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Growth depression of Rosa plants at sites previously used to cultivate the same or closely related species is a typical symptom of rose replant disease (RRD)
limited information is available on the causes and the etiology of RRD compared to apple replant disease (ARD)
this study aimed at analyzing growth characteristics
as well as microbial communities in the rhizosphere of the susceptible rootstock Rosa corymbifera ‘Laxa’ grown in RRD-affected soil from two sites (Heidgraben and Sangerhausen)
either untreated or disinfected by γ-irradiation
plants developed significantly more biomass in the γ-irradiated than in the untreated soils of both sites
corymbifera ‘Laxa’ roots were site- and treatment-dependent
Although aloesin was recorded in significantly higher concentrations in untreated than in γ-irradiated soils from Heidgraben
the concentrations of phenylalanine were significantly lower in roots from untreated soil of both sites
Rhizosphere microbial communities of 8-week-old plants were studied by sequencing of 16S rRNA
and cox gene fragments amplified from total community DNA
sequences affiliated to the bacterial genus Streptomyces and the fungal genus Nectria were identified as potential causal agents of RRD in the soils investigated
The relative abundance of oomycetes belonging to the genus Pythiogeton showed a negative correlation to the growth of the plants
the effects of soil treatments on the composition of the rhizosphere microbial community revealed striking similarities to findings related to ARD
only little information is available about phytoalexins in roses
These studies recorded changes in the soil microbiome
possibly indicating a lack of plant growth-promoting microbes in ARD soils
detailed scientific investigations of the soil
and root microbiome under RRD conditions have not yet been carried out
RRD is affecting rose rootstock production and field production of garden roses in tree nurseries
Changing sites of production is often not feasible due to specialization and a lack of virgin soil
Soil disinfection is not environmentally friendly and cost intensive
efforts are needed to better understand the etiology of RRD and to develop measures to overcome the problem
Motivated by the recently achieved insights into ARD
this study aimed at a concise characterization of rose rootstock responses to RRD soils by investigating the rhizosphere microbiome (bacteria
the root morphology and phenolic secondary metabolite profiles
The analyses were conducted using the RRD-susceptible rootstock Rosa corymbifera ‘Laxa’ grown in untreated and γ-irradiated RRD soils from two sites under greenhouse conditions
The following hypotheses were addressed in the present study
(1) The composition of the rhizosphere microbiome (bacteria
fungi and oomycetes) is significantly affected by the soil treatments
corymbifera ‘Laxa’ plants is linked to a reduction in the relative abundance of taxa known as potential plant pathogens
(3) Symptoms on RRD-affected roots are similar to those observed for ARD-affected apple roots
(4) The secondary metabolite profiles in RRD-affected roots differ from those detected in roots from disinfected RRD soil
Appearance (a) and biomass (b) at the end of the bioassay
after 8 weeks of growth in soil from Heidgraben (H) and Sangerhausen (S)
Data are means ± SD (n = 10 and 5 for shoot and root dry masses
Letters indicate significant differences between variants within sites (t-test
Symptoms were detected after eight weeks of cultivation in RRD soil from the site Heidgraben (H) in whole-mount samples (a–g) and thin sections stained with toluidine blue (f–m)
a Healthy fine root with intact root hair zone; b constricted root structure and brown necrotic zones with black cell clusters (arrows) in a toluidine blue-stained root segment; c
d black root tips and clusters (arrows) of necrotic rhizodermal and cortical cells; e fine root with intracellular cauliflower-like (CF) fungal structures (arrows) in necrotic rhizodermal and cortex cells; f CF structures and black cell inclusions (arrows); g green fluorescent Actinobacteria in cortex cells after FUN®1 cell vital staining
h Longitudinal thin section through healthy fine root tissues with rhizodermis R
and xylem X; i cross section of an infected fine root with fungal hyphae and CF structures (arrows) in necrotic cortex cells; j infected fine root with fungal CF structures and hyphae (arrows) in rhizodermal and cortical cells; k thick fungal hyphae
intercellular entry point and developing intracellular CF structure (black arrows) and Actinobacteria (white arrows); l mixed infection with a CF structure-forming fungus (black arrows)
and thread-like Actinobacteria (white arrows); m cortex cells with fungal CF structures and round-shaped fungal chlamydospores (arrows)
Contents of the secondary metabolites (catechin
and quercitrin) and a primary metabolite (phenylalanine) were detected by HPLC-HR-MS and GC-MS (epicatechin and gallic acid)
untreated RRD soils from Heidgraben and Sangerhausen
γ-irradiated RRD soils from Heidgraben and Sangerhausen
Letters indicate significant differences in the contents of compounds between untreated and γ-irradiated samples within a site
Except for a significant reduction in operational taxonomic unit (OTU) numbers in SG compared to SU soil, bacterial α-diversity indices were not significantly altered due to treatment effects at both sites (Supplementary Table S3)
Bacterial (a) and fungal (b) community compositions were revealed by principal coordinate analysis (PCoA) using Bray–Curtis distance metric
based on operational taxonomic units (OTUs)
The relative abundance of dominant bacterial (a) and fungal (b) phyla (>1%) is indicated
Letters indicate significant differences between treatments within a site and phylum
p < 0.05 and n = 4 and 5 for sites H (Heidgraben) and S (Sangerhausen)
Plants were grown for eight weeks in untreated (U) and γ-irradiated (G) RRD soils from two sites (S and H)
Asterisks (*) in red and green color indicate significantly decreased and increased relative abundances of bacteria
in γ-irradiated compared to untreated RRD soils at both sites (H and S)
n = 4 and 5 for sites H (Heidgraben) and S (Sangerhausen)
The relative abundance of the respective taxon below detection limit is indicated in black color
As members of Nectria contain a complex of several closely related species that are difficult to separate taxonomically
a BLAST search against the NCBI database (nucleotide collection
nr/nt) was applied and the OTUs belonging to the fungal genus Nectria could be annotated to Neonectria sp
Low OTU numbers of 6–12 and no significant treatment effects on oomycete α-diversities were obtained using the cox 2 sequence analyses (Supplementary Table S6)
the significantly increased growth in the G soils compared to the respective U soils clearly indicated that both soils were affected by RRD
The stronger relative reduction in growth of R
corymbifera ‘Laxa’ plants in soil from site H suggests that RRD in this soil was more severe than in the soil from site S
This could be due to the higher sand content
corresponding to a bigger pore volume or to the fourth rose replanting cycle at this site
the role of the Actinobacteria remains unsolved
as they might invade damaged tissue as primary colonizers or as successors of pathogenic fungi
The wide spectrum of beneficial and harmful Actinobacteria calls for further detailed studies on the species level
phenylalanine also showed a clear trend of higher concentrations in roots grown in G soils
which is the starting compound of the phenylpropanoid pathway
the massive damage of roots observed microscopically is likely to be associated with an increased deposition of lignin
which is a product of phenylpropanoid metabolism
the increased carbon flow from phenylalanine to phenylpropanoids leads to a decreased level of this amino acid
The remarkably high variation in the metabolite contents between single plants might be due to either fast dynamic responses or different responses in different parts of the root system
which is also reflected by the patchy appearance of RRD
aloesin might function as a phytoalexin in rose
it is a derivative of polyketide metabolism
it is the first time that aloesin was detected in rose
no studies have been reported so far that Peronospora sp
high relative abundance detected for members of Peronospora in RRD soil at both sites might be likely due to leaf residuals fallen onto the ground and to residing zoospores in the soils
a more detailed investigation of longer and other Peronospora gene sequences potentially based on rhizosphere RNA rather than DNA is needed
The phylogenetic tree showed that this species was closely related to P
the Pythiogeton members identified in the RRD soils of this study might have also contributed to RRD
Quantification by quantitative PCR is recommended for future studies dealing with RRD to confirm the relative abundance of several of the identified responders
isolations followed by inoculation experiments to investigate their effects on plant performance and root morphology will lead to a better understanding of their role in the development of RRD
Studies using the same plant materials for rhizosphere microbial community and metabolite analyses should be considered for future investigations
Correlation analyses then can be applied to reveal relationships between root metabolites and microbial community composition in the respective rhizospheres
Our study revealed pronounced similarities between ARD symptoms on apple roots and RRD symptoms on rose roots at the microscopic level
In contrast to the highly increased levels of phytoalexins in apple roots affected by ARD
the phenolic secondary metabolites detected in rose roots from RRD soil did not show a clear trend
a significantly higher level was detected in roots grown in RRD soil from site H
suggesting this compound as a phytoalexin candidate of rose roots
Changes in the rhizosphere microbiome composition due to treatment effects correlated with changes in plant growth and root integrity and revealed Nectriaceae and Streptomyces as potential causal agents of RRD
Twenty 1 L pots were filled per soil variant and one seedling was planted per pot
The length of the main (longest) shoot was recorded weekly
Five each of the 20 plants per variant were destined for the microscopic investigations and the secondary metabolite analyses (LC-MS)
the final evaluation took place to record fresh and dry mass of the shoots and root fresh mass from ten plants per treatment
blotted dry and fresh masses were determined
Rhizospheres were extracted from five of the ten plants per variant
whereas RDM data (after drying for five days at 70 °C) were collected from the remaining five plants which were then used for metabolite analysis by GC-MS
the primers additionally included sequencing adapters and individual sample tags were used
amplicon products were purified using a HighPrep™ PCR Clean Up System (AC-60500
USA) applying a 0.65:1 (beads:PCR reaction) volumetric ratio to remove DNA fragments below 100 bp in size
Samples were normalized by SequalPrep Normalization Plate (96) Kit (Invitrogen
The pooled sample libraries were concentrated using the DNA Clean and Concentrator™-5 kit (Zymo Research
The pooled library concentration was determined via the Quant-iT™ High-Sensitivity DNA Assay Kit (Life Technologies) and adjusted to 4 nM
Amplicon sequencing was performed on an Illumina MiSeq platform using Reagent Kit v2 [2 × 250 cycles] (Illumina
Rarefaction of 16S rRNA gene fragment sequence reads at 5,602 led to exclusion of two samples (HU14
and HG13 (546 reads) were excluded from subsequent analyses after rarefying sequence reads at 10,554
namely SG3 (57 reads) and HU6 (341 reads) were excluded from subsequent analyses due to rarefaction of data at 775 reads
The sequence contingency table was exported on genus levels
clustering sequences into the OTU using VSEARCH (defined threshold at 97% sequence similarity)
OTU abundance tables and the RDP taxonomic assignment table (using the UNITE fungal ITS reference data set) were generated by the PIPITS_PROCESS
where n was the numbers of sequences from each OTU and N was the total numbers of sequences in the sample. The Student’s t-test, function t.test() and a significant threshold of p < 0.05 was applied using the transformed data (eqs. (1) and (2)
above) to check the effects of the soil treatments on the relative abundances of the rhizosphere microbiome with the software R-3.5.2
fungal and oomycetes genera that presented significant differences in their relative abundances (>0.5%) between soil treatments were tested for correlation with SDM and root fresh mass of the plants using the Pearson’s correlation coefficient (r) by Past3 (3.02)
Induction and diagnosis of apple replant disease (ARD)
Diagnosis of apple replant disease (ARD): microscopic evidence of early symptoms in fine roots of different apple rootstock genotypes
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Evaluation of apple replant problems based on different soil disinfection treatments—links to soil microbial community structure
A multi-phasic approach reveals that apple replant disease is caused by multiple biological agents
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Impaired defence reactions in apple replant disease-affected roots of Malus domestica ‘M26’
Transcriptomic analysis of molecular responses in Malus domestica ‘M26’ roots affected by apple replant disease
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Investigations about the course of infestation of rootlets of apple seedlings by root pathogenic actinomycetes in soils with specific apple replant disease
Fine-root system development and susceptibility to pathogen colonization
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Cylindrocarpon species associated with apple tree roots in South Africa and their quantification using real-time PCR
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Genes involved in stress response and especially in phytoalexin biosynthesis are upregulated in four Malus genotypes in response to apple replant disease
metabolite profiling and expression analysis reveal Rosaceae roots as the site of flavan-3-ol biosynthesis
distribution and physiological relevance in plants
Biochemical and antimicrobial activity of phloretin and its glycosilated derivatives present in apple and kumquat
Phytochemical constituents and in vitro radical scavenging activity of different Aloe species
Soil indigenous microbiome and plant genotypes cooperatively modify soybean rhizosphere microbiome assembly
Plant species and soil type cooperatively shape the structure and function of microbial communities in the rhizosphere
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Analysis of phenol-adapted microbial community: degradation capacity
Phylogeny of the genus Pythium and description of new genera
a new species causing root and basal stalk rot of wild rice in the United States
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We are deeply grateful for financial support by the Mathias-Tantau-Stiftung
are also thankful to the Deutsche Forschungsgemeinschaft (DFG) for funding
We thank Florian Losch and Selahaddin Sezgin for instrumental analytical service
Andreas Wrede for providing us with both the stratified rose seeds originally obtained from Harald Klei nurseries and the soil from Heidgraben
and Thomas Hawel for providing us with the soil from Sangerhausen
We also thank our technicians for their help in the experimental setup and evaluations
Open access funding provided by Projekt DEAL
Institute of Horticultural Production Systems
Institute for Epidemiology and Pathogen Diagnostics
Faculty of Chemistry and Chemical Biology (CCB)
Cluster of Excellence on Plant Sciences (CEPLAS)
The authors declare that they have no conflict of interest
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DOI: https://doi.org/10.1038/s41438-020-00365-2
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Last month, sixteen members of the Trinity Boys’ Choir set off on a tour of Germany
giving performances in locations including Sangerhausen
The tour began in the historic town of Sangerhausen where our students took part in a fledging festival collaborating with students from the choirs of Rossleben’s Klosterschule and Sangerhausen’s Geschwister-School
The festival culminated in two spell binding ‘promenade performances’ with audiences walking from one historic church to another between concerts
“The highlight for me was singing Singet dem Herrn by Bach
It was very interesting hearing a wonderful different style of singing and when we came together to sing the Bach
My favourite part other than the music was bonding with the rest of the choir
I really enjoyed the experience and feel privileged to have gone
It will be something I will look back on.”
“We thoroughly enjoyed the diversity of music that we sang including renaissance music from Tallis and White in addition to more modern works by Graham Lack and Jonathan Dove
including ‘Seek him that maketh the Seven Stars’ which was arguably our best piece
The lower voices formed an amateur seven-part bell choir
which featured in Lack’s “Candlemas.” What made this tour even more enjoyable was being surrounded by friends and doing what we all loved
singing; even on the bus journeys home where Disney and Queen often featured in our karaoke
this has undoubtedly been the best tour we have ever been on.”
The tour was a fantastic opportunity for the students to perform a very challenging programme of music for audiences across Eastern Germany and establish lasting relations with schools and choirs across the region
They are followed by another tour of nine members of the Trinity Boys’ Choir who fly out to Germany in November to give performances in Marburg
Hamburg and Lüneburg with male voice ensemble
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Anne Frank’s 75th birthday is being marked by a series of exhibits in Europe and the United States that shine a spotlight on the famed Shoah victim’s prewar life
June 15 (JTA) — Like tens of millions of others
Siegfried Kuhn read Anne Frank’s diary — but he didn’t stop there
“I try to convince others that they should be interested
so that nothing like that can ever happen again,” he said
On Saturday — which would have been Anne’s 75th birthday — Kuhn
came to Berlin from the German town of Sangerhausen to see an exhibit of 40 family photos by Otto Frank
which is tucked inside a courtyard in the former East Berlin the photos shed new light on the Franks’ daily life before World War II
Similar exhibits are being mounted by the Anne Frank House in Amsterdam at the Foam Gallery
which was arranged by The Anne Frank Center USA in Manhattan
Hanging on the walls are pictures of Anne and her sister
All indicate that Otto and Edith Frank tried to preserve a sense of normalcy until they went into hiding in the attic of 263 Prinsengracht St
and the family was arrested and deported in August 1944
Anne and Margot Frank died of typhus in the Bergen-Belsen concentration camp in March 1945
One of the most striking photos is one of Anne sitting at a desk in Amsterdam
Anne “was a girl who loved life,” said Buddy Elias
as long as she was able to come to Switzerland,” said Elias
an actor who since 1996 has been president of the Anne Frank Foundation of Basel
“and her essays and fairytales were amazing
there was an Anne who belonged to me and an Anne that belonged to the world,” Elias said
“She had the same problems as any other teenager: parents
that a child had to hide because a government declared her to be less than human
came to the opening from Jerusalem to see photos of the girl whose voice she last heard over a barbed wire fence in Bergen-Belsen
“We went to the same kindergarten” in 1934
whose family also fled Germany for Holland
but in 1942 Pick came back to school after vacation “and I wanted to play and she was not there.” The exhibit
“Anne Frank and Her Family — Photographs by Otto Frank,” is one of several projects honoring the memory of the girl whose diary has been translated into 60 languages since its initial 1947 publication in Dutch
German schoolchildren will compose essays for a competition on how to combat prejudice; winning essays will become “A Book for Anne Frank.” The diary also will be translated into Arabic for the first time
head of the Anne Frank Center’s board
said the center decided to make it possible for Arabic-speaking children in Europe to read the diary “because we believe the book is a symbol of understanding and of human rights and understanding
a symbol against persecution and discrimination,” he said
realistic facsimiles of Frank family photo albums lie open in glass cases
looking as they might have when Anne filched a snapshot to slip into the pages of her diary
was an avid photographer who took both candid and posed shots of his family
He reportedly never picked up his camera after the war
One of the few photos of Otto Frank in the exhibit was taken by the American photographer Arnold Newman in the Prinsengracht attic
on the day it opened to the public in 1960
Otto Frank leans against one of the support beams in the attic
the church bells that Anne described in her diary began to ring
Both Otto Frank and the photographer cried
The center also contains a permanent exhibit about the fate of the family
including a video interview with Otto Frank
who survived Bergen-Belsen with her sister Rachel
Anne and Margot had been sent to the camp from Auschwitz
while Pick and her sister had come via the Westerbork transit camp in Holland
the two groups were separated by a barbed wire fence filled with straw
“I heard by chance that she was there behind the fence,” she said
A woman who had hidden with the Franks in Amsterdam brought Anne to the other side of the fence
“but I never saw her.” Once Anne “asked if I could throw over something to eat,” she said
Pick threw Anne a Red Cross package with bread and dried plums
Because another person caught it and ran away and gave her nothing,” Pick said
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