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Theme: Crop Improvement
Towards broad, stable and durable resistance against
Tospoviruses and Begomoviruses in Solanum Lycopersicon
BBE Workshop Brazil-NL programme 5-6 April 2017
University of Brasilia
Let me introduce
University of Brasilia
Renato O. Resende
Richard Kormelink
The team and collaborating partners
University of Brasilia
Embrapa Hortaliças
Wageningen University:Laboratory of VirologyDr. ir. Richard KormelinkProf. dr. Monique van Oersing. Dick LohuisLaboratory of Plant BreedingDr. Yuling BaiUniversity of Brasilia:Dr. Renato O. Resende
University of Goias-Goiania:Erico de Campos DianeseEMBRAPA-Virology:Dr. Alice Inoue-NagataMirtes Freitas Lima Simone da Graça Ribeiro EMBRAPA-Plant Breeding:Dr. Leonardo BoiteuxEMBRAPA-Plant Physiology:Maria Esther N. Fonseca
Students in the project:Ms. Corien Voorburg (Nl) PhD studentMr. Athos S. de Oliveira (Br) PhD studentMs Mariana Almeida (Br) PhD studentMs. Geane Fontes (Br) PhD student
Students associated to the project:Ms. Mikhail Leastro (Br) PhD student
Our shared interest: viruses
University of Brasilia
~20 plant viruses cause annual crop-losses of > $ 20 billion (Rybicki , 2015)
Cassava
Rice
Grapevine
Maize
Cotton
Tulip
Almost half (47%) of all emerging infectious diseases are caused by plant viruses. (Anderson et al., 2004): be on constant alert.
The inspirers.
Food – Vegetable Crops
University of Brasilia
Towards broad, stable and durable resistance against plant viruses
Our focus: Tospoviruses & Geminiviruses
University of Brasilia
TSWV
Geminivirus (DNA) Plant-infecting
bunyaviruses (RNA)
Top 10 most important plant viruses worldwide (scientific-economic)
1. Tobacco mosaic tobamovirus (TMV)
2. Tomato spotted wilt virus (TSWV; tospovirus)3. Tomato yellow leaf curl virus (TYLCV; geminivirus) 4. Cucumber mosaic cucumovirus (CMV)5. Potato virus Y (potyvirus, PVY)6. Cauliflower mosaic caulimovirus (CaMV)7. African cassava mosaic begomovirus (ACMV) 8. Plum pox potyvirus (PPV)9. Brome mosaic bromovirus (BMV)10. Potato virus X (potexvirus, PVX)
Scholthof et al. (2011)
Both viruses/related members are most
prevalent to the warmer ((sub)tropical)
climate regions
Tospoviruses and Geminiviruses in
Solanum Lycopersicon
University of Brasilia
Tospoviruses in Capsicum
University of Brasilia
Research focus of the collaborating PIs
University of Brasilia
• Tospovirus and geminivirus diversity• Mechanisms involved in tospoviruses and geminiviruses / host
interactions with focus on horticultural crops• Molecular breeding platform for marker assisted selection to broad
spectrum resistance against geminivirus and tospovirus species • Development and release of multi-resistant cultivars/hybrids
• Unravelling the mechanisms of host defence against tospoviruses and geminiviruses based on RNA silencing and resistance genes (PAMP-resp. Effector-Triggered Immunity)
• Modes of viral counterdefence against RNAi or R-gene-mediated resistance
Brasilia
Wageningen
Expertise
University of Brasilia
Virus Taxonomy & Evolution
(1992)
Thrips vector species
F. occidentalisF. fuscaF. schultzeiF. intonsaF. bispinosaF. cephalicaF. geminaT. setosusC. claratrisT. palmi S. dorsalisT. tabaciD. betaeN. variabilisF. zucchini
Most tospoviruses and thrips
vector species have a restricted
geographical distribution with
the exception of TSWV.
Many of them do not occur in
The Netherlands (quarantaine).
Expertise
University of Brasilia
Development of Diagnostics
Molecular (RT-PCR)
Hassani-Mehraban et al. (2016), J. Virol. Meth.
Serological (polyclonal antisera production)
De Avila et al. (1993), J. Gen.Virol.
Expertise
University of Brasilia
Tools (gene-constructs, protocols, assays, systems, plant resources) to support
studies onVirus resistance and virus-host
interactions
Fundamental & Applied
Objective
University of Brasilia
Resistance genes available for commercial resistance breeding
TSWV (tospoviruses):Sw5-Solanum LycopersiconTsw- Capsicum chinense
TYLCV (begomo- /geminiviruses):Ty-1 to Ty-6 from Solanum chilense, Solanum habrochaites, Solanumperuvianum
• Resistance mechanism is unknown• Resistance spectrum is (partially) unknown or very restricted• Resistance breaking isolates emerge• For many Tospoviruses – Begomo-Geminiviruses resistance genes are lacking• Emergence of new tospoviruses & begomoviruses (recombination and reassortant)• Many of these viruses and their pest insect vectors are listed as quarantine agents• Climate change enhances the risk of future introduction into the more temperate
climate regions (Western Europe)
Constant threat of these viruses, incl. new emerging strains and emerging
resistance breaking strains, urges the need to investigate and exploit current
resistance genes, as well as search for new ones to support development of
broad, stable and durable resistance
NWO-CNPq project
University of Brasilia
Advantages (mutual benefits):
• Financial support for (continuing) joint research
• Rapid exchange of expertise, knowledge, tools and biological resources
• Implementation of research findings into breeding programs (EMBRAPA, Brazil)
• Support the exhange and training of brazilian research students in Wageningen
(Athos de Oliveira, Mariana Almeida and Geane Fontes)
Highlights-Achievements
University of Brasilia
Resistance to geminiviruses (TYLCV, begomovirus):• Showed that the Ty-1 gene confers resistance by enhancing transcriptional gene silencing of the viral
DNA genome• Identified an Achilles’ heel of the Ty-1 resistance gene (mixed infection with the CMV RNA virus
compromises Ty-1 resistance)• Demonstrated that Ty-1 also confers resistance to a bipartite begomovirus• Partial elucidation of the resistance mechanisms of Tcm-1 gene to begomoviruses
Resistance to tospoviruses (TSWV and GRSV):• Identified the TSWV NSm (movement) protein as trigger for the Sw5 resistance gene in tomato• Identified the roles of the respective domains of the Sw-5 resistance gene (SD-CC-NBS-LRR) in
triggering the hypersensitive response • Mapped the dysfunctionality of Sw-5 homologs in susceptible tomato• Showed that Sw5 confers resistance against TSWV, GRSV, TCSV, CSNV, INSV, ANSV (broad spectrum)• Display of transcriptome analyses of compatible and resistant (conferred by Sw-5 gene) tomato
genotypes challenged by GRSV • Provided the transcriptome of Rey de Los Tempranos (showed broad field resistance to tospoviruses)• Partial elucidation of the genetic control of Rey de Los Tempranos resistance genes
Ongoing
University of Brasilia
Resistance to geminiviruses (TYLCV, begomovirus):
• Comparitive transcriptome analysis of susceptible and resistant tomatoes
• Small RNA profiling in Ty-1 tomato lines
• Analysis of co-replicating satellites on the durability of Ty-1 resistance against TYLCV
• Resistance spectrum analysis of Ty-1 against curtuviruses (another geminivirus genus)
• Functional/cell biological analysis of the Ty-1 protein and homologs from Arabidopsis
thaliana
Resistance to tospoviruses (TSWV):
• Screening for new tospovirus resistance genes in tomato and sweet pepper germplasm
(cultivated and wild accessions)
• Screening for Sw-5 analogs in solanaceous vegetables (Solanum and Capsicum species)
pyramiding resistance genes to provide broad and durable against tospoviruses
• Identification of TSWV-host protein interactions (new targets)
Output (I)
University of Brasilia
• Hallwass, M., De Olivera, A et al. (2014), Molecular Plant Pathology 15,
871–880.
• Almeida, M. et al. (2014), Plant Disease 98, 1285.
• Butterbach et al. (2014), Proceedings of the National Academy of
Sciences USA vol. 111, 12942–12947,
• Leastro et al. (2015), Virology 478, 39–49.
• Pereira-Carvalho et al. (2015), Viruses 7, 2518-2533.
• De Oliveira et al. (2016), Molecular Plant Pathology 17, 1442–1454
• Turina et al. (2016), Annual Review of Phytopathology 54, 347-371.
• Leastro et al. (2017), Virus Research 227, 57-68.
• Leastro, M., De Oliveira, A. et al. (2017), in preparation
Output (II)
University of Brasilia
Co-Tutelle – Double Degree: (UnB – WUR)1. Athos Silva de Oliveira (2015). The Sw-5 gene cluster: Unravelling the keys to tomato resistance against tospoviruses.
PhD Sandwich: (UnB – WUR)2. Mariana Martins Severo de Almeida (2016).Study of the Evolutionary Capacity of Tospovirus via Genetic Rearrangement and Its Implication in Viral Epidemiology, PhD Program in Molecular Biology
PhD Thesis - UnB3. Maria Geane Fontes (2017). Transcriptome of the response to Groundnut ringspot virus mediated by the Sw-5 gene and studies of the genetics and mechanisms of resistance to Tomato spotted wilt virus in the cultivar 'King of Los Tempranos‘, PhD Program in Phytopathology.
Related PhD Sandwich4. Mikhail Oliveira Leastro (2015). Functional Study of the Movement Protein (NSm) ofdifferent Tospovirus, PhD Program in Molecular Biology
Benefits for breeders-growers
University of Brasilia
• Knowlegde on resistance spectrum (screening for Ty-1 homologs in other crops)
• Knowlegde on resistance spectrum (screening for Sw-5 homologs in other crops)
• (A)Biotic stress factors that compromise resistance (mixed infections; reassortants)
• Implications for disease management practices
Continuation!
Project Contributions
University of Brasilia
The CNPq / NWO was based on long term research cooperation (Brazil – The Netherlands) and expertise in the virology field
Long history on training and developing human resources:• 10 Full PhD• 5 Sandwich PhDs• 4 Postdocs
The project meets the CNPq and CAPES policy – “Internationalization of Brazilian Science”
The project Significantly improved the Quality of Scientific Publications:• 9 International Papers in High Impact Factor Journals related to the project
Human resources training:• 4 PhDs students related to the project• First Double Degree agreement between UnB / WUR
Scientific Achievements – answering basic and applied questions
Displayed new perspectives and challenges for research lines and cooperation:• Ongoing research activities in tospoviruses and geminivirues• New Joint Projects including new partners in South America – (e.g. Universidad
Nacional de La Plata)• New Joint Project – Arboviruses (Zika)/Vectors - (UnB-WUR)
