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We are very happy to welcome you this year to the Dutch Annual Meeting Experimental Plant
Sciences. Also this year we have an excellent program with three well known keynote speakers
and plenty of opportunities to network. The poster session is back in the program. The EPS PhD
council is organizing the poster awards and will reward the three best posters with a prize. This
year we also introduce the EPS Awards for the first time. Members of the EPS community could
nominate their colleagues for the three following awards: EPS Spotlight, EPS Mentor and EPS
Young researcher. The EPS Award committee is really excited to announce the winners.
Organizing Committee
Prof. Guido van den Ackerveken Prof. Christa van Testerink Prof. Jian Xu
(Chair)
EPS Office
Gold Sponsor
Silver Sponsor
Bronze Sponsor
Venue
Address
Hotel De Werelt
Westhofflaan 2
6741 KH Lunteren
Travel information
You can reach the hotel by car or bike ( 45 mins from Wageningen). There is free parking next to
the hotel. The train station is in walking distance to the Venue (see map). It will take
approximately 15- 20 min to walk. For Monday morning we arranged a taxi bus to bring you to
the venue. The bus will stand outside of the train station from 8:30 until 09:45. On Tuesday, after
the meeting is finished, a taxi bus will bring you back to the train station. If you plan to use the
train please check the NS side for any updates before your travel.
Hotel Rooms
If you reserved a room ( single, double with twin beds or double with double bed), you can
check in from Monday 15:00 onwards. Please pick up the key at the reception of the hotel.
Breakfast will be served between 8:00 and 9:00. Please check out before the meeting starts on
Tuesday morning (not later than 10:00).
The hotel has arranged a room where you can store your luggage during the meeting.
Lactation room
There is a lactation room including fridge located in the D wing of the hotel. Please always check
with the reception if it is free to use.
We reserved a pray and calming room for both days. Please contact Juliane Teapal if you want
to make use of it.
Extra Information
Registration
The registration opens 08:30 on Monday morning. When you enter the hotel you will find the
registration area on the right side. Please pick up your badge before the meeting starts.
Catering
Coffee/ Tea and water will be available all day long throughout the whole venue. For lunch and
dinner all diet wishes are taken into account, but if something is missing please contact: Juliane
Teapal.
Each participant receives two consumption vouchers ( at the back of your badge). You can use
them on both days for soft drinks, beer and wine. Further drinks can be purchased on your own
costs. Please be aware that you can only pay with card, no cash possible.
PhD/postdoc/PI-hour
The PhD, postdoc hour and PI hour will take place on Monday from 17:30 until 19:00.
Poster Presentation
With this program you receive the number of your poster board. Please hand in your poster
during the registration. We will make sure your poster ends up at the correct board. The posters
can be visited from the first coffee break after the lunch on Monday until the last coffee break
before the lunch on Tuesday. The poster session takes place on Monday from 20:30 until 22:00.
Social activities
Party
This year we arranged a venue for a small party. On Monday evening the room FIRE will turn
into a party room including a dancefloor and a small bar, starting at 21:30. We are also very
happy to announce that Nanne Taks (UvA) will be our DJ for this evening. At midnight the sound
and lights will be turned off. Enjoy!
Running activity
Ready to get up early ? Join the morning run under the guidance of Jason Gardiner (PI at UU
Translational Plant Biology). Meet up in front of the conference center for the warming up at
07:00.
Programme
Monday 17 April 2023
08:30-09:25 Registration and Coffee
09:25 - 09:30 Welcome
Plenary Session 1 Chair: Guido v.d. Ackerveken
09:30-10:15 Jonathan Jones
10:15-10:45 Petra Bleeker (UvA)
10:45-11:15 Coffee break
11:15-11:45 Rashmi Sasidharan (UU)
11:45-12:15 Joris Sprakel (WUR)
12:15-12:45 Daan Weits (UU)
12:45-12:55 EPS Awards
12:55-14:15 Lunch
Parallel Session 1 Biotic Interactions I (Air) Genomics I (Water) Cell Biology (Fire)
14:15-14:35 Miguel Ramirez Gaona (WUR) Jillis Grubben (WUR) Yosapol Harnvanichvech (WUR)
14:35-14:55 Ava Verhoeven (UU) Meixin Yang (WUR) Enric Martínez-Calvó (UvA)
14:55-15:15 Thomas Aalders (UvA) Jorge Aleman Baez (WUR) Bas Jacobs (WUR)
15:15-15:45 Coffee break Coffee break Coffee break
Parallel Session 2 Biotic Interactions II (Air) Genomics II (Water) Physiology (Fire)
15:45-16:05 Arezoo Rahimi (LEI) Iqbal Maulana (WUR) Ludovico Caracciolo (WUR)
16:05-16:25 Gijs Selten (UU) Dong Zhang (WUR) Lisa Oskam (UU)
16:25-16:45 Valerie Buijs (KNAW) Mandy Ravensbergen (WUR) Phuong Nguyen (WUR)
16:45-17:05 Pedro Beschoren da Costa (WUR) Nam Hoang (WUR) Kees Ketting (WUR)
17:05-17:20 Flash Talks Flash Talks Flash Talks
17:30-19:00 PI/postdoc/PhD hour
19:00-20:30 Dinner
20:30-22:00 Poster Session
21:30-00:00 Party
Tuesday 18 April 2023
07:00-08:00 Running activity
Parallel Session 3 Development I (Air) Stress (Water) Novel Methods (Fire)
09:00 - 09:20 Judit Nadal-Bigas (WUR) Jasper Lamers (WUR) Mitja M. Zdouc (WUR)
09:20 - 09:40 Thalia Luden (LEI) Linge Li (UU) Sebastian Tonn (UU)
09:40 - 10:00 Honglei Wang (WUR) Martijn Jansen (RU) Peter Bos (WUR)
10:00 - 10:20 Flash Talks Flash Talks Flash Talks
10:20 - 10:50 Coffee break Coffee break Coffee break
Parallel Session 4 Development II (Air) Biotic Interaction III (Water) Non-Model Species (Fire)
11:00 - 11:20 Iris Zahn (WUR) Dario Ramirez (NIOO/UU) Manuel Aguirre Bolaños (UU)
11:20 - 11:40 Merijn Kerstens (WUR) Sietske van Bentum (UU) Francesco Garassino (WUR)
11:40 - 12:00 Nina Guarneri (WUR) Nanne Taks (UvA) Erbil Güngör (UU)
12:00 - 13:45 Lunch
Plenary Session 2 Chair: Christa Testerink
13:45-14:30 Peter Kuipers Munneke
Charlotte Gommers
14:30 - 15:00 (WUR)
15:00-15:30 Coffee break
Plenary Session 3 Chair: Jian Xu
15:30-16:00 Sandra Irmisch (LEI)
16:00 - 16:45 Caroline Dean
16:45 - 17:00 Poster Award
17:00 - 17:10 Closing remarks
Monday, 17 April 2023
08:30 - 09:25 Registration and Coffee/Tea
10:45-11:15 Break
12:50-14:15 Lunch
15:15-15:45 Break
17:30-19:00 PhD hour (room Air) /PI hour (room Water)/ Postdoc hour (room 21)
19:00-20:30 Dinner
10:20-11:00 Break
12:00-13:45 Lunch
15:00-15:30 Break
Parallel Session 01
Date: 17 April 2023
Time: 14:15 – 15:15
Parallel Session 02
Date: 17 April 2023
Time: 15:45 – 17:20
Parallel Session 04
Date: 18 April 2023
Time: 11:00 – 12:00
Session 01
Session 02
Date: 18 April 2023
Time: 10:00 – 10:20
Common scab, caused by Streptomyces spp., is worldwide one of the most important skin
blemish diseases in potato, leading to a significant reduction in economic value. Resistant
varieties are the most effective way of dealing with this disease, because control by
irrigation is expensive and unsustainable in the long term, if not contributing to yield.
The aim of this project is to identify the hereditary factors involved in resistance to
common scab. Several sources of resistance are known, such as the old potato varieties
Jubel and Hindenburg, as well as modern tetraploid and diploid varieties. These are being
used to create mapping populations for field trials. Specifically, we are trying to
understand whether resistance is controlled by a single locus or by multiple small-effect
QTL with additive effect. Because a vast body of historical data on scab resistance is
available, for ~ 500 varieties with SNP array genotypic data, a GWAS-analysis is being
carried out. Pot assays and in vitro assays will be developed to test for resistance gene by
isolate interactions with reference strains of Streptomyces spp.. Knowledge on the
spectrum of resistance is essential to breed for broad-spectrum scab resistant varieties.
Identification of molecular markers associated with alleles contributing to common scab
resistance will simplify the long-term objective of breeding resistant varieties with greater
efficiency and less expense than in conventional field screening.
People working in mushroom farms can become seriously ill (severe allergic reactions and
lung problems) when frequently exposed to high concentrations of spores. Especially P.
ostreatus is known to produce huge amounts of spores that are released from early stages
in mushroom development, Therefore, access to spore-less or low sporulating strains of
P. ostreatus is of great importance for commercial cultivation of these mushrooms. A
sporeless strain (SPOPPO), developed via a natural occurring sporeless mutant is already
on the market. However, cultivars with properties important for e.g., the Asian market are
not yet available. To obtain sporeless cultivars, germplasm with disrupted pathways
towards sporulation are needed.
For food, non-GMO strains are strongly preferred, requiring random mutagenesis. Two
methods to mutagenize P. ostreatus protoplasts were tested, EMS and UV. SNP analysis
of resequenced mutants showed that EMS only resulted in a low mutation rate (average
20 mutations), while UV irradiation resulted in much higher mutation rates (average 278
mutations). Currently, mutants are being analysed by 2-D pooled sequencing to screen for
mutations in genes that are known to be involved in sporelessness (e.g., the MSH4 gene).
Plant breeding heavily relies on genetic diversity in (wild) plant accessions that can be
crossed with a crop of interest. The integration of genomes from cultivated species and
their wild relatives in a super-pangenome allows for the discovery of genetic mechanisms
underlying traits of interest, such as pest resistance. For lettuce (Lactuca sativa L.) and its
wild relatives, a growing number of genetic resources enables the analysis of hitherto
unknown genetic mechanisms.
Vascular wilt disease, caused by the fungal pathogen Fusarium oxysporum (Fo), is a major
threat for many important crops. Dominant genetic resistance to Fo is rare and typically
overcome by the constantly evolving fungus. As an alternative Susceptibility (S) genes of
the host can be inactivated to potentially confer broad and durable resistance. Here, we
identify tomato S gene candidates required for susceptibility to Fo. The Fo effector SIX8,
which is present in many pathogenic isolates but absent in non-pathogenic ones, was
found to specifically interact with two members of the TOPLESS family from tomato: TPL1
and TPL2. SlTPL1 knockouts exert strongly reduced susceptibility to Fusarium wilt. An even
stronger resistance was observed in tpl1;tpl2 mutants, albeit with a mild pleiotropic
phenotype. The undesired phenotype can be alleviated by grafting, as tpl1:tpl2 mediated
resistance was found to be root-mediated. Single and double knockouts of Arabidopsis
TPL and/or TPR1 showed a similar reduction in Fo susceptibility. Therefore, we conclude
that TPLs are genuine S genes whose inactivation can confer resistance to F. oxysporum.
Fruit development and ripening in tomato involves coordination and tight regulation of
gene expression. MADS-domain transcription factors are important in many biological
processes of plants and interactions between MADS-domain proteins are essential for
their functions. In tomato (Solanum lycopersicum), several MIKC-type MADS-domain
proteins, such as FUL1, FUL2, MADS-RIN, playing a role in fruit development and ripening
have been identified, but an in-depth characterization of their unique and (partially)
redundant functions is lacking. In this project, we aim to elucidate further the tomato fruit
development and ripening regulation exerted by MADS-box TFs and their interactions
among each other and with other genes, particularly for FUL1, FUL2. We will use
CRISPR/Cas and RNAi to generate single mutants and combinations thereof to reveal any
redundant functions or cooperativity among these TFs during fruit development and
ripening. Altogether, this study will unveil the molecular regulatory network of MADS-BOX
together with other TFs and of FUL1/2 in fruit development and ripening.
6. “The role of chloroplast-derived signals in intercellular communication”
Chloroplasts are organelles with an endosymbiotic origin and possess their own genome
and protein synthesis machinery. Retrograde signals (RS) inform the nucleus about
chloroplast development and operational status under changing environmental
conditions. The developmental state of the chloroplast has an impact on whole plant
development via RS, including the development in distal organs. Moreover, a number of
recent discoveries have established that signals originating in the chloroplasts control
plasmodesmata, the direct cell-to-cell conduits for many physiologically and
developmentally relevant signals. Therefore, RS might be directly linked to developmental
signalling across cells and tissues through plasmodesmata. We aim to determine how RS
regulate local and distal plant cell development. In order to do so, we will use the
bryophyte Physcomitrium patens as a model. This moss grows long, filamentous chains of
cells called protonema. Despite its simple structure, this tissue exhibits cell-differentiation
accompanied by distinct chloroplast development. Thus, this model gives the opportunity
to study the interplay between overall plant development and chloroplast development
as well as conveniently track the intercellular communication during this process. By
investigating RS pathways in P. patens, we will also better understand the evolution of this
process in land plants. A better understanding of the maintenance of chloroplast function
during suboptimal conditions might prove essential when improving photosynthesis in
crops for yield-improvement in the future.
The allelic variation pattern in the autotetraploid potato is less explored. Here, we
identified 81 genes involved in starch metabolism and detected 65,245 allelic variants in
six haplotype-resolved autotetraploid potato genomes. Comparative analyses revealed
uneven distribution of allelic variants among gene haplotypes and that the vast majority
of deleterious mutations in these genes were retained in heterozygous state in
autotetraploid potato. Population genetic analyses identified starch biosynthetic genes
that have been possibly diverged between potato varieties with high or low starch
content. These results increase our understanding of haplotype diversity in
autotetraploids and will also facilitate functional characterization of genes contributing to
starch-related traits in potato.
The root exodermis is characterized by the deposition of two secondary cell wall polymers,
suberin and lignin in the outermost cortex layer. Many plant species can dynamically
develop the exodermis in response to abiotic stresses. The phytohormone abscisic acid
(ABA) is a core bridge between the abiotic stress perception and the exodermis
differentiation program. Yet very little is known on how the ABA signal is integrated into
the exodermis regulatory network. Our work aims to investigate the ABA regulatory
network involved in exodermis differentiation of roots in different plant species. We focus
on the legume family (Fabaceae) as it contains a diversity of exodermis forms with a wide
range of developmental responses to ABA.
We developed Cell Layer Adjusted Pixel Intensity (CLAPI) method to characterize the cell
type specific accumulation of suberin and lignin in roots. Using this method, we identified
a wide range of exodermis type in the legume family. We found a few species with
constitutively suberized and lignified exodermis, while others showed the unique ability
to promote the exodermal differentiation only in the presence of ABA. Interestingly, we
observed that the pattern of suberin and lignin deposition in the cortex differs across
distinct legume species in response to ABA. While a few species showed a more diffuse
deposition of suberin and lignin in the cortex, other species promoted the accumulation
of these polymers specifically in the outermost layer of the cortex. Our next steps are
building gene regulatory network models of the exodermis development across the
legume family.
9. “Dissecting the regulatory regions of the pleiotropic transcription factor FUL2 in tomato"
Kai Thoris, Plant Developmental Systems, Wageningen University and Research
Pleiotropic genes are genes that influence more than one trait and can therefore be
problematic for plant breeding. Unintended effects can occur when breeding for a single
trait, as one mutation in the coding sequence can disturb all traits at once. Molecular
breeding tools and phylogenetic analysis can be used to have a more modular look at
pleiotropic genes and subsequently dissect their functions, so that more specific breeding
can be achieved. An example of a pleiotropic gene is the transcription factor FUL2 in
tomato. FUL2 is one of the two co-orthologs of the Arabidopsis FRUITFULL (FUL) gene and
regulates fruit development, fruit ripening, flowering time and inflorescence architecture.
To understand how FUL2 is regulated and performs these different functions, we aim to
identify and characterize the cis-regulatory elements that are required for tissue-specific
activity. Phylogenetic footprinting was performed to identify conserved regulatory motifs,
which were then screened against transcription factor binding site (TFBS) databases to
reveal potential binding sites. The identified conserved motifs were then characterized
with various methods such as CRISPR/Cas12a mutagenesis, yeast one-hybrid and GUS
reporter lines.
The ornamental crop Tulipa gesneriana (tulip) is widely known for its beautiful flowers,
which are at the basis of its long history of cultural and economical value. Tulip can
reproduce through flowers and storage organs, the bulbs. It derives that two reproductive
ways are utilized, sexual and vegetative. Current knowkedge on the regulation of both
reproduction strategies (and, possibly, their cross-talk) is limited. Recent studies on other
plants that produce storage organs (such as potato and onion), have highlighted the role
of PHOSPHATIDYLETHANOLAMINE-BINDING PROTEINS (PEBPs) as key molecular switches
of both flowering and storage organ formation, through complex formation with
transcriptiona factors. Plant PEBPs constitute a very conserved gene family, which in
Angiosperms can be divided in three main clades: FLOWERING LOCUS T-like (FT-like),
TERMINAL FLOWER-like (TFL1-like) and MOTHER OF FT AND TFL1-like (MFT-like). In
Monocots, expansion of the FT-like clade resulted in numerous paralogs which sometimes
evolved into antagonistic pairs, competing for binding to the same transcription factors,
and therefore determining the timing of a developmental event. Our research is aimed at
characterizing the PEBP family in tulip, and ultimately understand their involvement in
orchestrating tulip’s dual reproduction.
Transcriptome mining resulted in the identification of eight full-length PEBPs in tulip.
Sequence comparisons show amino acid substitutions at key positions, pointing towards
functional diversification. Additional analyses such as RT-qPCR, yeast-two hybrid and
heterologous overexpression contributed to shape the hypothetical role of TgFT1 and
TgFT3 as antagonistic regulators of bulb formation, while the flowering regulators seem
to be the meristem-expressed TgFT4 and TgTFL1. Moreover, the leaf-born TgFT2 signal
could in principle affect both reproductive modesm opening the possibility of a PEBP-
mediated crosstalk between the two types of reproduction.
Heat waves are becoming more frequent and intense, causing a great threat to
agriculture. Due to the sessile lifestyle of plants and their necessity to endure such
unfavorable conditions, they have developed a remarkable capacity to establish a memory
of heat stress. With this so-called thermomemory, plants retain the experience of previous
moderate (non-lethal) heat stress exposure (priming) and survive a subsequent, otherwise
lethal heat stress. A previous study from our lab has demonstrated the existence of natural
variation between Arabidopsis thaliana accessions for the strength and duration of
thermomemory and identified a plastidial regulatory module (FtsH6-HSP21) to be
involved in superior thermomemory of accession N13. We now identified another
accession (ACC1) with a much higher thermomemory capacity than N13, Col-0, and its
reported mutants. Expression patterns of known thermomemory genes are virtually
identical in ACC1 and Col-0, suggesting that ACC1 employs memory mechanisms that are
conserved across accessions but also other, entirely unknown regulators that provide
superior thermomemory. We are applying genetic and systems biology approaches to
unravel the genes and mechanisms underlying ACC1’s high thermomemory capacity. The
progress and recent results will be presented.
12. “Concentration is key: unravelling mechanisms controlling Auxin Response Factor stability”
Martijn de Roij, Laboratory of Biochemistry, Wageningen University and Research
The signalling molecule auxin controls nearly all developmental processes in land plants
through a the nuclear auxin signalling pathway (NAP). The NAP consists of an auxin
receptor TIR1/AFB, its Aux/IAA degradation substrate, and the DNA-binding ARF
transcription factors. While an extensive qualitative understanding of the pathway and its
interactions has been obtained by studying the flowering plant Arabidopsis thaliana, it is
so far unknown how these translate to quantitative system behaviour in vivo, a problem
that is confounded by large NAP gene families in this species. Here, we used the minimal
NAP of the liverwort Marchantia polymorpha to quantitatively map NAP protein
accumulation and dynamics in vivo through the use ofusing knock-in fluorescent fusion
proteins. Beyond revealing the native accumulation profile of the entire NAP protein
network, we discovered that the two central ARFs, MpARF1 and MpARF2, are
proteasomally degraded. We mapped the degrons of both MpARFs to specific domains
and show that MpARF2 dimerization is an important factor for degron accessibility and
thus protein stability. In future work we aim to fully uncover the molecular mechanisms
which define MpARF stability through a variety of approaches which will allow us to study
functional implications of targeted MpARF proteolysis for plant development.
13. “Fine tuning desiccation tolerance in developing seeds of Arabidopsis thaliana using
controlled drying.”
Asif Ahmed Sami, Laboratory of Plant Physiology, Wageningen University and Research
Desiccation tolerance (DT) is the capacity of cells to withstand extreme losses of water
without incurring irreversible damage. DT played a decisive role in facilitating the
successful transition of plants from an aquatic to a terrestrial lifestyle. However, following
terrestrialization, DT was gradually lost from the vegetative tissues of most angiosperms
and became restricted to only seeds, spores, and pollens. Interestingly, in most seeds, DT
is acquired during the maturation phase of development, before the seeds experience a
gradual decrease in their total water content leading to an osmotic stress. Previous studies
have mostly focused on the role of ABA in DT acquisition, however, the role of controlled
drying or osmostress alone or in conjunction with ABA, on the regulation of seed DT has
not yet been deciphered. In this project, we aim at understanding how DT acquisition is
regulated and what is the effect of drying on seed survival. For that we optimized drying
treatments to seeds of Arabidopsis thaliana during several maturation stages by - (i) fast
drying the seeds outside the silique, and (ii) slow drying inside the silique. We found that
drying seeds at early and mid-maturation inside the siliques improves their DT acquisition.
From here on the plan is to perform RNA-seq and degradome-seq in these seeds to
investigate what transcriptome and the translatome changes, respectively, accompany
acquisition of DT during seed development.
14. “Exploring proteome interaction with immune receptors by using proximity labeling”
Tatiana Marti Ferrando, Laboratory of Plant Breeding, Wageningen University and
Research
The plant immune system comprises a complex network of mechanisms and it needs a
more complete understanding. In this context, pattern recognition receptors (PRRs) play
a key role in defense against pathogens. However, little is known about PRR-interacting
proteins and their regulation upon pathogen perception. We applied a TurboID-based
approach to study the interactors of the PRR ELicitin Receptor (ELR) in Nicotiana
benthamiana in presence or absence of the INF1, an elicitin from Phytophthora infestans.
We found that ELR-TurboID is able to biotinylate other PRRs at the plasma membrane
including the well-known co-receptor SUPRESSOR OF BIR1 (SOBIR1), as well as proteins
involved in vesicular trafficking. When INF1 is present, we reported less interaction with
these PRRs and more specific interaction with PAMP-triggered immune (PTI) related
proteins, such as the BRI1 ASSOCIATED RECEPTOR KINASE 1 (BAK1) and the RESPIRATORY
BURST OXIDASE HOLOMOG D (RBOHD). We further evaluated the application of this
approach using SOBIR1-TurboID in a wild potato Solanum microdontum. Enriched
proteins from SOBIR1-TurboID treatment showed interaction with proteins related to
defence and growth such as BOTYRIS-INDUCED KINASE1 (BIK1) homologs and
BRASSINOSTEROID-SIGNALLING KINASE (BSK) homologs. Interestingly, we indentified
common interactors of ELR in N. benthamiana and SOBIR1 in S. microdontum as
REMORIN, RPM1-INTERACTING PROTEIN 4 (RIN4) and PROTEIN PHOSPHATASE 2C (PP2C).
These findings illustrate the dynamics of the plant cell and reveal potential interactors to
be exploited in plant breeding. In conclusion, we successfully applied TurboID proximity
labeling in a non-model crop plant and created a roadmap for future research in protein-
protein interactions with PRRs among plant species.
15. “Achieving Durable Resistance Against Tomato Leaf Mold: Past, Current, and Future
Perspectives”
Mr. Christiaan R Schol1, Mr. Mats Bours1, Ms. Anne M Hilgers1, Ms. Anan Hu1, Ms. Ruifang
Jia1, Ms. Ángeles Ramos Peregrina1, Ms. Marjam Saleh1, Mr. Roeland F Seesink1, Mr.
Samuel L van Zwoll1, Dr. Silvia de la Rosa2, Ms. Hannah M McCarthy2, Dr. Like Fokkens1,
Dr. Matthieu HAJ Joosten1, Dr. Carl H. Mesarich2 and Dr. Yuling Bai1, (1)Wageningen
University, Wageningen, NETHERLANDS, (2)Massey University, Palmerston North, NEW
ZEALAND
Tomato leaf mold is caused by the fungus Fulvia fulva (syn. Cladosporium fulvum).
Resistant tomato recognizes F. fulva avirulence (Avr) effectors by means of Cf receptor
proteins, resulting in hypersensitive response-based resistance. So far, several Cf genes
have been characterized and deployed, however the durability of these genes has proven
to be limited. Durable resistance remains elusive since F. fulva readily overcomes Cf
resistance by loss or mutation of the corresponding Avr effector, after which the deployed
Cf gene becomes ineffective. To achieve a more durable form of genetic resistance, we
are introgressing, mapping and characterizing a significant number of novel Cf genes from
wild Solanum germplasm. In parallel, we are using CRISPR-Cas9 technology to generate
knockouts of the corresponding effectors in F. fulva to determine their contribution to
fungal virulence on susceptible tomato. We hypothesize that (combinations of) Cf genes
corresponding to effectors that strongly contribute to virulence will result in more durable
resistance. So far, we have taken steps towards mapping of the novel Cf genes, and we
have generated knockouts of the corresponding effectors in F. fulva. Additionally, we have
identified Avr9B matching the Cf-9 homolog, Cf-9B, explaining the sequential breakdown
of the widely used Cf-9 resistance locus, and currently we are working on identifying Avr6,
which matches the most recently deployed Cf protein, Cf-6.
16. “Lettuce Big Vein associated Virus ORF3 encodes a 30K-like protein that facilitates cell-to-
cell movement”
Viral movement proteins (MPs) are critical for the local spread of viruses in plants by
modulating plasmodesmata, membrane-lined pores connecting plant cells through cell
walls. Lettuce Big-Vein associated Virus, a member of the Rhabdoviridae, has been linked
to the Lettuce Big Vein Disease complex. This viral disease is characterized by vein clearing,
banding, and retarded growth of lettuce plants. Despite that this virus is known for
decades, its viral proteins have not been studied in detail yet. Using structural predictions,
we found that ORF3 of Lettuce Big-Vein associated Virus (LBVaV) appears to adopt the
structural topology of the 30K-like movement protein family. This is remarkable, since
classical protein multiple sequence alignment was unable to show such relationship. We
next showed that ORF3 locates at the plasmodesmata by expressing it in concert with the
marker protein PDCB1. To show that ORF3 from LBVaV facilitates cell-to-cell movement of
plant viruses, movement-impaired infectious clones of Potato Virus X (PVX) and Tomato
Mosaic Virus (ToMV) were co-expressed with LBVaV ORF3. LBVaV ORF3 indeed facilitated
movement of both plant viruses. The combined observations revealed that LBVaV ORF3
encodes a 30K-like movement protein that facilitates viral cell-to-cell movement.
17. “Combating Vascular Diseases: Identifying the “Guardians” of the Xylem Sap”
Margarita Simkovicova, Laboratory of Molecular Plant Pathology, University of
Amsterdam
The fungus Fusarium oxysporum (Fo) is a destructive pathogen of a wide variety of crop
species. Four different resistance (R) genes (I, I2, I3 and I7) are employed in cultivated
tomatoes to protect against Fo f.sp. lycopersici (Fol). These R-genes encode structurally
diverse immune receptors that recognize Fol effectors in different root tissue layers. Even
though R-protein activation prevents disease, Fol still colonizes the vessels of resistant
tomato varieties, the extent of colonization depending on the R-gene present. The
mechanisms by which the different R-genes control vascular Fol abundance are currently
unknown. Since Fo resides mostly in xylem tissues during infection, we hypothesize that
specific compounds in the xylem sap restrict Fol proliferation. In this study, the xylem sap
proteomes of Fol-inoculated resistant plants were compared to that of mock-treated
plants using label-free quantitative LC-MS/MS. The quantity of various Fol effector
proteins varied between R-gene lines corresponding to the observed differences in fungal
proliferation. Besides several unique proteins, all four R-receptors induced the
accumulation of the same set of five proteins (e.g. pathogenesis-related proteins,
glucosidases, endochitinases). To assess their involvement in Fol resistance, these
candidates are currently being knocked out or overexpressed. Preliminary data indicate
that these proteins are key players in resistance against Fol.
Seed priming is a pre-sowing treatment that enables a more efficient and uniform
germination. However, it negatively affects seed longevity. In this work, the mRNA
dynamics underlying a hydropriming treatment has been investigated. Polysome profiling
was performed on seeds during different stages of hydropriming. Ribosome nascent chain
complex sequencing (RNC-seq) elucidated transcriptomic and translatomic changes
during the treatment. In contrast to mature dry seeds, hydroprimed seeds contain
polysome complexes indicating that the mRNAs that need to be translated during
germination are already associated to ribosomes, leading to a quicker germination upon
re-imbibition. During priming, seeds lose part of their stress-related transcriptome. This
work highlights the genes that might play a central role in faster germination and/or
reduced longevity following the priming treatment.
19. “The eXodermis files: uncovering gene regulatory networks behind (dynamic) exodermis
development and evolution”
Rianne Kluck, Laboratory of Plant-Environment Signaling, Utrecht University
Since the transition out of the water, plants have had to evolve and adapt to the dry and
ever-changing environmental conditions on land. One of the adaptations angiosperms
evolved is the root barrier cell types: the endodermis and the exodermis. The endodermis
evolved earlier, its physical barriers being suberin and/or lignin depositions in the cell
walls. The exodermis evolved later and likely co-opted parts of the endodermis network
for deposition of lignin and/or suberin to the first sub-epidermal cortex layer(s) in order
to control water and solute uptake in stressed soil conditions. Despite the physiological
relevance of this adaptation, a consistent angiosperm-wide characterization of exodermis
that takes into account stress conditions, screening methods, developmental age, and
growth media is currently lacking. Therefore we are screening several phylogenetically
distinct angiosperms in control and stress conditions using abscisic acid (ABA) as a stress
signal. We have selected three major angiosperm families, Poaceae, Brassicaceae, and
Fabaceae, to investigate exodermis diversity, dynamics, regulation, and networks. To
eXpose whether a species has hidden characteristics in the exodermis cell file, we use
sectioning, histology, and confocal imaging. Moreover, we will study the spatiotemporal
aspects of transcription during ABA-induced exodermis development in various species.
Combining the morphological and physiological exodermis diversity with underlying
transcriptional responses on a multi-species level, we will infer which gene networks
regulate distinct exodermis cell file types in a phylogenetic context.
Fusarium wilt disease, caused by the fungus Fusarium oxysporum (Fo), affects over one
hundred plant species, resulting in significant crop losses globally. Pathogenic Fo strains
are often host specific, only able to infect one or a few related plant species. Fo. f. sp
melonis (Fom) and Fo f. sp. cucumerinum (Foc) are host-specific Fusarium strains that
cause disease in melon and in cucumber, respectively. In contrast, Fo f. sp. radicis-
cucumerinum (Forc) can infect three different hosts within the cucurbits: cucumber,
melon and watermelon. In previous research, the first ‘non-host’ avirulence gene was
found in Fom, Effector Candidate for Cucurbits (ECC1aFom). Transferring this gene, which
encodes a small secreted protein, into a Forc strain compromised its ability to infect
cucumber. Based on these findings, we aim to determine plant responses that control
compatibility with cucurbit-infecting Fo isolates and to uncover how Fo evolved
compatibility towards different cucurbit species. To identify the role of ECC1a and its
homologs in host compatibility, knock-out mutant strains of Fom and Forc were generated
using a CRISPR/Cas9-mediated genome editing approach. The mutants were tested for
altered virulence on cucurbits. Preliminary results indicate that ECC genes are promising
effector candidates to be involved in Fo virulence on different cucurbit species.
22. “Hijacking the plant DNA replication machinery - towards the structure of the viral
replication initiator protein and its interaction partners”
Sandra Eltschkner, Laboratory of Molecular Plant Pathology, University of Amsterdam
Problems with begomoviruses have been heavily aggravated in recent years; particularly
Tomato Yellow Leaf Curl Virus (TYLCV) causes severe damage on tomato and cucurbits
impacting growers worldwide. Climate change has allowed the spread of the main vector
for viral transmission, the whitefly Bemisia tabaci, which is now present in massive
populations in more moderate climate zones including the Mediterranean. With the ban
of pesticides and a lack of strong resistance genes in our crops, we urgently need
alternative strategies to halt viral spread.
Due to their small genome, these viruses rely on host factors for DNA replication. The viral
proteins need to reprogram the host cell cycle to stimulate entry of the S phase, and to
recruit the plant DNA-replication machinery to the viral DNA. Only one viral protein, the
Replication initiator protein (Rep), is essential and sufficient to promote viral DNA
replication inside plant cells. Rep interacts with a multitude of plant proteins including
PCNA, a central processivity factor for DNA polymerases. Our goal is to elucidate the
structure of Rep from TYLCV in complex with tomato PCNA. To this end, it will be critical
to trap Rep in a fixed conformation and stoichiometry with PCNA to obtain a complex
suitable for structural studies. Elucidating the Rep-PCNA interaction interface will provide
invaluable insight in how Rep orchestrates rolling-circle DNA replication – a likely
conserved mechanism among Circular Rep-encoding single-stranded DNA (CRESS-DNA)
viruses – and help to identify their weak spot.
23. “Dissecting salt-dependent regulation of the floral transition in Arabidopsis using promoter
editing”
Joram Dongus, Laboratory of Plant Physiology, Wageningen University and Research
Salt delays plant growth and development in Arabidopsis, including flowering & bolting
time. Using a Genome-wide association study (GWAS) we set out to identify regulators
that delay the floral transition exclusively during salt stress. This analysis revealed that the
absence of a highly variable region in the promoter of a UDP-GLYCOSYLTRANSFERASE
(UGT) correlates with reduced UGT expression and delayed bolting during salt stress. To
assess whether the absence of this variable region is causal for the reduced UGT
expression and delayed bolting time, we employed CRISPR/Cas9 editing to delete the
variable region from the Arabidopsis Col-0 reference genome. In line with our
observations in naturally occurring Arabidopsis accessions, UGT variable region deletion
mutants bolt and flower later in salt stress, but behave like Col-0 wild type in mock
conditions. This indicates that this region contains key cis-elements that regulate the floral
transition during salt stress. Currently, we are aiming to use Cas9-nickase (D10A) to make
more specific mutations inside the variable region to dissect which cis-elements are
responsible for the salt-dependent flowering & bolting time phenotype. Furthermore, we
aim to identify upstream and downstream regulators of UGT in the salt-dependent floral
transition.
Plants use light as their primary energy source. That light, converted to chemical energy
via photosynthesis, is essential for the survival of life on earth. To finetune the use of light,
plants continuously monitor their environment to optimize their growth and
development. In dense plant stands, there is a relative enrichment of green and far-red
light compared to the photosynthetically active blue and red light that is used in
photosynthesis. These spectral changes are followed by dedicated wavelength-specific
photoreceptors that cover most of the visible light spectrum. However, to date, no
dedicated green light receptor has been identified, although plants do respond to green
light perception by changing their growth and development. In this project, we aim to find
a green light photoreceptor that sheds new light on how plants perceive their
environment.
25. “Friend or Foe: Colonization of alternative hosts by the banana pathogen Fusarium
odoratissimum TR4”
Lisanne Kottenhagen, Laboratory of Phytopathology, Wageningen University and
Research
Seaweeds are currently an underused source of food, feed and for medicine, bioplastics
and bio stimulants. Cultivation in the North Sea consists mostly of kelp species seeded
onto ropes. Kelp has a biphasic life cycle with alternating gametophyte and sporophyte
life stages. To seed the commercially interesting sporophyte, a mix of gametophytes and
young sporophytes that are attached to the female gametophytes is used. Currently, the
seeding process is highly inefficient: up to 99% of the material is lost. We aim to improve
attachment of seeding material by identifying and understanding the factors that play a
role in attachment. We have developed a novel method to quantitatively assess kelp
attachment and have investigated the roles of the age of the seeding material and the
substrate. We show that after gametophyte seeding, they gradually increase their
attachment strength to a substrate over a period of two weeks. Attachment of young
sporophytes doesn't increase of attachment over two weeks. Our results suggest that the
majority of losses occur during the first days after seeding. The method that we developed
allows for high throughput screening of different substrates, strains, and treatments for
their performance in attachment to identify optimal seeding conditions to minimize
losses.
Plants may effectively tailor defenses by recognizing their attackers and reprogramming
their physiology. Although most plants are under attack by a large diversity of herbivores,
surprisingly little is known about the physiological capabilities of plants to deal with attack
by multiple herbivores. Studies on dual herbivore attack identified that defense against
one attacker may cause energetic and physiological constraints to deal with a second
attacker. How these constraints shape plant plasticity in defense to their full community
of attackers is a major knowledge gap in plant science. Our recent work identifies that
species richness of herbivores as well as their trait composition affect how plants deal with
simultaneous attack by multiple herbivores. Attack by a more species rich aphid
community result in stronger compromises of resistance to attack by a caterpillar. Leaf
chewing herbivores enhanced induced resistance to subsequent attack by a leaf chewer,
regardless of the number of species that were attacking the plant. When multiple
herbivores attack in sequence, the identity of earliest as well as most recent herbivore
attacking the plant determine how plants deal with current herbivore attack. Moreover,
by linking attacker patterns in the field and resistance to dual attack, we identified that
plant defense strategies may be adaptive to deal with more common patterns of herbivore
attack than to rare. This identifies that some of the physiological concepts developed by
studies on single and dual herbivore attack mismatch with the adaptive nature of plant
responses in community context. Identifying plant plasticity to deal with their full
community of attackers is key in understanding plant defense strategies and requires
unified terminology to illustrate the repertoire of plant plasticity matching ecological
patterns of attack.
28. “TRACK//GENE: a novel method of identifying induced and constitutive, insect resistance
and susceptibility genes by means of video tracking with EntoLab”
Maarten Jongsma, Business Unit Bioscience, Wageningen University and Research
Auxin Response Factors (ARFs) are the ultimate arbiters of the nuclear auxin signalling
pathway (NAP), since ARFs decide which genes are auxin response by virtue of their DNA-
binding. ARFs have a preference for TGTCNN motifs, consisting of a core, unchangeable
TGTC and two final, variable nucleotides. The ARF crystal structure revealed how a single
histidine residue, H146 in MpARF1, is the key for either high affinity motifs (TGTCGG) or
medium affinity motifs (TGTCTC). In this project, we aim to investigate how natural
variation for this histidine residue could affect DNA-binding and ultimately, the phenotype
of the plant. To that end, we use a complementation assay in Marchantia polymorpha, a
liverwort with a minimalistic NAP. We find that the mutations H146N and H146Y result in
partial complementation of the Mparf1-4 null mutant, that this is likely due to altered
DNA-binding affinity, and that lower DNA-binding affinity of MpARF1-H146N can be
compensated for by higher in vivo expression.
30. “Photosynthetic response to fluctuating light”
Anna Calabritto, Laboratory of Physics and Astronomy, Vrije University Amserdam
Photosynthesis allows plants to convert the energy of the sun into biomass. Crop yields
are subjected to fluctuations in irradiance that reduce photosynthetic light-use efficiency.
Our aim is to obtain an overview of short- and long-term physiological and developmental
responses of tomato and Arabidopsis to fluctuating light conditions.
Proteins encoded by the nuclear and organellar genomes interact to perform essential
biochemical processes such as respiration and photosynthesis. Genetic variation in this so-
called cyto-nuclear interactions can result in profound phenotypic differences. A well-
known phenotype associated with variation for cyto-nuclear interactions is cytoplasmic
male sterility (CMS). Most CMS systems are the result of transcription of a putative
mitochondrial gene (orf). Often the gene interferes with expression of genes encoding ATP
synthase subunits. Fertility can be restored via RNA editing, mediated by nuclear encoded
pentatricopeptide repeat proteins. CMS systems in different plant species are caused by
orfs that have a high degree of homology, but how such homologous orfs appear remains
largely unexplored. Recently, an Arabidopsis thaliana cybrid panel was constructed that
combined 60 species-representative organellar genomes with four divergent nuclear
genomes. Screening of this cybrid panel for sterility revealed a new CMS system induced
by the Staro-2 organellar genomes. Plants with the Staro-2 organellar genomes produce
viable pollen, but the pollen numbers are substantially reduced, likely due to a failure of
anther dehiscence. De novo assembly of the organellar genomes reveals a highly
rearranged mitochondrial genome, in which an insertion compared to Col-0 is found. The
insertion carries an orf predicted to encode a 225 amino acid sequence, referred to as
orf225Staro-2. Furthermore, we reveal a minimum of four Restorer of Fertility loci, that
only allow full restoration of fertility when homozygous alleles from both Col-0 and Staro-
2 are combined. The orf225Staro-2 shows a high degree of similarity with mitochondrial
orfs in B. napus and some other related Brassicaceae species, which suggests a common
ancestor. However, as a part of the chimeric orf225Staro-2 has a 100% similarity with the
Atp8 gene in A. thaliana, but seven SNPs compared to the orf in B. napus, we conclude
that orf225Staro-2 arose independently within A. thaliana. This suggests that CMS systems
can evolve independently, but result in orfs with high sequence homology.
32. “Understanding the role of insect herbivory on the balance between cross- and self-
pollination in Brassica rapa”
Hanneke Suijkerbuijk, Laboratory of Entomology, Wageningen University and Research
Individual plants can be associated with a very broad insect community; from antagonists
such as specialist and generalist herbivores, to mutualists such as natural enemies of pests
and pollinators. Apart from direct damage to flowers and seeds, herbivores affect plant
reproduction indirectly by changing plant-pollinator interactions or pollen acceptance
mechanisms. The aim of my research is to understand how herbivory affects the Brassica
rapa mating system through its effect on cross-pollination and self-incompatibility in
Brassica rapa.
To truly understand how herbivory affects plant reproduction, we need more insights in
the entire reproductive pathway: from pollen production, to pollen transfer by pollinators,
(self) pollen acceptance by the stigma and ultimately seed set. How herbivory affects the
full sum of female (seed formation) and male (pollen transfer) components of
reproduction is poorly understood.
So far, we have collected a large set of data on the pollination and fitness of herbivore-
treated and control plants of different genetic lines of Brassica rapa in common garden
field studies. For pollination, we looked at pollinator community composition, the number
of pollinators, the number of flowers visited per plant and the time spend by pollinators
on flowers and plants. The first look at the data indicates that for the attraction of
pollinators, the different genetic lines may respond differently to herbivory by chewing
herbivores (Pieris brassicae and Mamestra brassicae).
In a greenhouse pilot study we found an indication that herbivory may decrease the level
of self-incompatibility in a self-incompatible Brassicaceae (Sinapis arvensis). Plants were
individually enclosed in nets to ensure self-pollination and treated with larvae of the
mustard beetle (Phaedon cochleariae). On average, control plants produced fewer seeds
than herbivore treated plants (103 seeds and 172 seeds respectively). We are currently
investigating this further in a large greenhouse study on Brassica rapa, as well as with
microscopy and qPCR experiments.
33. “Remember or die: when plants face recurring heat stress events”
Priyanka Chopra, Department of Plant Sciences, Leiden University
Plants have an inherent ability to survive certain levels of heat stress (HS) called basal
thermotolerance. In nature, stresses are recurrent, and their intensity increases
occasionally. In addition to coping with a single event of HS, plants have also evolved the
ability to establish ‘thermomemory’ by retaining the experience from previous exposure
to HS. Establishing thermomemory helps plants to cope with recurrent and possibly more
escalated HS conditions. Thermomemory is an important molecular mechanism to ensure
optimal plant growth and survival after repetitive severe HS events. However, the
molecular mechanisms controlling plant thermomemory are still largely unknown. We aim
to unravel regulatory mechanisms and molecular components in establishing
thermomemory in different cellular compartments in Arabidopsis thaliana and crops such
as Solanum lycopersicum (tomato). Previous studies in our lab showed that sustained
thermomemory requires higher levels of HSP21 (a small plastidial heat shock protein). We
found that the plastid-localized metalloprotease FtsH6 regulates HSP21 abundance. Other
targets of FtsH6 (that might be potential novel memory components) are not yet known.
In addition to FtsH6, we found that autophagy (a conserved intracellular process involved
in protein degradation in eukaryotes) complements FtsH6 in degrading HSP21, thus
resetting the memory of heat stress (HS) in Arabidopsis. We are investigating 1) the
functional link between autophagy and FtsH6, 2) the proteins that are targeted and
degraded by them, and 3) their transcriptional regulators. Recent findings will be
presented.
34. “Effect of bacterial volatiles on A. thaliana and B. oleracea drought stress resilience”
Zulema Carracedo Lorenzo1,2, Marcel Dicke2, Karen Kloth2, Christa Testerink1, Rumyana
Karlova1
1 Wageningen University, Laboratory of Plant Physiology
Plant roots are in a close relationship with soil microorganisms, some of which can
promote plant growth and resilience to different (a)biotic stresses. Microbial volatile
organic compounds (VOCs) have been proven to play an important role in the interaction
between soil microorganisms and the host plant. In this project, we aim to understand
how three different Pseudomonas strains can increase the resilience to drought in
Arabidopsis thaliana and Brassica oleracea var. Rivera through their VOCs. Our results
show that in vitro, the Pseudomonas VOCs increased the resilience of Arabidopsis thaliana
to osmotic stress. RNA-seq analysis, revealed that the bacterial VOCs induced important
transcriptional changes in the plant root, that were more pronounced under drought
conditions compare to well-watered conditions. Additionally, an in vivo experiment in the
crop Brassica oleracea, shown that the Pseudomonas VOCs were also increasing plant
resilience to drought stress in non-sterile conditions. Altogether, our study contributes to
understand the mechanisms by which microbial VOCs increase plant resilience to drought
stress and suggests their potential to improve drought stress tolerance of crops
Plant pathogens are next to changing environmental and climatic conditions one of the
major threats to food security worldwide. A major aspect of plant breeding involves the
introgression of natural genetic resistances that have arisen from co-evolution of plants
with their respective pathogens. The process of identifying novel resistances and then
subsequently introgress them into elite cultivars is a time consuming and labor-intensive
process which in case of dominant resistance traits like NOD-like receptor genes (NLRs)
has only a limited durability in the field. Fast evolving pathogens like viruses can overcome
such resistances sometimes by a single amino-acid change in their effector proteins. One
example for this would be the interaction of the non-structural movement (NSm) protein
of the tomato spotted wilt virus (TSWV) with the coiled-coil NLR (CNL) Sw5b protein where
the amino-acid change Y118A causes breaking of the NLR mediated resistance. To
counteract the evolutionary adaption of pathogens we propose a pipeline on molecular
evolution via error prone PCR coupled with a high-throughput screening method for fast
discovery of gain of function mutations in domain regions corresponding to effector
recognition.
36. “How Plant Leaves Navigate the Canopy: Signals and Mechanisms”
Sanne Matton, Laboratory of Plant-Environment Signaling, Utrecht University
Plants are able to detect several different wavelengths of light, for example blue, red and
far-red . This enables them to adequately adjust their growth for optimal light capture and
thereby photosynthesis. Via their phytochrome photoreceptors plants are able to detect
a difference in red (R) and far-red light (FR) abundance. A low red to far-red light ratio (low
R:FR) is an indication of shade from a neighboring plants. Upon low R:FR perception A.
thaliana responds by, among other shade avoidance responses, shifting growth
investment towards increased petiole elongation and more upwards growth (hyponasty).
As shade within a canopy is almost never homogeneously distributed over the whole
plant, understanding responses towards local FR-enrichment is relevant to better
understand how plant leaves navigate the canopy.
Previous research has shown that the location of R:FR perception determines the
response. Local FR-enrichment on the leaf tip leads to hyponasty, and local FR-enrichment
on the petiole leads to petiole elongation. In this poster we describe the mechanisms
behind horizontal movement upon local FR-enrichment on one side of the leaf lamina
(FRside). The transcription factors PIF4, PIF5 and PIF7 and the auxin efflux carriers PIN3,
PIN4 and PIN7 have an important role in both FR induced vertical and horizontal leaf
movement. Horizontal petiole movement upon FRside can be inhibited by applying N-1-
naphthylphthalamic acid (NPA), a direct inhibitor of PIN activity, on the petiole-lamina
junction. Increase in auxin signaling visualized with the pDR5::GUS construct on one side
of the petiole upon FRside is absent in plants treated with NPA on the petiole-lamina
junction.
We recently discovered that plants under attack by fungal root pathogens can actively
recruit endophytic microbes inside their root tissues (endosphere) for protection. We
showed that Cupriavidus and Stenotrophomonas species were significantly enriched in the
root endosphere of sugar beet upon Rhizoctonia solani infection. In addition,
metagenomic analyses of the endosphere showed a high abundance of genes encoding
for phenazines, non-ribosomal peptide synthetases (NRPSs) and lanthipeptides associated
with these taxa, but their functional roles in plant colonisation and protection are yet
unknown. To characterise the taxonomic, genomic and functional traits of Cupriavidus and
Stenotrophomonas, we established a collection of 45 Cupriavidus and 310
Stenotrophomonas isolates from sugar beet root endosphere and selected five unique
isolates after dereplication based on BOX-PCR and 16S amplicon sequences. Genome
sequencing, in vitro and in vivo antagonistic activity and metabolomics experiments are
ongoing to investigate their functional potential and to resolve the role of the identified
BGCs in endophytic colonization and plant protection. We also analysed the genomic
sequences in MicroLIFE, a new bioinformatic pipeline to identify specific genomic features
associated with the endophytic lifestyle of bacteria. Our initial results from MicroLIFE
showed a large degree of genomic variation among the endophytic Cupriavidus isolates
and those isolated from other environments. In conclusion, our results and ongoing
experiments will shed light on microbiome assembly in the endosphere and which genes
and metabolites are expressed in plants under siege.
38. “Root flooding prepares the shoot for future low oxygen stress through systemic
signaling.”
Melissa Leeggangers, Laboratory of Plant Stress Resilience, Utrecht University
Flooding can negatively impact plant performance and development, and reduces crop
production in agricultural fields. Improving flood tolerance in plants is important for
limiting yield losses. Plants are impacted by flooding due to the slow diffusion of gases in
water which causes a reduction of oxygen availability. Limited gas diffusion also leads to
entrapment of the volatile hormone ethylene in flooded tissues. In nature, flooding events
start with water saturation of the soil (waterlogging). Thus, plant roots are the first to
experience low oxygen stress. When the rainfall persists, water levels rise and eventually,
the shoot is also submerged. It is unknown whether a root-derived signal can trigger a
survival mechanism in the shoot to protect the shoot meristem. Plants often use long-
distance signaling to survive suboptimal conditions. However, the underlying mechanisms
of a root-derived signal during a flooding event are uncharacterized. Our results
demonstrate that a period of waterlogging prior to hypoxia enhances shoot hypoxia
tolerance. Ethylene signaling and 1-aminocyclopropane-1-carboxylic acid (ACC) transport
are important components of the survival mechanism leading to this enhanced tolerance
of the shoot.
39. “A population derived from Solanum chacoense segregates for soft-rot resistance in a
detached petiole assay”
Elizabeth Yanez, Laboratory of Plant Breeding, Wageningen University and Research
Many efforts have been dedicated to potato to introgress resistance against soft rot
bacteria but success has been limited. One of the main obstacles has been the low
reproducibility of plant assays to identify resistant reliable phenotypes that can be
associated to genetic markers. In this study, we used a petiole test to characterize the
resistance to Pectobacterium parmenteri using a segregating population derived from a
Solanum chacoense accession showing resistance to petiole maceration. Due to the
population size and other constrains the experiment was conducted in two sub
experiments. Petiole tests were reproduced in assays independently performed. Disease
scores were integrated in one AUDPC value (area under the disease progress curve) for
each of the genotypes in each assay. To assess the agreement between result assays,
AUDPC values were adjusted to remove additional interference exerted by differences of
the disease pressure in the independent assays and a Lin’s concordance coefficient
correlation was calculated. To determine phenotype reproducibility the same susceptible
individual was used as common reference inside each of the assays to be compared to
allocate genotypes into a susceptibility scale. Result reproducibility using a petiole test
fluctuated between 7 to 49% and phenotype reproducibility between 54 to 78 %.
41. “Uncovering the genetic mechanisms of leaf senescence within a population of Lactuca
sativa”
Jelmer van Lieshout, Department of Plant Sciences, Leiden University
42. “The role of the C-terminal cytoplasmic tail of Cf proteins in determining the intensity of
the hypersensitive response”
Esranur Budak, Laboratory of Phytopathology, Wageningen University and Research
Plants only have an innate immune system to protect themselves against microbial
infection. The first layer of this immune system is mediated by extracellular plasma
membrane-associated receptors. These cell surface receptors perceive extracellular
immunogenic patterns and trigger the initiation of downstream defense signaling, which
finally leads to extracellularly-triggered immunity. Cf resistance proteins of tomato that
act against the fully extracellular pathogenic fungus Cladosporium fulvum are so-called
trans-membrane receptor-like proteins (RLPs) that localize at the cell surface. Cf proteins
require two co-receptors for the activation of downstream cellular responses, because of
the lack of a cytoplasmic kinase domain. In the resting state, Cf proteins constitutively
interact with the receptor-like kinase (RLK) SUPPRESSOR OF BIR1 (SOBIR1), whereas upon
recognition of the matching effector of C. fulvum by the Cf protein, the RLK BRI1-
ASSOCIATED KINASE (BAK1) is recruited. The overall structure of Cf proteins is typical for
LRR-RLPs and consists of an LRR ectodomain, an extracellular juxtamembrane domain
(eJM), a TM domain and a intracellular juxtamembrane (iJM) domain. This iJM domain, or
C-terminal tail, of Cf proteins is rich in basic amino acid residues, while the corresponding
juxtamembrane part of SOBIR1 has opposite charges. Possibly, these opposite charges
stabilize the interaction between the Cf protein and SOBIR1. Cf-4 and Cf-9 have identical
iJM domains, and Cf-5 and Cf-2 are also identical for this C-terminal tail. Interestingly, the
Cf-5/Avr5- and Cf-2/Avr2-triggered responses are slow and activate a less strong
hypersensitive response (HR) than the Cf-4/Avr4- and Cf-9/Avr9-triggered response. To
understand the role of the C-terminal tail of the Cf proteins in the intensity of the HR,
various domains of Cf-5 and Cf-9 were swapped and the chimeric proteins were checked
for the intensity of the HR that they trigger upon their activation. The results suggest that
the C-terminal tail of the Cf-proteins, and possibly all RLPs, has a specific role in
determining the intensity of the immune response.
Abstract- confidential
Increased demand and shortage of arable land necessitates growing crops such as Lettuce
in high density planting systems. Shaded plants in these high density planting systems
receive highly reflected light enriched in the far red (FR) part of the spectrum1,2. Shade
avoidance syndrome is a combination of responses to FR enriched light conditions under
shade, that include elongated hypocotyl, increased angle of leaves (hyponasty), lateral
root inhibition and primary root growth inhibition3. These light conditions can be
mimicked in the lab environment by treating plants with FR light in addition to white light.
As a part of the LettuceKnow consortium, this project will focus on understanding the
effect of WL + FR light conditions on domesticated Lettuce (L. sativa) shoot and root
architecture through both top-down (orthologs) and bottom-up (GWAS and Time series
RNA-Seq) approaches.
Plants produce a plethora of specialized metabolites (i.e. natural products), many of which
have great potential for human health applications. Montbretin A (MbA), found in the
ornamental plant montbretia (Crocosmia x crocosmiiflora), is such a promising metabolite,
as it can serve as an improved therapeutic for the treatment of type 2 diabetes. To date,
MbA, a complex acylated flavonol glycoside, has only been found in small quantities in the
underground storage and reproductive organs (the corms) of montbretia. The chemical
complexity of the metabolite and the low quantities found within the plant make scalable
production through chemical synthesis or conventional agriculture, respectively, not
feasible and unsustainable. Hence, we are exploring ways to enable large-scale MbA
production for drug development and application.
Plant tissue culture technologies offer a promising strategy to obtain specialized
metabolites in greater quantities from their native sources. However, little attention has
been given to the development of a tissue culture system for montbretia thus far, and it
remains unclear whether in vitro-grown tissues would produce MbA. Furthermore, the
cellular and subcellular localization of MbA production and storage within the corm is
unknown but might provide valuable insights for the establishment of a successful tissue
culture production system.
As part of my thesis work, I am setting up an in vitro montbretia tissue culture system,
using state-of-the-art plant tissue culture techniques. By tailoring available knowledge of
tissue culture to montbretia, I have successfully grown plants from seeds using optimized
sterilization protocols. Additionally, the induction of callogenesis and/or organogenesis in
various explant types was achieved by supplementing the culture medium with growth-
promoting phytohormones. In my future research, I plan to explore the effects of stress-
related phytohormones and study MbA production in vitro. To elucidate the storage
location of MbA within the corm or MbA-producing tissues, my objective is to utilize and
adapt state-of-the-art imaging technologies, such as confocal microscopy and desorption
electrospray ionization mass spectrometry (DESI-MS). My work aims to advance our
understanding of specialized metabolite biosynthesis in specialized plant organs, to
contribute to the establishment of a scalable plant-based MbA production system.
Oxygen levels in plants vary depending on environment conditions, tissue morphology and
metabolic activity. Indeed, meristems experience hypoxia as a chronic condition while
normally well oxygenated tissues may experience acute hypoxia as a consequence of
either abiotic or biotic stress. The signaling cascade initiated by the impoverished supply
of oxygen was shown to be implicated in development, stress response, defense activation
and gall formation. Current tools to monitor changes in oxygen levels and its 3D-
distribution in tissue have clear limitations. Invasive electrode-based oxygen sensors are
unsuitable for microscopic and hard tissues, and oxygen-sensitive dyes are not well
absorbed by plant tissue. To bridge this gap we engineered a set of biosensors that detect
oxygen levels without wounding or altering the plant. Here we present a state-of-the-art
toolbox of ratiometric, genetically encoded oxygen biosensors that may be employed to
investigate oxygen dynamics and signaling at cellular resolution.
Fast and unpredictable climate changes, low rainfall and high land evapotranspiration
rates are leading to high soil salinization. Plants respond to salt stress very fast, starting at
the molecular/gene expression, to metabolome change and post-transcriptional
regulation, ultimately leading to undesirable reduced growth and yield losses. Current
crop varieties are not adapted to the high soil salinity and the demand to develop salt-
tolerant varieties is high. To discover new potential solutions aimed at reducing the
negative impact of soil salinization on plants and their fruits, it is first necessary to
understand the biomolecular mechanisms involved in the plant responses to salt stress. In
this study, we focused on an economically important crop tomato. We combined
transcriptomic and metabolomic approaches to pinpoint potential markers in molecular
pathways regulated by salt stress. Clear metabolome and transcriptome-wide effects of
salt stress in tomato roots were observed. We identified metabolite- guanosine as one of
the molecules involved in salt responses and verified its role in root architecture changes
upon exogenous application only under salt stress. Upon correlation of the omics datasets,
we identified a potential marker, WRKY80 to regulate salt stress responses. Finally, we
confirmed the role of WRKY80 in salt stress resilience using an ectopic expression system
in two different species, tomato and Arabidopsis.
48. “Biological control in a circular agriculture: Improving crop growth and resistance using a
residual stream from insect production”
Els van de Zande, Laboratory of Entomology, Wageningen University and Research
The production of insects as food and feed is increasing rapidly. Therefore, also the
resulting residual streams, exuviae and faeces, are increasing. To create a more circular
food production system, these residual streams can be used as soil amendment for
agricultural crops. Besides adding extra nutrients, the side-streams may also induce
defences in the crops and thereby help in crop protection against pest insects. In this study
we tested whether soil amendment with mealworm exuviae affects the rhizosphere
bacterial community, plant growth, herbivore performance and parasitoid recruitment in
the field.
49. “Thrips tabaci: the cryptic and dominant thrips species in Allium hosts”
Bettina Porta, Laboratory of Plant Breeding, Wageningen University and Research
Thrips are small insects that encompass around 6000 species, some of which are serious
pests. Thrips tabaci is reported as the most harmful in onion cultivation, but other species
are found as well. Thrips tabaci is a cryptic species with two different phylogenetic groups
and reproductive modes; arrhenotokous (sexual and parthenogenetic) and thelytokous
(only parthenogenetic). This research investigated; 1) which thrips species affect onion
and related Allium species, 2) the genetic variation present within and between 14
locations worldwide and over time, and 3) the reproductive modes present in the Thrips
tabaci populations. Thrips samples, consisting of 33 individuals, were obtained from 14
locations of which three locations were sampled two or three times during the season.
Species and haplotypes were determined through DNA barcoding of the COI gene. The
reproductive mode of T. tabaci haplotypes was detected using COI specific primers. A
Neighbour Joining tree was built using Nei’s genetic distance. As expected, T. tabaci was
the main species (93%). In addition, we also found Scirtothrips dorsalis, Thrips palmi,
Frankliniella intonsa, F. occidentalis and F. tenuicornis on specific locations. Sites sampled
multiple times showed that at the end of the season only T. tabaci was present, while
other species were present early in the crop season. The haplotypes of T. tabaci grouped
into two phylogenetic groups each of which linked to the arrhenotokous and telytokous
mode of reproduction respectively. Arrhenotokous T. tabaci were present at certain
locations while thelytokous T. tabaci are globally distributed. Genetic diversity was
observed between and within T. tabaci populations, except in Mexico where only one
haplotype was found. That haplotype was found in 10 of the 14 locations. Some other
haplotypes also showed worldwide distribution. Gene flow by global trading of onions or
garlic may be the cause of the worldwide distribution of some T. tabaci haplotypes.
The southern green shield bug Nezara viridula is an invasive piercing and sucking pest
insect, feeding on important crop plants and posing a threat to worldwide food
production. Since insects are known to harbor unique support by microorganisms, our
study aims at providing insights into N. viridula-associated microbiota and their effect on
host plant defenses. We determined the core bacterial microbiota with 16S rRNA gene
amplicon sequencing throughout different development stages and distinguishing gut
systems from salivary glands in adult animals. Our results confirms that part of N. viridula
microbiota is vertically transmitted, changes throughout insect development and includes
a core set of five abundant microbial genera, including Sodalis and Pantoea symbionts. In
contrast to prior studies, we confirmed that N. vriridula transmits microbiota during
feeding, suggesting that microbes may play a role in interactions between N. viridula and
its host plant. We tested the effect of isolated N. viridula microbiota (including yeast) on
Arabidopsis thaliana plant defense gene expression via qPCR and found that Sodalis and
Pantoea were able to repress plant defenses directed towards insects. Our findings
demonstrate that insect-associated microbiota play an important role in interactions
between insects and plants, and could therefore be considered a valuable target for the
development of sustainable pest control strategies in the future.
51. “Mapping out the effect of ethylene on the root tip during hypoxic conditions”
Gyöngyi Macias Honti, Laboratory of Plant Stress Resilience, Utrecht University
The mechanisms behind meristem tolerance to stress conditions have yet remained
underexplored. Previous experiments have shown that, in Arabidopsis, an ethylene pre-
treatment following subsequent hypoxic stress can enhance root survival. While it is clear
that the activating mechanism caused by ethylene accumulation boosts the regrowth
capacity of roots after hypoxia, it is yet unknown how meristem protection is conferred.
Is ethylene mediated by specific cell layers in the root? Do specific cell layers communicate
signals in order to coordinate meristem protection? In order to address these questions
different lines expressing EBF2 in a cell-type specific manner are used to disrupt ethylene
and elucidate on what cells are more important for root survival after a hypoxia treatment.
Somatic embryogenesis (SE) is a form of induced plant cell totipotency where embryos
develop from vegetative cells without fertilization. SE can be induced in vitro by ectopic
expression of embryo identity transcription factors like the BABY BOOM (BBM)
AINTEGUMENTA-LIKE AP2/ERF domain protein. We found that genes in the indole-3-
acetaldoxime (IAOx) metabolic pathway (PAD3, FOX1, CYP82C2) are transcriptionally
down-regulated by BBM at a very early timepoint during 35S:BBM-mediated somatic
embryogenesis. IAOx is an intermediate in tryptophan-dependent auxin (IAA) and defense
compound biosynthesis.
Given the role of auxin in promoting BBM-induced cell proliferation, we hypothesized
that, similar to superroot mutants, down regulation of IAOx pathway gene expression by
BBM would enhance IAA biosynthesis to promote somatic embryogenesis. However, the
pad3, fox1 and cyp82c2 mutants reduced 35S:BBM somatic embryo formation in favor of
increased ectopic shoot formation. Moreover, exogenous IAOx application to 35S:BBM
pad3 and 35S:BBM fox1 lines partly complemented the reduced somatic embryo
formation phenotype. Assuming that down-regulation of IAOx pathway genes is positively
related to BBM-induced SE, our results suggest that 1) BBM-mediated down-regulation of
IAOx-pathway genes does not increase the flux of IAOx to IAA and 2) that down-regulation
of IAOx-pathway gene expression is tightly controlled temporally and/or spatially.
53. “Maroon rice diversity preserves a genomic record of 350 years of colonial history”
Marieke S. van de Loosdrecht1, Nicholaas M. Pinas1,2, Frank F.M. Becker3, Robin van
Velzen1, Harro Maat4, Tinde van Andel1,2,5*, M. Eric Schranz1*
The Maroons in Suriname and French Guiana descend from enslaved Africans that
escaped the plantations during colonial times. In Maroon farming, women have
continued the maintenance of a large diversity of rice landraces, their oldest staple crop.
Landraces are cultivated crops that have been locally adapted by farmers. Genomic
analyses of the Maroon rice landraces may hence clarify from where certain rice varieties
were introduced, and as such be a powerful tool in reconstructing historical events in the
Maroon past.
Here, we sequenced the genomes of 144 Maroon rice landraces to investigate their
geographical origins and the historical contexts associated with their introduction to the
Guianas. Using various population genomic methods based on genome-wide SNPs, we
show that a subset of the Maroon rice diversity is closely related to some landraces in
West Africa and hence may trace back to the trans-Atlantic slave trade (1660-1825). Other
subsets of rice diversity can be linked to interactions with diverse ethnic-cultural groups
that post-date the era of slavery. These include landraces with long stiff awns brought by
indentured laborers from Java (1890-1930), the USA early cultivar Rexoro from rice
breeders in Louisiana (1932 onwards), and a more recent cultivar from Hmong refugees
from Laos who fled the Vietnamese War (1977 onwards). All in all, our results underline
that the Maroon’s past is dynamically related to a global history of slavery and
colonization. Moreover, the inclusive selection methods of Maroon farmers make their
rice variety collections unique cultural heritage. Maroon farmers have been preserving
their community’s past, and today are important custodians of global rice diversity.
55. “Phloem Fight: exploring wild tomato phloem to identify whitefly resistance”
Lissy Denkers, Laboratory of Plant Physiology, University of Amsterdam
Azolla is an aquatic fern able to triple its biomass every week while relying solely on
atmospheric dinitrogen gas thanks to its cyanobacterial endosymbiont Nostoc azollae.
Combined with its high protein content of ∼20% it has potential to serve as a novel plant
based protein crop in delta regions by harvesting excess nutrients of flooded agricultural
lands. The limiting step for protein extraction is the accumulation of polyphenolic
secondary metabolites, such as proanthocyanidins, binding to proteins and interfering
with digestion. Severe stress also triggers 3-deoxyanthocyanin accumulation resulting in
the characteristic red Azolla mats. Whilst these molecules mainly have been ascribed
defensive roles, they also seem to be involved in symbiotic crosstalk.
Nostoc fixes dinitrogen gas inside the leaf pocket: a specialized organ present in each leaf.
A permanent Nostoc colony, however, resides at the shoot apex, serving as inoculum for
each new leaf developing. If the fern undergoes sexual reproduction, Nostoc also
colonizes the developing spores for transfer to the next generation. A structure
reoccurring at locations where Nostoc is present are trichomes. We therefore think these
trichomes are crucial to regulate the synchronized development of both organisms by
secreting signaling molecules.
Cornicinine, a glycosylated triketide delta lactone isolated from the crane fly Nephrotoma
cornicina, distorts the Azolla-Nostoc symbiosis. Already at nanomolar concentrations, it
turns all Nostoc cells into resting stages called akinetes resulting in chlorotic ferns, most
probably because of nitrogen starvation. An apparent effect on sterile Arabidopsis
thaliana or free-living filamentous cyanobacteria could not be seen. Why the crane fly
accumulates cornicinine, and if it is directly related to Azolla, remains a mystery.
Weakening plants by switching off symbioses could, however, be a powerful tool to have
for grazers.
57. “HeatGenes: towards a generic genetic framework for reproductive heat tolerance in
plants”
Max Frencken, Laboratory of Plant Systems Physiology, Radboud University Nijmegen
AGC kinases are very conserved kinases in eukaryotes. PDK1 is one of the AGC kinases and
considered as a master regulator of other AGC kinases. There are 39 AGC kinases in
Arabidopsis thaliana, including two PDK1 genes, we named them PDK1 and PDK2. The
pdk1 pdk2 double mutant shows short and twisted pollen tubes and precocious pollen
germination in anther. We found another AGC kinase, AGC1.5, can rescue these pollen
phenotypes of pdk1 pdk2.
The global issue of food shortages, exacerbated by population growth, has stimulated
scientific efforts to increase crop production. In pursuit of higher crop yields, farmers often
sow more crops in a limited area, leading to inevitable competition for limited resources,
including light. Many plants respond to this competition by growing taller to access
sufficient light and avoid shading, a phenomenon known as the shade avoidance response.
Our research focuses on commercial tomato cultivars M82 and Moneymaker, aiming to
investigate similarities and differences in their mechanisms-shade avoidance responses.
To explore this phenomenon, we began by measuring tomato growth under control white
light normal and far-red supplemented conditions. Using microscopy and statistical
analyses, we observed a significant elongation of the first internode, where pith cells had
generated additional layers and increased in length. Subsequently, we harvested
internode and pith cell samples from both treatments and conducted tissue-specific RNA-
seq experiments.
Our RNA sequencing results revealed a role for the auxin signaling in the internode
response to far-red. We proceeded to perform a range of detailed physiological
experiments, including, IAA application, and IAA inhibitor application. We also analyzed
DR5:GUS expression pattern. These experiments suggested a complex model of shade
avoidance response signaling in internode elongation, highlighting the intricate
involvement of the auxin signaling pathway.
Using genomic selection I aim to show that targeting selection on rapid photosynthetic
protection mechanism (non-photochemical quenching(NPQ)) rates, will increase
photosynthesis in a magic population of Arabidopsis thaliana. The poster shows how I
intend to do this and why this is important to do. Come by to have a chat!
61. “PHYC mutations decelerated the circadian clock in cultivated lettuce while breeding for
delayed flowering time.”
Cèlia Anton Sales, Laboratory of Plant Breeding, Wageningen University and Research
Although the use of synthetic pesticides helps to maintain crop yields, their use has
detrimental effects on ecosystems and human health and led to pathogen resistance.
Natural products are an excellent alternative to such pesticides, provided they are
environmentally friendly, economical, targeted and biodegradable and their use does not
significantly affect crop yields compared to conventional pesticides. Several studies have
reported that essential oils (EOs) have a broad spectrum of activity against plant
pathogens, including oomycetes and fungi. But before EOs can be used as pesticides, many
fundamental and practical questions need to be answered. Our interdisciplinary research
team, that consists of biologists and chemists, focusses mainly on two questions: I) is the
effect of EOs direct, meaning the effect is either fungistatic/fungicidal, or is it indirect,
meaning EOs have an effect on the plant immune system, thus “priming” the plant for
future microbial attack, and II) can we overcome the disadvantages of using EOs in the
field, like low rain fastness, volatility and degradation, through nanoencapsulation of the
substance. With the expectation that the results can be extrapolated to other crops, a case
study in viticulture (Grapevine-downy mildew) was chosen for this short-term project.
63. “Morphological and Cytological Analyses for Leaf Curving in Cabbage Development”
Zihan Liu, Laboratory of Plant Breeding, Wageningen University and Research
Leafy heads are representing the harvested organs of cabbage (Brassica oleracea), formed
by the wrapping and overlapping of leaves. Cabbage vegetative growth goes through
seedling, rosette, folding and heading stages. The expanded leaves of each of these stages
are characterized by morphological differences. The leaf blade of seedling and rosette
leaves are flat while the leaf blade of folding and heading leaves are in/upward curving.
We are interested in the genetics of this typical domestication trait and set out to analyse
leaf development. Besides leaf curvature, essential to form a leafy head, leaf shape is also
associated with leafy head shape (pointed, flat, round). In this study, we assess leaf tissue
growth between flat rosette and curved heading leaves both at whole leaf and cytological
levels. We compared cellular organization focusing on adaxial palisade cells and abaxial
spongy parenchyma cells at different positions of the leaf. The results demonstrated two
trends: a) a higher growth rate of the leaf centre compared to the marginal leaf tissues
leads to a bowl-like leaf and b) a higher growth rate of the leaf abaxial compared to the
leaf adaxial side leads to an inward curving leaf of heading leaves. Besides, we noticed
that leaf shape contributes to leafy head shape in addition to the specific leaf curvature.
Overall, differential growth between the leaf ad/abaxial and central/marginal parts shape
the heading leaves resulting in leafy head formation.
64. “The potential of volatile organic compounds (VOCs) from bacterial seed endophytes on
Arabidopsis thaliana seed germination and seedling growth under abiotic stresses”
Sasiwimon Siricharoen, Laboratory of Plant Physiology, Wageningen University and
Research
Seeds harbor various microorganisms within their tissues without causing any harm, the
so-called seed endophytes. The seed endophytes have been shown to influence plant
performance at the early developmental stage, including seed germination and seedling
establishment, by producing volatile organic compounds (VOCs). We investigated the
effect of potential VOCs from a bacterial seed endophyte isolated from wild cabbage seeds
(E44) on seed germination and seedling growth of Arabidopsis thaliana under salinity and
osmotic stresses We found that seeds sown under salinity (125mM) or osmotic stress
(300mM mannitol) showed higher and faster germination when sown in the presence of
E44. Similarly, we observed that Arabidopsis seedlings showed a higher growth (fresh
weight and root length) under 75mM NaCl or 150mM sorbitol in the presence of E44. In
our setup the E44 endophytic bacteria is cultivated in a separate plate than the seed and
seedling, indicating that their VOCs are potentially stimulating germination and growth.
We are now investigating if different Arabidopsis ecotypes respond differently to E44, and
in the future we will identify what specific VOCs emitted by E44 are stimulating
Arabidopsis seed germination and seedling growth under abiotic stresses.
65. “R gene-induced selection pressure affects the prevalence of the virulence effector RBP-1
of Globodera pallid”
Vera Putker, Laboratory of Nematology, Wageningen University and Research
The potato cyst nematode (PCN) is considered a dominant threat to yield for major food
crops such as potato and tomato. Current control strategies mainly depend on crop
rotation and the use of resistant cultivars. In potato the single dominant resistance (R)
gene Gpa2 activates effective host-specific resistance to the PCN Globodera pallida
population D383. More specifically, the matching effector gene eliciting Gpa2-mediated
immune responses is the effector gene RBP-1. However, resistance breaking populations
in the field such as Rookmaker harbour specific RBP-1 alleles evading resistance. It is
however unknown how these alleles behave in natural populations under selection. Here
we aim to understand the genetic dynamics of RBP-1 during infection under selection
pressure of an R gene.
We monitored nematode development during 21 days in avirulent (D383) and virulent
(Rookmaker) G. pallida populations in potato plants with or without the R gene Gpa2. In
these in vitro infection assays, after 21 days, Rookmaker juveniles developed into females
in both potato lines. D383 juveniles however were not able to develop into females in the
resistant potato line, and a strong plant immune response was visible, while in the
susceptible potato line D383 was able to develop into females. In parallel, we conducted
the same experiment but harvested infected root segments at 6 dpi for RNA isolation and
transcriptomics. In total, we revealed 326 differentially expressed genes (DEGs) upon
nematode infection, of which 13 DEGs were specifically regulated in Rookmaker infection
on the resistant potato line. Amongst them, we found that in D383 RBP-1 is upregulated
compared to Rookmaker, irrespective of the presence of the R gene Gpa2.
Our data show the strong selection pressure exerted by Gpa2, as well as differences in
expression between avirulent and virulent populations. We are currently using two long-
read reference genomes of both D383 and Rookmaker to investigate the genomic
organisation of RBP-1. We will also map the allelic variation of RBP-1 before infection and
in young females during infection to reveal the effect of the Gpa2 selection pressure on
RBP-1.