Embed Size (px)
Citation preview
Plexor multiplex qPCR tool as a way to quantify esca-associated
fungal agents in grapevine wood.
Plexor multiplex qPCR tool as a way to quantify esca-associated
fungal agents in grapevine wood.
Photo: Nancy Wenner, Penn State
Alban JACQUES, INP-EI PURPAN, Toulouse Lisbon, 2014, 20th February
ESCA Without symptoms Chronic symptoms Apoplectic symptoms
a.k.a. « tiger stripes »
5-10 yrs
Death occurs within days
Introduction: esca
Epidemiology of P.ch and P.al
Penetration Pruning wounds Roots
Sporulation All the year or only in vegetative period Perithecia located in crevices
Inoculum sources Air Soil Plant material grafted in nurseries Insects
Introduction: esca
Rolshausen et al. (2010); Whiting et al. (2006); Larignon et al. (1997)
Goals
Development of tools to understand plant microbes interactions in an early model of esca
Diagnosis development
Introduction
Goals
Development of tools to understand plant microbes interactions in an early model of esca
Introduction
Development of a molecular tool to detect and quantify esca-associated fungal agents
Introduction: available tools
In vitro plate screening PCR (qPCR ?)
Speed - +
False negative + (competition) +
False positive - +
Exhaustive + -
Identification complex Known before
Sensitivity -/+ +
Our developped tool
qPCR multiplex
Quantitative Multi-fungal Specific
Chemistry ? Primers ? Test ? Purposes we developped
Plexor chemistry
Plexor chemistry
Plexor chemistry
i.e. P.al
Plexor chemistry
i.e. P.al
Plexor chemistry
i.e. P.al
copies
Plexor chemistry
i.e. P.al
copies
Plexor chemistry
Primers
Targeted gene
ITS ? rDNA ? Encoding gene ?
b-tubulin
Primers
Targeted species
Species for specificty tests
Mostert et al (2006) Taxonomy and Pathology of Togninia (Diaporthales) and its Phaeoacremonium Anamorphs. Studies in mycology 56
Ex. Phaeoacremonium aleophilum
Phylogenetic studies
Primers
b-tubulin
Test
In lab infected plants Nursery samples
Test
In lab infected plants
Test
Nursery samples
TAMRA P.ch
ROX P.al
FAM VvActin
Test
Nursery samples
Test
Nursery samples
Test
Nursery samples
RBE are more contaminated than GU
More contamination by P.al than P.ch
Fungal pressure is higher in GU
In few words
Purposes
Conclusion
Plexor multiplex qPCR esca pathosystem Plexor multiplex qPCR BDA pathosystem (unpublished) Plexor qPCR Eutypa dieback In lab model to test early efficiency of putative solutions Tool to measure defence efficiency of grapevine
Conclusion
Team
R.Pierron J.Pouzoulet N.Mailhac C.Couderc F.Violleau M.Gaucher
Thanks for your attention!!!
