At Sweden’s annual life science conference
and industry discussed implementing the country’s new life science strategy
Key topics included patient involvement and clinical trials
with an emphasis on moving from strategy to action
particularly in terms of promoting uptake of advanced precision medicines
While these have improved personalized care
implementing these innovations across the healthcare system remains a challenge
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USA: Danfoss Turbocor has celebrated the opening of its new $62m
145,000ft2 production facility in Tallahassee
The ribbon-cutting ceremony was attended by over 100 people
Danfoss president and CEO Kim Fausing and descendants of Danfoss founder Mads Clausen.
The additional facility will host state-of-the-art manufacturing for TTS/TG/TH lines for Danfoss Turbocor compressors
magnetic bearing technology to more than 14,000 compressors per year.
“The increased production will help meet the growing market demand for cooling and heating high-efficiency compressors in North America
North America is the largest market and region for Danfoss
and we couldn’t be prouder of this latest chapter in Turbocor’s growth.”
“As part of Danfoss’ green growth strategy
we are regionalising our supply chains in order to manufacture products closer to our customers
improve service and decarbonise our operations
Expanding our capacity in Tallahassee is key to our growth strategy in North America,” commented Danfoss Turbocor president Ricardo Schneider
Turbocor celebrated its 10th anniversary last November as a wholly-owned Danfoss company
Initially founded in 1994 and later becoming a joint venture with Danfoss in 2004
Danfoss Turbocor is continuing to grow its footprint
and the establishment of a configuration centre at Danfoss’ corporate headquarters in Nordborg
The Nordborg site currently handles sales and service for Turbocor’s European customers
A new production facility is scheduled for completion in 2026
Danfoss marks 10 years with Turbocor – 13 November 2023USA: Danfoss has marked 10 years as owner of the pioneering Turbocor oil-free, magnetic bearing compressor. Read more…
Danfoss plans Turbocor production in Europe – 12 November 2022DENMARK: Danfoss is to establish a Turbocor service centre at its headquarters in Nordborg, with plans to start full production of its oil-free compressors in Denmark by 2026. Read more…
Danfoss to expand Turbocor facility – 22 December 2017USA: Danfoss is to build a new $12m, 44,000ft² facility at its Turbocor Compressor site in Tallahassee. Read more…
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titled "Southwestern Strength," premiered on June 25 and featured four chefs from the Southwest: two from New Mexico
Using ingredients such as prickly pear and mesquite flour
the chefs were tasked with creating an appetizer first
One chef is eliminated after each round until one winner remains
taking home the grand prize of $10,000 after wowing the judges with his mesquite-spiced date clafoutis
white chocolate chips and cactus-shaped macarons
He celebrated his win at a watch party at Wren House Brewing in Phoenix on Tuesday night
It was his second time appearing on a Food Network show
Christensen appeared on an episode of "Cutthroat Kitchen," which he said was "the most chaotic show to be on." In contrast
he said the "Chopped" set was full of good energy and a generally more relaxed experience
"Watching a few episodes before I went out there to film
I'll have all of my plates ready and plated with five minutes left," Christensen told The Arizona Republic
Where to find Derek Christensen's food in PhoenixYou can try Christensen's cooking every Thursday and Friday from 6 to 10 p.m. at his Nordborg pop-up inside Sauvage wine bar in Phoenix
He said Nordborg has evolved over the last few months from experimenting with Nordic and Scandinavian food to his current "summer holiday" series
which takes inspiration from a different part of the Mediterranean each week
"Southwestern Strength" was the 12th episode of Season 58 of "Chopped." The episode aired on Food Network on June 25
The best places to dine in 2024: 100 essential restaurants in metro Phoenix
Reach the reporter at endia.fontanez@gannett.com. Follow @EndiaFontanez on X
Support local journalism. Subscribe to azcentral.com today.
Juni eröffnet Center Parcs das neue Nordborg Resort auf der dänischen Insel Nordals
Das Nordborg Resort ist der erste Ferienpark in Dänemark
Sie müssen sich einloggen oder registrieren
Spielregeln
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Autoradiographic and histological evidence of postnatal hippocampal neurogenesis in rats
Time of neuron origin in the hippocampus and dentate gyrus of normal and reeler mutant mice: an autoradiographic analysis
Subgranular zone of the dentate gyrus of young rabbits as a secondary matrix
Neurogenesis in the dentate gyrus of the adult rat: Age-related decrease of neuronal progenitor proliferation
Proliferation of granule cell precursors in the dentate gyrus of adult monkeys is diminished by stress
Mitotic neuroblasts in the 9-day-old and 11-month-old rodent hippocampus
Neurogenesis in the adult rat: electron microscopic analysis of light radioautographs
Evidence that granule cells generated in the dentate gyrus of adult rats extend axonal projections
Differentiation of newly born neurons and glia in the dentate gyrus of the adult rat
Bromodeoxyuridine: a diagnostic tool in biology and medicine
Genetic influence on neurogenesis in the dentate gyrus of adult mice
Immunocytochemical detection of 5'-bromodeoxyuridine incorporation in the central nervous system of the mouse
nucleator and point sampled intercepts and their use in pathological research and diagnosis
Regionally specific loss of neurons in the aging human hippocampus
Methods for determining numbers of cells and synapses: a case for more uniform standards of review
More hippocampal neurons in adult mice living in an enriched environmen Nature 386
a neuronal specific nuclear protein in vertebrates
NeuN: A useful neuronal marker for diagnostic histopathology
Calcium-binding protein (calbindin-D28k) and parvalbumin immunocytochemistry: localization in the rat hippocampus with specific reference to the selective vulnerability of hippocampal neurons to seizure activity
Brain enolases as specific markers of neuronal and glial cells
In vitro neuronal production and differentiation by precursor cells derived from the adult human forebrain
Nature and fate of proliferative cells in the hippocampal dentate gyrus during the life span of the rhesus monkey
Experience-induced neurogenesis in the senescent dentate gyrus
Survival and differentiation of adult neuronal progenitor cells transplanted to the adult brain
Differentiation of adult hippocampus-derived progenitors into olfactory neurons in vivo
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van Praag for the contribution of rat tissue
This study was supported by grants from the Swedish Medical Research Council (project no
the John and Brit Wennerströms Stiftelse for Neurologisk Forskning
the Rune and Ulla Amlövs Stiftelse for Neurologisk and Reumatologisk Forskning
Stiftelsen Göteborgs MS förenings forsknings och byggnadsfond
Stiftelsen Handlanden Hjalmar Svenssons Forskningsfond
Stiftelsen Assar Gabrielssons Fond and the Edit Jacobssons Fond and from NIA and NINDS and the Alzheimer's Association (F.H.G.) and the American Federation for Aging Research (D.A.P.)
Institute of Neurology Sahlgrenska University Hospital
Ekaterina Perfilieva & Ann-Marie Alborn
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a supplier of mobile and industrial hydraulics as well as electric powertrain systems
has invested in an advanced 3D scanner robot for its joystick manufacturing lines in Nordborg
The company says the new device is boosting inspection routines by measuring component dimensions to “micron-level precision
providing customers with even higher product quality”
Danfoss Power Solutions’ Nordborg facility manufactures thousands of joystick variants for off-highway vehicles
operators can tell immediately if a joystick component is within specifications
This capability has eliminated the need to outsource time-critical metrology requirements to a third-party coordinate measuring machine technology provider
he investment is thus saving considerable time and providing greater in-house control over the manufacturing process
mechanical engineer in the Danfoss’ connect and controls solutions business unit in Nordborg
is leading the 3D scanner project alongside his colleague
Dukai says: “The scanner is so fast and easy to use that we can literally verify component dimensions while we manufacture
We now know immediately whether part dimensions are 100 percent identical to the computer-generated design files
so the 3D scanner has taken our quality testing capability to a new level.”
Dukai adds that he was not dissatisfied with the third-party metrology specialist and will continue to use the company for tasks where speed is less critical
The innovative GOM 3D scanner robot uses narrow-band blue light to measure up to 12 million points on the component surface within a few seconds
Colleagues at Danfoss Power Solutions then compare this image to the original CAD file
“We are talking extreme accuracy,” he says
“And it can even scan highly complex shapes and forms
It’s incredible technology that’s already gaining prevalence in the automotive industry
For more information: www.danfoss.com
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is opening its first Denmark location next year at the CenterParcs Nordborg Resort near the German border incorporating 440 holiday homes featuring views of the forest
"We continuously strive to bring our product offering to new markets; the brand has been well-received in the Irish and the German market and we are delighted to announce its expansion into Denmark," Rocket Restaurants Group Business Development Director Tobi Lukaschek said in a company press release
"Customer feedback has been overwhelmingly positive for our Irish burgers
The restaurant anticipates serving at least 300,000 customers annually
"We are delighted to partner again with Rocket's to bring their popular hamburger offering to a new group of customers in Denmark
following the successful launch of Rocket's with us in Germany," franchise partner Anne-Pierre de Cosnac
Areas Deutschland designs dining facilities at train stations
Construction of the Nordborg Resort began in May 2022 and completion is expected in Q1 2025
CenterParcs Nordborg will be the 29th resort for CenterParcs Europe and the first in Scandinavia
Rocket Restaurants Group includes 45 outlets trading under the Eddie Rocket's and Rockets brands
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Join us on a journey around Europe to see why protecting ecosystems matters
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is designed to optimise energy flow to save operational costs and to be climate-friendly
A ‘Smart store’ system was set up in collaboration with a Danish engineering giant
Brugsforeningen for Als and Sundeved (BALS)
At the back of the 750m2 shop is a 250m2 technology centre where gigantic machinery to control the temperatures of the supermarket can be seen through a glass window
Danfoss says the shop functions as a "live" test centre for energy efficiency technology
Shoppers can learn how the shop reuses energy through the glass and the facility allows engineers to experiment with an advanced compressor pack in a real environment
“The beauty about this is that this is a real realistic test facility
There are not two stores that are the same because every day customers come in and some forget to close the close to cover (…) There are new foodstuffs put in,” Henry Steffensen
the director of strategic marketing at Danfoss Climate Solutions
“So putting additional load on the refrigeration system and that's what makes it a realistic and a test facility for us”
Danfoss says the tech installed in this supermarket can help save operating costs while reducing food waste
this one is designed to be about 50 per cent more energy efficient
Supermarkets produce and lose heat as they operate multiple refrigerators and cooling rooms
Supermarket refrigerators use a closed-loop system with refrigerants that absorb heat from inside refrigerated display cases and transfer it through the condenser often located outside or on the building's roof
this supermarket captures the ‘waste heat’ from its cooling systems and reuses it to provide space heating and water heating
“When you cool you also create heat and often this is wasted
We accumulate all the heat in the heat tanks here,” Steffensen said
The heat recovery unit which consists of two tall tanks processes excess heat from the cooling systems and turned into warm water to provide space heating for the entire shop
Danfoss said the supermarket sells back any residual heat if it produces too much
Metres from a local heating company at the facility show the shop has so far bought about 90 kWh from the local district heating grid while selling 33,380 kWh to the grid since it opened in May 2023
“The tests so far have shown that we don't even need to buy additional heat from our energy company
and we also have more heat to sell,” Steffensen said
“We are self-sufficient here with heating and it's all heat that would typically be wasted on the roof out to the birds,” he added
about 150 per cent of the heat needed to operate the shop was produced in the heat recovery unit
The store has 18 cabinets and two cold rooms which are tracked and controlled by a ‘smart store’ system through a monitoring panel
If temperatures of the units drop below a certain point
the system sends an alert to notify maintenance staff
The supermarket also has solar panels on the roof
providing 100,000 kWh per year that can help with shop operations
can be used to lower the temperature of the freezers below the usual temperature to help accumulate solar energy while saving battery storage
When the solar panels are less efficient due to no sun
the temperature returns back to normal settings
a 1 km-long ground loop filled with brine is buried under the ground and can create additional heat
“So we can actually use both to create additional heat for the store
So we run our refrigeration pack as a kind of a big heat pump
Danfoss says the installations and technologies in the supermarket can be applied to various sizes of shops from the smallest shop up to the biggest hypermarket and in different climates from Jordan to Denmark
a standard one needed to run a regular shop in Denmark and an advanced version that can be used in warmer climates
Steffensen said that a typical payback time for investing in energy-efficient products is three to four years but the heat recovery unit only takes less than one year
“You can kick out your gas boiler in the store and get all your heat from waste heat from the refrigeration system”
Amme Travelling in comfort with the Nordborg 42 from DenmarkSleek lines
good sailing characteristics and the finest Danish boat building skills below deck: the largest Nordborg put to the test
Campe Crack drawing of the Nordborg 42With the new 42
the Danish shipyard has developed a beautifully crafted cruising yacht
A striking feature is the large bow platform
which makes it easy to get off the jetty - just one of many good details
We test sailed the Nordborg 42 in the Alsenfjord
Nordborg 42 (pdf)
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Much of what we know about eukaryotic transcription stems from animals and yeast; however
plants evolved separately for over a billion years
leaving ample time for divergence in transcriptional regulation
Here we set out to elucidate fundamental properties of cis-regulatory sequences in plants
Using massively parallel reporter assays across four plant species
we demonstrate the central role of sequences downstream of the transcription start site (TSS) in transcriptional regulation
Unlike animal enhancers that are position independent
plant regulatory elements depend on their position
as altering their location relative to the TSS significantly affects transcription
We highlight the importance of the region downstream of the TSS in regulating transcription by identifying a DNA motif that is conserved across vascular plants and is sufficient to enhance gene expression in a dose-dependent manner
The identification of a large number of position-dependent enhancers points to fundamental differences in gene regulation between plants and animals
we show that the basic property of the majority of animal enhancers
data are smoothed using a 100-bp rolling window
These findings suggest that downstream regulatory regions are enriched between the TSS and the start codon and affect transcription rather than mRNA stability
given that the clear dip in TF binding sites is centered on annotated TSSs
These analyses support the notion that many Arabidopsis genes have a transcriptional regulatory region downstream of the TSS
originating from upstream or downstream of the TSS of Arabidopsis genes
pooled and inserted upstream of the TSS or within the intron of a reporter gene and tagged with barcodes
Following transient transformation into one of four species
barcoded RNA sequencing was used to quantify expression
High reproducibility in MPRA experiment replicates
demonstrated here for CaMV 35S minimal promoter-based libraries
Pearson’s correlation coefficient r and number of fragments n are indicated
Comparison of construct expression with control enhancer fragments and no-insert constructs for upstream (top) and downstream (bottom) insertions
tomato (red) and maize (yellow); construct backgrounds are coded by symbol shape
Left: Pearson’s correlation coefficients across all libraries
Construct background and insertion position are indicated
n = 11,817) libraries of Arabidopsis and tomato and between upstream and downstream libraries of Arabidopsis (bottom
Comparison of mRNA steady-state levels and synthesis rates in the downstream MPRA with pTRP1 constructs
Correlation and fragment counts are indicated as in b
Comparison of activity of upstream-derived (blue
3,966 fragments) and downstream-derived (red
7,928 fragments) fragments relative to no-insertion constructs when fragments are positioned upstream (left) or downstream (right) of the TSS
unlike the position independence seen for animal enhancers
the activity of flowering plant enhancers is strongly dependent on their position relative to the TSS
along with the strong correlation among the relative activity of fragments across all libraries
while absolute levels are strongly influenced by backbone and species
the relative effects of different fragments in the same position are similar across species
benthamiana with pTRP1-based constructs (right)
Relative activity of downstream fragments when inserted downstream as a function of the number of YVGATCBR consensus motifs in the tested fragments
Error bars in b and c represent mean ± 1 s.d
The numbers in parentheses in b and c indicate the number of fragments
Differences in activity of the p35S-downstream library between original and mutated fragments
Effect of removing GATC motifs on the activity of 823 fragments initially containing the motif (top) and a specific example (fragment 7,571) with four motifs (bottom)
Deep mutagenesis of fragment 7,080: effects of 10-bp deletions (top)
1-bp deletions (X) or 1-bp mutations (bottom)
Effects of adding GATC motifs for 221 fragments with incremental motif additions (top) and four specific examples (bottom)
GATC motif-gain or motif-loss alleles in the 1001 Genomes population of accessions linked to nearby gene expression
The bar graph showcases the ratio of the number of significant associations with higher versus lower expression in the GATC motif allele
GATC motif’s enrichment in proximity to the TSS for all Arabidopsis genes
Gene expression response to overexpression of GATA TFs (individual graphs) versus GFP
2 or ≥3 GATC motifs within 500 bp downstream of the TSS
Overexpression of GATA TFs or GFP was driven by double 35S promoters in Arabidopsis protoplasts
which were collected 8 h after transformation
Box plots in a and c display the median (center line)
whiskers (minimum and maximum within 1.5 IQR) and outliers (points beyond whiskers)
This finding suggests that the GATC motif and other activity-enhancing sequences may act by the same mechanism to increase expression of the reporter constructs
reinforcing its role in enhancing gene expression when located downstream of the TSS
the GATC motif likely acts as a widespread and conserved regulatory signal in diverse biological functions
whiskers (minimum and maximum within 1.5 IQR) and outliers (points beyond whiskers); the number of genes per category is also indicated
P values in a–c and f were calculated using a two-sided t-statistic
This suggests that the GATC motif functions like a general rheostat
modulating gene expression of thousands of genes across plant cell types
we have identified the 500-bp region downstream of the TSS as a prominent site for transcription regulation for a large fraction of plant genes
We demonstrate that the function of regulatory sequences near the TSS is dependent on their position relative to the TSS
making them distinct from animal enhancers
We further examined a specific downstream GATC motif that modulates transcription in a dose-dependent manner through GATA TFs
the effect size of the GATC motif surpassed that of any other short DNA motif
The motif apparently acts as a regulatory module
operating much like a rheostat in tuning gene expression between cell types throughout vascular plants
Although this contradicts the common view of the role of the upstream region in controlling expression
the different ways in which enhancers are ‘read’ on either side of the TSS may account for these contrasting results
might create different local environments on either side of the TSS
but many other scenarios can be imagined as well
the GATC motif regulatory program exerts a widespread influence
modulating the gene expression of a substantial proportion of genes throughout the plant body
The adaptive advantages this mechanism offers and how it has evolved across different lineages promise to be a fertile ground for future exploration
Additional methods section can be found in Supplementary Information
Plasmids pPSup_iGFP (149420) and pPSint (149421) were obtained from Addgene and were modified to generate six new plasmids
and a 1-bp mutation was introduced to eliminate an extra BsmBI site in both plasmids
A ccdB lethal cassette was inserted between the two BsaI sites
These alterations produced pPSup_iGFP_v2 and pPSint_v2
and pPSint_v2_rmBsaI was created by removing the BsaI insertion sites with the ccdB cassette from pPSint_v2
a 633-bp genomic region was cloned into both pPSup_iGFP_v2 and pPSint_v2
This region was lifted from the TRP1 gene’s promoter
extending up to 40 bp upstream of the main TSS
The pTRP1 was positioned upstream of the BsaI site in pPSup_iGFP_v2 and upstream of the BsmBI site in pPSint_v2
generating pPSup_iGFP_v2_pTRP1 and pPSint_v2_pTRP1
the BsaI site and the ccdB cassette were removed from pPSint_v2_pTRP1 to create pPSint_v2_pTRP1_rmBsaI
25 µl KAPA HiFi HotStart ReadyMix (KK2601) and 14 µl double-distilled water (DDW)
The reactions were run using the following thermal cycling protocol: 95 °C
The amplified oligonucleotide pool was then purified using 1.4× AMPure XP (A63880) beads
and amplification was verified with an Agilent Fragment Analyzer
A dilution series was plated on LB–spectinomycin medium for both reaction and control to estimate cloning complexity
The rest of the transformation was grown overnight and purified with the Qiagen Plasmid Plus Midi Kit (12943)
diluted to 3.13 ng µl−1 and inserted into pPSup_iGFP_v2
A second insert contained the minimal promoter and the 5′ UTR of the TRP1 gene and was amplified using P6–P8 primers
diluted to 5 ng µl−1 and inserted into pPSup_iGFP_v2_pTRP1
Fifteen Golden Gate reactions and one control reaction were carried out for pPSup_iGFP_v2
pPSup_iGFP_v2_pTRP1 and pPSint_v2_pTRP1 as described above
substituting the restriction enzyme with BsmBI-v2 (R0739L) and adding the corresponding insert to each reaction
coli and efficiency calculation followed the same procedure
For pPSint_v2_rmBsaI and pPSint_v2_pTRP1_rmBsaI
two modifications were made: only ten Golden Gate reactions and one control were performed
The labeled MIX1 contained six libraries: pPSup_iGFP_v2 (p35S.SynJ-OP1)
pPSint_v2_pTRP1_rmBsaI (pTRP1) and pPSint_v2_rmBsaI (p35S.SynJ) in 50:50:50:50:1:1 proportions
with four libraries: pPSint_v2 (p35S.SynJ-OP1 + OP2)
pPSint_v2_rmBsaI (p35S.SynJ) and pPSint_v2_pTRP1_rmBsaI (pTRP1) in 100:100:1:1 proportions
contained two libraries: pPSint_v2_pTRP1 (pTRP1-OP1) and pPSint_v2_pTRP1_rmBsaI (pTRP1) in 100:1 proportions
These three mixes were transformed into MegaX DH10B T1R Electrocomp Cells with an efficiency of >108 and then purified with the Qiagen Plasmid Plus Giga Kit (12191)
To connect barcodes to tested fragments and libraries
the relevant region from the cloned libraries was sequenced with next-generation sequencing
To avoid the same initial bases in reads 1 and 2 in the Illumina run
due to amplification using the constant sequence around the barcode and the enhancer
which is detrimental to the imaging analysis of sequence signal
primers were chosen to create variation in read start
Every forward or reverse primer used was a combination of four primers that each had a different
0–3 nucleotides of shift before the constant sequence
Primers P9–P12 and P13–P16 were used for pPSup_iGFP_v2 (p35S.SynJ-OP1)
P17–P20 and P21–24 were used for pPSint_v2 (p35S.SynJ-OP1) and pPSint_v2 (p35S.SynJ-OP1 + OP2)
P17–P20 and P25–P28 were used for pPSint_v2_rmBsaI (p35S.SynJ)
P13–P16 and P29–P32 were used for pPSup_iGFP_v2_pTRP1 (pTRP1-OP1)
P33–P36 and P21–P24 were used for pPSint_v2_pTRP1 (pTRP1-OP1) and pPSint_v2_pTRP1 (pTRP1-OP1 + OP2)
and P33–P36 and P25–P28 were used for pPSint_v2_pTRP1_rmBsaI (pTRP1)
additional shotgun Tn5-based DNA-seq libraries
were prepared from each of the library mixes and sequenced in 50-bp or 100-bp paired-end mode on an Illumina NovaSeq 6000 machine
thaliana Col-0 seeds were sterilized with 70% (vol/vol) ethanol and 6.5% (vol/vol) bleach and sown on circular plates containing 0.5× MS
The plants were grown under long-day conditions (21 °C
M82 (sp−/sp−) seeds were sterilized with 70% ethanol and 3.25% bleach and sown in Magenta boxes (6 cm × 6 cm × 9.5 cm) containing 62.5 ml 0.217% (wt/vol) Nitsch medium (Duchefa Biochemie
Tomato plants were grown under long-day conditions (21 °C
B73 seeds were grown in soil (4:1 Klasmann Substrate 2:perlite) in the greenhouse (long-day photoperiod
150 μmol m−2 s−1) until 1–2-cm shoots were visible
The pots were covered and grown in the dark for 7–9 d
LAB seeds were sown on soil (4:1 Gramoflor 2006:perlite)
stratified for 4 d in the dark (4 °C) and transferred to short-day conditions (21 °C
The protocol was adapted from ref. 59
five to six leaves from 30–35 Arabidopsis seedlings or the true leaves from 20–25 tomato seedlings were cut into strips 0.5–1 mm wide using a razor blade and immediately submerged in 15 ml enzyme solution (0.4 M mannitol
1.5% (wt/vol) Cellulase R-10 (Duchefa Biochemie
0.4% (wt/vol) Macerozyme R-10 (Duchefa Biochemie
The enzyme solution was incubated overnight at 25 °C in the dark with gentle agitation (25 rpm)
The solution was strained through a 100-μm filter and centrifuged at 100g for 10 min at room temperature
This and all other centrifugation steps on protoplasts were performed with a soft start and end
The pellet was resuspended in 3 ml W5 solution (154 mM NaCl
and healthy protoplasts were isolated using a sucrose gradient (23% (wt/vol)) by centrifugation at 450g for 3 min at room temperature
The protoplast fraction was resuspended in 14 ml W5 solution and counted on a hemocytometer
The suspension was centrifuged at 100g for 10 min at room temperature
and the pellet was resuspended in MMG solution (0.4 M mannitol
pH 5.7) to a concentration of 1 million cells per ml
These were placed into two distinct 1.5-ml tubes
which codes for the Clover protein with a nuclear localization signal sequence expressed with the pUBI promoter
The rest of the suspension was split into 50-ml Falcon tubes with 4–6 ml suspension each and mixed with 50 μg plasmid library per million cells
MW 4000) equal to that of the protoplast–DNA suspension was added to each tube
and the protoplasts were incubated for 20 min in the dark at room temperature
W5 solution (0.95 ml) was added to the controls
and 4.75 ml per million cells was added to the samples
After 15 min of incubation at room temperature
the protoplasts were centrifuged at 450g for 5 min at room temperature
and the pellets were resuspended in 1 ml W1 solution (0.5 M mannitol
pH 5.7) for the controls and 5 ml per million cells for the samples
Each protoplast suspension was transferred to a separate sterile Petri dish and incubated at 25 °C under constant light (85 μmol m−2 s−1) for 6 h (samples) or overnight (controls)
the samples were centrifuged at 450g for 5 min at room temperature
and the pellets were flash frozen in liquid nitrogen
The samples were stored at −70 °C until RNA extraction
The controls were imaged under a microscope to check the transformation efficiency: typically around 70–80% in the positive control were successfully transformed
Testing the efficiency of transformation on a large scale
This experiment was carried out with four replicates for Arabidopsis
10 and 14 million protoplasts and with 3 replicates for tomato
The protocol was adapted from ref. 60
The middle parts (6–8 cm) of the second leaf from 30 maize plants were used
and both halves were placed on top of each other
followed by cutting into 0.5–1-mm-wide strips perpendicular to the veins using a razor blade
Strips were immediately submerged in 60 ml enzyme solution (0.6 M mannitol
0.3% (wt/vol) Macerozyme R-10 (Duchefa Biochemie
5 mM β-mercaptoethanol) split into four Petri dishes with 15 ml solution each
The enzyme solutions were covered and vacuum infiltrated for 1 h at room temperature and then incubated for 2 h in the dark at room temperature with gentle agitation (40 rpm)
The protoplasts were released by shaking at 80 rpm for 10 min
The solutions were strained through a 100-μm filter and combined in two tubes with 30 ml solution each
The tubes were centrifuged (all centrifugations with protoplasts were carried out with a soft start and end) at 70g for 3 min at room temperature
and the pellets were resuspended in 10 ml 0.6 M mannitol
The two protoplast suspensions were combined in one tube and counted on a hemocytometer
The suspension was centrifuged at 70g for 3 min at room temperature
and the pellet was resuspended in MMG solution (0.6 M mannitol
The protoplast suspension was split into several 50-ml Falcon tubes with 4–6 ml suspension each and mixed with 200 μg plasmid library per million cells
and the protoplasts were incubated for 15 min in the dark at room temperature
W5 solution (5 ml per million cells) (the same as for Arabidopsis and tomato protoplasts) was added
The samples were centrifuged at 70g for 3 min at room temperature
and the pellets were resuspended in 5 ml incubation solution (0.6 M mannitol
Each suspension was transferred to a separate sterile Petri dish and incubated at 25 °C in the dark for 12 h
and the rest of the samples were centrifuged at 70g for 3 min at room temperature
The pellets were flash frozen in liquid nitrogen and stored at −70 °C until RNA extraction
The 1-ml aliquot was imaged under a microscope to check the transformation efficiency: 77% and 93% of protoplasts were successfully transformed in the first and second replicates
while the third replicate was not quantified
The protoplast counts for the experiments were 21
25 μl ACC-110 competent cells was mixed with 1 μl plasmid
transferred to a prechilled 0.1-cm cuvette (Bio-Rad) and electroporated (1,800 V
1 ml of prewarmed (30 °C) LB medium was added
and the mixture was incubated at 30 °C for 160 min at 200 rpm
Agrobacterium was then plated in a dilution series on LB plates containing spectinomycin
gentamicin and rifampicin to estimate transformation efficiency
The remaining Agrobacterium was grown overnight in 50 ml LB with the same antibiotics
Agrobacterium was centrifuged for 5 min at 3,000g
resuspended to an OD600 of 0.5 in infiltration medium (50 mM NaH2PO4
pH 5.6–5.7) and incubated for 1–2 h in the dark at room temperature with gentle agitation
Agrobacterium was then infiltrated into 17–20 plants (two leaves per plant
Plants were not watered for 2 d before infiltration and were watered immediately after
plants were covered with a transparent cover for 24 h
and infiltrated leaves were harvested in liquid nitrogen 48 h after infiltration
This experiment was carried out with four replicates
tomato and maize protoplasts was extracted using the Monarch Total RNA Miniprep Kit (New England Biolabs
The frozen pellets were thawed shortly on ice
and then the protocol for ‘Cultured Mammalian Cells’ in part 1 of the kit manual was followed by resuspending each sample with 400–600 μl lysis buffer and proceeding to part 2
The Arabidopsis and tomato samples were treated with DNase I as recommended in the kit manual
Due to the larger amount of plasmid used for transformation of maize protoplasts
DNase I treatment was repeated three times for maize samples
Harvested samples were ground using a mortar and pestle in liquid nitrogen and then mixed with TRIzol LS (up to a 1:3 tissue:TRIzol volume ratio)
vortexed for 10 min at room temperature and centrifuged for 5 min at 12,000g and 4 °C
The clear fraction was transferred to a new tube
and 0.2 ml chloroform per 1 ml TRIzol LS was added for lysis
incubated for 3 min at room temperature and then centrifuged for 15 min at 16,000g and 4 °C
The aqueous phase was combined (1:1) with chloroform
incubated for 3 min and centrifuged for 15 min at 16,000g and 4 °C
The resulting aqueous phase was mixed with 0.5 ml isopropanol per 1 ml TRIzol used for lysis
incubated for 10 min on ice and centrifuged for 60 min at 21,000g and 4 °C
and the sample was washed twice with 70% ethanol (1:1 ratio of TRIzol used for lysis)
vortexed and centrifuged for 5 min at 7,500g and 4 °C
and residual ethanol was allowed to evaporate for 7 min at room temperature
RNA was eluted by adding 500 μl DDW and incubating for 10 min at 42 °C
followed by resuspension in 0.5× volume of 2× binding buffer
Total RNA was mixed with the washed beads and incubated for 10 min on a rolling shaker
The tubes were placed on a magnetic separator and washed twice with the same volume of washing buffer as the starting volume of the beads
the beads were resuspended in 60 μl 10 mM Tris-HCl (pH 7.5) and incubated at 80 °C for 3 min at 750 rpm
The tubes were placed immediately on a magnetic separator and incubated for >1 min
The eluted mRNA was transferred to a new RNase-free tube
The beads were re-eluted with new 30 μl 10 mM Tris-Cl (pH 7.5) buffer
ten reactions with 11 μl mRNA each and a construct-specific primer (P37–P40) were prepared for complementary DNA (cDNA) synthesis using SuperScript IV reverse transcriptase (RT; Thermo Fisher Scientific
One of the reactions was used as a no-reverse transcription control
in which the RT enzyme was replaced with RNase-free water
1 μl RNase A (200 μg ml−1) was added to each reaction
and the samples were incubated at 37 °C for 1 h
The nine RT reactions were pooled and purified with 1.8 volumes of AMPure XP beads (Beckman Coulter
The no-RT control was processed separately in the same way as the RT reactions
the RT reactions and the no-RT control were eluted in 146 μl and 49 μl 10 mM Tris-HCl (pH 8) buffer
The purified cDNA was split into two 72-μl aliquots
and each aliquot was used for the preparation of three PCR reactions amplifying the p35S-based transcripts (P17–P20 and P41) and the pTRP1-based transcripts (P33–P36 and P41)
The no-RT control was also split in two 24-μl aliquots
and each aliquot was used for one PCR reaction of the p35S and pTRP1 transcripts
All PCR reactions were prepared with 24 μl template
25 μl KAPA HiFi HotStart ReadyMix (Roche Molecular Systems
0.5 μl forward primer (100 μM) and 0.5 μl reverse primer (100 μM)
benthamiana tissue were amplified with 21 cycles
tomato and maize protoplasts were amplified with 24 cycles
The three reactions from each library (p35S and pTRP1) were pooled and purified with an equal volume of AMPure XP beads
The two no-RT reactions (p35S and pTRP1) were processed separately in the same way
all samples were analyzed with the 5200 Fragment Analyzer (Agilent
M5310AA) and sequenced on an Illumina NovaSeq 6000 or NextSeq 2000 system in paired-end configuration
The protocol was adapted from ref. 23 and the manual of the Click-iT Nascent RNA Capture Kit (Invitrogen)
Arabidopsis protoplasts were transformed with MIX3 following the same procedure as for the MPRA in Arabidopsis protoplasts
the samples were incubated for 5 h and 40 min at 25 °C under constant light (85 μmol m−2 s−1)
A 200 mM stock solution of 5-EU (Click-iT Nascent RNA Capture Kit
C10365) was added to each sample to a final concentration of 200 μM
and the samples were incubated for an additional 20 min before collection
Total RNA was extracted from the frozen pellets as described above
DNase I treatment was repeated three times for each sample
mRNA was isolated from the total RNA as described above
Dynabeads Oligo(dT)25 (Thermo Fisher Scientific
61005) were resuspended in 55 μl 10 mM Tris-HCl (pH 7.5) and then re-eluted with the first eluate
Five to eight microliters of mRNA was set aside for preparation of libraries from total mRNA
whereas the remaining mRNA was split into three aliquots and each aliquot was used for one Click reaction with 0.25 mM biotin azide following the Click-iT kit manual
Biotinylated mRNA was precipitated from each Click reaction following the manual and resuspended in 25 μl RNase-free water
A bead suspension (3 μl) was added to each aliquot of biotinylated mRNA
and samples were incubated for 30 min at room temperature with rotation at 30 rpm
the three aliquots were pooled and washed five times with wash buffer 1 and five times with wash buffer 2 (wash buffers from the Click-iT kit)
The bead suspension was resuspended in 50 μl wash buffer 2 and used immediately for cDNA synthesis
Libraries were prepared from the total mRNA and 5-EU-labeled mRNA bead suspension samples following the procedure for MPRA libraries with a few modifications
The total mRNA samples were diluted to a final volume of 50 µl with RNase-free water
four reactions with 11 μl mRNA each and a construct-specific primer (P37–P40) were prepared for cDNA synthesis using SuperScript IV RT (Thermo Fisher Scientific
An additional reaction with the remaining mRNA (5.5 µl) was prepared as a no-reverse transcription control
The cDNA reactions with 5-EU-labeled mRNA were incubated on a shaker at 1,500 rpm to prevent settling of the beads on the bottom of the tube
whereas the reactions with total mRNA were incubated without mixing
Following the final step of cDNA synthesis
the reactions with 5-EU-labeled mRNA were immediately placed on a magnetic rack and the supernatants were transferred to new tubes
the total mRNA and 5-EU-labeled mRNA samples were handled identically
RNase A treatment and purification with AMPure XP beads was performed as described above
the RT reactions and the no-RT control were eluted in 73 μl and 24.5 μl 10 mM Tris-HCl (pH 8) buffer
The purified cDNA from the RT reactions was used to prepare three PCR reactions amplifying the pTRP1-based transcripts (P33–P36 and P41)
The no-RT control was used for one PCR reaction of the pTRP1 transcripts
All PCR reactions were incubated for 24 cycles
The second purification with AMPure XP beads was performed as described above
This experiment was carried out with two replicates
To estimate the specificity of the Click-iT Nascent RNA Capture Kit in our system
Arabidopsis protoplasts were transformed with a plasmid encoding the Clover protein under the control of the Petroselinum crispum ubiquitin (PcUbi) promoter
the protoplast suspension was split into three samples of 4.3 million protoplasts
and the samples were incubated at 25 °C under constant light (85 μmol m−2 s−1) for 6 h
200 mM 5-EU stock solution was added to one of the samples (‘2 h 5-EU’) to a final concentration of 200 μM
the same amount of 5-EU stock solution was added to the second sample (‘20 min 5-EU’)
and an identical volume of DMSO was added to the third sample (‘no 5-EU’)
all three samples were collected and stored at −70 °C
Total RNA was extracted from the samples as described above
RNA labeled with 5-EU was isolated from the total RNA as described above
Biotin azide (0.5 mM) was used for each Click reaction
cDNA was prepared from both the total RNA and 5-EU-labeled RNA samples using the SuperScript VILO cDNA Synthesis Kit (Invitrogen
No-reverse transcription controls were prepared for each sample
in which the enzyme mix was heat inactivated at 65 °C for 10 min following the kit manual
qPCR reactions were prepared from all samples using an in-house qPCR mix and oligonucleotides specific for ACTIN2 (AT3G18780) mRNA and Clover mRNA
The results were analyzed using the LightCycler 96 system (Roche Diagnostics)
the estimated amount of nonlabeled mRNA in the ‘20 min 5-EU’ and ‘2 h 5-EU’ samples was less than 9% and 2% for ACTIN2 mRNA
The plasmids containing the coding sequences of the three GATA TFs were synthesized by Twist Bioscience
The CDS of GATA1 was modified to remove the internal BsaI restriction site with a synonymous mutation at the sequence for Gly46 (GGT → GGA)
The four final plasmids were transformed into DH5α competent E
coli and purified using the Qiagen Plasmid Plus Midi Kit (12943)
A plasmid (10 μg) encoding the GATA TF or GFP was transformed into 200,000 Arabidopsis leaf protoplasts
Each construct was transformed in two replicates into two independent protoplast preparations
resulting in four replicates per construct
An additional two samples per protoplast production were transformed with 10 μg GFP control vector and 10 μl elution buffer
serving as positive and negative imaging controls
After 8 h of incubation at 25 °C under constant light (85 μmol m−2 s−1)
and the pellets were flash frozen in liquid nitrogen and stored at −70 °C
The controls were imaged shortly before collection under a microscope to check the transformation efficiency: 60% and 70% of protoplasts were successfully transformed in the first and second replicates
with each library constructed in multiple technical replicates and sequenced on a NovaSeq X or NovaSeq 6000 system with paired-end configuration
All experiments were performed in at least triplicate as described in Methods. No data were excluded from analysis. The statistical tests used for data analysis are described in the main text or in Methods
No statistical method was used to predetermine sample size
and the investigators were not blinded to allocation during the experiments and outcome assessment
Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article
Integrated genomic and fossil evidence illuminates life’s early evolution and eukaryote origin
The multiple origins of complex multicellularity
The dynamic regulatory genome of Capsaspora and the origin of animal multicellularity
Plants compared to animals: the broadest comparative study of development
Genome-wide computational prediction and analysis of core promoter elements across plant monocots and dicots
Transcription factor families have much higher expansion rates in plants than in animals
Plant-TFClass: a structural classification for plant transcription factors
evolution and chromatin signatures of plant regulatory elements
Cis-regulatory sequences in plants: their importance
Advances in understanding cis regulation of the plant gene with an emphasis on comparative genomics
Epigenomic diversity in a global collection of Arabidopsis thaliana accessions
1,135 genomes reveal the global pattern of polymorphism in Arabidopsis thaliana
High-resolution mapping of expression-QTLs yields insight into human gene regulation
target reporter transcripts for rapid decay in tobacco
Genome-wide analysis of mRNA decay rates and their determinants in Arabidopsis thaliana
Cistrome and epicistrome features shape the regulatory DNA landscape
Identification of plant enhancers and their constituent elements by STARR-seq in tobacco leaves
Intron DNA sequences can be more important than the proximal promoter in determining the site of transcript initiation
The intron of Arabidopsis thaliana polyubiquitin genes is conserved in location and is a quantitative determinant of chimeric gene expression
Metabolic labeling of RNAs uncovers hidden features and dynamics of the Arabidopsis transcriptome
Synthetic promoter designs enabled by a comprehensive analysis of plant core promoters
Arabidopsis transcription factors: genome-wide comparative analysis among eukaryotes
Integrative analysis from the epigenome to translatome uncovers patterns of dominant nuclear regulation during transient stress
Direct measurement of transcription rates reveals multiple mechanisms for configuration of the Arabidopsis ambient temperature response
A high resolution map of the Arabidopsis thaliana developmental transcriptome based on RNA-seq profiling
The embryonic transcriptome of Arabidopsis thaliana
A single-cell Arabidopsis root atlas reveals developmental trajectories in wild-type and cell identity mutants
A single-cell analysis of the Arabidopsis vegetative shoot apex
The GATA family of transcription factors in Arabidopsis and rice
Spatiotemporal transcriptome provides insights into early fruit development of tomato (Solanum lycopersicum)
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Differential expression and co-localization of transcriptional factors during callus transition to differentiation for shoot organogenesis in the water fern Ceratopteris richardii
The Chinese pine genome and methylome unveil key features of conifer evolution
Conserved gene expression programs in developing roots from diverse plants
Transcriptome-wide profiling and expression analysis of transcription factor families in a liverwort
Intron-mediated regulation of gene expression
Genomic editing of intronic enhancers unveils their role in fine-tuning tissue-specific gene expression in Arabidopsis thaliana
Introns as gene regulators: a brick on the accelerator
Identification of cis-regulatory motifs in first introns and the prediction of intron-mediated enhancement of gene expression in Arabidopsis thaliana
An intron-derived motif strongly increases gene expression from transcribed sequences through a splicing independent mechanism in Arabidopsis thaliana
Genome-wide enhancer identification by massively parallel reporter assay in Arabidopsis
Global quantitative mapping of enhancers in rice by STARR-seq
Regulatory enhancer–core-promoter communication via transcription factors and cofactors
Genome-wide analysis of chromatin packing in Arabidopsis thaliana at single-gene resolution
Accessible gene borders establish a core structural unit for chromatin architecture in Arabidopsis
The G-box transcriptional regulatory code in Arabidopsis
A gene expression map of Arabidopsis thaliana development
Potential targets of VIVIPAROUS1/ABI3-LIKE1 (VAL1) repression in developing Arabidopsis thaliana embryos
STARR-seq and UMI-STARR-seq: assessing enhancer activities for genome-wide-
A 28 nt long synthetic 5′UTR (synJ) as an enhancer of transgene expression in dicotyledonous plants
Uncovering the dynamics of precise repair at CRISPR/Cas9-induced double-strand breaks
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and efficient cloning system for plant transgenesis
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Ben-Tov for help establishing the protoplast system; Y
Bindics for sharing seeds; and the Plant Sciences and Next Generation Sequencing facilities at the Vienna BioCenter Core Facilities
This work was supported by core funding to M.N
from EU Horizon 2020 via the VIP2 program and Marie Skłodowska-Curie individual fellowships (101028014)
Almudena Mollá-Morales & Magnus Nordborg
conceived and designed the experiments and wrote the paper
The authors declare no competing interests
This study did not require any specific ethical approval
Nature Genetics thanks the anonymous reviewers for their contribution to the peer review of this work. Peer reviewer reports are available
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations
with data aligned to the ATG and not the TSS
with the upstream insertion placed just before the minimal core promoter
(b) The pTRP1-based construct consists of the 791 bp genomic sequence preceding the coding region of the TRP1 gene (purple)
The upstream insertion site is situated within this sequence
118 bp upstream of the major TSS and 40 bp upstream of the TSS annotation in the TAIR10 database
Data represent average signal from two replicates of mRNA synthesis MPRA experiments for total mRNA and newly synthesized mRNA
Error bars represent the mean ± 1 standard deviation
Examination of the effect of GATC motif’s impact on MPRA expression levels using 6-mers
For all 26 unique 6-mer sequences containing a GATC
the average log2 expression difference between sequences having the 6-mer and those lacking the 6-mer is plotted
6-mers are ordered based on their median effect across the eight experimental setups
Shape indicates the construct backbone (p35S or pTRP1)
and AgatcG) with the highest median effect were used to establish the consensus YVGATCBR sequence defined as the GATC motif
which was then used in downstream analyses
MPRA experiments were performed in Arabidopsis protoplasts using synthetically mutated sequences inserted only in the downstream position
Both p35S- and pTRP1-based libraries were used
The libraries contained 30,000 fragments: 12,000 from the initial pool and an additional 18,000 fragments
each being a variant of one of the original fragments
(a) Comparison between each pair from the three replicates
displaying results with the p35S-based library at the top and the pTRP1-based library at the bottom
Pearson’s correlation coefficients (r) and numbers of compared fragments (n) are indicated on each graph
(b) Log2 expression ratio between mutated fragments and their original fragment
Mutations encompass: addition of a GATC motif
which include both deletions and nucleotide substitutions
either p35S- or pTRP1-based; numbers below the x-axis indicate the number of fragments in each category
Error bars depict the mean with ±1 standard deviation
Effects across 221 fragments (top) and 4 specific examples (bottom)
(c) Pearson’s correlation coefficients between expression of 221 fragments with varying GATC-motif additions in p35S- and pTRP1-based libraries
(d) Influence of GATC-motif additions depends on initial sequence expression
Depicted is the average log2 expression difference upon motif addition relative to original fragments
for different copy numbers of the added motif
Color indicates original fragment (log2) expression level ranges
Plotted for p35S- (top) and pTRP1-based libraries (bottom)
Boxplots in a,b represent the median (center line)
Genes showing at least a 50% change in expression with an adjusted p-value < 0.001 were defined to be differentially expressed and are shown in blue (downregulated) and red (upregulated); all other genes are shown in gray
(d) The percentage of genes with a GATC motif within 500 bp downstream of the TSS is shown
or unchanged expression (labeled ‘Unchanged’) as defined in c
Further separation is provided for genes with 1
or 3 or more GATC motifs for each GATA TF OE experiment
with effect sizes of 0.01 (p-value: 0.4) at 17 °C and −0.01 (p-value: 0.6) at 27 °C
p-values are calculated using a two-sided t-statistic
presented as in their original study for comparison
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DOI: https://doi.org/10.1038/s41588-024-01907-3
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Recent work has shown that Arabidopsis thaliana contains genetic groups originating from different ice age refugia
with one particular group comprising over 95% of the current worldwide population
relicts of other groups can be found in local populations along the Mediterranean Sea
Here we provide evidence that these ‘relicts’ occupied post-glacial Eurasia first and were later replaced by the invading ‘non-relicts’
which expanded through the east–west axis of Eurasia
leaving traces of admixture in the north and south of the species range
The non-relict expansion was likely associated with human activity and led to a demographic replacement similar to what occurred in humans
Introgressed genomic regions from relicts are associated with flowering time and enriched for genes associated with environmental conditions
such as root cap development or metal ion trans-membrane transport
which suggest that admixture with locally adapted relicts helped the non-relicts colonize new habitats
In this study we investigate this post-glacial expansion in greater detail
We first identify the traces of hybridization between relicts and non-relicts and investigate whether the pattern of introgression appears to have been associated with adaptation to the local environment
We then explore the geographic patterns of relict introgression
focusing in particular on finding traces of extinct relicts in non-relict genomes
and on inferring the possible origin of non-relict expansion
thaliana of today is the product of a dramatic series of range expansions
thaliana genomic variation is very different from that before the last ice age
(a) ADMIXTURE with K=2 separates Iberian relicts and French non-relicts and shows the hybrid origin of Iberian non-relicts
the SNP with highest climate and phenology GWAS scores
The colour gradient is based on climate PC2
separating southern hot and dry (yellow) versus northern cold and wet (red) environments
Two alleles are labelled as X (reference) or solid dots with black outline (alternative allele)
(c) 1 Mb haplotype around this SNP (vertical black line)
Columns are SNPs whose major allele is different between Iberian relicts and French non-relicts
and rows are accessions with black horizontal lines separating populations
The ‘reference’ or ‘alternative’ labels below each population denote the allele each population has in this SNP
Red and blue: major allele in Iberian relicts and French non-relicts respectively
Iberian non-relicts with the alternative allele mostly contain haplotypes from Iberian relicts and some from Italy
(d) SNP LD with chr4:10999188 across the 1 Mb region
Solid vertical line is the location of chr4:10999188
and dashed vertical lines mark every 100 kb away from that SNP
The map was created with data from package ‘rworldmap’ of R
The QQ-plot shows the enrichment of high flowering-time GWAS scores of ‘SNPs with high Iberian relict introgression into Iberian non-relicts’ versus ‘genomic SNPs with similar allele frequencies’
This analysis excludes the 1 Mb region near SNP chr4:10999188
Blue dots and line represent true value distribution
and the grey area denotes 5% significance thresholds based on 1,000 permutations
suggesting that the Iberian relicts might have been better adapted than the early invaders with respect to traits other than flowering
and that non-relicts currently inhabiting Iberia received the adaptive allele from relicts
(d) The density of outlier haplotypes across Eurasia
using thinned samples to take uneven sampling into account
and with relicts and Iberian non-relicts excluded due to their exceptionally high number of outliers (see ‘Methods’)
The maps were created with data from the package ‘rworldmap’ of R
The origin of each outlier haplotype in each local population (a horizontal bar) was assigned to one of the four known relict groups or an unknown relict
For each relict group (the four lower bar)
the comparison was made only with the three other relict groups without to itself
serving as positive controls (origin of most outlier haplotypes unknown)
Iberian non-relicts (the top bar) were used as negative control
where the origin of outlier haplotypes is mostly known
The only significant difference of proportion of unassigned outlier haplotypes (black) is between Karelia and Kashmir (P<0.001
Fisher’s exact test from random resampling 1,000 outlier haplotypes in each population)
We used the Iberian non-relicts as a negative control
the source of introgression is known (mainly Iberian relicts)
and thus the fraction of unassigned outlier haplotypes (in the sense given above) serves as lower bound for what we might expect to see if there is no introgression from an unknown source
we use the known relict population as a form of ‘positive control’
we assign outlier haplotypes to the other three relict populations (that is
not allowing them to be assigned to their own populations—their true origin)
The proportion of unassigned outliers after this procedure should give us an idea of what to expect when the true source of the outlier haplotypes is missing
The different results for Kashmir suggest that an unknown relict population may have contributed to the present-day polymorphism in this region
Grey denotes area with 1% lowest values
and the red cross is the most likely origin
Populations and accessions with high number of outlier haplotypes
were excluded to minimize relict influence
Blue dots represent non-relict populations
Colour scale represents Spearman’s rank correlation between population polymorphisms π and the geographic distances from all populations to each location
(a) Geographical distribution of accessions with the derived (grey dots) or ancestral (coloured dots) haplotypes
The five different colours denote five ADMIXTURE groups among the latter
Black dots are admixed ancestral haplotypes
(b) Pairwise distance distribution for the whole genome (solid line) and the transposition region (dashed line)
showing that differences among ancestral haplotypes are comparable to the whole-genome difference between relicts and non-relicts
(c) Ancestral haplotypes show stronger pattern of isolation by distance (red dots) than derived haplotypes do (blue dots)
Grey denotes regions with 1% lowest value
(a) Estimate based on population polymorphism of the derived haplotypes
(b) Estimate based on the genetic distances
Black dots represent accessions with the ancestral haplotype
Colour scale represents the genetic distance between local ancestral and derived haplotypes
The maps were created with data from package ‘rworldmap’ of R
populations in different refugia diverged into separate groups
one group from northern Balkan and eastern Europe (the non-relicts) quickly expanded east–west
generating the present-day pattern that (d) relict genomic regions are mainly found in the south and north of species range
We emphasize that, although we have done our best to rule out alternative models, the model space in these types of historical analyses is infinite, and it is therefore impossible to ‘prove’ that the scenario in Fig. 8 is the right one
It is a model that fits the current data well (and we welcome attempts to find better ones—in particular as more samples from under-sampled regions become available)
and relicts were defined as those who have exceptional high median distance to others
We performed 10 independent runs with different random number seeds and reported the one with lowest cross validation error
Note that we did not intend to estimate the best K value and the accessions’ corresponding population assignments
given the high divergence between relicts and non-relicts
unsupervised ADMIXTURE K=2 would separate Iberian relicts from French non-relicts and estimate the proportional ancestry of Iberian non-relicts from either parental populations
A significant negative value suggests Iberian non-relicts being hybrids from the two other populations
For both ABBA–BABA and the three-population test
results are regarded as significant if the absolute value of Z scores is higher than three
To visualize the extent and size of introgression
for each 10 kb window we estimated the probability that each individual descended from either parental population
A SNP was regarded informative if the major allele differed between Iberian relicts and French non-relicts
For each informative SNP in each individual
we calculated the probability of the individual’s allele descending from the relict population as pr*(pr+pn)−1
where pr and pn are the frequencies of the target individual’s allele in the relict and non-relict populations respectively
This value was then averaged across all informative SNPs within each window
Using the multiplication rather than the average of conditional probabilities produced similar results
Windows with <5 informative SNPs were excluded
using a kinship matrix in a mixed model framework to correct for population structure
and we suspect this may be caused by the non-independence among SNPs
To compare the magnitude of relict introgression into Iberian non-relicts in the 300 kb high-LD region around SNP chr4:10999188 with the rest of the genome
we calculated the average ancestry index of all diagnostic SNPs inside the 300 kb region and compare the observed value to those calculated from 1,000 randomly drawn 300 kb window from the genome
We used similar logic to estimate the ancestry index for 10 kb windows
using SNPs from coding regions of the 20,186 genes
for each 10 kb window in each accession we calculated the probability that its haplotype descended from Iberian relicts (as opposed to from French non-relicts)
Here in each window the probability was averaged among accessions within each of the three populations
and a diagnostic window was defined as one with mean probability ≥0.75 in Iberian relicts and ≤0.25 in French non-relicts
Ancestry index was calculated with the same equation above
and therefore the top 25% windows with highest ancestry index represented ones where haplotypes from Iberian relicts and French non-relicts were highly differentiated and where Iberian non-relict haplotypes were mostly descended from the relicts
We then compared the enrichment of GO terms between these high-introgression windows versus background ones
significance was estimated by Fisher’s exact test with the same permutation method described above
and haplotypes of relict origin behaved as outliers
This method did not require the relict parental information as the Iberia–France comparison and may identify residual relict haplotypes from any unknown relict group that did not exist among our samples
Note that a genomic region with highly diverged haplotypes could be caused by either long-term balancing selection or recent relict introgression
should have much shorter haplotype length than the latter due to generations of recombination and was less likely to be identified by our 10 kb window size
The whole outlier haplotype analysis was applied on this resampled data set
and we compared accessions’ number of outlier haplotypes with latitudinal origin among the 1,000 resampling trials
for each accession we averaged the numbers of outlier haplotypes identified across the resampling trials it appeared
Using different grid sizes did not change our conclusion
the outlier grids were identified in each 10 kb window
and we plotted number of outlier windows of each grid on the map
We performed this analysis to test whether outlier haplotypes within non-relicts were introgressed from the four previously defined relict groups (Cape Verde
Sicily and Lebanon) or other unknown relicts not present in our samples
with emphasis on regions where outlier haplotypes were enriched (northern Sweden
For every outlier haplotype in an accession
we estimated its average genetic distance to the four known relict groups and assigned the relict group with closest genetic distance as its origin
We did not include non-relicts as a potential origin because the haplotype was already identified as outlier from all non-relicts
If the genetic distance to the closest relict is still higher than the threshold used to define it as an outlier (see the previous section)
the origin was estimated from all outlier haplotypes in all accessions
and we compared the proportion of unknown origin among populations to infer possible introgression from an unknown relict
Since previous results showed strong evidences of introgression from Iberian relicts (and possibly other Mediterranean relicts) into Iberian non-relicts
we estimated the origin of outlier haplotypes in Iberian non-relicts as a negative control
representing the proportion of unknown origin if the origin of all outlier haplotypes were known
The four relicts groups were also analysed while respectively excluding themselves from the reference
thus serving as positive controls when a significant proportion of outlier haplotypes had an unknown origin
If all outlier haplotypes in a population were descended from the four known relicts
the proportion of unknown origin should be significantly different from positive controls but not from negative controls
and the four target populations based on their proportion of unknown origin and tested for significant difference of proportional unknowns between neighbouring populations
Since the total number of outlier haplotypes varied among populations
a naïve test might tend to assign higher significance to comparisons with larger sample size
We therefore randomly resampled 1,000 outlier haplotypes with replacement from each population and use Fisher’s exact test on this data set where all populations had the same sample size
and the opposite is true if this specific geographic location is far from the origin
Populations were defined from geographic regions18
To minimize the influence of relict introgression
we excluded all accessions from Iberia and Africa
we also exclude parts of their genome showing evidences of relict introgression: In an accession
if a 10 kb window was identified as a outlier haplotype from the previous analyses
all sites in this region of this accession were treated as missing
Since the relict introgression only constituted a small portion of non-relict genomes
considering these regions as missing effectively removed the interference from relict introgression without heavily biasing the estimate of pairwise genomic distances
Mean pairwise distances π within each population were calculated from this filtered data
we examined the distribution of this transposition in the 1,002 accessions
Accessions were determined to have the ancestral or derived arrangement based on: (1) Illumina paired-end reads spanning breakpoints of the ancestral or derived haplotypes
(2) accessions’ position in the neighbor-joining tree and (3) pairwise distances between accessions
we reported accessions’ population assignment from the run with lowest cross-validation error
An accession was assigned admixed (in this transposition region) if its proportional ancestry was below 0.5 in all populations
The ADMIXTURE assignment was also compared with results from principal coordinate analysis of pairwise genetic distances of the ancestral haplotypes
All data are available at http://1001genomes.org/ or available from the authors upon request
On the post-glacial spread of human commensal Arabidopsis thaliana
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations
Evidence for archaic adaptive introgression in humans
An early modern human from Romania with a recent Neanderthal ancestor
Genetic history of an archaic hominin group from Denisova Cave in Siberia
The genomic landscape of Neanderthal ancestry in present-day humans
The complete genome sequence of a Neanderthal from the Altai Mountains
Altitude adaptation in Tibetans caused by introgression of Denisovan-like DNA
Genetic isolation by distance in Arabidopsis thaliana: biogeography and post-glacial colonization of Europe
Demographic history of European populations of Arabidopsis thaliana
Native range genetic variation in Arabidopsis thaliana is strongly geographically structured and reflects Pleistocene glacial dynamics
The pattern of polymorphism in Arabidopsis thaliana
The extent of linkage disequilibrium in Arabidopsis thaliana
Natural genetic variation of Arabidopsis thaliana is geographically structured in the Iberian Peninsula
The genetic structure of Arabidopsis thaliana in the south-western Mediterranean range reveals a shared history between North Africa and southern Europe
Genome-wide patterns of genetic variation in worldwide Arabidopsis thaliana accessions from the RegMap panel
Whole-genome sequencing of multiple Arabidopsis thaliana populations
Massive genomic variation and strong selection in Arabidopsis thaliana lines from Sweden
1,135 Genomes reveal the global pattern of polymorphism in Arabidopsis thaliana
Ecological Imperialism: The Biological Expansion of Europe
900-1900 1st edn Cambridge University Press (1986)
Exposito-Alonso, M. et al. The rate and effect of de novo mutations in a colonizing lineage of Arabidopsis thaliana. Preprint at http://biorxiv.org/content/early/2016/11/22/050203 (2016)
The scale of population structure in Arabidopsis thaliana
Very high resolution interpolated climate surfaces for global land areas
Altitudinal and climatic adaptation is mediated by flowering traits and FRI
Genome-wide association mapping of flowering time in in nature: genetics for underlying components and reaction norms across two successive years
Linkage and association mapping of Arabidopsis thaliana flowering time in nature
Lasky, J. R. & Reimherr, M. Uncovering the genomic basis of local adaptation by coherent synthesis of associations with phenotypes and home environments. Preprint at http://biorxiv.org/content/early/2016/11/03/051110 (2016)
Excavating Neandertal and Denisovan DNA from the genomes of Melanesian individuals
Support from the relationship of genetic and geographic distance in human populations for a serial founder effect originating in Africa
Common sequence polymorphisms shaping genetic diversity in Arabidopsis thaliana
Keeping it local: evidence for positive selection in Swedish Arabidopsis thaliana
Fast model-based estimation of ancestry in unrelated individuals
Massive migration from the steppe was a source for Indo-European languages in Europe
Surfing during population expansions promotes genetic revolutions and structuration
The fate of mutations surfing on the wave of a range expansion
Mutations arising in the wave front of an expanding population
VAT: a computational framework to functionally annotate variants in personal genomes within a cloud-computing environment
Testing for ancient admixture between closely related populations
Evaluating the use of ABBA-BABA statistics to locate introgressed loci
The Arabidopsis lyrata genome sequence and the basis of rapid genome size change
Using environmental correlations to identify loci underlying local adaptation
Adaptation to climate across the Arabidopsis thaliana genome
A mixed-model approach for genome-wide association studies of correlated traits in structured populations
Mathematical model for studying genetic variation in terms of restriction endonucleases
R Core Team. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austriahttp://www.R-project.org/ (2014)
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a collaborative grant from Austrian Science Fund and DFG (SPP ADAPTOMICS; M.N.
BIO2013-45407-P from the Ministerio de Economia y Competitividad of Spain (C.A.-B.)
and 105-2311-B-002-040-MY2 from Ministry of Science and Technology of Taiwan (C.-R.L.)
Institute of Ecology and Evolutionary Biology & Institute of Plant Biology
Max Planck Institute for Developmental Biology
Departamento de Genética Molecular de Plantas
Consejo Superior de Investigaciones Científicas (CSIC)
conceived the study and wrote the manuscript
performed the analyses with contributions from all authors
The authors declare no competing financial interests
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DENMARK: Danfoss has officially opened its latest Application Development Centre (ADC)
this one within its new Smart Store supermarket near the company’s headquarters in Nordborg
The new Application Development Centre will offer the cooling and heating industry the opportunity to access state-of-the-art test facilities and expert support
for field testing new components and cloud technologies for both small and large applications
It supplements the existing 1,200m2 ADC located at the Nordborg headquarters and joins additional Danfoss ADCs in India
Described as one of the world’s most energy efficient supermarkets
the 1,500m2 Smart Store was opened at the end of June
It is said to be approximately 50% more energy efficient than a typical supermarket with a first generation CO2 refrigeration system
and 20-30% more efficient than an equivalent local store already fitted with multiple energy efficiency solutions
The supermarket has two refrigeration systems that run independently
ensuring that product testing does not interfere with the operations of the supermarket
The 750m2 ADC is split over the Smart Store’s ground and first floors and also includes a 334m2 outside test area
It houses a standard CO2 refrigeration pack with Bock compressors providing 29kW MT and 13kW LT to the 800m2 Smart Store sales area.
Energy Pack for test and innovation purposes
It uses a variety of Danfoss products and Bock compressors
These supply eight LT cabinets and10 MT cabinets
“The new ‘Smart Store showcases the incredible possibilities we have ready today with existing solutions for natural refrigerants
and sourcing renewables — all-in-one installation,” said Danfoss Climate Solutions president Jurgen Fischer.
The ADC opening was celebrated with an open house event for Danfoss partners and customers who have contributed to the site
Peder Gabrielsen from the European Environment Agency offered a keynote speech
followed by a site tour of the event led by Danfoss leadership.
Metrics details
Coral reefs are at risk of exposure to petroleum hydrocarbons from shipping spills and uncontrolled discharges during extraction
The toxicity of petroleum hydrocarbons can substantially increase in the presence of ultraviolet radiation (UVR)
therefore spills in shallow coral reef environments may be particularly hazardous to reef species
Here we investigated the sensitivity of coral larvae (Acropora tenuis) to dissolved hydrocarbons from heavy fuel oil (HFO) and diesel in the absence and presence of UVR
Larval settlement success decreased with increasing concentrations of dissolved HFO
and co-exposure to UVR doubled the toxicity: 50% effect concentrations (EC50) decreased from 96 (−UVR) to 51 (+UVR) total petroleum aromatic hydrocarbons (TPAH)
Toxic thresholds for HFO were similar to concentrations reported during marine spills: EC10s of 24 (−UVR) and 15 (+UVR) µg l−1
diesel also reduced settlement and exhibited phototoxicity: EC10s of 122 (+UVR) and 302 (−UVR) µg l−1
This study demonstrates that the presence of UVR increases the hazard posed by oil pollution to tropical
Further research on the effects of oils in the presence of UVR is needed to improve the environmental relevance of risk assessments and ensure appropriate protection for shallow reef environments against oil pollution
so the impacts of hydrocarbon pollution on coral reefs may be significantly underestimated in the context of likely high UVR exposure in situ
To assess the potential for UVR to increase the sensitivity of coral larvae to spills of heavy fuel oil (HFO) and diesel we: (i) assessed UVR irradiance on one inshore and one mid-shelf reef on the Great Barrier Reef (GBR; Australia); (ii) characterised the chemical composition of the two fuels and their water accommodated fractions (WAFs); and (iii) predicted their narcotic toxicity to marine species using the NTLM
We then (iv) exposed larvae of the reef building coral Acropora tenuis (Dana
1846) to HFO and diesel WAFs in the absence and presence of UVR (±UVR)
at intensities similar to those encountered on the GBR
and assessed the ability of exposed larvae to successfully complete settlement and metamorphosis into sessile polyps following each treatment
Penetration of ultraviolet radiation (UVR) on Trunk Reef (a) and Esk Reef (b) as well as a comparison (c) of exposure intensity and spectrum of artificial UVR used during settlement toxicity assays and UVR observed in situ on the Great Barrier Reef (GBR; Australia) during spring
Full spectrum measurements of UVR in air and at 0.1
and 3.8 m depth at a mid-shelf (Trunk reef) and inshore (Esk reef) reef on the GBR in October 2016
Total irradiance values calculated using the percentage reductions in light intensity recorded for each wavelength at each depth (in relation to measurements in air) and total irradiance measurements in air on a clear day (cloud coverage <5%)
Comparison of UVR intensity and spectrum emitted from fluorescent tubes used in settlement toxicity assays
calculated UVR exposure inside scintillation vials and UVR exposures observed at 1 m depth on the GBR during spring
tenuis planulae larvae exposed to water accommodated fractions (WAFs) of heavy fuel oil (HFO) in the absence (−UVR) or presence (+UVR) of ultraviolet radiation as well as juvenile polyps (following settlement)
Larvae exposed to (a) filtered seawater (0 µg TPAH l−1)
(b) approximately 900 µg TPAH l−1 after 48 h of exposure as well as juvenile polyps
attached and unattached larvae treated with (c) FSW (0 µg TPAH l−1)
(d) 115 µg TPAH l−1 and (e) approximately 900 µg TPAH l−1 HFO WAF after 48 h of exposure
introduction of settlement inducer and a 24 h settlement period (a total ~72 h after experiment start)
TPAH = total petroleum aromatic hydrocarbons
Concentration-response curves for coral larval settlement following exposure to heavy fuel oil (a and c) and diesel (b and d) water accommodated fractions (WAF) in the presence (blue) and absence (green) of ultraviolet radiation (µg TPAH l−1)
Model mean (solid line) and 95% confidence intervals (shaded area) for quasibinomial GLMs fitted for the settlement success data of each treatment combination as well as observed settlement success for each replicate (open ring) used in model fitting
At moderate to high TPAH concentrations (≥200 µg l−1) underdeveloped juvenile polyps were observed in the presence of UVR
and the few attached juvenile polyps observed in the highest concentration (782 µg l−1 TPAH) were either underdeveloped or abnormal
some underdeveloped and malformed attached juvenile polyps were also observed; however
fully metamorphosed polyps were still present at the highest concentration tested (758 µg l−1 TPAH)
bends and lumps) were observed in unattached larvae from ± UVR treatments but with no apparent relationship to TPAH concentration
The total narcotic toxicity for undiluted diesel WAF indicated that relatively low mortality would be expected following exposure
when calculated using predicted WAF concentrations (TUNeat fuel: 0.19)
when measured WAF concentrations were applied (TUWAF: 0.05 −UVR to 0.06 +UVR)
These results indicate that by ignoring phototoxicity
the hazards posed by oil spills to coral larvae may be substantially underestimated in shallow-water tropical reef systems
very large increases in oil or fuel WAF toxicity in the presence of UVR are unlikely
The development of abnormal morphologies in coral larvae may indicate narcosis or more specific toxic effects of petroleum hydrocarbons on cellular developmental processes
emphasising the need to accurately estimate the intensity and wavelengths experienced by each species or ecosystem when investigating the influence of UVR on the toxicity of pollutants
embryos and larvae developing at the water surface may also be exposed to substantially higher UVR intensities in situ than applied in the present study
potentially reducing the toxic threshold values further
these results suggest that the hazard (hence risk) posed by aqueous petroleum hydrocarbons to shallow-water tropical coral reefs will be underestimated if phototoxic activation by UVR is not taken into consideration
its use is currently limited to estimation of the acute toxicity of individual PAHs
and further development is necessary before it can be applied to complex hydrocarbon mixtures or chronic exposures
The increase in toxicity of dissolved aromatics from HFO by UVR exposure resulted in low toxic thresholds
underscoring the potential hazard to corals posed by phototoxic compounds found in petroleum oils and fuels
Previous assessments may therefore have substantially underestimated the risks posed by oil and petroleum product spills on shallow-water
tropical coral reefs by not accounting for interactions with environmental factors such as UVR
Further research into the effects of petroleum hydrocarbons on more tropical reef organisms
including potential interactions with UVR and other stressors
is needed to more effectively quantify these risks
a reef-building coral common throughout the Pacific Ocean
were collected by hand on SCUBA from Magnetic Island (October 2016
under Great Barrier Reef Marine Park Authority Permit G12/35236.1
Colonies were placed in flow-through seawater of ambient temperature and transported to the National Sea Simulator
On arrival colonies were transferred to 70% shaded flow-through outdoor holding tanks and kept at temperatures equivalent to the collection site (27 °C) until spawning
When showing signs of setting colonies were isolated and gametes collected by gentle scooping
500 l fibreglass rearing tanks with cone-shaped bases
Flow-through seawater (1.5 turnovers per day) was 1 µm-filtered and a round air stone at the base of each tank provided aeration and created a gentle curtain of bubbles to keep larvae from a submerged cylindrical mesh filter (15 h × 6 d cm
light attenuation of vials used for experimental exposures and in situ intensities of UVR on the GBR)
PAR was provided on a 12:12 h L:D cycle and UVR on a 6:18 h L:D cycle (total irradiance 16.1 W cm−2 d−1)
An additional 30 vials containing undiluted WAF
were also placed in each incubator to allow for the collection of chemical samples at the end of the 48 h exposure
The positions of vials within each incubator were exchanged randomly throughout each experiment to minimize variation in light exposure
Temperature was continuously logged (Onset HOBO temperature logger
salinity and dissolved oxygen concentrations were measured at the beginning and end of each experiment
At the end of the 48 h exposure approximately 600 ml of undiluted WAF was pooled
from the 30 additional vials (not containing larvae)
for chemical analysis (see Chemical analysis below)
EC10 and EC50 values with 95% confidence intervals (CI) were interpolated from model mean values and 95% CI (adapted from Venables & Ripley59)
Predicted model mean values and 95% CI were exported and graphical outputs produced using GraphPad Prism (version 7.02
The high levels of replication used allowed the identification of outliers in the dataset which were excluded
These comprised three HFO FSW controls and one diesel low concentration replicate where CCA chips induced 0% and 7% settlement
likely due to misidentification of a few of the live CCA chips
WAFs were analysed directly for BTEX using Purge and Trap GC-MS in full scan mode (USEPA method 8260)
The 500 ml WAF samples were extracted three times with dichloromethane (DCM) and the combined extracts analysed for PAH and alkylated PAH using GC-MS in SIM or scan mode
Neat HFO and diesel were diluted in DCM and analysed for TRH
PAH/alkylated PAH and phenols using the same methodology
except additional surrogate (2-fluorophenol
phenol-d5 and 2,4,6-tribromophenol) and internal (1,4-dichlorobenzene-d4) standards were added to the samples prior to analysis
the neat oils (1 µl) were subjected to whole oil analysis using GC-MS and hydrocarbons were identified through comparison with a pre-characterised reference oil
In situ UVR irradiance on the Central GBR during spring was assessed through full spectrum measurements of UVR in air and at five depths at two reefs
was selected as a representative inshore reef site while Trunk Reef (18.329°S
146.846°E) was selected as a representative clear-water
Three replicate light intensity measurements for wavelengths between 300–400 nm were performed on SCUBA using a Jaz handheld spectrometer (Ocean Optics
USA) and a 5 m fibre optic cable (CPATCH-5074768
USA) with a planar irradiance collector for underwater use (HOBI Labs
Measurements were made with the sensor positioned vertically at 0
Measurements were performed close to solar noon on the mid-shelf (14:17–14:20 on 12 October 2016) and inshore reefs (13:55–14:04 on 14 October 2016)
Intensity data collected was used to calculate the average relative decrease in radiation for wavelengths between 300–400 nm with depth
Total UVA and UVB radiation above the surface was recorded (Solarmeter Model 5.0 UVA + UVB meter
USA) and turbidity measurements were performed (90FL-T
Cloud coverage was low (<5%) with 17 km/h E winds (BOM
2016) and turbidity of 0.1 NTU during mid-shelf measurements with medium-high cloud coverage (~80%)
33 km/h ENE winds (maximum gusts 46 km/h ENE; BOM
2016b) and 0.8 NTU turbidity during inshore reef measurements
Theoretical irradiance at each depth was calculated using full spectrum measurements of natural sunlight close to solar noon on a low cloud coverage day (<5%)
Measurements were made using a calibrated Jaz spectrometer and a 250 mm UVR compatible fibre optic cable (QP600-025-UV
Z10% values at 305 and 340 nm were estimated by calculating the irradiance corresponding to 10% of surface irradiance (in air) for each reef site and the negative linear relationship between irradiance and measurement depth of measurements made at Trunk Reef and Esk Reef
Full spectrum measurements of radiation emitted by the UVR fluorescent tubes (Deluxlite Black Light Blue 18W; Reptile One UVB 5.0 18W)
were performed using the same calibrated Jaz spectrometer and 250 mm UVR compatible fibre optic cable used to quantify the UV radiation of natural sunlight
Full spectrum measurements were made at approximately the same distance as sample vials during experimental exposures (170 mm) in five separate positions relative to the three sets of fluorescent tubes
Total UVA and B radiation measurements were also performed (Solarmeter Model 5.0 UVA + UVB meter
emitted from fluorescent tubes by scintillation vial glass was estimated (calibrated Jaz spectrometer)
Measurements were made 200 mm from UVR fluorescent tubes (Deluxlite Black Light Blue 18W; Reptile One UVB 5.0 18W) through the base of a 20 ml scintillation vial
The average total UVA and UVB exposure of larvae inside scintillation vials was calculated using measurements performed with the Solarmeter model 5.0 and the average attenuation of scintillation vial glass between 300–400 nm
An average marine CTLBB was used as no CTLBB currently exist for acroporid corals
observed TUs (TUWAF) for each light treatment were calculated using measured concentrations of individual compounds in undiluted HFO and diesel WAFs and the same CTLBB value
Oil pollution on coral reefs: a review of the state of knowledge and management needs
Turner, N. R. & Renegar, D. A. Petroleum hydrocarbon toxicity to corals: A review. Mar. Pollut. Bull. 119, 1–16, https://doi.org/10.1016/j.marpolbul.2017.04.050 (2017)
and prioritisation of emerging contaminants present in the GBR and Torres Strait marine environments
Report to the National Environmental Science Programme
Reef and Rainforest Research Centre Limited
Effects of weathering on the toxicity of three offshore Australian crude oils and a diesel fuel to marine animals
Assessment of polycyclic aromatic hydrocarbon exposure in the waters of Prince William Sound after the Exxon Valdez oil spill: 1989–2005
AMSA. Montara well release - Operational monitoring study O2: Monitoring of oil character, fate and effects. (Australian Maritime Safety Authority, Australian Government, https://www.amsa.gov.au Accessed on 13.04.2017
Crude oil contamination interrupts settlement of coral larvae after direct exposure ends
Characterization of subsurface polycyclic aromatic hydrocarbons at the Deepwater Horizon site
The North Cape oil spill: hydrocarbons in Rhode Island coastal waters and Point Judith pond
Subsurface distributions of petroleum from an offshore well blowout
Redman, A. D. Role of entrained droplet oil on the bioavailability of petroleum substances in aqueous exposures. Mar. Pollut. Bull. 97, 342–348, https://doi.org/10.1016/j.marpolbul.2015.05.068 (2015)
The effects of near ultraviolet radiation on the toxic effects of polycyclic aromatic hydrocarbons in animals and plants: a review
Roberts, A. P., Alloy, M. M. & Oris, J. T. Review of the photo-induced toxicity of environmental contaminants. Comp. Biochem. Physiol. C 191, 160–167, https://doi.org/10.1016/j.cbpc.2016.10.005 (2017)
Validation of the target lipid model for toxicity assessment of residual petroleum constituents: monocyclic and polycyclic aromatic hydrocarbons
Guidance for improving comparability and relevance of oil toxicity tests
Phototoxicity of individual polycyclic aromatic hydrocarbons and petroleum to marine invertebrate larvae and juveniles
Phototoxicity in a coral reef flat community
Overmans, S. et al. Phototoxic effects of PAH and UVA exposure on molecular responses and developmental success in coral larvae. Aquat. Toxicol. 198, 165–174, https://doi.org/10.1016/j.aquatox.2018.03.008 (2018)
Institue for Environment and Sustainability
Quantifying solar spectral irradiance in aquatic habitats for the assessment of photoenhanced toxicity
PAH phototoxicity—an ecologically irrelevant phenomenon
Tedetti, M. & Sempere, R. Penetration of ultraviolet radiation in the marine environment. A review. Photochem. Photobiol. 82, 389–397, https://doi.org/10.1562/2005-11-09-IR-733 (2006)
Attenuation coefficients of ultraviolet and photosynthetically active wavelengths in the waters of Heron Reef
Barron, M. G., Vivian, D. N., Yee, S. H. & Santavy, D. L. Methods to estimate solar radiation dosimetry in coral reefs using remote sensed, modeled, and in situ data. Environmental Monitoring and Assessment 151, 445–455, https://doi.org/10.1007/s10661-008-0288-4 (2009)
Yee, S. H., Santavy, D. L. & Barron, M. G. Comparing environmental influences on coral bleaching across and within species using clustered binomial regression. Ecological Modelling 218, 162–174, https://doi.org/10.1016/j.ecolmodel.2008.06.037 (2008)
Review of flow rate estimates of the Deepwater Horizon oil spill
Photoinduced toxicity of the polycyclic aromatic hydrocarbon
Effects of natural gas condensate–water accommodated fraction on coral larvae
Negri, A. P. et al. Acute ecotoxicology of natural oil and gas condensate to coral reef larvae. Sci. Rep. 6, 21153, https://doi.org/10.1038/srep21153 (2016)
Phototoxic target lipid model of single polycyclic aromatic hydrocarbons
The effects of oil pollutants on fertilisation rates in the scleractinian coral Acropora tenuis
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Management and Sustainability of Marine Habitats in the 21st Century
Predicting the toxicity of neat and weathered crude oil: toxic potential and the toxicity of saturated mixtures
Negri, A. P. & Heyward, A. J. Inhibition of fertilization and larval metamorphosis of the coral Acropora millepora (Ehrenberg, 1834) by petroleum products. Mar. Pollut. Bull. 41, 420–427, https://doi.org/10.1016/S0025-326x(00)00139-9 (2000)
Metamorphosis of Heteroxenia fuscescens planulae (Cnidaria: Octocorallia) is inhibited by crude oil: a novel short term toxicity bioassay
The ecotoxicology of vegetable versus mineral based lubricating oils: 3
Water contamination reduces the tolerance of coral larvae to thermal stress
Redman, A. D., Parkerton, T. F., McGrath, J. A. & Di Toro, D. M. PETROTOX: An aquatic toxicity model for petroleum substances. Environ. Toxicol. Chem. 31, 2498–2506, https://doi.org/10.1002/etc.1982 (2012)
Toxicity of Deepwater Horizon source oil and the chemical dispersant
Acute lethality of polycyclic aromatic hydrocarbons to Daphnia magna
Effects of two petroleum products on Pocillopora damicornis planulae
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Harmful effects of chronic oil pollution on a Red sea scleractinian coral population
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The chemistry and toxicity of sediment affected by oil from the North Cape spilled into Rhode Island Sound
Effects of sediments on the reproductive cycle of corals
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Natural inducers for coral larval metamorphosis
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Sensitivity of juvenile Macomona liliana (bivalvia) to UV‐photoactivated fluoranthene toxicity
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The authors would like to thank the Australian Institute of Marine Science Townsville (AIMS) and the National Sea Simulator (SeaSim)
Tristan Lever and Edith Strecker for logistical support as well as Gerard Ricardo and Patricia Menendez for their advice regarding statistical analysis
This work was supported by a research grant set up as part of the collaborative research project between the Red Sea Research Center (RSRC) at King Abdullah University of Science and Technology (KAUST) and the AIMS
King Abdullah University of Science and Technology
Biological Environmental Science and Engineering Division
James Cook University and Australian Institute of Marine Science
designed the experiment with input from F.F.
wrote the manuscript with input from A.P.N
Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations
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DOI: https://doi.org/10.1038/s41598-018-26972-7
Photochemical & Photobiological Sciences (2023)
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DENMARK: Danfoss has officially opened its new commercial heat pump test facility at its Application Development Centre in Nordborg
The state-of-the-art test chamber is capable of testing air-to-water heat pumps and air-cooled water chillers according to European standard EN14511:2022.
The 1,200m2 Application Development Centre is designed to help customers test new solutions and improve performance on applications in a live environment
The facility is capable of testing a wide range of products from split-system air conditioning to transport refrigeration units and supermarket packs
Already set up to test residential heat pumps
the facility can now test commercial units to a wide range of temperatures
including propane and other A3 refrigerants up to 250kW
The companies are promising greater efficiency for clients
HPE is working with engineering company Danfoss on a new package to help data center operators recover waste heat
It will see HPE’s scalable modular data center (MDC) products combined with Danfoss heat reuse solutions
HPE IT Sustainability Services – Data Center Heat Recovery
is an “off-the-shelf heat recovery module” designed to help organizations “manage and value excess heat as they transition towards more sustainable IT facilities,” according to the vendors
“Our strategic partnership with HPE is a great example of how we revolutionize building and decarbonizing the data center industry together with customers,” said Jürgen Fischer
“With this latest cross-industry partnership we’re building the blueprint for the next generation of sustainable data centers - using technologies available today.”
HPE says its MDC units are small-footprint
high-density containers that can be deployed nearly anywhere in the total absence of heavy industry
They incorporate direct liquid cooling and other technologies
which HPE says can reduce overall energy consumption by 20 percent
The units can apparently achieve a power usage effectiveness (PUE) of 1.14
These can now be paired with a range of Danfoss solutions
such as heat reuse modules that capture excess heat from data centers to provide renewable heating onsite and to neighboring buildings
and Turbocor oil-free compressors that can enhance data center cooling efficiency by up to 30 percent
vice president and general manager for WW Advisory
professional services and managed services at HPE said the companies can “multiply value” for their clients by “harnessing the typically untapped resource of waste heat
showing the future of energy usage is efficient
Danish business Danfoss is also a client of HPE
and uses HPE GreenLake for SAP S/4HANA Cloud
Its heat reuse technology scored a high-profile client earlier this year when it was revealed it was providing cooling and heat reuse systems for Google data centers
Data Centre Dynamics Ltd (DCD), 32-38 Saffron Hill, London, EC1N 8FH Email. [email protected]DCD is a subsidiary of InfraXmedia
News & Analysis on Food & Beverage Development & Technology
30-Jun-2023 Last updated on 03-Jul-2023 at 13:09 GMT
The 1,500 sqm flagship store – leased to the discount retailer COOP 365 and built next to the Danfoss headquarters in Nordborg – runs off sustainable energy sources and reuses the excess heat created by cooling cases to cut heating costs by up to 90%
is expected to be around 50% more energy efficient compared to a typical supermarket with a first-generation CO2 refrigeration system and no energy efficiency solutions
It is also expected to be approximately 20-30% more efficient than an equivalent local store already fitted with multiple energy efficiency solutions
Using advanced heating and cooling technology makes good business sense too
These savings come with a typical payback time of under three years
The store operates both as a functioning supermarket and a development centre for testing new technology
Danfoss hopes it will ‘inspire food retailers in a world of rising energy costs
“This supermarket is purpose-built for the world ahead of us; a world of more urbanization
and efficient food storage,” said Jürgen Fischer
“Danfoss has reimagined what food retail stores could look like in the 21st century,” he said
all of Danfoss’ most cutting-edge technology and energy efficient food retail solutions are being brought together into one retail site
But the new Smart Store supermarket is only the beginning
Because it will also serve as an Application Development Center
a ‘live’ testing site for new technologies which we hope will inspire food retailers around the world to move towards zero emissions supermarkets – while making economic sense.”
The 1,500 sqm flagship 'Smart Store' in Nordborg is now open to the public
Image: DanfossThe efficiency solutions employed to reduce the overall energy needs of the store
Supermarkets in industrialised countries consume around 3% of a nation’s electricity production
Energy is therefore an area where significant savings can be achieved by food retailers with relatively low investment and good payback times
The supermarket will utilize solar power as its primary energy source
with 100 kW solar panels on the building’s roof providing green energy to support the supermarket operations
Heat capture and reuse is also key to the energy efficiency of the supermarket
Excess heat is the world’s largest untapped source of energy
The store is therefore fitted with state-of-the-art heat recovery units
designed to recover the waste heat from all the refrigeration systems: refrigeration systems represent by far the highest share of the total energy consumed in supermarkets
The recovered heat is reused to heat up the store and produce domestic hot water
with any additional heat shared with residents of the surrounding town through a district energy network
Danfoss expects up to a 90% reduction in supermarket heating costs as a result
there are about 30,000 supermarkets which have low temp cooling of minus 18 to minus 24 degrees Celsius,” Fischer revealed
“If you use that as energy storage that corresponds to 10 nuclear power plants.”
such as installing doors on refrigerator and freezer cases
while the choice of LED lighting uses up to 85% less electricity than incandescent bulbs
Automation and monitoring of the ‘Smart Store’ adds another layer of energy saving
Danfoss claimed there is 3% energy leak rate in Europe
which is significantly more in the developing world
it’s important to match capacity to demand
and you waste energy and risk system damage; too little
Adapting energy consumption can further allow supermarkets to benefit from cheaper tariffs
effectively storing or borrowing cooling capacity in the store freezers while energy is cheap or solar electricity is plentiful
then temporarily switching off the compressors during high-cost peak times until the peak has passed
Industry is increasingly seeking ways to save energy via temperature controls
The issue of refrigeration systems, energy and the environment is timely. Unilever, for example, is looking to ‘warm up’ ice cream freezers to help tackle emissions. The downside to this plan is that products will have to withstand the higher storage temperatures.
Danfoss said 'Smart Store' products won’t need to be reformulated. “There is no reason why you should not have several temperatures in the supermarket organised in a good way with good technology and digital systems in place,” Fischer said.
The company does though believe that timely responses to issues in cooling units are critical to preventing food losses. The resources needed to produce the food that becomes lost or wasted have a carbon footprint of about 3.3 billion tonnes of CO2, it claimed.
The company is therefore employing solutions at various stages of the value chain to address the problem. In India, for example, solar powered refrigeration units installed in fields can ensure produce lasts up to three days longer and doesn’t spoil by the time it reaches market.
The store is fitted with state-of-the-art heat recovery units, designed to recover the waste heat from all the refrigeration systems. Image: DanfossIts solutions also promise to control temperature during transportation to food retailers. Once in the ‘Smart Store’, meanwhile, it has installed digital, real-time monitoring technology on refrigeration systems and individual products which will alert store managers to problems before they occur.
For example, if a supermarket freezer case was to fail overnight then it’s probable that the entire line of frozen foods stocked in that freezer would need to be disposed of as food loss. “Preventing food loss at such scale is critical to preventing lost revenue and reducing emissions,” Danfoss said. “Immediately when there is something wrong on the temperature side an alarm will out and we can fix it remotely,” elaborated Fischer.
Danfoss also takes care of 24/7 technical support, mostly remotely, but crucial amid a shortage of qualified heating engineers and a significant green energy skills gap across Europe.
Danfoss said the fixed fee for supermarkets will 'not be more or less' than the rents supermarkets pay already. But the building the 'Smart Store' cost over £10 billion (EUR 11.6 billion), around three times the cost of standard stores with no green technology.
“The global financial world is very interested in green technology and there is much more money than opportunity,” added Fischer. “That’s why the service model will be a super accelerator.”
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The notion of species as reproductively isolated units related through a bifurcating tree implies that gene trees should generally agree with the species tree and that sister taxa should not share polymorphisms unless they diverged recently and should be equally closely related to outgroups
It is now possible to evaluate this model systematically
We sequenced multiple individuals from 27 described taxa representing the entire Arabidopsis genus
corresponding to described species that capture the structure of the genus
only the separation of Arabidopsis thaliana from the remaining species was universally supported
the amount of shared polymorphism demonstrated that reproductive isolation was considerably more recent than the estimated divergence times
We uncovered multiple cases of past gene flow that contradict a bifurcating species tree
we showed that the pattern of divergence differs between gene ontologies
In total the alignment contains 7.5 million SNPs
Because the BEAGLE algorithm requires phased data, only diploid individuals were used. Heatmap colors represent the total length of IBD blocks for each pairwise comparison (Online Methods)
the ancient African subdivision would have had to persist for a very long time while still allowing sufficient gene flow for modern humans to be most closely related at most loci (and evolve into anatomically modern humans)
Admixture thus seems a more likely explanation
the greater number of shared alleles between A
arenosa could be due to ancient population subdivision
with the same subpopulation or region giving rise to A
the argument against the likelihood of ancient subdivision in humans is even stronger in Arabidopsis
because the species are so highly diverged
Whereas average coalescence times in modern humans are greater than the supposed split from Neanderthals
the average coalescence times between the common outcrossing Arabidopsis species is less than half the divergence between these species and A
regardless of whether they are maintained by selection
genes identified in the morphologically highly divergent A
cebennensis were overrepresented in the 'organ morphogenesis' and 'tissue development' categories
We performed correction of raw reads for the A. halleri ssp. halleri 3 sample (Supplementary Data Set 1)
The ErrorCorrectReads.pl module of ALLPATHS-LG version 44837 was used for correction of raw reads (34.0 million reads)
and the other parameters were set to default according to the ALLPATHS-LG manual
24.9 million corrected reads were used in downstream data analyses
Raw reads were uploaded to NCBI SRA (numbers of BioProjects and BioSamples are available in Supplementary Data Set 1). Chloroplast assemblies were uploaded to ENA (Supplementary Data Set 1)
kamchatica sample as two different diploid individuals
Population structure analysis was performed using all the samples of A
thaliana accessions from different geographical locations and C
We focused on genic regions where at least half of the total exon length was covered in each individual analyzed
Then we calculated copy number for those genes in each individual on the basis of total exon coverage depth normalized by mean coverage depth and filtered for genes with copy number between 0.4 and 1.6 in all individuals
and number of trimmed markers were roughly doubled compared to the default parameters
which were optimized for human SNP array data; input data in our case are more dense in number of SNPs per cM
and this was satisfied using a minimum LOD score of 10
we would expect to see shared haplotypes of length 2 kb if t = 20,000 generations
Note that all the species were mapped to A
which can be considered an outgroup for the other Arabidopsis species
This approach makes mapping bias unlikely when considering admixture between A
thaliana and the remaining species (whereas mapping to A
Complete chloroplast genomes were assembled in CLC Genomics Workbench version 6.0.4 (CLC bio)
Reads were trimmed for adapters as well as a minimum quality of 0.001 (Phred score 30) and a minimum length of 50 bp
Paired reads were assembled using the legacy version of the CLC de novo algorithm with length fraction 0.9
similarity 0.9 and appropriate distance settings
Contigs belonging to the chloroplast were identified using blastn and aligned manually to the closest related published complete chloroplast sequence (A
rubella and Camelina sativa) using PhyDE version 0.9971
A preliminary pseudo-reference was created by filling gaps between nonoverlapping contigs from the reference sequence
The complete chloroplast genome was obtained by repeated cycles of mapping back to the pseudo-reference and variant detection in CLC (minimum coverage 1
variant probability 0.1) as well as manually
Misalignments and mismatches were adjusted at every step
thus accounting for rate heterogeneity among genes
Only models implemented in BEAST were tested
with BIC used for model selection in a 'greedy' search and unlinked branch lengths
The best partitioning scheme comprised two subsets
one (55,892 bp) evolving under the generalized time-reversible model of evolution with gamma model of rate heterogeneity and invariant sites (GTR+I+Γ) and the other (62,184 bp) under the GTR+I model
An additional constraint was set on the root height by truncation to 4–12 Myr
The expected pairwise coalescence time is E(T2) = 2Ne generations within each species
and E(T2) = 2Ne + tsplit generations between species
where Ne in the latter equation is that of the ancestral species
we will assume that Ne of the ancestral species equaled the larger of the Ne of the descendant species
Simple moments estimators for all relevant parameters can be obtained by noting that the expectation of the pairwise sequence divergence d is proportional to E(T2)
We can thus obtain an upper bound for the probability of a trans-specific polymorphism as
where dA is the estimated pairwise sequence divergence (nucleotide diversity) within species A
and dAB is the estimated pairwise sequence divergence between species
the per-site probability of a trans-specific polymorphism under neutrality between A
lyrata is less than e−(11.57–3.04)/0.69 × e−(11.6–3.04)/3.04 = 2.5 × 10−7
because it ignores the probability of the right order of coalescences in the ancestral species
The pairwise sequence divergence at aligned fourfold degenerate sites was used for the calculation
if we were to choose two alleles at random from each these two species
we would expect <1 out of the 2,909,657 fourfold degenerate sites to be trans-specific by chance
and we conclude that either there was gene flow more recently than what is implied by the high divergence under the simple model of splitting or polymorphisms were maintained by selection
Fst estimates for pairwise common species (A. thaliana, A. halleri, A. lyrata, A. arenosa), comparisons were calculated at biallelic synonymous SNPs (Supplementary Table 1) as (Htotal – Hsubsp) / Htotal
where Htotal is heterozygosity in total population (for example
Hsubsp is average of heterozygosities in subpopulations (for example
where p and q are derived and ancestral allele frequencies
We searched for SNPs segregating in all four common Arabidopsis species
Even though we filtered out potential duplicated genes on the basis of coverage before calling variants
we noticed that some of the shared sites show: (i) only heterozygous genotypes (Aa) or (ii) heterozygous and only one of the homozygous genotypes (Aa and AA or Aa and aa) among diploid A
which can be explained by the presence of a duplicated region
we ended up with 3,818 sites spanning 2,365 genes
Additional filtering based on linkage was applied to avoid potential back mutations among shared SNPs
we selected genes with shared haplotypes such that a pair of shared SNPs were in linkage disequilibrium (r2 > 0.3) in A
thaliana and in at least two out of the three other common species
we required that at least one of the shared SNPs was nonsynonymous
and for some comparisons we also considered nucleotide diversity as this resulted in more interesting results
The reported P values do not take this implicit multiple testing into account and should thus be interpreted with caution
There is no obvious way of correcting for this kind of overfitting
and there is also no way of knowing a priori what parameters might be appropriate to detect selection in a given species
divergent genes were identified as those with Fst values in the upper eighty-fifth percentile in comparisons between this clade and A
we used the upper eightieth percentile threshold for Fst and the additional criterion of nucleotide diversity in the lower fiftieth percentile in A
divergent genes were identified as having Fst values in the upper eighty-fifth percentile
and we also required nucleotide diversity in the lower forty-fifth percentile and Tajima's D values in the lower thirty-fifty percentile
Individuals that showed signs of introgression between A
arenosa in the ADMIXTURE analysis we excluded
which were not deeper than the third node on a GO categories graph for 'biological process' root category (136 categories in total) and also used only the first node categories of INTERPRO database (627 categories)
We also required a minimum of five genes from a category to be present in a candidate set to perform a test
Gene categories enrichment was tested by Fisher's exact test: all P values were adjusted for multiple testing (using R function 'p.adjust' with method = 'fdr')
Accession codes are listed in Supplementary Data Set 1
Taming the wild: resolving the gene pools of non-model Arabidopsis lineages
Patterns of population epigenomic diversity
The allopolyploid Arabidopsis kamchatica originated from multiple individuals of Arabidopsis lyrata and Arabidopsis halleri
A time-calibrated road map of Brassicaceae species radiation and evolutionary history
Ecology meets molecular genetics in Arabidopsis
The genetic basis of zinc tolerance in the metallophyte Arabidopsis halleri ssp
halleri (Brassicaceae): an analysis of quantitative trait loci
Improving the accuracy and efficiency of identity-by-descent detection in population data
gene trees and selfing: an ancestral recombination graph with partial self-fertilization
The transition to self-compatibility in Arabidopsis thaliana and evolution within S-haplotypes over 10 Myr
Evolution of selfing: recurrent patterns in molecular adaptation
Subdivision in an ancestral species creates asymmetry in gene trees
Fixation indices in subdivided populations
Polymorphism at the self-incompatibility locus in Solanaceae predates speciation
Trans-specific Mhc polymorphism and the origin of species in primates
Statistical method for testing the neutral mutation hypothesis by DNA polymorphism
Multiple instances of ancient balancing selection shared between humans and chimpanzees
Roles of Arabidopsis cyclin-dependent kinase C complexes in cauliflower mosaic virus infection
Arabidopsis protein kinases GRIK1 and GRIK2 specifically activate SnRK1 by phosphorylating its activation loop
A novel nucleocytoplasmic traffic GTPase identified as a functional target of the bipartite geminivirus nuclear shuttle protein
A domain swap approach reveals a role of the plant wall-associated kinase 1 (WAK1) as a receptor of oligogalacturonides
Molecular mechanisms of metal hyperaccumulation in plants
A recent local sweep at the PHYA locus in the Northern European Spiterstulen population of Arabidopsis lyrata
coalescent theory and the analysis of genetic polymorphisms
Retroposon analysis and recent geological data suggest near-simultaneous divergence of the three superorders of mammals
Mammalian evolution may not be strictly bifurcating
Genome sequencing reveals complex speciation in the Drosophila simulans clade
Genomic identification of founding haplotypes reveals the history of the selfing species Capsella rubella
Genome-wide evidence for speciation with gene flow in Heliconius butterflies
Speciation with gene flow in equids despite extensive chromosomal plasticity
Mailund, T., Munch, K. & Schierup, M.H. Inferring the process of human–chimpanzee speciation. eLS. http://dx.doi.org/10.1002/9780470015902.a0020833.pub2 (2014)
Extensive introgression in a malaria vector species complex revealed by phylogenomics
Evolution of Darwin's finches and their beaks revealed by genome sequencing
The dynamics of incomplete lineage sorting across the ancient adaptive radiation of neoavian birds
Phylogenomics reveals three sources of adaptive variation during a rapid radiation
Is gene flow the most important evolutionary force in plants
Denisova admixture and the first modern human dispersals into Southeast Asia and Oceania
A rapid DNA isolation procedure for small quantities of fresh leaf tissue
Fast and accurate short read alignment with Burrows-Wheeler transform
The Sequence Alignment/Map format and SAMtools
A framework for variation discovery and genotyping using next-generation DNA sequencing data
The Genome Analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data
A program for annotating and predicting the effects of single nucleotide polymorphisms
SnpEff: SNPs in the genome of Drosophila melanogaster strain w1118; iso-2; iso-3
The 1001 Genomes Consortium. 1,135 genomes reveal the global pattern of polymorphism in Arabidopsis thaliana. Cell http://dx.doi.org/10.1016/j.cell.2016.05.063 (2016)
Genome-wide quantification of homeolog expression ratio revealed nonstochastic gene regulation in synthetic allopolyploid Arabidopsis
APE: Analyses of Phylogenetics and Evolution in R language
DendroPy: a Python library for phylogenetic computing
Population genetic inference from genomic sequence variation
ANGSD: Analysis of Next Generation Sequencing Data
MAFFT multiple sequence alignment software version 7: improvements in performance and usability
Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis
Partitionfinder: combined selection of partitioning schemes and substitution models for phylogenetic analyses
Selecting optimal partitioning schemes for phylogenomic datasets
RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies
Bayesian phylogenetics with BEAUti and the BEAST 1.7
On incomplete sampling under birth-death models and connections to the sampling-based coalescent
Relaxed phylogenetics and dating with confidence
The probability and chromosomal extent of trans-specific polymorphism
Inferring the joint demographic history of multiple populations from multidimensional SNP frequency data
Gowinda: unbiased analysis of gene set enrichment for genome-wide association studies
Genome-wide comparison of nucleotide-binding site-leucine-rich repeat-encoding genes in Arabidopsis
msABC: a modification of Hudson's ms to facilitate multi-locus ABC analysis
Download references
de Meaux for discussions and comments on the manuscript; R
Savolainen for help with collections; and P
This work was supported by the German Research Foundation (DFG SPP 1529 to M.A.K.
by the Swiss National Science Foundation and URPP Evolution in Action (to K.K.S.)
by the Czech Science Foundation (P506/12/0668 to K.M.)
and by the Austrian Science Funds (FWF DK W1225 to C.S.)
Present address: Current address: Department of Biology
Vienna Graduate School of Population Genetics
Department of Evolutionary Biology and Environmental Studies
Karelian Research Center of the Russian Academy of Sciences
National Institute of Advanced Industrial Science and Technology
contributed plant material or contributed or generated sequence data
Two local minima (K=5 and K=8) were chosen for further analysis shown on Figure 1
(a) Neighbor-joining tree based on a genetic distances
Nodes are labeled with percentage of individual gene trees supporting the split
(b) Overlapped neighbor-joining trees for 300 randomly chosen genes
Divergence times are indicated with 95% confidence intervals
Bootstrap support from 1000 replicates: ** 99-100%
(a) Distribution of IBD blocks length identified within common species: A
(b) Distribution of IBD blocks length identified within local endemic Arabidopsis species: A
(c) Distribution of all identified IBD blocks between different Arabidopsis clades
(a) D statistics from ABBA-BABA tests shows that both A
Pink dashed line connects individuals sharing IBD blocks identified with BEAGLE
Tajima’s D value in common Arabidopsis species for all analysed genes (white)
genes with at least one shared SNP in all 4 species (green)
genes with at least 2 linked shared SNP in all 4 species (orange); * indicates p-values <<0.01 (‘Mann-Whitney’ test)
Supplementary Figures 1–8 and Supplementary Tables 1 and 2 (PDF 1492 kb)
List of analysed samples with accession codes
containing at least 2 linked SNPs segregating in all 4 common Arabidopsis species (A
Download citation
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Danfoss has opened a new Application Development Centre at flagship energy-efficient 'smart store' supermarket near Danfoss’ headquarters in Nordborg
This collaborative test environment will empower original equipment manufacturers (OEMs)
and Danfoss engineers to develop new technologies and solutions to enhance energy and operational efficiency for food retail
The D anfoss ‘smart store’ is a functioning supermarket
providing the unique opportunity to understand how new technology will operate in the real world
while empowering the store managers to focus on their business while saving energy and costs
The store uses heating and cooling technology and automation solutions with payback times said t be less than 3-4 years
The Application Development Centre within the smart store supermarket
which is part of a full Decarbonisation Park including several innovation centres for applications such as heat pumps
Danfoss has built the smart store supermarket at its headquarters and it aims to lead the way for climate-friendly food retail with energy-efficient heating and cooling technologies
The store is expected to be 50% more energy efficient than a traditional store and 90% of the space heating needs for the entire store will be provided by a heat recovery unit that captures excess heat produced by the cooling systems
Saving energy while decreasing upfront costs
If you use too much cooling you waste energy
With smart controls and digital monitoring
retailers can optimise capacity and demand
allowing them to respond to anomalies in a timely manner preventing energy and food losses
The store will be managed by Danfoss and ANEO Retail’s partnership
which allows grocery stores to subscribe to technical facilities as a service
reducing their operational expenses and time spent on issue management
The concept allows supermarkets to implement the most energy-efficient equipment without large investments and high up-front costs
The store’s refrigeration and comfort cooling systems run exclusively on natural refrigerants (CO2)
which have the lowest possible global warming potential score
“The new smart store showcases the incredible possibilities we have ready today with existing solutions for natural refrigerants
and sourcing renewables – all in one installation,” said Jürgen Fischer
president of Danfoss Climate Solutions
“We are proud to officially welcome customers and partners to the Application Development Center today
to take the next steps together to reimagine the future and develop new heating and cooling technologies that pave the way towards zero emission food retail.”
The occasion was celebrated with an open house event for Danfoss partners and customers who have contributed to the site
Peder Gabrielsen from the European Environment Agency offered a keynote speech where he said:“With the fluorinated gas (F-gas) Regulation in Europe we are seeing a reduction of F-gas emissions and the Kigali Amendment to the Montreal Protocol is driving the refrigerant transition at a global level
The example we see here today is a good example of movement in the right direction
When energy efficiency and low global warming potential refrigerants work in tandem
we can vastly cut emissions from heating and cooling
The need to use energy more efficiently and to reduce costs is constantly growing
Innovation like what we see here has a key role to play in finding the best solutions.”
Delegates to the IOR Annual Conference taking place from 21 to 22 April will get the chance to access the event live and all sessions and recordings for up six months afterwards providing fantastic value and allowing anyone registering for the event ..
Parts Town UK has formed a new partnership with York
Parts Town UK stocks a wide range of York spare parts
The ability of plants to self-pollinate — a big factor in the spread of weeds — is much older than previously thought in one widely studied species
The findings show that at least in plant evolution
sex with others may be more trouble than it’s worth
The results contradict a 2004 estimate from North Carolina State University that A
thaliana began self-pollinating in the last 400,000 years
“We can rule out a very recent change to self-fertilization,” said Chris Toomajian, USC College research associate in molecular and computational biology and co-author of two new papers on A. thaliana in Science Express and Nature Genetics
confers a major advantage to weedy species
A selfing plant can invade new territory by itself and colonize it alone
The potential downside — a nasty case of inbreeding depression — is averted by rare sexual breeding
thaliana have received pollen from other plants of the species
“A little sex goes a long way,” Nordborg said
The researchers arrived at their findings by studying common combinations of genetic variants
Certain variants at different points on the genome tend to go together
is important because it helps predict what an individual’s genome looks like based on information from selected locations
the researchers used the genome-wide pattern of L.D
to estimate the time at which selfing evolved
Since cross pollination mixes two plants’ genomes and disrupts some combinations of variants
a recent transition to selfing should have left a footprint in the L.D
the researchers expected to find a relatively high level of L.D.
the disruption of variant combinations would have slowed dramatically
pattern instead suggested that selfing evolved “on the order of a million years ago or more,” and perhaps soon after the evolution of the species itself
pattern to design a method for mapping the associations between genetic variants and corresponding physical traits in A
as Arabidopsis is likely to become an important model for identifying the genetic basis of evolutionary change,” Nordborg said
Nordborg was the corresponding author on both studies
Cambridge University and the Max Planck Institute for Developmental Biology
Funding came from the National Science Foundation, the National Institutes of Health and the Max Planck Society
Copyright © 2025 University of Southern California
The TimesAbove the dairy aisle in a supermarket 20 miles north of the Danish-German border
a cheery yellow-on-black sign reads: “The right place to save.” There is more truth to the slogan than most of the shoppers crowding in on the first day of trading realise
dials and digital schematics designed to make this the greenest food store on the planet
This branch of the Coop 365 discount chain in the town of Nordborg is the prototype of a “smart store”
in which the majority of the energy that goes into heating and cooling is recycled
By Maria Gonçalves2023-08-12T17:50:00+01:00
A new ‘smart’ supermarket in Denmark has been touted as the future of energy-efficient retail
And its creators believe the technology has the potential to go worldwide
From the outside, the 365 Discount supermarket in Nordborg, southern Denmark, looks like any other grocery store. Located on a busy road – just across from the HQ of Danish engineering giant Danfoss – it’s around the same size as a Tesco Metro and follows a typical supermarket layout
It’s all part of a partnership between Danfoss
which is using the store as a testbed for its energy-efficient solutions
and Bals (Brugsforeningen for Als and Sundeved)
Denmark’s largest independent supermarket association
which reopened last month to queues of customers
Solar power is the primary source: 100kW solar panels power the supermarket operations year-round
Danfoss will use the store as a testing site for energy-efficient technologies
But the reuse of excess heat created by fridges and freezers has proven the real game-changer
The excess is not only used to provide heating and hot water in the store – heating costs have fallen by up to 90% – but any residual energy is shared with residents of the surrounding town through a district energy network
“the world’s largest untapped source of energy”
says Danfoss Climate Solutions president Jürgen Fischer
this project is about implementing a range of solutions
are set up to take advantage of changes in electricity prices
Whenever the systems recognise a drop in prices through the local grid
they lower the temperature in the freezers to take advantage
Other energy-saving initiatives include doors on refrigerators and freezers
while LED lighting uses up to 85% less electricity than incandescent bulbs
but its application is: the Smart Store aims to provide a model for energy-efficient retail in the future
The heat recovery unit manages and buffers the heat from the CO2 refrigeration pack
which is reused for the store’s heating and hot tap water
A comfort cooling system uses glycol as a coolant
the 365 Discount branch is approximately 50% more energy-efficient than a typical supermarket with a first-generation CO2 refrigeration system and no real energy efficiency solutions
And it’s around 20% to 30% more efficient than an equivalent local store already fitted with multiple energy efficiency solutions
A typical supermarket can cut its energy bill by up to 40% by implementing all this tech
which claims the capital investment has a payback time of only three to four years
The Nordborg prototype cost a little over £10m to build
but Danfoss expects costs to become lower as the concept expands
Fischer says ROI will be the selling point for many grocery businesses in Europe and beyond
the Environmental Protection Agency has estimated that every dollar a retailer saves on energy is equivalent to increasing sales by $59 (£46)
The store is run primarily on solar energy
Shoppers queued for the reopening of the store in Nordborg
Fischer is particularly looking forward to bringing the concept to supermarkets in regions such as Latin America and Asia
where the warm climate requires stores to spend more money on refrigeration
but a lot of energy is wasted due to dated technology
“The Smart Store supermarket is only the beginning,” he says
“It will also serve as an application development centre – a live testing site for new technologies that we hope will inspire food retailers around the world to move towards zero-emissions supermarkets
As energy costs soar and grocers look to hit net zero targets
the Smart Store proposition certainly couldn’t come at a better time
Its chances of wider adoption look high – and until then
the store provides a glimpse as to what a sustainable retail environment could look like in the not-so distant future
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A BRAND new Center Parcs resort is set to open in Europe - and it will be the first of its kind
Center Parcs Nordborg will open in Denmark in 2025
Also the first in Scandinavia
the holiday resort had hoped to open by the end of next year although this has since been pushed back
The first phase of the park will include 440 holiday lodges sleeping between two and eight people
Also included in the first phase will be an activity centre and bike hire as well as a 7,200m2 indoor waterpark
restaurant and grocery store will also be on-site
PARK UP Holiday park 'better than Center Parcs' has 10% off this summer with bargain dealsThe second phase will include the creation of 160 private holiday homes
It is hoped the resort could welcome up to 160,000 tourists per year
The resort is being designed by European architect Thierry Huau, who also designed Les Villages in Paris as well as the botanical theme park Terra Botanica
Construction for the holiday park has already started, and when completed will be the 28th resort for Center Parcs in Europe
which means that we strive to have as little impact as possible on the surrounding nature when building the accommodation and leisure facilities
"Of course, typical Danish elements of architecture and design will be an integral part of the holiday park
The facilities will be available all year round and will offer great experiences for all age groups from young to old."
Managing Director of Center Parcs Germany & Scandinavia
added: "We are convinced that the holiday park with direct beach access and a wide range of indoor and outdoor as well as water sports activities will be an enrichment for this attractive region at the Baltic Sea and the local community."
While ticket prices are yet to be revealed
it is likely to be cheaper than the UK Center Parcs resorts which are some of the most expensive in Europe
Earlier this year, a couple said they paid just £447 for a holiday in Center Parcs in Belgium
with a similar break in the UK Center Parc costing £1,699
Another woman said she saved hundreds on her family holiday by going to the Center Parcs in the Netherlands
Catherine Lofthouse explained: "We're paying about £400 less at Zandvoort Center Parcs in Holland, just a short train ride outside Amsterdam compared to staying at one of the five Parcs in the UK
€30 (£26.22) of activities vouchers and the choice of which lodge to stay in
And here's what you can expect from a holiday to Center Parcs in France.
The holiday resort announced plans for a sixth holiday resort in the UK
However, this was scrapped earlier this year
If you don't want to leave the country to visit Center Parcs, here are some clever hacks to still save money
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The movie itself may not have been that frightening
but the idea behind the monster — that he resurfaces to replace his parts with yours
paired with the “Jeepers Creepers” tune — did get spooky
Especially when you found out he stuffed and skinned his victims in his basement
https://www.youtube.com/watch?v=bJRp1dHOAYs
It may be a giant lesson wrapped into a moral
tied with a “don’t be a bad boy” bow
but watching the little tykes plead for their mamas while turning into donkeys and being sold to the salt mines was a horrific experience as a child
The donkey boy continues to haunt my dreams
Alfred Hitchcock’s thriller pins people against every single bird in a local seaside village
The Wicked Witch of The West – The Wizard of Oz
The Wicked Witch is also overlooked but let’s not forget that she lights one character on fire and threatens Dorothy with death
Why have people ever imagined the keeper of the gold to be anything but evil and conniving
The Leprechaun proves to us that we were kidding ourselves when we thought he could be anything but a shape-shifting
teleporting ugly creature who can smell his gold
and you better believe he’ll kill to get it back
The Crypt Keeper – Tales From The Crypt
The only thing actually frightening in all of the Tales of The Crypt movies is the Crypt Keeper himself
He’s almost too real looking with his half decomposed faced and shrill laugh
Mary Shelley might have created Frankenstein’s monster
but it’s Boris Karloff who brought the modern version to life
His portrayal of the lumbering grotesque golem made the monster one of the great horror movie icons
and made the name Frankenstein synonymous with spooks
Rat Man In The Wall – Mulberry Street
Indie film Mulberry Street shows you what would happen if your apartment was infested with rat people — that’s right
While I couldn’t find the picture of him
there was one particular Mulberry Street tenant who bounded through the walls of the poorly constructed apartment complex like a rabid beast
blasting out of the paper thin plaster whenever he was hungry for a snack
Michael Rooker gets possessed by a lonely space alien
who then forces the entire town to mate/bond with him
And that’s a lot of naked people no one wanted to see
Judge Alvin Valkenheiser – Nothing But Trouble
Dan Aykroyd was terrifying as Judge Alvin ‘J.P’ Valkenheiser
because he decided who got to leave and who got to ride the flesh ripping roller coaster
who could forget those slimy disgusting babies and his nose was a penis
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