Carrie’s hectic schedule has been made somewhat easier by the fact that oldest daughter Allison has been home for the summer
“Having her home has been a blessing as it gave us an extra driver to get everyone to practices and games,” Carrie said
Again the family and friends thing has really helped make that possible
we have had so many relatives help us with the organizational part of these events that parents are supposed to help with.”
Carrie says she is happy that her “normal” school schedule is about to begin
our season is our busiest time of the year
I’m one of the few who can say it’s almost the most relaxing time of the year for me,” she says with a laugh
The athletic prowess of the Palmi children comes by naturally
Celebrating their 20th wedding anniversary this week
Carrie and Brett met at Jacksonville University where she was a star volleyball player and he was an all-conference football player
“Sports have always been a big part of our lives and it has become a huge part of our children’s lives
&copy;&nbsp;2025&nbsp;Osceola News Gazette
The city of Palmi is preparing to experience an unforgettable Christmas with a rich calendar of events scheduled from December to January.
Music, cinema, theatre, shows, entertainment and gastronomy will intertwine to create a unique atmosphere, made even more special by the splendid illuminations that will light up every corner of the city.
Street view</a>Interested<a href="https://calabriastraordinaria.it/en/events/armonie-darte-festival">Multiple eventsArmonie D&#x27;Arte FestivalAug 2, 2024
</a><a href="https://calabriastraordinaria.it/en/places/michele-guerrisi-plaster-collection-palmi">Point of interestPalmi
</a>Last update: Jan 6, 2025 6:00 AMContent owned by the Calabria Region | Tourism and Commerce Department issued under CC-BY License
Microbe and Virus Interactions with Plants
Volume 14 - 2023 | <a class="ArticleLayoutHeader__info__doi" href="https://doi.org/10.3389/fmicb.2023.1254246">https://doi.org/10.3389/fmicb.2023.1254246</a>
Thrips palmi (Thysanoptera: Thripidae) is a major agricultural pest infesting over 200 plant species
Along with direct injury caused by feeding
Groundnut bud necrosis orthotospovirus (GBNV
genus Orthotospovirus) is the predominant orthotospovirus in Asia
It is responsible for almost 89 million USD losses in Asia annually
and apoptosis are responsive to the infection of orthotospoviruses in thrips
palmi tyrosine kinase Btk29A isoform X1 (Btk29A) and collagen alpha-1(III) chain-like (COL3A1) are significantly regulated post-GBNV and capsicum chlorosis orthotospovirus infection
palmi Btk29A and COL3A1 were silenced and the effect on virus titer and fitness was assessed
The expression of Btk29A and COL3A1 was significantly reduced by 3.62 and 3.15-fold
Oral administration of Btk29A and COL3A1 dsRNAs induced 60 and 50.9% mortality in T
palmi significantly dropped post-silencing Btk29A
the silencing of COL3A1 led to an increase in GBNV concentration in T
this is the first report on the effect of silencing Btk29A and COL3A1 on the fitness and GBNV titer in T
palmi Btk29A and COL3A1 were silenced to study the effect on thrips fitness and orthotospovirus titer in thrips
The study employed a homogenous population of T. palmi derived from a single adult female. The isofemale line has been maintained on eggplant since 2018. The identity of the population was further verified by morphometric keys (<a href="#ref6">Bhatti, 1980</a>; <a href="#ref10">Cluever and Smith, 2017</a>) and mitochondrial cytochrome oxidase subunit I (mtCOI) nucleotide sequencing
The initial GBNV inoculum was collected from a pure culture maintained at the Advanced Center for Plant Virology, Indian Agricultural Research Institute (IARI), New Delhi. Sap inoculation of healthy cowpea plants at the two-leaf stage was done with GBNV under insect-proof conditions as described by <a href="#ref15">Ghosh et al. (2021)</a>. The presence of GBNV infection in cowpea was verified using RT-PCR using GBNV-specific primers AG109F-AG110R (<a href="#tab1">Table 1</a>)
The selected regions were 1,166&#x2013;1,444 for Btk29A and 213&#x2013;469 for COL3A1 genes
The dsRNA stretches were amplified using primer pairs listed in <a href="#tab1">Table 1</a>. The PCR conditions were optimized using a gradient PCR. Total DNA was isolated from T. palmi using a CTAB extraction buffer as described by <a href="#ref24">Jangra and Ghosh (2022)</a> and used for amplification in a thermal cycler
A 25&#x2009;&#x03BC;L PCR mixture comprised of 1X PCR buffer (Thermo Fisher Scientific
0.4&#x2009;&#x03BC;M each forward and reverse primer (GCC Biotech
0.26&#x2009;mM dNTP mix (Thermo Fisher Scientific)
and 2&#x2009;U of DreamTaq DNA polymerase (Thermo Fisher Scientific) were used
PCR was performed in a T100 thermocycler (Bio-Rad
United States) with an initial denaturation at 94&#x000B0;C for 5&#x2009;min
35&#x2009;cycles of denaturation at 94&#x000B0;C for 40&#x2009;s
followed by a final extension at 72&#x000B0;C for 10&#x2009;min
The PCR products were resolved on a 2% agarose gel
and visualized under a gel documentation system (MaestroGen Inc.
The amplified PCR products were sequenced for confirmation purposes
palmi based on nucleotide homology screening was used as a negative control
The purified Btk29A and COL3A1 dsRNAs were administered to T. palmi orally using an artificial feeding setup as reported by <a href="#ref35">Priti et al. (2022)</a>
a diet was prepared by mixing 50&#x2009;mg/mL pine pollen (Lost Empire Herbs
United States) extract with 1% sucrose solution and 0.0001% methylene blue tracker dye (Bio Basic
The diet was supplemented with 3&#x2009;&#x03BC;g/&#x03BC;L dsRNA (total volume 300&#x2009;&#x03BC;L)
The diet with dsRNA was filled into UV-sterilized detachable caps of 2&#x2009;mL microcentrifuge tubes and covered with stretched parafilm
Around 20 adults or larvae were taken in the perforated microcentrifuge tubes
The cap filled with diet was closed gently
palmi were allowed to feed for 24&#x2009;h at 28&#x2009;&#x000B1;&#x2009;1&#x000B0;C temperature
60&#x2009;&#x000B1;&#x2009;10% relative humidity
and 16&#x2009;h light&#x2013;8&#x2009;h dark
a diet without dsRNA and a diet with BtTOB1 dsRNA (non-specific to T
Several such replicates were maintained to get enough dsRNA-treated and untreated thrips to assess the mortality
The statistical analysis was performed and graphs were generated in Microsoft Excel 2019
palmi Btk29A and COL3A1 in BtTOB1 dsRNA-fed thrips served as the negative control
palmi individuals that fed on the artificial diet were identified by a blue tinge in their abdomen and were exclusively considered to assess the effect of dsRNA on survival
palmi was recorded 24&#x2009;h post-dsRNA feeding
The whole experiment was replicated nine times for each treatment
palmi was recorded and compared with the thrips fed on the dsRNA-free diet and diet with BtTOB1 dsRNA
Tukey&#x2019;s test was performed with XLSTAT 2014.5.03 to differentiate means across categories with a 95% confidence interval
post-dsRNA feeding were also checked under a stereo microscope (M205 FA
The viral titer was estimated by fitting the mean CT values in the standard curve
The following formula was used to calculate the virus copy number in Microsoft Excel 2019
N&#x2009;=&#x2009;(x&#x2009;&#x000D7;&#x2009;6.022&#x2009;&#x000D7;&#x2009;1023)/ (n&#x2009;&#x000D7;&#x2009;340&#x2009;&#x000D7;&#x2009;109)
where N&#x2009;=&#x2009;number of viral copies
x&#x2009;=&#x2009;amount of amplicon in ng
and n&#x2009;=&#x2009;length of linearized plasmid
One-Way ANOVA was performed to determine the mean differences in virus copies among treatments at p&#x2009;&#x02264;&#x2009;0.05 in XLSTAT 2014.5.03
The effect of Btk29A and COL3A1 dsRNAs feeding on survival and mRNA expression at this stage (L1) was also recorded as mentioned above
T. palmi adults had quadrangular heads with seven segmented antennae. At the top of the head, three brick-red ocelli were present in a triangular formation. A pair of interocellar setae originated outside the ocellar triangle. Besides, the nucleotide sequence of a 595&#x2009;bp portion of mtCOI was amplified using primer pairs LCO 1490 and HCO 2198 (<a href="#ref11">Folmer et al., 1994</a>) and was 100% identical with T
The sequence can be retrieved from GenBank with the accession number OP223495
RT-PCR with GBNV-specific primers produced ~1767&#x2009;bp desired amplicon from all the sap-inoculated plants
The nucleotide sequence was found to be more than 97% identical to other GBNV isolates
The sequence can be retrieved from GenBank with the accession number MN566913
(A) A 279&#x2009;nt conserved region of Thrips palmi Btk29A was selected for designing Btk29A dsRNA
palmi COL3A1 was selected for designing COL3A1 dsRNA
The putative siRNAs were marked in different colored boxes
(C) A 167&#x2009;nt conserved region of Bemisia tabaci TOB1
palmi was used to design the negative control dsRNA
PCR with primer pair AG507F-AG508R specific to Btk29A yielded an amplicon of 279&#x2009;bp (accession no. OP346052). Whereas a 257&#x2009;bp (accession no. OP345816) fragment of COL3A1 was amplified in PCR with the primer pair AG188F-AG189R. On a 2% agarose gel, the Btk29A and COL3A1 dsRNAs purified from the total RNA of recombinant E. coli HT115 cells generated ~279 and&#x2009;~&#x2009;257&#x2009;bp bands, respectively (<a href="#fig2">Figure 2</a>)
The concentration of the purified dsRNA was 621.5&#x2009;ng/ &#x03BC;L for Btk29A and 531.0&#x2009;ng/ &#x03BC;L for COL31A
It was 779.9&#x2009;ng/&#x03BC;L in the case of BtTOB1 dsRNA
(A) Purified Btk29A dsRNA (1) and COL3A1 dsRNA (2)
BtTOB1 dsRNA (3) used as a negative control
palmi fed on artificial diet for 24&#x2009;h indicated by blue tinge on abdomen (red arrow)
palmi COL3A1 mRNA expression level was recorded in T
palmi Btk29A and COL3A1 post-exposure to BtTOB1 dsRNA was statistically non-significant with untreated control
There was no significant difference in the expression of the endogenous control
indicating that the Btk29A and COL3A1 dsRNAs had a specific action on the target mRNAs
and &#x03B2;-tubulin produced no secondary peaks in the RT-qPCR melting curve analysis
indicating that the reactions were specific
Effect of silencing Btk29A and COL3A1 on survival of T
palmi Btk29A and COL3A1 mRNA post-dsRNA exposure
palmi 24&#x2009;h post-Btk29A and COL3A1 dsRNA feeding at 3&#x2009;&#x03BC;g/mL
The error bars are the standard error of the mean (SEM)
Mean denoted by an asterisk (&#x0002A;) indicates a significant difference (p&#x2009;&#x0003C;&#x2009;0.05)
(C) Btk29A and COL3A1 dsRNA-exposed and unexposed T
No morphological deformities were recorded post-dsRNA exposure
palmi Btk29A and COL3A1 was downregulated by 5.23 and 2.04-fold
palmi larvae as compared to larvae fed on the diet without dsRNA
There was no significant difference in GBNV titer following BtTOB1 dsRNA exposure when compared to the untreated control
palmi Btk29A and COL3A1 on groundnut bud necrosis orthotospovirus (GBNV) titer in T
(A) Mean GBNV copy numbers acquired by Btk29A and COL3A1 dsRNA-exposed T
palmi Btk29A and COL3A1 mRNA in larval stage post-dsRNA exposure
palmi Btk29A and COL3A1 were silenced using exogenous uptake of dsRNA
no such morphological deformities were recorded in adults of T
Either the exposure was too short to produce any obvious immediate effects on the structural shape or Btk29A and COL3A1 might not be associated with morphogenesis of T
its role in antiviral resistance has never been explored
The depletion of COL3A1 mRNA probably aided the virus to evade the innate immune response which led to an increase in virus copies in T
The current study is the first to validate that silencing Btk29A and COL3A1 affects the fitness and orthotospovirus titer in T
Btk29A and COL3A1 would be novel genetic targets for sustainable thrips management and help elucidate the thrips-orthotospovirus relationships
The datasets presented in this study can be found in online repositories
The names of the repository/repositories and accession number(s) can be found in the article/Supplementary material
Ethical review and approval were not required for the study on animals in accordance with the local legislation and institutional requirements
The fellowship of VR was supported by the Indian Council of Agricultural Research
The authors are thankful to the advisories of Vinay Kalia
Advances in exogenous RNA delivery techniques for RNAi-mediated pest control
Optimization of dietary RNA interference delivery to western flower thrips Frankliniella occidentalis and onion thrips Thrips tabaci
<a name="ref3" id="ref3"></a>Badillo-Vargas
RNA interference tools for the western flower thrips
is conferred by G275E mutation in &#x03B1;6 subunit of nicotinic acetylcholine receptor and cytochrome P450 detoxification
<a href="https://doi.org/10.1111/j.1365-3113.1980.tb00404.x" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=J.+S.+Bhatti&#x00026;publication_year=1980&#x00026;title=Species+of+the+genus+Thrips+from+India+(Thysanoptera)&#x00026;journal=Syst.+Entomol.&#x00026;volume=5&#x00026;pages=109-166" target="_blank">Google Scholar</a>
<a href="https://doi.org/10.1016/j.cropro.2006.10.023" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=R.+J.+C.+Cannon&#x00026;author=L.+Matthews&#x00026;author=D.+W.+Collins&#x00026;publication_year=2007&#x00026;title=A+review+of+the+pest+status+and+control+options+for+Thrips+palmi&#x00026;journal=Crop+Prot.&#x00026;volume=26&#x00026;pages=1089-1098" target="_blank">Google Scholar</a>
Topical spray of dsRNA induces mortality and inhibits chilli leaf curl virus transmission by Bemisia tabaci Asia II 1
RNAi in insects: a revolution in fundamental research and pest control applications
<a href="https://doi.org/10.1653/024.100.0208" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=J.+D.+Cluever&#x00026;author=H.+A.+Smith&#x00026;publication_year=2017&#x00026;title=A+photo-based+key+of+thrips+(Thysanoptera)+associated+with+horticultural+crops+in+Florida&#x00026;journal=Fla.+Entomol.&#x00026;volume=100&#x00026;pages=454-467" target="_blank">Google Scholar</a>
Transcriptomic identification and characterization of genes responding to sublethal doses of three different insecticides in the western flower thrips
Exposure to watermelon bud necrosis virus and groundnut bud necrosis virus alters the life history traits of their vector
<a href="http://scholar.google.com/scholar_lookup?author=A.+Ghosh&#x00026;author=D.+Dey&#x00026;author=B.+Mandal&#x00026;author=R.+K.+Jain&#x00026;publication_year=2017&#x00026;title=Thrips+as+the+vectors+of+tospoviruses+in+Indian+agriculture&#x00026;journal=A+century+of+plant+virology+in+India&#x00026;pages=537-561" target="_blank">Google Scholar</a>
<a name="ref16" id="ref16"></a>Gnanasekaran
Geminivirus Betasatellite-encoded &#x03B2;C1 protein exhibits novel ATP hydrolysis activity that influences its DNA-binding activity and viral pathogenesis
<a name="ref17" id="ref17"></a>Gnanasekaran
A geminivirus betasatellite encoded &#x03B2;C1 protein interacts with PsbP and subverts PsbP-mediated antiviral defence in plants
Selection of lethal genes for ingestion RNA interference against western flower thrips
Strategies for the production of dsRNA biocontrols as alternatives to chemical pesticides
<a href="https://doi.org/10.1007/s13337-019-00521-w" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=S.+S.+Jagdale&#x00026;author=A.+Ghosh&#x00026;publication_year=2019&#x00026;title=In+silico+analyses+of+molecular+interactions+between+groundnut+bud+necrosis+virus+and+its+vector+Thrips+palmi&#x00026;journal=VirusDisease&#x00026;volume=30&#x00026;pages=245-251" target="_blank">Google Scholar</a>
<a href="http://scholar.google.com/scholar_lookup?author=M.+Jahani&#x00026;author=O.+Christiaens&#x00026;author=G.+Smagghe&#x00026;publication_year=2018&#x00026;title=Analysis+of+artificial+diet+to+deliver+dsRNA+in+the+western+flower+thrips+(Frankliniella+occidentalis)&#x00026;journal=IOBC-WPRS+Bulletin&#x00026;volume=131&#x00026;pages=41-50" target="_blank">Google Scholar</a>
Rapid and zero-cost DNA extraction from soft-bodied insects for routine PCR-based applications
A novel binding site in collagen type III for integrins alpha1beta1 and alpha2beta1
Type III collagen (COL3A1): gene and protein structure
Receptor tyrosine kinase inhibitors block multiple steps of influenza a virus replication
MEGA X: molecular evolutionary genetics analysis across computing platforms
<a href="http://scholar.google.com/scholar_lookup?author=H.+H.+Karny&#x00026;publication_year=1925&#x00026;journal=The+Thysanoptera+found+on+tobacco+in+Java+and+Sumatra.+Bulletin+van+het+Deli+Proefstation+te+Medan-Sumatra&#x00026;volume=23&#x00026;pages=1-55" target="_blank">Google Scholar</a>
<a href="https://doi.org/10.1006/meth.2001.1262" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=K.+J.+Livak&#x00026;author=T.+D.+Schmittgen&#x00026;publication_year=2001&#x00026;title=Analysis+of+relative+gene+expression+data+using+real-time+quantitative+PCR+and+the+2&#x0394;&#x0394;CT+method&#x00026;journal=Methods&#x00026;volume=25&#x00026;pages=402-408" target="_blank">Google Scholar</a>
Groundnut bud necrosis virus modulates the expression of innate immune
and cuticle development-associated genes to circulate and propagate in its vector
RNAi technology: a new platform for crop pest control
<a href="https://pubmed.ncbi.nlm.nih.gov/28878489" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.1007/s12298-017-0443-x" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=B.+Mamta&#x00026;author=M.+V.+Rajam&#x00026;publication_year=2017&#x00026;title=RNAi+technology:+a+new+platform+for+crop+pest+control&#x00026;journal=Physiol.+Mol.+Biol.+Plants&#x00026;volume=23&#x00026;pages=487-501" target="_blank">Google Scholar</a>
Feeding-based RNA Intereference of a gap gene is lethal to the pea aphid
is induced by the HTLV-1 Oncoprotein tax and impacts virus transmission
Silencing of Thrips palmi UHRF1BP1 and PFAS using antisense Oligos induces mortality and reduces Tospovirus titer in its vector
<a href="https://doi.org/10.55446/IJE.2023.1113" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=V.+Rajesh&#x00026;author=V.+Rakesh&#x00026;author=S.+Jangra&#x00026;author=A.+Ghosh&#x00026;publication_year=2023&#x00026;title=RNA+interference+in+thrips+vectors:+a+step+forward+toward+sustainable+management&#x00026;journal=Indian+J.+Entomol.&#x00026;volume=1&#x2013;10&#x00026;" target="_blank">Google Scholar</a>
RNA interference suppression of v-ATPase B and juvenile hormone binding protein genes through topically applied dsRNA on Tomato leaves: developing biopesticides to control the south American pinworm
<a href="http://scholar.google.com/scholar_lookup?author=D.+V.+R.+Reddy&#x00026;author=A.+A.+M.+Buiel&#x00026;author=T.+Satyanarayana&#x00026;author=S.+L.+Dwivedi&#x00026;author=A.+S.+Reddy&#x00026;author=A.+S.+Ratna&#x00026;publication_year=1995&#x00026;title=Recent+studies+on+Peanut+bud+necrosis+disease&#x00026;journal=ICRISAT+Asia+Centre&#x00026;volume=1995&#x00026;pages=3-7" target="_blank">Google Scholar</a>
<a href="https://doi.org/10.1603/IPM10020" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=D.+G.+Riley&#x00026;author=S.+V.+Joseph&#x00026;author=R.+Srinivasan&#x00026;author=S.+Diffie&#x00026;publication_year=2011&#x00026;title=Thrips+vectors+of+tospoviruses&#x00026;journal=J.+Integrat.+Pest+Manag.&#x00026;volume=2&#x00026;pages=I1-I10" target="_blank">Google Scholar</a>
Evaluation of a cuticle protein gene as a potential RNAi target in aphids
Variable resistance to spinetoram in populations of Thrips palmi across a small area unconnected to genetic similarity
Association between susceptibility of Thrips palmi to spinetoram and frequency of G275E mutation provides basis for molecular quantification of field-evolved resistance
selection of reference genes and demonstration of feeding RNAi in Thrips tabaci (Lind.) (Thysanoptera: Thripidae)
<a href="http://scholar.google.com/scholar_lookup?author=S.+J.+Singh&#x00026;author=M+Krishnareddy&#x00026;publication_year=1995&#x00026;title=Watermelon+bud+necrosis:+a+new+tospovirus+disease&#x00026;journal=Tospoviruses+and+Thrips+of+Floral+and+Vegetable+Crops&#x00026;volume=431&#x00026;pages=68-77" target="_blank">Google Scholar</a>
<a href="http://scholar.google.com/scholar_lookup?publication_year=2018&#x00026;" target="_blank">Google Scholar</a>
Effect of silencing Bemisia tabaci TLR3 and TOB1 on fitness and begomovirus transmission
Collagen fragments modulate innate immunity
src64 and tec29 are required for microfilament contraction during Drosophila cellularization
RNA interference in insects: protecting beneficials and controlling pests
Efficient control of western flower thrips by plastid-mediated RNA interference
Delta and jagged are candidate target genes of RNAi biopesticides for the control of Nilaparvata lugens
Delivery of dsRNA for RNAi in insects: an overview and future directions
expression analysis and functional verification of two genes encoding small heat shock proteins in the western flower thrips
Induced resistance combined with RNA interference attenuates the Counteradaptation of the Western flower Thrips
Jangra S and Ghosh A (2023) Effect of silencing Thrips palmi Btk29A and COL3A1 on fitness and virus acquisition
Received: 06 July 2023; Accepted: 29 September 2023; Published: 19 October 2023
Copyright &#x000A9; 2023 Rajesh, Jangra and Ghosh. This is an open-access article distributed under the terms of the <a rel="license" href="http://creativecommons.org/licenses/by/4.0/" target="_blank">Creative Commons Attribution License (CC BY)</a>
&#x0002A;Correspondence: Amalendu Ghosh, <a id="encmail">YW1hbDRlbnRvQGdtYWlsLmNvbQ==</a>
&#x2020;Present address: Sumit Jangra,Tropical Research and Education Center, Institute of Food and Agricultural Sciences, University of Florida, Homestead, FL, United States
Disclaimer:  All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher. 
94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish.
Volume 15 - 2024 | <a class="ArticleLayoutHeader__info__doi" href="https://doi.org/10.3389/fpls.2024.1361276">https://doi.org/10.3389/fpls.2024.1361276</a>
This article is part of the Research TopicOmics in Plant-Insect Interactions<a class="Link Link--linkType Link--maincolor Link--medium Link--icon Link--chevronRight Link--right" aria-label="View all 17 articles" href="https://www.frontiersin.org/research-topics/53294/omics-in-plant-insect-interactions/magazine" target="_self" data-event="customLink-linkType-a_viewAll17Articles">View all 17 articles</a>
Thrips are serious pests of Hemerocallis citrina Baroni (daylily)
daylily has evolved a set of sophisticated defense mechanisms
induction of systemic resistance in Hemerocallis citrina &#x2018;Datong Huanghua&#x2019; by Thrips palmi feeding was investigated at both biochemical and molecular levels
The soluble sugar content of daylily leaves was significantly lower than that in control check (CK) at all time points of feeding by T
whereas the amino acid and free fatty acid contents started to be significantly lower than those in CK after 7 days
which are harmful to the growth and reproduction of T
The activities of defense enzymes such as peroxidase (POD)
and polyphenol oxidase (PPO) were significantly increased
and the degree of damage to plants was reduced
The significant increase in protease inhibitor (PI) activity may lead to disrupted digestion and slower growth in T
1,894 differentially expressed genes (DEGs) were identified between control and treatment groups at five timepoints
DEGs were mainly enriched in secondary metabolite synthesis
and other defense hormone signal transduction pathways
698 DEGs were predicted to be transcription factors
including bHLH and WRKY members related to biotic stress
WGCNA identified 18 hub genes in four key modules (Purple
and Red) including MYB-like DNA-binding domain (TRINITY_DN2391_c0_g1
The results indicate that biosynthesis of secondary metabolites
and defense hormones pathways are involved in the induced resistance to T
the safest and most effective strategy for thrips prevention and control is to utilize the insect resistance of the host plant
investigation of the physiological mechanisms of thrips resistance in daylily provides a basis for breeding insect-resistant plants
Herbivorous insect feeding initiates the inducible defense mechanism of plants, triggering a series of signal transduction and gene expression events, and the generation of defense substances. Inducible defense plays an more important role in the self-protection of plants (<a href="#B34">Maleck and Dietrich, 1999</a>)
there are few reports on the physiological responses and omics differences of daylily in response to thrips feeding
citrina &#x2018;Datong Huanghua&#x2019; inoculated with T
palmi was used to determine the content of nutrients and secondary metabolites and defense enzyme activities in leaves to elucidate the physiological changes that induce pest defenses
Transcriptome analysis of thrips-infested leaves was performed with healthy leaves as controls
Differentially expressed genes (DEGs) were identified
and the main transcription factors and their expression patterns were analyzed
Key insect resistance genes were identified to elucidate the induced defense mechanism of daylily in response to T
palmi individuals naturally occurring in daylily fields at the Horticultural Station of Shanxi Agricultural University were used as the source of test insects
The daylily variety used in the study was Datong Huanghua
which was planted at the Horticultural Station of Shanxi Agricultural University
The study was performed from March to June 2023 at the Horticultural Station of Shanxi Agricultural University
a 60-mesh insect-proof net was used to set up a net room
from west to east in the field to establish the experimental plot
To establish the treatment group with induction of T
palmi were collected in the field and brought to the laboratory in a cage (118.7 &#xd7; 100 &#xd7; 100&#xa0;cm) made of 60-mesh insect-proof net
palmi was starved for 12&#xa0;h prior to the test
to ensure adequate feeding induction on plants
palmi were transported to the net room in 50-mL centrifuge tubes
palmi from one tube were released onto 3&#x2212;4 plants such that there were ~90 individuals per plant; at least 15 plants were treated overall to ensure that three biological replicates could be sampled at each point
palmi concentrated on the upper&#x2013;middle position of young leaves
Datong Huanghua plants in this treatment group (TF) were individually covered with a 60-mesh insect-proof net to prevent T
daylily plants were allowed to grow normally without any treatment in the net room
and each plant was individually covered with insect-proof net
Each treatment group included three biological replicates
and leaves of CK group plants collected at the same time served as controls (named CK1&#x2212;CK5)
Three replicates were included at each stage
yielding five extractions with 30 samples in total
which were frozen in liquid nitrogen and stored at -80&#xb0;C until future use
All kits were purchased from Beijing Solarbio Science &amp; Technology Co.
Data were summarized and processed using Microsoft Excel 2010 and statistically analyzed with SPSS software v20.0
The significance of the difference in nutrients between healthy daylily leaves and leaves fed on by T
palmi was tested by Tukey&#x2019;s test (p &lt; 0.05) and graph plotting using SigmaPlot 14.0 software
Data processing for secondary matter content and defense enzyme activities of daylily leaves before and after feeding by T
palmi was done in the same way as data processing for nutrient content determination
RNA was extracted from TF1&#x2212;TF5 and CK1&#x2212;CK5 samples using the TRIzol method (<a href="#B57">Wang et&#xa0;al., 2022</a>)
RNA integrity was assessed using 1% agarose gel electrophoresis
and the RIN value was determined using an Agilent 2100 bioanalyzer (Agilent Technologies Inc.
the cDNA library was constructed and high-throughput sequencing was performed on an Illumina Hiseq platform (Shanghai Majorbio Bio-Pharm Technology Co.
Raw data obtained by sequencing were filtered to remove adapters and low-quality reads
The base quality score (Q30) of clean data was determined
Trinity software was used to assemble the clean data obtained by sequencing to construct the UniGene library
We constructed a transcriptome expression matrix of leaves from the TF1-TF5 samples and screened for genes with TPM values &lt;1
we used the WGCNA package (version 1.6.6) in R software (version 3.4.4) to construct a gene co-expression network
We selected &#x3b2; = 16 as the soft threshold for subsequent analysis and used the &#x2018;blockwiseModules&#x2019; function to construct the gene network
with the following parameter settings: power = 6
nThreads = 0; all other parameters were set to default values
and module feature genes for each module were calculated
We used the &#x2018;exportNetworkToCytoscape&#x2019; function in the WGCNA package to export network relationships between genes in relevant modules
and Cytoscape software (version 3.7.1) was used to create graphs
Figure&#xa0;1 Determination of plant nutrients
(A) Amino acids content; (B) free fatty acids content; (C) soluble sugars content
Different letters indicate significant differences in nutrient composition between healthy leaves and leaves after feeding by T
Figure&#xa0;2 Determination of plant secondary metabolites
(A) Tannins content; (B) flavonoids content; (C) total phenols content
Different letters indicate significant differences in the content of secondary metabolites between healthy leaves and leaves after feeding by T
Figure&#xa0;3 Determination of plant defense enzymes
(A) Peroxidase (POD) activity; (B) phenylalanine ammonia lyase (PAL) activity; (C) polyphenol oxidase (PPO) activity; (D) protease inhibitor (PI) activity
Different letters indicate significant differences in defense enzymes activities between healthy leaves and leaves after feeding by T
To study the changes in transcription levels of daylily under T. palmi stress, using Illumina 2&#xd7; 150 bp paired-end sequencing, 141.58 Gb of clean data was obtained from 10 samples. Clean data from each sample reached &gt;6.03 Gb, the percentage of Q30 bases was &gt;94.18%. The percentage in brackets in the last column of <a href="#T1">Table&#xa0;1</a> is the comparison rate for clean reads; clean reads comparison efficiency ranged between 78.76% and 82.94%
The results showed that the quality of the sequencing output data was good
and the data could be used for further analysis
Table&#xa0;1 Transcriptome sequencing data statistics
Figure&#xa0;4 Differentially expressed genes (DEGs) in Datong Huanghua exposed to Thrips palmi feeding
(A) Number of DEGs at different stages; (B) venn diagram analysis of DEGs; (C) gene ontology (GO) enrichment analysis of DEGs; (D) Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of DEGs
Based on the findings from DEGs, and GO enrichment and KEGG pathway analyses, 787 potential candidate genes related to T. palmi resistance were subjected to differential expression analysis (<a href="#f5">Figure&#xa0;5</a>)
These genes could be divided into two expression patterns
Its functions include the synthesis of secondary substances such as flavonoids
the synthesis of defense enzymes such as POD
the signal transduction of defense hormones such as JA and SA
which are mainly upregulated after feeding by T
and are more significant in the TF2 period
Cluster 2 contains 108 genes whose functions include amino acid metabolism
which are mainly downregulated after feeding by T
Figure&#xa0;5 Analysis of gene expression patterns related to T
and Cluster 3 were upregulated after feeding by T
The transcription factors in Cluster 1 were mainly bHLH and WRKY
and were significantly upregulated at the TF4 stage
Cluster 2 was significantly upregulated at the TF5 stage
and Cluster 3 was significantly upregulated at the TF2 stage
The transcription factors in Cluster 4 were mainly bZIP and NAC
Cluster 3 and Cluster 1 included the highest numbers
with 107 and 104 upregulated transcription factors
indicating that they play an important role in the resistance of daylily to T
Figure&#xa0;6 Expression of transcription factors
(A) Number of transcription factors; (B) transcription factors&#x2019; expression patterns; (C) expression pattern clustering results
TRINITY_DN6738_c0_g2 plays a major regulatory role in the secondary material synthesis pathway
which influences pest feeding; TRINITY_DN21120_c0_g1 promotes the synthesis of PIs and hinders the digestive function of pests; TRINITY_DN167_c0_g1 regulates nutrient redistribution by plant amino acid metabolism to reduce the nutrients available to pests while ensuring normal plant growth; TRINITY_DN855_c0_g2 regulates defense hormone signaling
three transcription factors were annotated
namely MYB-like DNA-binding domain (TRINITY_DN2391_c0_g1
and regulatory protein NPR1 OS (TRINITY_DN13045_c0_g1
Figure&#xa0;7 Weighted gene co-expression network analysis (WGCNA) of plant defense-related genes
(A) Scale-free network model index under different soft thresholds; (B) gene clustering tree based on the topological dissimilarity matrix; (C) heatmap of correlations between modules and traits; (D) gene co-expression network in the plant defense-related gene module; hub genes are colored pink
Table&#xa0;2 Hub genes and predicted functions
To confirm the reliability of the transcriptome data, seven genes were selected for qRT-PCR verification. Comparison of transcriptome sequencing data and qRT-PCR data indicated very similar expression trends, with a Pearson correlation coefficient (R2) of 0.838 (<a href="#f8">Figure&#xa0;8</a>; <a href="#hsm">Supplementary Figure&#xa0;1</a>)
demonstrating good reliability for the RNA-seq data
Figure&#xa0;8 Correlation point map between RNA-Seq and qRT-PCR expression patterns
PI activity was significantly increased in plants exposed to insect damage
resulting in the obstruction of insect digestion and slow growth
and total phenols content in daylily leaves were significantly higher in plants exposed to insect damage
which were not conducive to colonization by T
WGCNA and DEGs analysis demonstrated that MYB-like DNA-binding domain (TRINITY _ DN2391 _ c0 _ g1
zinc-finger of the FCS-type C2-C2 (TRINITY _ DN21050 _ c0 _ g2)
and regulatory protein NPR1 (TRINITY _ DN13045 _ c0 _ g1
TRINITY _ DN855 _ c0 _ g2) are closely related to the synthesis of anti-stress compounds such as antioxidant enzymes
These results suggest that these genes play an important role in the defense responses of daylily to T
the present findings elucidate the potential mechanism and hub genes of the resistance of daylily to T
and defense enzymes increased the resistance of daylily to T
The mechanisms include reducing the nutrients available to T
catalyzing defense enzymes to produce secondary metabolites that are toxic to T
and other defense hormones signal transduction pathways
improving the resistance of daylily plants
The results of this study expand our the understanding of the mechanisms of insect resistance in daylily
and inform the development of effective strategies to control T
palmi by inducing exogenous factors to enhance insect resistance
The data presented in the study are deposited in the NCBI repository
The author(s) declare that financial support was received for the research
The research was supported by the Development and Promotion of Green Prevention and Control Technology of Hemerocallis citrina Baroni Diseases and Insect Pests (2021YFD1600301&#x2013;5) and Screening of olfactory Attractants for Frankliniella intonsa in Hemerocallis citrina Baroni (2022&#x2013;104)
Wang Chen Zhu for project discussion and manuscript suggestions
The Supplementary Material for this article can be found online at: <a href="https://www.frontiersin.org/articles/10.3389/fpls.2024.1361276/full#supplementary-material">https://www.frontiersin.org/articles/10.3389/fpls.2024.1361276/full#supplementary-material</a>
Herbivory-induced stress: leaf developmental instability is caused by herbivore damage in early stages of leaf development
<a href="https://doi.org/10.1016/j.ecolind.2015.09.036" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=E.+Alves-Silva&amp;author=K.+Del-Claro&amp;publication_year=2016&amp;title=Herbivory-induced%20stress%3A%20leaf%20developmental%20instability%20is%20caused%20by%20herbivore%20damage%20in%20early%20stages%20of%20leaf%20development&amp;journal=Ecol.+Indicators.&amp;volume=61&amp;pages=359" target="_blank">Google Scholar</a>
Infestation of sucking insect pests on five cotton cultivars and their impacts on varietal agronomic traits
An overview of plant defense-related enzymes responses to biotic stresses
<a href="https://doi.org/10.1016/j.plgene.2021.100302" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=M.+Appu&amp;author=P.+Ramalingam&amp;author=A.+Sathiyanarayanan&amp;author=J.+Huang&amp;publication_year=2021&amp;title=An%20overview%20of%20plant%20defense-related%20enzymes%20responses%20to%20biotic%20stresses&amp;journal=Plant+Gene.&amp;volume=27&amp;" target="_blank">Google Scholar</a>
<a name="B4" id="B4"></a> Badenes-P&#xe9;rez
<a href="https://pubmed.ncbi.nlm.nih.gov/35567140/" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.3390/plants11091140" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=F.%20R.+Badenes-P%C3%A9rez&amp;publication_year=2022&amp;title=Plant-insect%20interactions&amp;journal=Plants&amp;volume=11&amp;pages=1140" target="_blank">Google Scholar</a>
Recent advances in plant-insect interactions
<a href="https://pubmed.ncbi.nlm.nih.gov/37511097/" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.3390/ijms241411338" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=F.+Barbero&amp;author=M.%20E.+Maffei&amp;publication_year=2023&amp;title=Recent%20advances%20in%20plant-insect%20interactions&amp;journal=Int.+J.+Mol.+Sci.&amp;volume=24&amp;pages=11338" target="_blank">Google Scholar</a>
Comparative transcriptomic and metabolic analysis of wild and domesticated wheat genotypes reveals differences in chemical and physical defense responses against aphids
Sequestration of plant defense compounds by insects: from mechanisms to insect-plant coevolution
doi:&#xa0;10.1146/annurev-ento-062821-062319
Morphological and biochemical resistance to Helicoverpa armigera (Lepidoptera: Noctuidae) in tomato
<a href="https://doi.org/10.1016/j.cropro.2022.106080" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=K.+Bisht&amp;author=S.%20K.+Yadav&amp;author=S.+Bhowmik&amp;author=N.+Singh&amp;publication_year=2022&amp;title=Morphological%20and%20biochemical%20resistance%20to%20Helicoverpa%20armigera%20%28Lepidoptera%3A%20Noctuidae%29%20in%20tomato&amp;journal=Crop+Protect.&amp;volume=162&amp;" target="_blank">Google Scholar</a>
<a href="https://doi.org/10.1016/j.cropro.2006.10.023" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=R.%20J.%20C.+Cannon&amp;author=I.+Matthews&amp;author=D.%20W.+Collins&amp;publication_year=2007&amp;title=A%20review%20of%20the%20pest%20status%20and%20control%20options%20for%20Thrips%20palmi&amp;journal=Crop+Protect.&amp;volume=26&amp;pages=1089" target="_blank">Google Scholar</a>
Phloem nutrition of detached cabbage leaves varies with leaf age and influences performance of the green peach aphid
Differential transcription and alternative splicing in cotton underly specialized defense responses against pests
Does low nutritional quality act as a plant defence
doi:&#xa0;10.1111/j.0307-6946.2006.00752.x
<a href="https://doi.org/10.1111/j.0307-6946.2006.00752.x" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=T.+Cornelissen&amp;author=P.+Stiling&amp;publication_year=2006&amp;title=Does%20low%20nutritional%20quality%20act%20as%20a%20plant%20defence%3F%20An%20experimental%20test%20of%20the%20slow-growth%2C%20high-mortality%20hypothesis&amp;journal=Ecol.+Entomol.&amp;volume=31&amp;pages=32" target="_blank">Google Scholar</a>
and vector competence of Thrips palmi (Thysanoptera: Thripidae)
Plant secondary metabolites as defense tools against herbivores for sustainable crop protection
Protease inhibitors: An induced plant defense mechanism against herbivores
and molecular control of insect resistance in rice
<a href="https://doi.org/10.1007/s11032-020-1103-3" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=B.+Du&amp;author=R.+Chen&amp;author=J.+Guo&amp;author=G.+He&amp;publication_year=2020&amp;title=Current%20understanding%20of%20the%20genomic%2C%20genetic%2C%20and%20molecular%20control%20of%20insect%20resistance%20in%20rice&amp;journal=Mol.+Breed.&amp;volume=40&amp;pages=24" target="_blank">Google Scholar</a>
RNA-Seq analysis reveals transcript diversity and active genes after common cutworm (Spodoptera litura Fabricius) attack in resistant and susceptible wild soybean lines
Molecular interactions between plants and insect herbivores
doi:&#xa0;10.1146/annurev-arplant-050718-095910
<a href="https://pubmed.ncbi.nlm.nih.gov/30786233/" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.1146/annurev-arplant-050718-095910" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=M.+Erb&amp;author=P.+Reymond&amp;publication_year=2019&amp;title=Molecular%20interactions%20between%20plants%20and%20insect%20herbivores&amp;journal=Annu.+Rev.+Plant+Biol.&amp;volume=70&amp;pages=527" target="_blank">Google Scholar</a>
Plant serpins: potential inhibitors of serine and cysteine proteases with multiple functions
Effects of multiple insect pests on squaring of daylily
doi:&#xa0;10.19518/j.cnki.cn11&#x2013;2531/s.2021.0030
A functional genomics approach to dissect spotted alfalfa aphid resistance in Medicago truncatula
H2O2 and phenolics in resistance of pigeonpea towards Helicoverpa armigera (Hubner)
<a href="https://doi.org/10.1007/s11738-014-1528-6" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=R.+Kaur&amp;author=A.%20K.+Gupta&amp;author=G.%20K.+Taggar&amp;publication_year=2014&amp;title=Role%20of%20catalase%2C%20H2O2%20and%20phenolics%20in%20resistance%20of%20pigeonpea%20towards%20Helicoverpa%20armigera%20%28Hubner%29&amp;journal=Acta+Physiol.+Plant.&amp;volume=36&amp;pages=1513" target="_blank">Google Scholar</a>
Constitutive and herbivore-induced plant defences regulate herbivore population processes
Comparative assessment of trypsin inhibitor vis-&#xe0;-vis kunitz trypsin inhibitor and bowman-birk inhibitor activities in soybean
A reliable and reproducible method for the lipase assay in an AOT/isooctane reversed micellar system: modification of the copper-soap colorimetric method
Integrated transcriptome and metabolome analysis to identify sugarcane gene defense against fall armyworm (Spodoptera frugiperda) herbivory
Effect of l-arginine on maintaining storage quality of the white button mushroom (Agaricus bisporus)
Sensitivity intensified ninhydrin-based chromogenic system by ethanol-ethyl acetate: application to relative quantitation of gaba
Comparative transcriptome analysis of defense response of rice to Nilaparvata lugens and Chilo suppressalis infestation
Small brown planthopper nymph infestation regulates plant defenses by affecting secondary metabolite biosynthesis in rice
Herbivory may mediate the effects of nutrients on the dominance of alien plants
<a href="https://doi.org/10.1111/1365-2435.14019" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=Y.+Li&amp;author=Y.+Gao&amp;author=M.+Van%20Kleunen&amp;author=Y.+Liu&amp;publication_year=2022&amp;title=Herbivory%20may%20mediate%20the%20effects%20of%20nutrients%20on%20the%20dominance%20of%20alien%20plants&amp;journal=Funct.+Ecol.&amp;volume=36&amp;pages=1292" target="_blank">Google Scholar</a>
Phenolic profiles and antioxidant activity in different organs of Sinopodophyllum hexandrum
<a href="https://doi.org/10.3389/fpls.2022.1037582" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=W.+Liu&amp;author=Z.+Zhang&amp;author=T.+Zhang&amp;author=Q.+Qiao&amp;author=X.+Hou&amp;publication_year=2022&amp;title=Phenolic%20profiles%20and%20antioxidant%20activity%20in%20different%20organs%20of%20Sinopodophyllum%20hexandrum&amp;journal=Front.+Plant+Sci.&amp;volume=13&amp;" target="_blank">Google Scholar</a>
<a href="https://pubmed.ncbi.nlm.nih.gov/28545146/" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.1371/journal.pcbi.1005457" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=R.+Lowe&amp;author=N.+Shirley&amp;author=M.+Bleackley&amp;author=S.+Dolan&amp;author=T.+Shafee&amp;publication_year=2017&amp;title=Transcriptomics%20technologies&amp;journal=PloS+Comput.+Biol.&amp;volume=13&amp;" target="_blank">Google Scholar</a>
Defense on multiple fronts: how do plants cope with diverse enemies
doi:&#xa0;10.1016/s1360&#x2013;1385(99)01415&#x2013;6
High frequency in vitro regeneration of adventitious shoots in daylilies (Hemerocallis sp) stem tissue using thidiazuron
Metabolic engineering of plant secondary metabolites: prospects and its technological challenges
<a href="https://doi.org/10.3389/fpls.2023.1171154" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=D.%20A.+Mipeshwaree&amp;author=D.%20K.+Khedashwori&amp;author=D.%20P.+Premi&amp;author=D.%20M.+Lakshmipriyari&amp;publication_year=2023&amp;title=Metabolic%20engineering%20of%20plant%20secondary%20metabolites%3A%20prospects%20and%20its%20technological%20challenges&amp;journal=Front.+Plant+Sci.&amp;volume=14&amp;" target="_blank">Google Scholar</a>
<a name="B37" id="B37"></a> Misiukevi&#x10d;ius
Assessing the genetic diversity of daylily germplasm using ssr markers: implications for daylily breeding
Host plant resistance to thrips (Thysanoptera: Thripidae) - current state of art and future research avenues
Contents of tannins of cultivars of sorghum cultivated in Brazil
as determined by four quantification methods
Genome-wide studies of PAL genes in Sorghum and their responses to aphid infestation
Intergenerational effects of early-life starvation on life history
and transcriptome of a holometabolous insect
Behavioral evidence for local reduction of aphid-induced resistance
<a href="https://pubmed.ncbi.nlm.nih.gov/20345289/" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.1673/031.007.4801" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=E.+Prado&amp;author=W.%20F.+Tjallingii&amp;publication_year=2007&amp;title=Behavioral%20evidence%20for%20local%20reduction%20of%20aphid-induced%20resistance&amp;journal=J.+Insect+Sci.&amp;volume=7&amp;" target="_blank">Google Scholar</a>
Gene expression analysis of different organs and identification of ap2 transcription factors in flax (linum usitatissimum l.)
Effect of elevated CO2 on the interaction between invasive thrips
Role of phenylpropanoids and flavonoids in plant resistance to pests and diseases
Combinations of waste seaweed liquid fertilizer and biochar on tomato (Solanum lycopersicum l.) seedling growth in an acid-affected soil of Jiaodong Peninsula
and whole plant of byttneria herbacea roxb
The effectiveness of physical and chemical defense responses of wild emmer wheat against aphids depends on leaf position and genotype
Dynamics of Zea mays transcriptome in response to a polyphagous herbivore
Thrips advisor: exploiting thrips-induced defences to combat pests on crops
Characterization of induced resistance in tomato plants
doi:&#xa0;10.1111/j.1570-7458.1996.tb00835.x
<a href="https://doi.org/10.1111/j.1570-7458.1996.tb00835.x" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=M.%20J.+Stout&amp;author=S.%20S.+Duffey&amp;publication_year=1996&amp;title=Characterization%20of%20induced%20resistance%20in%20tomato%20plants&amp;journal=Entomol.+Experiment.+Applicata.&amp;volume=79&amp;pages=273" target="_blank">Google Scholar</a>
Transcriptional networks in plant immunity
<a href="https://pubmed.ncbi.nlm.nih.gov/25623163/" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.1111/nph.13286" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=K.+Tsuda&amp;author=I.%20E.+Somssich&amp;publication_year=2015&amp;title=Transcriptional%20networks%20in%20plant%20immunity&amp;journal=New+Phytol.&amp;volume=206&amp;pages=932" target="_blank">Google Scholar</a>
Current opinions about herbivore-associated molecular patterns and plant intracellular signaling
Virus-induced gene silencing of WRKY53 and an inducible phenylalanine ammonia-lyase in wheat reduces aphid resistance
TPMcalculator: one-step software to quantify mrna abundance of genomic features
Spraying chitosan on cassava roots reduces postharvest deterioration by promoting wound healing and inducing disease resistance
An optimized method to obtain high-quality RNA from different tissues in Lilium davidii var
Transcriptomic analysis of tobacco plants in response to whitefly infection
and genome editing for insect pest resistance in field crops
Defensive responses in groundnut against chewing and sap-sucking insects
<a href="https://doi.org/10.1007/s00344-012-9294-4" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=A.%20R.+War&amp;author=M.%20G.+Paulraj&amp;author=S.+Ignacimuthu&amp;author=H.%20C.+Sharma&amp;publication_year=2013&amp;title=Defensive%20responses%20in%20groundnut%20against%20chewing%20and%20sap-sucking%20insects&amp;journal=J.+Plant+Growth+Regulat.&amp;volume=32&amp;pages=259" target="_blank">Google Scholar</a>
Plant defence against herbivory and insect adaptations
<a href="https://doi.org/10.1093/aobpla/ply037" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=A.%20R.+War&amp;author=G.%20K.+Taggar&amp;author=B.+Hussain&amp;author=M.%20S.+Taggar&amp;author=R.%20M.+Nair&amp;author=H.%20C.+Sharma&amp;publication_year=2018&amp;title=Plant%20defence%20against%20herbivory%20and%20insect%20adaptations&amp;journal=AoB+PLANTS.&amp;volume=10&amp;pages=37" target="_blank">Google Scholar</a>
Whiteflies weaponize a plant defense via horizontal gene transfer
<a href="https://pubmed.ncbi.nlm.nih.gov/33798437/" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.1016/j.cell.2021.03.017" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=N.%20K.+Whiteman&amp;author=R.%20L.+Tarnopol&amp;publication_year=2021&amp;title=Whiteflies%20weaponize%20a%20plant%20defense%20via%20horizontal%20gene%20transfer&amp;journal=Cell&amp;volume=184&amp;pages=1657" target="_blank">Google Scholar</a>
Dormancy in Tilia miqueliana is attributable to permeability barriers and mechanical constraints in the endosperm and seed coat
<a href="https://doi.org/10.1007/s40415-021-00749-1" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=Y.+Wu&amp;author=Y.+Shen&amp;publication_year=2021&amp;title=Dormancy%20in%20Tilia%20miqueliana%20is%20attributable%20to%20permeability%20barriers%20and%20mechanical%20constraints%20in%20the%20endosperm%20and%20seed%20coat&amp;journal=Braz.+J.+Bot.&amp;volume=44&amp;pages=725" target="_blank">Google Scholar</a>
Physiological and biochemical response of different resistant alfalfa cultivars against thrips damage
Whole genome wide expression profiles of vitis amurensis grape responding to downy midew by using solexa sequencing technology
Comparative transcriptome analysis provides insights of anti-insect molecular mechanism of Cassia obtusifolia trypsin inhibitor against Pieris rapae
Combined transcriptome and metabolome analysis of alfalfa response to thrips infection
Roles of phytohormones and their signaling pathways in leaf development and stress responses
Insect response to plant defensive protease inhibitors
doi:&#xa0;10.1146/annurev-ento-010814-020816
<a href="https://pubmed.ncbi.nlm.nih.gov/25341101/" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.1146/annurev-ento-010814-020816" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=K.+Zhu-Salzman&amp;author=R.+Zeng&amp;publication_year=2015&amp;title=Insect%20response%20to%20plant%20defensive%20protease%20inhibitors&amp;journal=Annu.+Rev.+Entomol.&amp;volume=60&amp;pages=233" target="_blank">Google Scholar</a>
Liu H and Li J (2024) Physiological responses and transcriptome analysis of Hemerocallis citrina Baroni exposed to Thrips palmi feeding stress
Received: 25 December 2023; Accepted: 01 May 2024;Published: 14 May 2024
Copyright &#xa9; 2024 Sun, Shen, Chen, Ma, Yang, Liu and Li. This is an open-access article distributed under the terms of the <a rel="license" href="http://creativecommons.org/licenses/by/4.0/" target="_blank">Creative Commons Attribution License (CC BY)</a>
*Correspondence: Jie Li, <a id="encmail">bGlqaWUzMDNAeWVhaC5uZXQ=</a>
Volume 13 - 2022 | <a class="ArticleLayoutHeader__info__doi" href="https://doi.org/10.3389/fmicb.2022.773238">https://doi.org/10.3389/fmicb.2022.773238</a>
Thrips palmi (Thysanoptera: Thripidae) is the predominant tospovirus vector in Asia-Pacific region
It transmits economically damaging groundnut bud necrosis virus (GBNV
family Tospoviridae) in a persistent propagative manner
and virus reservoirs making tospovirus management very challenging
Insecticides and host plant resistance remain ineffective in managing thrips&#x2013;tospoviruses
Recent genomic approaches have led to understanding the molecular interactions of thrips&#x2013;tospoviruses and identifying novel genetic targets
most of the studies are limited to Frankliniella species and tomato spotted wilt virus (TSWV)
Amidst the limited information available on T
the present study is the first report of the transcriptome-wide response of T
The differential expression analyses of the triplicate transcriptome of viruliferous vs
palmi identified a total of 2,363 (1,383 upregulated and 980 downregulated) significant transcripts
The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses showed the abundance of differentially expressed genes (DEGs) involved in innate immune response
and receptor binding and signaling that mediate the virus invasion and multiplication in the vector system
the gene regulatory network (GRN) of most significant DEGs showed the genes like ABC transporter
were enriched with higher degrees of interactions
the expression of the candidate genes in response to GBNV infection was validated in reverse transcriptase-quantitative real-time PCR (RT-qPCR)
This study leads to an understanding of molecular interactions between T
palmi and GBNV and suggests potential genetic targets for generic pest control
Several management strategies have been adopted around the world
with insecticides and host plant resistance being the core components for thrips and tospovirus management
Farmers worldwide spend more than US$ 50&#x2009;million annually to manage this vector
Insecticides are largely ineffective due to the emergence of resistant thrips population
Genetic host plant resistance is not available for all the crop species that are infected by thrips&#x2013;tospoviruses
Understanding the interactions between thrips and tospovirus will be helpful to identify novel genetic targets to interrupt the interrelationship and restrict the epidemics of thrips&#x2013;tospoviruses
Little is known about the differentially expressed genes (DEGs)
The present study reports the candidate genes of T
palmi responsive to GBNV infection through a comparative transcriptomic approach that would be novel targets for generic management of the thrips&#x2013;tospovirus complex
An isofemale population of T. palmi maintained at Advanced Centre for Plant Virology, Indian Agricultural Research Institute (IARI), New Delhi, was used in the study. The identification of T. palmi was based on the morphological keys and mitochondrial cytochrome oxidase subunit I (mtCOI) sequences amplified in PCR using primer pairs, LCO 1490 and HCO 2198 (<a href="#ref19">Folmer et al., 1994</a>; <a href="#ref36">Jangra et al., 2020</a>)
The population was established from a single adult female of T
The population has been maintained at the thrips rearing facility under controlled environmental conditions at 28&#x2009;&#x000B1;&#x2009;1&#x000B0;C temperature
and 16&#x2009;h of light&#x2013;8&#x2009;h of the dark since 2018
Fresh virus-free healthy eggplants were supplied as and when required
The isofemale population has been used in replicate throughout the experiment
The total RNA was isolated from cowpea plants using RNeasy Plant Mini Kit (Qiagen
Complementary DNA (cDNA) was synthesized with random primers using FIREScript RT cDNA synthesis kit (Solis Biodyne
The reaction mixture contained 1X RT reaction buffer
The reverse transcription was carried out in a T100 thermocycler (Bio-Rad
United States) by primer annealing at 25&#x000B0;C for 10&#x2009;min
reverse transcription at 50&#x000B0;C for 30&#x2009;min and followed by enzyme inactivation at 85&#x000B0;C for 5&#x2009;min
PCR was carried out in 25&#x2009;&#x03BC;l reactions containing 2&#x2009;&#x03BC;l cDNA
2.5&#x2009;&#x03BC;l 10X PCR buffer (Thermo Fischer Scientific)
0.4&#x2009;&#x03BC;M each of forward and reverse primers
260&#x2009;&#x03BC;M dNTP mix (Thermo Fischer Scientific)
and 2&#x2009;U DreamTaq polymerase (Thermo Fisher Scientific)
PCR was performed in a T100 Thermal Cycler with 95&#x000B0;C for 5&#x2009;min followed by 30&#x2009;cycles of 94&#x000B0;C for 30&#x2009;s
and 72&#x000B0;C for 1&#x2009;min followed by a final extension at 72&#x000B0;C for 7&#x2009;min
RT-PCR products were resolved on 1% agarose gel stained with GoodView (BR Biochem
India) and observed in a gel documentation system (MasteroGen Inc.
The purified product was sequenced bidirectional
and BLASTn was performed to check the species homology
The consensus sequence was submitted to GenBank
The freshly emerged first instar larvae (L1, less than 24&#x2009;h old) of T. palmi were collected using a Camel hairbrush from the isofemale population on eggplants and used for GBNV acquisition in three replicates to generate the viruliferous population. The virus acquisition setup was established as described by <a href="#ref23">Ghosh et al. (2021)</a>
palmi L1s in each replicate were placed on a GBNV-infected leaf and allowed to feed for 24&#x2009;h at 28&#x2009;&#x000B1;&#x2009;1&#x000B0;C and 60&#x2009;&#x000B1;&#x2009;10% relative humidity
The setup was continuously monitored to confirm the feeding of larvae on the leaf
the larvae were transferred on a detached healthy cowpea leaf
The petiole of the detached leaf was inserted in a slant of 0.8% dextrose agar within an insect breeding dish (10&#x2009;cm diameter
palmi were reared up to the adult stage on cowpea leaves at 28&#x2009;&#x000B1;&#x2009;1&#x000B0;C
and 16&#x2009;h of light&#x2013;8&#x2009;h of dark
The breeding dishes were regularly monitored and supplied with fresh leaves as and when required
the L1s were released on the healthy cowpea leaves placed on the agar slant within the insect breeding dishes in three replicates as described above
Adults were collected immediately after their emergence and used in the study
Three sets of viruliferous (designated as TpTrI1
One part was used for transcriptome sequencing and another part was preserved at &#x02212;80&#x000B0;C for gene expression analysis in reverse transcriptase-quantitative real-time PCR (RT-qPCR)
palmi was confirmed by randomly collecting five individual adults from each of the viruliferous and nonviruliferous insect populations and testing in RT-PCR
palmi was isolated using NucleoSpin RNA XS (Macherey-Nagel
and RT-PCR was performed as described above
palmi were subjected to total RNA isolation using TRIzol (Invitrogen
Each set comprised 10 adult individuals that included both females and males (1:1)
RNA quality was checked using RNA 6000 Nano Kit (Agilent Technologies
United States) on 2100 Bioanalyzer (Agilent Technologies) with a minimum RNA Integrity Number (RIN) value of 7
RNA concentrations were determined with a NanoDrop ND-8000 spectrophotometer (Thermo Fischer Scientific)
RNA-Seq libraries for all samples were prepared using NEBNext UltraII RNA library preparation kit for Illumina (New England Biolabs
and sequencing was done in a single HiSEQ 4,000 (Illumina
United States) lane using 150&#x2009;bp paired-end chemistry
The library preparation and sequencing were done by commercial service providers (NxGen Bio Life Sciences
total RNA was used to purify poly(A) messenger RNA (mRNA) using oligo-dT beads
Magnetic beads were used for two rounds of purification
During the second elution of the poly-A RNA
the RNA was also fragmented into 200&#x2013;500&#x2009;bp pieces in the presence of divalent cations at 94&#x000B0;C for 5&#x2009;min using an ultrasonicator
The cleaved RNA fragments were copied into first-strand cDNA using SuperScript-II Reverse Transcriptase (Thermo Fischer Scientific) and random primers
The products were purified and enriched with PCR to create the final cDNA library
The tagged cDNA libraries were pooled in equal ratios and used for 2&#x2009;&#x000D7;&#x2009;150&#x2009;bp paired-end sequencing on a single lane of the Illumina HiSeq4000
Illumina clusters were generated and were loaded onto Illumina Flow Cell on Illumina HiSeq 4000 instrument and sequencing was carried out
the samples were demultiplexed and the indexed adapter sequences were trimmed using the CASAVA v1.8.2 software (Illumina
palmi with corrected value of p&#x2009;&#x0003C;&#x2009;0.05
KEGG pathway enrichment was performed by calculating the enrichment factor for all the identified KEGG pathways
The top 20 highly enriched pathways were plotted based on the rich factor
The top 20 genes each from up- and downregulated DEGs were selected for the construction of gene regulatory networks (GRNs). These DEGs were filtered based on the log fold change values. The network was analyzed and visualized using Cytoscape v3.7.2, where gene correlation was computed using the Pearson&#x2019;s correlation coefficient of the normalized expression values (<a href="#ref65">Shannon et al., 2003</a>)
Network Analyzer plug-in was used to estimate the network centrality and topology
The &#x0394;CT was calculated as CT of targeted gene&#x2014;CT of endogenous control
&#x0394;&#x0394;CT value was calculated as average &#x0394;CT of nonviruliferous replicates&#x2014;&#x0394;CT of each replicate
The average 2-(&#x0394;&#x0394;CT) was calculated for both the viruliferous and nonviruliferous populations and transformed to log2 2-(&#x0394;&#x0394;CT)
The log2-fold change of the target gene expression in viruliferous T
palmi population was estimated by normalizing the fold change values of the nonviruliferous population
The relative expressions of target genes post-GBNV exposure in RT-qPCR assay were compared with log2-fold changes obtained through RNA-Seq analysis
and Pearson&#x2019;s correlation coefficient was calculated using CORREL function in MS Excel
The nucleotide sequence of the amplified product showed 100% homology to other T
The sequence can be retrieved from the GenBank with the Accession number MW020349
The initial inoculum of GBNV was collected from the pure culture (tomato isolate) maintained at Advanced Centre for Plant Virology
Healthy cowpea plants (Vigna unguiculata var
Characteristic symptoms of GBNV were recorded on sap-inoculated cowpea plants 10&#x2013;14&#x2009;days post-inoculation (dpi)
Local lesions appeared on inoculated leaves and yellow necrotic spots with multiple rings-like structures were visible on systemic leaves
RT-PCR with GBNV-specific primers produced an amplicon of 1.8&#x2009;kb for all symptomatic samples that confirmed the GBNV infection in inoculated plants
No amplification was observed in RT-PCR for healthy plant samples
the nucleotide sequence of the amplified product showed more than 97% identity to GBNV
The sequence can be retrieved from the GenBank with Accession No
and TpTrI3) exposed to GBNV-infected cowpea plants during the L1 stage were tested in RT-PCR
A 1.8&#x2009;kb product was visualized on agarose gel that confirmed the infection of GBNV in T
and TpTrH3) that were exposed to healthy cowpea plants did not produce any GBNV-specific amplification in RT-PCR
palmi under the stringent filtering criteria in response to GBNV-infection
and ubiquitin-like containing PHD and RING finger domains 1 (UHRF-1)-binding protein 1
were found highly upregulated after exposure to GBNV
Significant downregulations of ABC transporter
elongation of very-long-chain fatty acid (ELOVL)
tyrosine&#x2013;protein kinase receptor Tie-1 (TIE1)
and endocuticle structural glycoprotein SgAbd-2-like
The proportion of differentially expressed genes (DEGs) in RNA-Seq analysis
(A) A total of 1,383 DEGs were found upregulated while 980 were downregulated
The x axis shows the fold change in gene expression between different samples
and the y axis shows the statistical significance of the differences
Significantly up- and downregulated genes with log2 FC&#x2009;&#x02265;&#x2009;2 are highlighted in green
The black dots represent insignificant differentially expressed genes
Gene ontology (GO) enrichment analysis of differentially expressed genes (DEGs)
(A) DEGs were characterized under cellular components
and biological processes on the basis of GO analysis
GO of differential expressed (B) up- and (C) downregulated genes of viruliferous vs
and blue bars represent molecular functions
The pathway enrichment of DEGs showed the DEGs were significantly enriched in ascorbate and aldarate metabolism (map00053)
and metabolism of xenobiotics by cytochrome P450 (map00980) pathways
The pathways identified in GO enrichment analysis were consistent with the findings of the KEGG pathway study
Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs of Thrips palmi in response to groundnut bud necrosis virus (GBNV) infection
The rich factor is defined as the ratio of the number of DEGs annotated in a pathway to the number of all genes annotated in this pathway
Y axis indicates the pathway name; x axis indicates the enriched factor in each of the pathways
The bubble size indicates the number of DEGs
The color bar indicates the corrected value of p
A total of 40 nodes comprised the top up- and downregulated DEGs were considered in network analysis. The degree and betweenness centrality in the interaction network of the top DEGs were calculated. Interaction networks corresponding with the highly regulated DEGs demonstrated a total of 438 edges. The degree of certain nodes was markedly higher compared with the average degree in the networks (<a href="#fig4">Figure 4</a>)
The upregulated DEGs with significantly higher degrees were calphotin-like isoform X2
cAMP-specific 3&#x2032;,5&#x2032;-cyclic phosphodiesterase isoform X1
phosphoribosylformylglycinamidine synthase (PFAS)
a group of DEGs associated with cuticle development such as larval cuticle protein A2B-like
endocuticle structural glycoprotein SgAbd-2-like
and ABC transporter F family member 4-like were with higher degrees of interactions
proline-rich protein HaeIII subfamily 1-like
alpha-tocopherol transfer protein (TTPA)-like
and dynamin-1-like protein showed significant interactions with higher degrees
and blue squares represent downregulated DEGs
The relative expression of target genes was estimated by a 2&#x02212;&#x0394;&#x0394;CT method. The variation among the biological replicates was normalized with respect to the CT value of endogenous control, &#x03B2;-tubulin. Among the selected highly expressed genes, the expression of UHRF1-binding protein 1 of T. palmi was upregulated by log2 5.5-fold in RT-qPCR in response to GBNV infection (<a href="#fig5">Figure 5</a>)
mRNA expression of nephrin was upregulated by log2 4.8-fold in viruliferous T
palmi adults in comparison to nonviruliferous adults
and GABA receptor were upregulated by log2 3.37
the expression of serpin was downregulated by log2 8.5-fold
log2 4.3-fold downregulation of tyrosin kinase was recorded in RT-qPCR in response to GBNV infection
Expression of ABC transporter F family member 4-like gene was also downregulated by log2 3.57-fold
while it was log2 1.65-fold downregulations for ARRDC3 in viruliferous T
Some other genes such as elongation of fatty acid chain
Expression of Thrips palmi putative genes in response to GBNV infection in RNA-Seq and reverse transcriptase-quantitative real-time PCR (RT-qPCR)
The values of log2-fold changes calculated in RNA-Seq analysis were in accordance with the RT-qPCR fold change values
Pearson&#x2019;s correlation coefficient value was 0.81 as calculated using the CORREL function in MS Excel
The fold change values estimated from RNA-Seq were plotted against fold change values obtained in RT-qPCR assays of the selected genes (<a href="#fig5">Figure 5</a>). The Pearson&#x2019;s correlation coefficient of 0.81 indicated the acceptability of DEGs through RNA-Seq (<a href="#SM1">Supplementary Table 3</a>)
The next generation of thrips again needs to acquire the virus during their early larval stage
palmi adults in response to GBNV infection
Pathological effects such as reduced survival and adult longevity of T
palmi were also recorded post-GBNV exposure in the present study
The role of PFAS in triggering such pathological effects in T
palmi may be worth for further investigation
CYP-mediated metabolism of xenobiotics pathway was also enriched in viruliferous T
An in-depth study of the mechanism involved would be necessary to shed further light on the role of GABA receptor and CYP in tospovirus replication in T
The innate differences among the thrips species
and virus isolates could be the reason behind the variation in response
Inhibition of ELOVL prevents the loss of plasma membrane integrity and cell death
palmi helps maintain the integrity of cells and supports viral growth
the molecular mechanisms underlying the downregulation of these genes in response to GBNV infection are not known and need in-depth study
These candidate genes would be potential targets for novel generic pest control
and other enzymes-mediated metabolism of xenobiotics
and mannose metabolism were highly enriched
GRN analysis showed that DEGs associated with innate immune response
and endocytotic pathways are involved in possible manipulation in host invasion by GBNV
we have assembled a whole-body transcriptome of adult T
palmi and reported the DEGs in response to GBNV infection
The majority of DEGs are involved in innate immunity
and endocytosis that facilitate the invasion
palmi that are highly regulated in response to GBNV infection would be potential targets for the effective management of tospoviruses
data generated in this study will enrich genomic information of thrips and will enable functional studies
The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: <a href="https://www.ncbi.nlm.nih.gov/">https://www.ncbi.nlm.nih.gov/</a>
and SC conceived and designed the research
and Priti carried out the wet laboratory experiments and wrote the draft manuscript
AG and VB reviewed the results and edited the final manuscript
The scholarship of DM was supported by Indian Council of Agricultural Research
This is a part of thesis work of DM submitted to Post Graduate School
The initial virus culture was supplied by Basavaraj YB (IARI)
The authors are thankful to Heena (IARI) for her help in maintenance of virus and insect culture
The Supplementary Material for this article can be found online at: <a href="https://www.frontiersin.org/articles/10.3389/fmicb.2022.773238/full#supplementary-material">https://www.frontiersin.org/articles/10.3389/fmicb.2022.773238/full#supplementary-material</a>
1. <a class="footnotetextanchor" href="#xfn0004" title="">^</a><a href="https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/">https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/</a>
2. <a class="footnotetextanchor" href="#xfn0005" title="">^</a><a href="http://www.bioconductor.org/packages/release/bioc/html/DESeq.html">http://www.bioconductor.org/packages/release/bioc/html/DESeq.html</a>
3. <a class="footnotetextanchor" href="#xfn0006" title="">^</a><a href="http://geneontology.org/">http://geneontology.org/</a>
4. <a class="footnotetextanchor" href="#xfn0007" title="">^</a><a href="https://www.ebi.ac.uk">https://www.ebi.ac.uk</a>
5. <a class="footnotetextanchor" href="#xfn0008" title="">^</a><a href="http://www.genome.jp/kegg">http://www.genome.jp/kegg</a>
<a name="ref2" id="ref2"></a>Badillo-Vargas
Proteomic analysis of Frankliniella occidentalis and differentially expressed proteins in response to tomato spotted wilt virus infection
Interaction of tomato spotted wilt tospovirus (TSWV) glycoproteins with a thrips midgut protein
Herbivore benefits from vectoring plant virus through reduction of period of vulnerability to predation
Functional analysis of the ATP-binding cassette (ABC) transporter gene family of Tribolium castaneum
and viral attachment: true receptors or adaptation bias
Present situation of Thrips palmi Karny (Thysanoptera: Thripidae) in Venezuela
<a href="http://scholar.google.com/scholar_lookup?author=M.+Cermeli&#x00026;author=A.+Montagne&#x00026;publication_year=1993&#x00026;title=Present+situation+of+Thrips+palmi+Karny+(Thysanoptera:+Thripidae)+in+Venezuela&#x00026;journal=Manejo+Integr.+Plagas.&#x00026;volume=29&#x00026;pages=22-23" target="_blank">Google Scholar</a>
A novel Laccase gene from Litopenaeus vannamei is involved in the immune responses to pathogen infection and oxidative stress
<a name="ref11" id="ref11"></a>Christianson
Heparan sulfate proteoglycan as a cell-surface endocytosis receptor
Heparan sulfate proteoglycan: An arbovirus attachment factor integral to mosquito salivary gland ducts
Blast2GO: a comprehensive suite for functional analysis in plant genomics
Impaired immunological synapse in sperm associated antigen 6 (SPAG6) deficient mice
Infestation of the southern yellow thrips Thrips palmi
<a href="http://scholar.google.com/scholar_lookup?author=B.+Cooper&#x00026;publication_year=1991&#x00026;title=Infestation+of+the+southern+yellow+thrips+Thrips+palmi+in+vegetables&#x00026;journal=J.+Agri.+Soc.+Trinidad+Tobago&#x00026;volume=88&#x00026;pages=37-38" target="_blank">Google Scholar</a>
and reproduction in western flower thrips (Thysanoptera: Thripidae) exposed to impatiens necrotic spot virus
The &#x03B1;-arrestin ARRDC3 mediates ALIX ubiquitination and G protein-coupled receptor lysosomal sorting
DNA primers for amplification of mitochondrial cytochrome oxidase subunit I from diverse metazoan invertebrates
<a href="http://scholar.google.com/scholar_lookup?author=O.+Folmer&#x00026;author=M.+Black&#x00026;author=W.+Hoeh&#x00026;author=R.+Lutz&#x00026;author=R.+Vrijenhoek&#x00026;publication_year=1994&#x00026;title=DNA+primers+for+amplification+of+mitochondrial+cytochrome+oxidase+subunit+I+from+diverse+metazoan+invertebrates&#x00026;journal=Mol.+Mar.+Biol.+Biotechnol.&#x00026;volume=5&#x00026;pages=294-299" target="_blank">Google Scholar</a>
&#x201C;Thrips as the vectors of tospoviruses in Indian agriculture,&#x201D; in A Century of Plant Virology in India
<a href="http://scholar.google.com/scholar_lookup?publication_year=2017&#x00026;title=Thrips+as+the+vectors+of+tospoviruses+in+Indian+agriculture&#x00026;journal=A+Century+of+Plant+Virology+in+India.&#x00026;pages=537-561" target="_blank">Google Scholar</a>
Genetics of Thrips palmi (Thysanoptera: Thripidae)
<a href="https://doi.org/10.1007/s10340-019-01160-2" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=A.+Ghosh&#x00026;author=S.+S.+Jagdale&#x00026;author=Y.+B.+Basavaraj&#x00026;author=R.+G.+Dietzgen&#x00026;author=R.+K.+Jain&#x00026;publication_year=2020&#x00026;title=Genetics+of+Thrips+palmi+(Thysanoptera:+Thripidae)&#x00026;journal=J.+Pest.+Sci.&#x00026;volume=93&#x00026;pages=27-39" target="_blank">Google Scholar</a>
Transcriptome changes in Culex quinquefasciatus (Diptera: Culicidae) salivary glands during West Nile virus infection
<a name="ref25" id="ref25"></a>Gonz&#x000E1;lez-Santoyo
Phenoloxidase: a key component of the insect immune system
<a href="https://doi.org/10.1111/j.1570-7458.2011.01187.x" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=I.+Gonz&#x000E1;lez-Santoyo&#x00026;author=A.+C&#x000F3;rdoba-Aguilar&#x00026;publication_year=2012&#x00026;title=Phenoloxidase:+a+key+component+of+the+insect+immune+system&#x00026;journal=Entomol.+Exp.+Appl.&#x00026;volume=142&#x00026;pages=1-16" target="_blank">Google Scholar</a>
&#x201C;Insect symbiotic bacterial GroEL (chaperonin 60) and plant virus transmission,&#x201D; in Moonlighting Cell Stress Proteins in Microbial Infections
<a href="http://scholar.google.com/scholar_lookup?author=R.+Gorovits&#x00026;author=H.+Czosnek&#x00026;publication_year=2013&#x00026;title=Insect+symbiotic+bacterial+GroEL+(chaperonin+60)+and+plant+virus+transmission&#x00026;journal=Moonlighting+Cell+Stress+Proteins+in+Microbial+Infections.+Heat+Shock+Proteins.&#x00026;volume=7&#x00026;pages=173-187" target="_blank">Google Scholar</a>
The involvement of heat shock proteins in the establishment of tomato yellow leaf curl virus infection
<a name="ref28" id="ref28"></a>G&#x000F6;tz
Implication of Bemisia tabaci heat shock protein 70 in Begomovirus-whitefly interactions
Chromosome-level assembly of the melon thrips genome yields insights into evolution of a sap-sucking lifestyle and pesticide resistance
Review and first observation in Guadeloupe on Thrips Palmi Karny
<a href="https://doi.org/10.1051/agro:19880701" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=J.+Guyot&#x00026;publication_year=1998&#x00026;title=Review+and+first+observation+in+Guadeloupe+on+Thrips+Palmi+Karny&#x00026;journal=Agronomie&#x00026;volume=8&#x00026;pages=565-575" target="_blank">Google Scholar</a>
Integration of transcriptomics and network analysis reveals co-expressed genes in Frankliniella occidentalis larval guts that respond to tomato spotted wilt virus infection
Molecular characterization and expression of laccase genes in the salivary glands of the green rice leafhopper
Nephotettix cincticeps (Hemiptera: Cicadellidae)
Modulation of potassium channels inhibits bunyavirus infection
Plant viruses alter insect behavior to enhance their spread
In silico analyses of molecular interactions between groundnut bud necrosis virus and its vector
A multiplex PCR assay for rapid identification of major tospovirus vectors reported in India
Population trends of a newly introduced species
on commercial watermelon plantings in Hawaii
<a href="https://doi.org/10.1093/jee/79.3.718" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=M.+W.+Johnson&#x00026;publication_year=1986&#x00026;title=Population+trends+of+a+newly+introduced+species+Thrips+palmi+(Thysanoptera:+Thripidae)+on+commercial+watermelon+plantings+in+Hawaii&#x00026;journal=J.+Econ.+Entomol.&#x00026;volume=79&#x00026;pages=718-720" target="_blank">Google Scholar</a>
Evaluation of the function of a type I peritrophic matrix as a physical barrier for midgut epithelium invasion by mosquito-borne pathogens in Aedes aegypti
Population management of Thrips palmi Karny
<a href="https://doi.org/10.1303/jjaez.2001.39" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=A.+Kawai&#x00026;publication_year=2001&#x00026;title=Population+management+of+Thrips+palmi+Karny&#x00026;journal=Jpn.+J.+Appl.+Entomol.+Zool.&#x00026;volume=45&#x00026;pages=39-59" target="_blank">Google Scholar</a>
Receptor tyrosine kinase inhibitors block multiple steps of influenza A virus replication
Importance and genetic diversity of vegetable-infecting tospoviruses in India
Constitutive activation of toll-mediated antifungal defense in serpin-deficient drosophila
The role of peritrophic membrane in the resistance of Anticarsia Gemmatalis larvae (Lepidoptera: Noctuidae) during the infection by its nucleopolyhedrovirus (AgMNPV)
clone sequences and assembly contigs with BWA-MEM
<a href="https://doi.org/10.6084/M9.FIGSHARE.963153.V1" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=H.+Li&#x00026;publication_year=2013&#x00026;" target="_blank">Google Scholar</a>
Fast and accurate short read alignment with burrows-wheeler transform
<a href="https://pubmed.ncbi.nlm.nih.gov/19451168" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.1093/bioinformatics/btp324" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=H.+Li&#x00026;author=R.+Durbin&#x00026;publication_year=2009&#x00026;title=Fast+and+accurate+short+read+alignment+with+burrows-wheeler+transform&#x00026;journal=Bioinformatics&#x00026;volume=25&#x00026;pages=1754-1760" target="_blank">Google Scholar</a>
Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method
Protein interactome of the deamidase phosphoribosylformylglycinamidine synthetase (PFAS) by LC-MS/MS
Calphotin: a Drosophila photoreceptor cell calcium-binding protein
<a href="https://doi.org/10.1073/pnas.90.4.1531" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=J.+H.+Martin&#x00026;author=S.+Benzer&#x00026;author=M.+Rudnicka&#x00026;author=C.+A.+Miller&#x00026;publication_year=1993&#x00026;title=Calphotin:+a+Drosophila+photoreceptor+cell+calcium-binding+protein&#x00026;journal=Proc.+Natl.+Acad.+Sci.&#x00026;volume=90&#x00026;pages=1531-1535" target="_blank">Google Scholar</a>
Thripidae) in the state of S&#x000E3;o Paulo
<a href="http://scholar.google.com/scholar_lookup?author=R.+C.+Monteiro&#x00026;author=R.+A.+Zucchi&#x00026;author=L.+A.+Mound&#x00026;publication_year=1995&#x00026;title=Record+of+Thrips+palmi+Karny+1925+(Thysanoptera+Thripidae)+in+the+state+of+S&#x000E3;o+Paulo+Brazil&#x00026;journal=Revue+de+l&#x2019;Agri.&#x00026;volume=70&#x00026;pages=53-55" target="_blank">Google Scholar</a>
Transmission mode of watermelon silver mottle virus by Thrips palmi
Replication of tomato spotted wilt virus after ingestion by adult Thrips setosus is restricted to midgut epithelial cells
The genus Tospovirus: emerging bunyaviruses that threaten food security
doi: 10.1146/annurev-virology-100114-055036 
<a name="ref51" id="ref51"></a>O&#x2019;Neal
ATP-sensitive inwardly rectifying potassium channel regulation of viral infections in honey bees
Thrips (Thysanoptera) from Pakistan to the Pacific: a review
<a href="http://scholar.google.com/scholar_lookup?author=J.+M.+Palmer&#x00026;publication_year=1992&#x00026;title=Thrips+(Thysanoptera)+from+Pakistan+to+the+Pacific:+a+review&#x00026;journal=Bull.+Br.+Mus.+Nat.+Hist.+Entomol.+Ser.&#x00026;volume=61&#x00026;pages=1-76" target="_blank">Google Scholar</a>
Thrips palmi (Thysanoptera: Thripidae): a new insect pest for Puerto Rico
<a href="http://scholar.google.com/scholar_lookup?author=A.+Pantoja&#x00026;author=A.+Segarra&#x00026;author=H.+Ruiz&#x00026;author=S.+Medina-Gaud&#x00026;publication_year=1988&#x00026;title=Thrips+palmi+(Thysanoptera:+Thripidae):+a+new+insect+pest+for+Puerto+Rico&#x00026;journal=J.+Agric.+Univ.+P.+R.&#x00026;volume=2&#x00026;pages=327-329" target="_blank">Google Scholar</a>
Global status of Tospovirus epidemics in diverse cropping systems: successes achieved and challenges ahead
Very long-chain fatty acids are functionally involved in necroptosis
An expanded family of arrestins regulate metabolism
<a href="https://pubmed.ncbi.nlm.nih.gov/22520962" target="_blank">PubMed Abstract</a> | <a href="https://doi.org/10.1016/j.tem.2012.03.003" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=P.+Patwari&#x00026;author=R.+T.+Lee&#x00026;publication_year=2012&#x00026;title=An+expanded+family+of+arrestins+regulate+metabolism&#x00026;journal=Trends+Endocrinol.+Metab.&#x00026;volume=23&#x00026;pages=216-222" target="_blank">Google Scholar</a>
Phosphorylation of Nephrin triggers its internalization by raft-mediated endocytosis
<a href="https://doi.org/10.1603/IPM10020" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=D.+G.+Riley&#x00026;author=S.+V.+Joseph&#x00026;author=R.+Srinivasan&#x00026;author=S.+Diffie&#x00026;publication_year=2011&#x00026;title=Thrips+vectors+of+tospoviruses&#x00026;journal=J.+Integr.+Pest+Manag.&#x00026;volume=2&#x00026;pages=I1-I10" target="_blank">Google Scholar</a>
De novo transcriptomics approach reveals genes involved in Thrips Tabaci resistance to spinosad
Thrips developmental stage-specific transcriptome response to tomato spotted wilt virus during the virus infection cycle in Frankliniella occidentalis
The ESCRT-associated protein Alix recruits the ubiquitin ligase Nedd4-1 to facilitate HIV-1 release through the LYPXnL L domain motif
Cytoscape: a software environment for integrated models of biomolecular interaction networks
Transcriptome changes associated with tomato spotted wilt virus infection in various life stages of its thrips vector
Direct and indirect effects of a thrips-transmitted Tospovirus on the preference and fitness of its vector
&#x201C;Status and control strategy of peanut bud necrosis disease in Uttar Pradesh.&#x201D; in Recent Studies on Peanut Bud Necrosis Disease: Proceedings of a Meeting
India: International Crop Research Institute for Semi-Arid Tropics
<a href="http://scholar.google.com/scholar_lookup?author=A.+B.+Singh&#x00026;author=S.+K.+Srivastava&#x00026;publication_year=1995&#x00026;" target="_blank">Google Scholar</a>
Assessment of yield losses due to potato stem necrosis disease
<a href="http://scholar.google.com/scholar_lookup?author=R.+B.+Singh&#x00026;author=K.+K.+Srivastava&#x00026;author=S.+K.+Pandey&#x00026;author=S.+M.+Khurana&#x00026;publication_year=1997&#x00026;title=Assessment+of+yield+losses+due+to+potato+stem+necrosis+disease&#x00026;journal=Indian+J.+Virol.&#x00026;volume=13&#x00026;pages=135-137" target="_blank">Google Scholar</a>
Interaction between intracellular bacterial pathogens and host cell mitochondria
<a name="ref71" id="ref71"></a>Stafford-Banks
Analysis of the salivary gland transcriptome of Frankliniella occidentalis
Ebola virus triggers receptor tyrosine kinase-dependent signaling to promote the delivery of viral particles to entry-conducive intracellular compartments
Vibrio cholerae T3SS effector VopE modulates mitochondrial dynamics and innate immune signaling by targeting Miro GTPases
&#x201C;Thrips-tomato spotted wilt virus interactions: morphological
behavioral and cellular components influencing thrips transmission,&#x201D; in Advances in Disease Vector Research 9
<a href="http://scholar.google.com/scholar_lookup?publication_year=1992&#x00026;title=Thrips-tomato+spotted+wilt+virus+interactions:+morphological+behavioral+and+cellular+components+influencing+thrips+transmission&#x00026;journal=Advances+in+Disease+Vector+Research&#x00026;volume=9&#x00026;pages=195-240" target="_blank">Google Scholar</a>
<a href="https://doi.org/10.1016/S0065-2296(02)36061-0" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=D.+Ullman&#x00026;author=R.+Meideros&#x00026;author=L.+R.+Campbell&#x00026;author=A.+E.+Whitfield&#x00026;author=J.+L.+Sherwood&#x00026;author=T.+L.+German&#x00026;publication_year=2002&#x00026;title=Thrips+as+vectors+of+tospoviruses&#x00026;journal=Adv.+Bot.+Res.&#x00026;volume=36&#x00026;pages=113-140" target="_blank">Google Scholar</a>
&#x201C;Thrips as vectors of plant pathogens,&#x201D; in Thrips as Crop Pests
<a href="http://scholar.google.com/scholar_lookup?publication_year=1997&#x00026;title=Thrips+as+vectors+of+plant+pathogens&#x00026;journal=Thrips+as+Crop+Pests.&#x00026;pages=539-565" target="_blank">Google Scholar</a>
&#x201C;Pest status,&#x201D; in Thrips palmi: A Literature Survey
International Institue of Biological Control
UK: International Institute of Biological Control)
<a href="http://scholar.google.com/scholar_lookup?publication_year=1992&#x00026;title=Pest+status&#x00026;journal=Thrips+palmi:+A+Literature+Survey+International+Institue+of+Biological+Control.&#x00026;pages=3-6" target="_blank">Google Scholar</a>
Tomato spotted wilt tospovirus ingestion by first instar larvae of Frankliniella occidentalis is a prerequisite for transmission
<a href="https://doi.org/10.1094/Phyto-86-900" target="_blank">CrossRef Full Text</a> | <a href="http://scholar.google.com/scholar_lookup?author=F.+Wetering&#x00026;author=R.+Goldbach&#x00026;author=D.+Peters&#x00026;publication_year=1996&#x00026;title=Tomato+spotted+wilt+tospovirus+ingestion+by+first+instar+larvae+of+Frankliniella+occidentalis+is+a+prerequisite+for+transmission&#x00026;journal=Phytopathology&#x00026;volume=86&#x00026;pages=900-905" target="_blank">Google Scholar</a>
doi: 10.1146/annurev.phyto.43.040204.140017 
Transcriptome-wide responses of adult melon thrips (Thrips palmi) associated with capsicum chlorosis virus infection
Propagation of tomato spotted wilt virus in Frankliniella occidentalis does neither result in pathological effects nor in transovarial passage of the virus
Dengue virus impairs mitochondrial fusion by cleaving mitofusins
Smoking-associated DNA methylation biomarkers and their predictive value for all-cause and cardiovascular mortality
De novo transcriptome sequencing in Frankliniella occidentalis to identify genes involved in plant virus transmission and insecticide resistance
Expression of heat shock protein genes in insect stress responses
<a href="http://scholar.google.com/scholar_lookup?author=L.+Zhao&#x00026;author=W.+A.+Jones&#x00026;publication_year=2012&#x00026;title=Expression+of+heat+shock+protein+genes+in+insect+stress+responses&#x00026;journal=Invertebr.+Surviv.+J.&#x00026;volume=9&#x00026;pages=93-101" target="_blank">Google Scholar</a>
Blood meal acquisition enhances arbovirus replication in mosquitoes through activation of the GABAergic system
Kalia VK and Chander S (2022) Groundnut Bud Necrosis Virus Modulates the Expression of Innate Immune
and Cuticle Development-Associated Genes to Circulate and Propagate in Its Vector
Received: 05 November 2021; Accepted: 04 February 2022; Published: 17 March 2022
Copyright &#x000A9; 2022 Mahanta, Jangra, Priti, Ghosh, Sharma, Iquebal, Jaiswal, Baranwal, Kalia and Chander. This is an open-access article distributed under the terms of the <a rel="license" href="http://creativecommons.org/licenses/by/4.0/" target="_blank">Creative Commons Attribution License (CC BY)</a>
&#x0002A;Correspondence: Amalendu Ghosh, <a id="encmail">YW1hbDRlbnRvQGdtYWlsLmNvbQ==</a>; <a id="encmail">YW1hbGVuZHUuZ2hvc2hAaWNhci5nb3YuaW4=</a> <a href="http://orcid.org/0000-0001-6634-5771">orcid.org/0000-0001-6634-5771</a>
 https://www.facebook.com/rds.grandisuccessi
RDS announces the return for the third consecutive year of the RDS Summer Festival, the most anticipated music event of the summer. For the 2024 edition there will be a double stage in Palmi, in Piazza Primo Maggio, on 19 and 20 July 2024. Some of the names on the Italian and international music scene presented by Anna Pettinelli.
</a>Last update: Jun 5, 2024 6:00 AMContent owned by the Calabria Region | Tourism and Commerce Department issued under CC-BY License
two aces and a block to lead Osceola to a straight set (25-22
25-10) win over Tohopekaliga to capture the 2022 Orange Belt Conference championship on Friday night
 Harmony won the third place game with a win over Gateway (25-22
“Winning the Orange Belt Conference title was one of our team goals coming into this season,” Osceola coach Carrie Palmi said
 “This will hopefully give us some momentum heading into the final week of the regular season and then districts.”
The win was the 13th in a row for Osceola (20-2) and gave the Kowboys a third consecutive win over 2021 OBC champion Tohopekaliga (12-7) – a team that beat them three times last year
Osceola’s win was even more impressive given the fact that the Kowboys were without the services of injured libero Eimy Negron
Negron was the team leaders in both digs (301) and aces (43)
 “We have some pretty good depth at that position
but she is such a valuable asset to this team,” Palmi said
but the Tigers did rally to go up briefly at 15-13
including three on kills from Kaitlyn Taylor
 The teams would battle down the stretch but two service errors by the Tigers gave Osceola the three-point win
Game two was also tight before Osceola went on a 9-3 run
 A thunderous Palmi spike off capped the run that gave the Kowboys a 20-15 lead
The would trade points the rest of the way with Sophie Spivey ending the game with a spike down the right side for a 25-20 win
taking leads of 5-0 and 11-1 before settling in for a 25-10 win
Palmi would end the match with cross court kill
 She would end up with two service aces and a block to go with her 20 kills
 Taylor finished with nine kills in the match; Ahnika Johnson had six kills and three blocks
“I would give our overall effort a B or B+,” Palmi added
 “We made some mistakes at the net and gave them a chance to get back in both the first and second game
they are a scrappy team and did a lot of good things defensively.”
“We fought hard the first two games and lost them both
 There’s no question that zapped us of our energy and they took full advantage of that in the third,” Tohopekaliga coach Eric Capestany said
 “You have to stay focused and play with high energy against a team like that for every game or you will get rolled
 We lost a lot of talent last year and got off to a rough start
 We had our chances to win both the first and second games.”
Both teams will close out the regular season next week with three games
 Osceola will host Timber Creek on Monday and Holy Trinity Episcopal on Wednesday; before closing out the regular season at Lake Mary
who is currently ranked second in Class 7A
  “It will be a great test for us heading into districts,” Carrie Palmi said
 “If we are fortunate to win those games in the final week
it could give us a lot of momentum headed into the post-season.”  
Tohopekaliga plays at Harmony on Monday before closing out with a pair of home games against St
Cloud (10/12) and Southland Christian (10/13)
 The Kowboys will host the 7A District 6 Tournament
 Perennial state power Lake Nona (9-4) and Cypress Creek also will participate in the tournament
District 5 with several Orlando schools; while St
Cloud is scheduled to participate in the 6A
District 7 tournament and Gateway will play in 5A
Designed by <a href="http://www.elegantthemes.com" title="Premium WordPress Themes">Elegant Themes</a> | Powered by <a href="http://www.wordpress.org">WordPress</a>
And that is the case out in Osceola County as one local athlete set a new record just three matches into the school year
Our Despina Barton introduces us to Osceola High School volleyball player
and what she is doing to stay at the top of her game… in today’s Athlete of the Week:
Allison Palmi has been playing volleyball since she was 7-years old
“I got into it because my mom played collegiate volleyball and I had a couple of aunts played the sport so when my mom took the job here at Osceola I kind of grew up in the gym
I was a little gym rat,” Palmi said of her exposure to the sport
Flash forward 10 years and she is a senior at Osceola High School and shattering a county record
the energy from the girls give me and going up there and scoring points for your team
it’s an adrenaline rush the best part of it,” Palmi added
the four-year varsity letterman passed Aspen Dangren for career kills in Osceola County
“I wasn’t focused on it at all I kind of just went out and played and they told me at the end of the game—and it was really exciting,” Palmi said of breaking the record
Palmi now tops the list with 1,189 kills and that number will keep growing as we are so early into the season
&nbsp;“I think she actually broke it in the first set
she had 9 kills that first set,” Osceola Head Coach and Alli’s mom
“So that was really kind of crazy but it was awesome and I am proud of her
Just to accomplish it when there has been so many strong players that have come through our program that’s really big.”
so she took the necessary steps away from the volleyball court to beef up her vertical this off-season and it’s already paying out as she embraces that underdog role
“I think I was touching about 9’4 and just recently I’m just under 10 foot so it’s a big goal of mine to get there by the end of the year,” Palmi added
work in different attack shots and help guide this Kowboys team to a deep playoff run
Something her mom thinks could be a reality
“This is one of the best teams that Osceola has had in probably the last decade,” Coach Palmi said
“So I want to go far and I feel like with Ally’s volleyball IQ is really high
On the court I feel like we have almost an assistant coach on the court.”
“I made a lot of friends throughout the program so knowing I get to leave a bit of an imprint here is a really great feeling.”
Alli Palmi is committed to play volleyball at the University of Incarnate Word in San Antonio
Tomorrow kicks off Merchant’s Day weekend (Verslunamannahelgi) and we’ve got some fresh new jams for your road trip playlist
Some lucky folks have already knocked off early to turn it into a sweet-ass four-day vacay
but not us — we’re here doing the good Óðin’s work of making sure the new tunes are delivered to the people
set up some folding chairs in a field and crack open a cold one
If you’re unfamiliar with general Nordic languages, you may be a little puzzled by that slashed-O (which Rex and Jói have discussed on <a href="https://podcasts.apple.com/is/podcast/66-degrees-of-sound/id1679547490">66 Degrees of Sound</a> as being on par with band names’ use of Ö in the 80s)
it makes this name to sound like “bad stuff”
That self-deprecating moniker is hilarious
emotive soundscapes with the gentle twisting of knobs
this four-track EP is the perfect soundtrack to this season of blue nights
The true gift of a talented singer-songwriter is being able to convey profound
complex stories and emotions with the simplest of tools
lush synthpop debut single “Skylines” earlier this year
newcomer Lea strips everything back to voice and piano on her new deeply earnest
Describing the helpless sadness of witnessing one’s partner gripped by depression
it’s a true call to allies of those suffering from the affliction — no shaming the one they love
just desperately hoping they can help them get to the other side together
As I sit in the office trying to finish everything before my vacation
I couldn’t have opted for a better soundtrack than Ultraflex’s latest track “Digg Digg Deilig” from their upcoming album Chill Pill Vol
Ultraflex has always been an equivalent of something chill
dreamy and nostalgic and so is their new track
“Digg Digg Deilig,” which is the band’s first single in Norwegian and translates as “Yummy Yummy Delicious” is an ode to good food
ice lollies on the beach and sunkissed cheeks
An ode to those hazy club nights and situationships that are forged on uneven grounds
“Elska” has popstars Daniil and Lil Binni (of ClubDub fame) repeat the statement
You don’t love me.” Presumably while pulling petals off an ox-eye daisy
Although a flower ultimately cannot make that prediction
Daniil and Lil Binni still yearn for that special someone
This is a track for people who just wanna know what love is
This no-wave-inspired hip-hop track is the newest from the underground label Heavy Knife Records
newcomer Öngþveiti – a project by artist Frumburður and rapper Axel – stumble over their lyrics under a sluggish
Heavy Knife label founder Lord Pusswhip joins the track
spitting seemingly non-sensical rhymes – until you listen to it again
which you should do because the track doesn’t even reach two minutes
the one and only Pálmi Gunnarsson is back with a brand-new single
The Great Sir Pálmi tells us that he will change the world
and all we wanna tell him is: you already have 😍
Between taking us to the Bank of Joy with ICY at Eurovision ‘86
his canon smash-hit &#8220;Þorparinn&#8221;
Icelandic music simply wouldn’t be what it is without him
This sweet mid-tempo ballad gives that late 80s heartwarming vibe that makes you want to group-singalong
Very 3 am at the countryside wedding when you’re about to have your first kiss with your future spouse
<a href="https://grapevine.is/news/2025/05/02/tourist-held-hostage-on-may-day/">Tourist Held Hostage On May Day</a>
<a href="https://grapevine.is/news/2025/05/05/tourist-sexually-harassed-in-blue-lagoon/">Tourist Sexually Harassed In Blue Lagoon</a>
<a href="https://grapevine.is/news/2025/05/02/culture-night-killer-sentenced-to-prison/">Culture Night Killer Sentenced To Prison</a>
<a href="https://grapevine.is/news/2025/03/26/starbucks-to-open-in-reykjavik-soon/">Starbucks To Open In Reykjavík Soon</a>
<a href="https://grapevine.is/travel/2025/04/30/an-ode-to-flateyri/">An Ode to Flateyri</a>
<a href="https://grapevine.is/news/2025/05/02/grapevine-events-the-clock-other-grounds-film-festival-icelandic-sorcery-festival-more/">Grapevine Events: The Clock, Other-Grounds Film Festival, Icelandic Sorcery Festival &#038; More</a>
<a href="https://grapevine.is/news/2025/04/30/police-officers-spied-on-citizens-for-icelands-richest-man/">Police Officers Spied On Citizens For Iceland’s Richest Man</a>
<a href="https://grapevine.is/mag/2025/04/26/from-army-brats-to-luxury-flats/">From Army Brats To Luxury Flats</a>
Italy (Reuters) - Most southern Italian businessmen do not cross the &#x27;Ndrangheta mafia
and paid a high price.The owner of a successful cement business near the small town of Palmi
Saffioti in 2002 became one of only 30 people to turn state&#x27;s witness against Calabria&#x27;s &#x27;Ndrangheta.That year
and he and his family have lived under 24-hour armed guard ever since.&quot;Most businessmen learn to live with the &#x27;Ndrangheta,&quot; Federico Cafiero de Raho
chief prosecutor in the region&#x27;s largest city
said of the crime syndicate with a global reach and deep pockets thanks to narcotics.&quot;It is the arbiter of who can do what in the economy,&quot; added Cafiero de Raho
whose court resides in a city that even saw its local government dissolved in 2012 because it had been infiltrated by the group.While the &#x27;Ndrangheta (pronounced en-DRANG-eta) flourishes
the poorest of Italy&#x27;s 20 regions with a population of almost 2 million
has seen no benefit for its local economy.Calabria is a natural and historical treasure
It has almost 500 miles (800 km) of pristine beaches
The Apennine mountains rise thousands of feet in the interior
covered in cacti and century-old olive groves.Once at the heart of the Greek and Roman empires
considered two of the most spectacular sculptures of the ancient world
were pulled out of the crystal clear waters of the Ionian coast.But like the rest of the Mezzogiorno -- Italy&#x27;s six southern regions plus the islands of Sicily and Sardinia -- Calabria has suffered seven straight years of recession and is challenging Prime Minister Matteo Renzi&#x27;s efforts to fuel a recovery.From 2008 to 2014
Unemployment is three times that of the north and annual per capita output is 15,800 euros ($17,600)
Svimez research institute said 744,000 people left the Mezzogiorno between 2001 and 2014
and more than 70 percent of the emigrants were under the age of 34
The body warned of a &quot;a permanent state of underdevelopment&quot; in a region also home to separate mafia groups in Sicily
Campania and Puglia.Renzi has promised to tackle the mob and offer a &quot;master plan&quot; for the withered south in coming weeks
But filing cabinets in Rome are full of failed economic initiatives for the south and well-meaning anti-mob plans that have achieved little.For Saffioti
the reason growth has stubbornly failed to take root in Calabria is because the &#x27;Ndrangheta chokes it off.&quot;The &#x27;Ndrangheta wants a hand in everything,&quot; Saffioti
there would be no need for the &#x27;Ndrangheta
Real growth would marginalise it.&quot;&quot;LIKE GUANTANAMO&quot;Thanks to Saffioti&#x27;s testimony and closed circuit video recordings he made when he paid the mob
48 &#x27;Ndrangheta members from nine different crime families went to jail
According to his own records and testimony
he paid the equivalent of 2.5 million euros in extortion over 18 years.He could have fled and assumed a new identity as part of the witness protection programme
but he chose to stay.Now both his home and adjacent business are surrounded by 4-metre (13-foot) concrete walls
towering spotlights and dozens of video cameras
Four police stand on duty.&quot;It looks like Guantanamo,&quot; quips the bearded and bear-like Saffioti
&quot;But I&#x27;m very happy to have rid my life of that scum
the Italian mob evokes images from the fictional &quot;Godfather&quot; movies or &quot;The Sopranos&quot; TV series
but the &#x27;Ndrangheta&#x27;s power is real and thriving on the euro zone&#x27;s southern periphery in the 21st century.Over the past two decades
which takes its meaning from &#x27;strong man&#x27; in ancient Greek
has eclipsed its more storied Sicilian cousin Cosa Nostra by becoming Europe&#x27;s biggest cocaine broker and establishing criminal colonies across the globe
prosecutor Cafiero de Raho told Reuters.But the &#x27;Ndrangheta business model
requires it to be a local power broker with broad consensus
and the Church.The &#x27;Ndrangheta&#x27;s role as an intermediary -- from job provider to lender of last resort -- dates back to the creation of Italy 150 years ago
The mob has long cultivated a warped sort of colonial mentality where the state is considered a foreign occupier.&quot;Whoever is born here must follow the unwritten rules of a parallel state
To buy or sell a property or open a business
not the bureau of commerce,&quot; Saffioti said
adding that no deal was too small.Before he turned state&#x27;s witness
he had a job pouring concrete in the nearby town of Polistena
Though he was going to earn only some 250 euros ($280) for the work
demanded his cut.&quot;He told me it wasn&#x27;t a question of money
He said: &#x27;It&#x27;s like when you go visit someone&#x27;s home
a mafia hit man shot Longo dead.DAVID VS GOLIATHDavid Bumbaca
whose seaside restaurant and bathing area in Locri is just the kind of economic activity the area needs
is weighing up whether his future lies there.Over the past year because he refused to pay extortion &quot;as a matter of principle&quot;
two men wearing ski masks tried to beat him up in front of his home
and he received an anonymous letter with a death threat.&quot;My problems began when I started to be visibly successful,&quot; Bumbaca said
sitting in a shaded corner of his restaurant
which specialises in fresh seafood salad and other local treats.The 46-year-old Bumbaca got a business degree in northern Italy
but he says he returned out of love for Calabria.&quot;Now I don&#x27;t know how long my love of this land will hold out
I&#x27;m thinking more about selling and moving away than investing at this point,&quot; he said
&quot;It&#x27;s not a good situation for my family
and these things weigh on you.&quot;For now
in part because magistrates and police are among his regular clients
but he does not want to become a state&#x27;s witness like Saffioti.&quot;I admire the people who make those choices
I want to be with my family and live a safe life
I don&#x27;t want to be anyone&#x27;s hero,&quot; he said.($1 = 0.8963 euros)Editing by Crispian Balmer and Mark John
Our Standards: <a data-testid="Link" target="_blank" href="https://www.thomsonreuters.com/en/about-us/trust-principles.html" referrerPolicy="no-referrer-when-downgrade" rel="noopener" class="text__text__1FZLe text__dark-grey__3Ml43 text__medium__1kbOh text__small__1kGq2 link__link__3Ji6W link__underline_default__2prE_ link__with-icon__3x3oD">The Thomson Reuters Trust Principles., opens new tab</a>
, opens new tab</a> Browse an unrivalled portfolio of real-time and historical market data and insights from worldwide sources and experts.
, opens new tab</a>Screen for heightened risk individual and entities globally to help uncover hidden risks in business relationships and human networks.
© 2025 Reuters. <a data-testid="Link" href="https://www.thomsonreuters.com/en/policies/copyright.html" referrerPolicy="no-referrer-when-downgrade" class="text__text__1FZLe text__inherit-color__3208F text__inherit-font__1Y8w3 text__inherit-size__1DZJi link__link__3Ji6W link__underline_on_hover__2zGL4">All rights reserved</a>
HIBBING — Iron Range trio MorningBird will debut their new album with a release show at Wussow’s Concert Cafe at 7 p.m
Rob Wheeler and Josh Palmi are independent singer-songwriters and full-time musicians playing traditional and original folk and Americana music across the Iron Range
The three spent several weeks working their way across the west in fall 2023
The transformative experience of playing shows
trekking the open road with a backdrop of grand mountain scenery graced by windswept meadows
completed a journey that led to songwriting in harmony
The musicians were weekly artists in residence earlier this year at Smoke on the Water Winery in Coleraine
Once Wheeler kicks down on the stomp box and the fiddles and strings from Burkes and Palmi dance in frantic unison
She was born with a deformed left hand that required multiple childhood surgeries
who played in the Mesabi Community Orchestra
was instrumental in helping Burkes overcome her self-consciousness
Burkes' mother connected her with Helina Pakola
hoping she could teach Burkes to play the instrument in a mirrored fashion
who moved from Finland to Virginia in 1962
was the founder of the Mesabi Community Orchestra and a violinist with the Duluth symphony
traveling to Pakola's native country of Finland to play for the president
Burkes was traveling professionally at age 11
Pakola taught Burkes that for success you must “demand from yourself” and practice daily
After performing with Youth Duluth Superior Symphony Orchestra in high school
Burkes turned 18 and toured in England with recognized music educator and director Michelle Gribbon
Burkes graduated from Hibbing High School and earned her occupational therapy degree at the College of St
She was drawn to holistic behavioral health
and for two decades worked her career before stepping away to fulfill her dream of being a full-time musician
“You recognize life will be lived differently when you make this choice," Burkes said
It is the freedom of creativity and happiness.”
Paul native grabbed his guitar and his dog and began hitchhiking cross-country
Wheeler said that prompted him to start strumming — learning cowboy chords
He traveled the states performing at local venues while picking up construction jobs along the way
Wheeler and Burkes met while Wheeler was working at a resort on Crane Lake
It is still one of their favorite places to perform
Burkes and Palmi met five years ago while playing together in a band
Palmi played electric bass with Burkes on violin
Palmi joined MorningBird in 2022 and the threesome have been on the move
touring and writing preparing to drop their own first album
The trio recently returned from Kansas City
where they performed at Folk Alliance International
They have been debuting original songs from the new album
Special guests on the album include the brother duo Jesse and Ryan Dermody of The Brothers Burn Mountain
Ryan is a producer and sound engineer at off-the-grid Diarmada Studios in Cotton
Some other songs on the album were recorded with Rich Mattson of Sparta Sound
By <a href="https://thebulletin.org/biography/filippa-lentzos-4/">Filippa Lentzos</a>, <a href="https://thebulletin.org/biography/jez-littlewood/">Jez Littlewood</a> | July 8
The agricultural pest Thrips palmi can wreak havoc on crops
Cuba once accused the United States of unleashing an infestation on the island
bringing the matter to a meeting of the Biological Weapons Convention
Credit: Florida Division of Plant Industry Archive/Florida Department of Agriculture and Consumer Services/Bugwood.org
The move to trigger Article V of the bioweapons treaty represents a significant escalation of Russia’s efforts to promote claims of illegal <a href="https://thebulletin.org/2018/11/the-russian-disinformation-attack-that-poses-a-biological-danger/">US “biolabs” ringing its borders</a>
Now the treaty’s 184 member states will have to hold a special session this summer to hear the Russian allegations and the US response
the coming meeting of bioweapons treaty members will have important implications for a major upcoming review of the treaty
discussions about strengthening the agreement
and prospects for increasing peaceful international cooperation on biological issues
The meeting is an indicator of just how the scope of Russia’s biological weapons disinformation has grown since it invaded Ukraine
the insects were ravaging crops on the island in an infestation
including allies of the United States like Canada
said they did not see a link between the US flight over Cuba and the country’s subsequent infestation
“found it impossible to make a determination.” According to Tucker
only North Korea supported the Cuban position
In its December 1997 report, which was <a href="http://www.sussex.ac.uk/Units/spru/hsp/occasional%20papers/HSPOP_3.pdf">not made public</a>
the committee said that the technical complexity of Cuba’s allegation
the time between the alleged incident and Cuba&#8217;s request for an Article V meeting (known as a formal consultative meeting)
and the lack of detailed information related to the matter
made it impossible to draw any consensus conclusions
worked as intended; it had provided a forum to bring a compliance issue to the attention of other treaty members
the experience was further evidence that treaty members needed to agree to an effective means of strengthening the ability to assess compliance
Moscow actively laid the <a href="https://www.chathamhouse.org/2022/06/disinformation-fight-goes-beyond-ukraine-and-its-allies">groundwork</a> for taking its disinformation campaign to new heights before invading Ukraine by pre-positioning its false narrative—making statements to the media
Russia has provided documentation to the Security Council and circulated Notes Verbales (diplomatic memos) to national missions to the UN in both New York and Geneva
supposedly providing evidence to back up its allegations of biological weapons activities in Ukraine
the consultation procedures under Article V of the bioweapons treaty are generally viewed as a way to discuss issues at a lower level before escalating to the Security Council
Russia has reversed this logic and is invoking Article V after failing to gain any substantive support in the Security Council or in the UN General Assembly for its allegations
Calling a bioweapons treaty meeting appears to be simply another airing of its allegations
Few experts take Russia’s claims seriously
but they are potentially more damaging to the bioweapons treaty than Cuba’s in 1997 for five reasons
First, they will re-energize false portrayals of biological labs from earlier this year, undermine biosafety and biosecurity efforts, further erode confidence in critical public and animal health infrastructure, and distract attention away from Russia’s own actions in Ukraine which include <a href="https://www.who.int/news/item/07-04-2022-who-records-100th-attack-on-health-care-in-ukraine">attacks on healthcare facilities</a>
Second, many countries have called for development aid along the lines of what US assistance, through its <a href="https://www.google.com/url?sa=t&amp;rct=j&amp;q=&amp;esrc=s&amp;source=web&amp;cd=&amp;ved=2ahUKEwiI9cmVyuj4AhVkOX0KHXaCCE4QFnoECCgQAQ&amp;url=https%3A%2F%2Fcrsreports.congress.gov%2Fproduct%2Fpdf%2FIN%2FIN11886&amp;usg=AOvVaw3EuIBLOPkoCVjByqT-cnTH">Biological Threat Reduction Program</a>
has provided to dozens of Ukrainian labs on public and animal health research and disease surveillance
Many countries want assistance and support to conduct this work
the World Health Organization requires its members (every country in the world save one) to monitor diseases within their borders
If Russia is effective at defining US development assistance as a form of non-compliance with the bioweapons treaty
the upcoming Article V process could seriously undermine peaceful cooperation under the treaty
Some countries might actually believe the Russian allegations
but others will view them as an opportunity to put the United States on the back foot and use the Article V meeting as another venue to challenge the US-led global order
The Article V deliberations could stall progress on that front if relations between treaty members sour as a result of Russia’s move
Russia could sway countries to its position at the upcoming Article V meeting about alleged US bioweapons programs
That’s already happened several times during the Ukraine conflict
comes from countries who are unconvinced by the submitted evidence and information but who remain silent and effectively abstain from being caught in a confrontation between the United States and Russia
and Vietnam were among those who voted against while some countries who are important players in the future of the bioweapons treaty such as India
Russia’s decision to trigger a special session of the Biological Weapons Convention now runs the risk of co-opting the treaty
and using it as a platform to spread and legitimize disinformation—particularly if important countries stay silent when these allegations are aired
It will take hard work and ingenuity from disarmament diplomats to ensure the meeting does not descend into acrimony and that any report from it is accepted as an accurate record of the deliberations
there is an urgent need to counter Moscow’s destructive narrative and to protect the norm and institutions that prohibit biological weapons from spurious and unsubstantiated allegations that could polarize the bioweapons treaty even more than it already is
The Bulletin elevates expert voices above the noise. But as an independent nonprofit organization, our operations depend on the support of readers like you. Help us continue to deliver quality journalism that holds leaders accountable. <a href="https://thebulletin.org/giving-donate/?utm_source=website&amp;utm_medium=article&amp;utm_campaign=giving%20button" target="_blank" rel="noopener">Your support of our work at any level is important</a>
we promise our coverage will be understandable
&#916;document.getElementById( "ak_js_2" ).setAttribute( "value"
Perhaps the patent absurdity of operating facilities in 2022 which originated under the Nunn-Lugar Cooperative Threat Reduction program 31 years earlier might have something to do with Russian skepticism…but then
Filippa Lentzos is an associate professor in Science &amp; International Security at King’s College London, where she leads research projects on... <a href="https://thebulletin.org/biography/filippa-lentzos-4/">Read More</a>
Jez Littlewood is a policy analyst in Alberta.  He previously worked at Carleton University (Ottawa) and the University of Southampton (United... <a href="https://thebulletin.org/biography/jez-littlewood/">Read More</a>
Copyright © 2025 Bulletin of the Atomic Scientists. All rights reserved. Registered 501(c)(3). EIN: 36-2136497 <a href="/terms-of-use" class="copy-link">Terms of Use</a><a href="/privacy-policy" class="copy-link">Privacy Policy</a>
Street view</a><a class="relative block aspect-video w-full overflow-hidden rounded-2xl sm:aspect-[16/7] lg:aspect-[16/6] xl:aspect-[16/5]" href="https://calabriastraordinaria.it/en/destinations/palmi-and-the-legend-of-the-olive-tree-rock">Discover more on Palmi</a>Around<a href="https://calabriastraordinaria.it/en/places/de-rosa-antiquarium-palmi">MuseumPalmi
</a><a href="https://calabriastraordinaria.it/en/places/leonida-e-albertina-repaci-art-gallery-palmi">MuseumPalmi
</a>Last update: Oct 25, 2023 9:47 AMContent owned by the Calabria Region | Tourism and Commerce Department issued under CC-BY License
the Wiwi Jury – our in-house panel of music unprofessionals – gathered in Reykjavik to discuss the contestants participating in Söngvakeppnin 2016
the national selection for Iceland at Eurovision 2016
Today we look at Pálmi Gunn with his song &#8220;I Lead You Home&#8221;
or should we call the police because we have an old
Josh: All of the delegations for Eurovision around the world need to understand that rehashing a former star almost never pays off
I&#8217;m sure Pálmi is a lovely man and good on him for giving Eurovision another crack
but I think that the grandfather of Iceland&#8217;s Eurovision alumni should maybe take a little nap &#8211; and I know a decent song to help him sleep
It&#8217;s called &#8220;I Lead You Home&#8221;
Luis: I hate to say this about a song, but I was falling asleep while hearing it. &#8220;I Lead You Home&#8221; is obviously dated, yet well constructed and well sung. Pálmi&#8217;s voice is charming, but the whole act sounds as if it was taken from a Sunday afternoon show for old people. And in a way it&#8217;s sad that while his mate at ICY, <a href="https://www.youtube.com/watch?v=IS69tr4_JbE">Eiríkur Hauksson</a>
brought an absolute masterpiece to ESC in 2007 (still crying about that not qualifying)
Pálmi can only give us a bland ballad with no risk at all
Robyn: Iceland is doing the Melfest trick of bring back a star from earlier years, with Pálmi part of <a href="https://youtu.be/eGegovbGTOg" target="_blank">Iceland&#8217;s debut act, ICY</a> in 1986
The disco sailor&#8217;s jacket is gone and instead Pálmi is delivering a very gentle
I don&#8217;t think it&#8217;s going to win &#8211; and I don&#8217;t think it would do well at Stockholm &#8211; but it&#8217;s lovely to have this as part of the national final
In the Icelandic jury we have seventeen jurors but only room for four reviews
The highest and lowest scores are dropped prior to calculating the average score
This is to remove outliers and reduce potential bias
We have removed a low of 1.5 and a high of 8
Josh is from Australia and has been a huge Eurovision fan since 2010. You can follow him on Twitter <a href="https://twitter.com/josh_salmon">@josh_salmon</a>. Keep up with the latest Eurovision news and gossip by liking <a href="https://www.facebook.com/pages/Wiwi-Bloggs-Eurovision-News-with-Attitude/236584169725787">our Facebook page</a> and by following the team on Twitter <a href="https://twitter.com/wiwibloggs">@wiwibloggs</a>
and -unlike most other songs-  it isn&#8217;t trying to cling to whatever pop-fad-sound feels relevant this year
but this one is to boring for the eurovision song contest
<a href="https://wiwibloggs.com/2025/05/04/house-of-helvetia-the-schedule-for-our-eurovision-2025-fan-event-in-basel/284646/"></a>
<a href="https://www.myeurovisionscoreboard.com/"></a>
<a href="https://wiwibloggs.com/about-2" target="_blank" rel="noopener">Overview</a>
<a href="https://wiwibloggs.com/meet-team" target="_blank" rel="noopener">Meet the team</a>
<a href="https://wiwibloggs.com/press" target="_blank" rel="noopener">Press</a>
<a href="https://wiwibloggs.com/?p=28689" target="_blank" rel="noopener">Write for us</a>
Input your search keywords and press Enter
Gísli Pálmi is one is Iceland&#8217;s foremost young rappers
known for his Icelandic-language flow and his melding of beats and electronic textures
We went round to his place to talk about 90s hip-hop
rapping in the language closest to Old Norse
and his first ever show &#8211; playing to a packed house at Iceland Airwaves
Follow the Gísli Pálmi&#8217;s Glacier Mafia on <a href="http://www.facebook.com/pages/G%C3%8DSLI-P%C3%81LMI-GLACiER-MAFiA/233327726691977">Facebook</a>
<a href="/articles/41846/metrics" data-track="click" data-track-action="view metrics" data-track-label="link" rel="nofollow">Metrics details</a>
The countries that have signed the 1975 Biological Weapons Convention will meet next week in Geneva to hear Cuban allegations that last year the United States discharged the insect pest Thrips palmi over Cuba to damage the country's agriculture
The US State Department dismisses the allegations as “outrageous”
Prices may be subject to local taxes which are calculated during checkout
<a data-track="click" data-track-action="view rights and permissions" data-track-label="link" href="https://s100.copyright.com/AppDispatchServlet?title=US-Cuba%20row%20over%20insects%20goes%20to%20weapons%20meeting&amp;author=Declan%20Butler&amp;contentID=10.1038%2F41846&amp;copyright=Macmillan%20Magazines%20Ltd.&amp;publication=0028-0836&amp;publicationDate=1997-08-21&amp;publisherName=SpringerNature&amp;orderBeanReset=true">Reprints and permissions</a>
<a data-test="citation-link" data-track="click" data-track-action="download article citation" data-track-label="link" data-track-external="" rel="nofollow" href="https://citation-needed.springer.com/v2/references/10.1038/41846?format=refman&amp;flavour=citation">Download citation</a>
Sign up for the Nature Briefing newsletter — what matters in science
Team Groupama-FDJ's French rider Arnaud Demare reacts after he crossed the finish line to win the 6th stage of the Giro d'Italia 2022
French rider Arnaud Demare made back-to-back stage wins in the Giro d'Italia bicycle race on Thursday
and pipped Australian Caleb Ewan in a photo finish on the line after the 192km ride from Palmi to Scalea in the southern Calabria region
Diego Rosa led for most of the 192-kilometre ride from Palmi to Scalea following a solo attack
but the Italian was reeled in by the peloton inside the final 30 kilometers
Cavendish kicked off the ensuing sprint finish
and the 'Manx Missile" was passed by Ewan in the final meters before Demare swung past the Australian just before the finishing line
becoming the most successful French rider in the race around Italy ahead of legends Jacques Anquetil and Bernard Hinault
Juan Pedro López of Spain and Team Trek - Segafredo Pink Leader Jersey celebrates at podium during the 105th Giro d'Italia 2022
Stage 6 a 192km stage from Palmi to Scalea
I had the feeling that I had won but it was hot so I waited for the photo-finish," said Demare
I felt like I was in front but you never know
I threw the bike and that was the difference."
The Frenchman sits down in 133rd place in the general classification despite his heroics
more than half-an-hour adrift of overall leader Juan Pedro Lopez of Spain
The seventh stage is a grueling 196km ride between Diamante and Potenza including four climbs between the Calabrian coast inland to Basilicata with a summit finish
Disinformation report hotline: 010-85061466
a journey through history and landscape on 12 May 2024.Guided Tour
Sunday 12 May 2024 the Trekking Train initiative will take place
in collaboration with the Department of Tourism of the City of Palmi; an experience under the banner of Sweet and Sustainable Mobility that combines tourist transport by train and nature excursion and urban trekking
The Trekking Train initiative is dedicated to discovering the natural beauty of the Costa Viola and specifically of Palmi
guided by the guides of the "Fogghi di Luna - Identità e paesaggio" Association
part urban (Palmi's historic centre) and part naturalistic (Sentiero Leonida)
The Leonida Trail is characterised by its many points of interest: history
also suitable for children from 12 years of age
and does not involve significant physical effort
For information and reservations: <a target="_blank" rel="noopener noreferrer" href="https://www.railbook.it/events/ferrovie-in-calabria">https://www.railbook.it/events/ferrovie-in-calabria</a>
Street view</a>Interested<a href="https://calabriastraordinaria.it/en/events/the-path-of-st-francis-of-paola-the-way-of-the-young-man">Guided TourThe Path of St Francis of Paola - The Way of the Young ManMay 31 - 13 Jul other dates: Jun 1, 2025, Jun 2, 2025, Jul 11, 2025, Jul 12, 2025 and Jul 13, 2025
May 9 - 22 Jun other dates: May 10, 2025, May 11, 2025, Jun 20, 2025, Jun 21, 2025 and Jun 22, 2025
</a>Last update: Mar 29, 2024 6:07 AMContent owned by the Calabria Region | Tourism and Commerce Department issued under CC-BY License
Deciphering Icelandic rap is difficult enough
but GP manages to put native speakers to task with his use of slang only understood by himself and the rest of the Glacier Mafia
his self-titled album could be heard blasting through car stereos all around Reykjavík
GP managed to do the supposedly impossible: get young people to buy CDs (for those of you who forget what a CD is
it&#8217;s a physical form of storing music called a Compact Disc)
GP will be performing at the Secret Solstice music festival
There is no news on whether <a href="http://grapevine.is/news/2015/06/21/bam-margera-gets-punched-in-the-face-by-icelandic-rapper-video/">Bam Margera will be attending again.</a>
Tonight&#8217;s edition of our monthly Húrra Grapevine
The Reykjavík Grapevine and Húrra are proud to announce that after long and arduous bout of negotiations
we have managed to enlist four local hip hop greats to join forces and throw all of Reykjavík all a monster hip hop party that will keep the city shakin&#8217; well into the small hours of Saturday morning
Get ready for an all out lyrical assault from emcees Bent
as the incomparable Logi Pedro brings a steady soundtrack of today&#8217;s freshest beats (some of which haven&#8217;t even been invented at the time of writing)
The crew will be joined on stage by a hand-picked gourmet selection of Reykjavík&#8217;s best
Including (but not limited to): LORD PU$$WHIP
Get your tickets <a href="http://www.tix.is/is/event/661/gisli-palmi%2c-emmsje-gauti-%26-bent/">here</a> (or at the door
<a href="http://listings.grapevine.is/events/hurra-grapevine-3-logi-pedro-bent-gisli-palmi-emmsje-gauti/"></a>
We met up with Gísli Pálmi to make some ice cold liquid nitrogen ice c.r.e.a.m with caramel chocolate
We figured who better to talk to about the joys of sci-fi ice cream than Mister Glacier Mafia himself
See GP bam and knock it up a notch with some well placed fruits in the first of an ongoing series of Reykjavík Grapevine food videos
You are using an outdated browser. Please <a href="http://browsehappy.com/">upgrade your browser</a> to improve your experience
spoke openly on drug use on Rás2 national radio yesterday
He says that he's never seen as many drugs on the street in Iceland as right now
The lyrics on his new album describe drug use and in an interview with radio host Ólafur Páll Gunnarsson
Gísli Pálmi says that this is today's reality
a reality that older generations don't know about
"Noone dares to tell the truth about it," he adds
"Unfortunately the people who have been so heavily into this are not talking about it." 
Gísli Pálmi describes his friends who were heavily into drugs as some having turned over a new leaf,and some ending up in jail
the high school and college kids were against drugs
But I'm not the one selling drugs anymore." When asked whether he still deals drugs he replied," That's totally unrelated to my music."  
Gísli Pálmi was asked whether he feels any responsibility as a role model he denis that."Not at all
I'm not trying to be any sort of role model."
Messy affair Icelandic rapper Gísli Pálmi (camouflage jacket) about to throw a punch at Jackass-member Bam Margera
Three Icelandic rap musicians attacked Jackass-member Bam Margera at the Secret Solstice music festival
The star has pressed charges against his three assailants
Read more: <a href="http://icelandmag.com/article/rapper-gisli-palmi-icelander-attitude">Rapper Gísli Pálmi: An Icelander with attitude</a>
According to eye witnesses Bam had tried to force his way into the festival’s production room and harassed two female employees when the three Icelandic rappers attacked him
punching him numerous times in the face and head
claims he had not been trying to force his way into the room
he had merely wanted to speak to one of the festival’s employees
Read more: <a href="http://icelandmag.com/article/photos-festival-goers-enjoyed-good-music-and-good-weather-secret-solstice-music-festival">Photos: Festival goers enjoyed good music and good weather at the Secret Solstice Music Festival</a>
rapper Gísli Pálmi (in camouflage jacket) can be seen throwing the first punch
After security and bystanders manage to break up the fight
Bam was arrested when he refused to pay for damages after having destroyed a rental car
Do you want to know more about this subject? Please send us a line at <a href="mailto:icelandmag@365.is">icelandmag@365.is</a>
Have you had an experience related to the contents of this article?  <a href="mailto:icelandmag@365.is">Let us know!</a>
Join our weekly hand curated newsletter to have all the latest news from Iceland sent to you
2022 Torg ehf. <a href="/terms-and-conditions">Terms </a>&amp; <a href="/our-privacy-policy-0">Privacy</a>
A video has been released on YouTube of the brawl between Jackass star Bam Margera and Icelandic musicians at the Secret Solstice outdoor music festival
The footage shows Icelandic rapper Gísli Pálmi punching the Jackass star
According to spokespeople of the festival, as reported by visir.is, Margera tried to break in to the festival's production room and harassed two female members of staff. They said that he was in a very inebriated state and that the Icelandic musicians were coming to the aid of the women in question. In an interview with <a href="http://www.visir.is/atokin-a-myndbandi-thverneitar-ad-hafa-areitt-starfsfolk/article/2015150629896">visir.is</a>
Margera denies harassing the women and said that matter revolved around an unpaid debt to him by one of the festival directors