Fabienne Moreau ADNid company Montpellier, France Objective of MARS - - PowerPoint PPT Presentation

Fabienne Moreau

ADNid company – Montpellier, France

Objective of MARS program

Markers Assisted Selection Technology

SMEs

TRANSFERT

To speed up the selection of new apricot cultivars resistant to Sharka

Objective of MARS program

Biotechnology SMEs

to use the molecular data for the selection of varieties resistant to Sharka

Genetic knowledge

• n apricot

Molecular tools as molecular markers linked to the resistance

TRANSFERT

Stone Fruit Breeder SMEs

Objectives of MARS program

• 1. The implementation of the MAS for

Sharka resistance in Apricot all over Europe

• 2. The finalization of the agronomical

evaluation of PPV resistant breeding material by the stone fruit breeders.

Workflow of the organisation and the role of partners

Partner P17: FAL, Greece Partner P13: BETA, Turkey Partner P10 *: ADNid, France

* 50 first sample plates

Partner P10: ADNid, France

Shipment of leaf samples to biotech SMEs

• 1. DNA extraction in plates of 96 samples
• 2. DNA quality verification
• 3. PCR amplification of the markers in multiplex
• 4. PCR fragment verification on agarose gels

Shipment of PCR products to partner P10

• 5. PCR fragment verification and validation
• 6. Quality checking
• 7. Capillary electrophoresis of the PCR products
• 8. Quality checking of genotyping data
• 9. Genotype scoring on Genemapper
• 10. Preparation of datafile for MARS partners

and for the genotyping database

Data transfer to MARS partners (breeders) and to the genotyping database (partner P02)

Different stages for implementation of MAS

• Stage 1 : Screening of the material with a first set of

markers developed by Sharco program

– Three markers associated to PPVres locus

• Stage 2 : Analysis of the resistant samples with a second

set of markers

– Four markers targeted also genetic factors closely linked to the resistance

Information about the different markers

Set 1 : PGS1-21/ZP002/PGS1-24 Set 2 : PGS1-10/AMMPG016 AMMPG021/SSRLG1-11m52

PGS1-21 ZP002 PGS1-24

Chromosome 1

PGS1-10 AMMPG016 AMMPG021 SSRLG1-11m52 PPVres locus

Stage 1 : screening of material

Screening of material

Plant adapted to various European environments Sources of resistance: Stark Early Orange or Goldrich but also Harlayne, Stella Continental climate Mediterranean basin Oceanic climate

Stage 1 : Collect of samples

43 370

France Italy Romania Bulgaria Turkey Greece Spain Czech rep

Stage 1 : Interpretation of results with the set 1

• Presence of 3 resistant alleles : resistant genotype
• Presence of 0 resistant allele : susceptible genotype

Conclusion of the stage 1

About half of the samples analysed with set 1 were resistant START OF THE STAGE 2 The resistant samples have been analysed with set 2  These samples have been transferred to the nurseries in charge of the agronomical evaluation, multiplication and characterization before registration

Stage 2 : Samples number

24 096

France Italy Romania Bulgaria Turkey Greece Spain Czech rep

Stage 2 : Molecular results with the set 2

Each marker = resistant alleles

Stage 2 : Results for set 2

By combining the molecular information obtained with 7 markers, we have a genotype, resistant or susceptible which were validated by a phenotypic information

CONCLUSION

At the end of MARS program we are able to offer to the nurseries a method: 1- to screen sensitive apricot progenies in breeding programs 2- to reduce by half the time and the work to produce new varieties resistant to Sharka