Introduction to the use of molecular genotyping techniques Gregorio López-Ortega, Almudena Bayo-Canha, Emma Skipper and Felicidad Fernández Budapest 3 rd -5 th of March
STSM (Spain to UK) Pomological characterization and introduction to the use of molecular genotyping techniques Pomological characterization Fingerprinting of 10 varieties S-alleles of 10 varieties Checking two molecular markers for fruit size Felicidad Fernández Fernández East Malling Research, United Kingdom COST STSM Reference Number: ECOST-STSM-FA1104-150713-034253
! "# $%&'( )'* +$&,-. ' Pomological characterization C210-7 X Colney 300 individuals Phenotypical and Pomological Evaluation of the Variation Present in a Sweet Cherry Mapping Population. E. S. Skipper 1, G. Lopez-Ortega 2, A. Bayo-Canha 2, and F. Fernández-Fernández 1 1 East Malling Research, New Road, East Malling ME19 6BJ UK Sweet September C210-7 X X Lapins Colney 2 IMIDA, C/ Mayor, Murcia 30150 Spain gregorio.lopez2@carm.es Introduction: A group of 125 individuals of the F1 progeny Cherry (Colney x C210-7 [Lapins x Sweet September]) has been evaluated at East Malling Research (EMR). This contribution shows the natural variation in flowering, harvest date and fruit quality traits. Harvesting date: Harvesting date records: Estimate ripening date each two-three days. Quality assessments: Standard quality assessments: good, rots, cracks, soluble solids, acidity, colour, fruit size, firmness, weight. Aims: Characterise phenotypical variation in the progeny to enable QTL analysis. Identify individuals with extreme phenotypes with a potential to extend harvest season due to delayed ripening/maturity. Results: The harvest period was between 16 th of July and 21 th of August. Half of the population had white flesh while the other half had black flesh. The average weight of the fruits were between 6-10 grams. There were only three hybrids with an average weight higher than ten. The 80% of the progeny had sugar content between 19-25 ºbrix and a 60% of the hybrids firmness was among 350-450 (g/mm). Period: 16th of July and 21st of August Average weight: 6-10 gr 80% 19-25 ºbrix 60% 350-450 g/mm2 50% white flesh/50 black flesh '! " &#" &! " %#" %! " $#" $! " #"! " %!! (%#! " %#! (&!! " &!! (&#! " &#! ('!! " '!! (' #! " ' #! (#!! " #!! (##! " Future Work 3,&# 4%.. ' /01# # 2 ' Develop a linkage map using the Illumina 6K SNP chip in collaboration with IASMA Record phenotype data over subsequent years and in different environments for the mapping of QTLs relating to flowering characteristics and quality traits. This work was conducted within the framework of COST Action 1104
Fingerprinting Objective: Fingerprinting ten varieties Methodology: DNA extraction Dneasy kit Quiagen PCR amplification in two multiplex Sizing with ABI
Molecular Markers
Results
S-Alleles Objective: S-alleles determination of ten varieties Methodology: PCR with two primer pairs Sizing with ABI Analyse the results
S-allele Markers *6-FAM-F-BOX5 A (Vaughan 2006): TTK SCH ATT RYC AAC CKC AAA AG *F-Boxintron R: CWG GTA GTC TTD SYA GGA TG *VIC-PaConsI-F (Sonneveld 2003): MCT TGT TCT TGS TTT YGC TTT CTT C *PaConsI-R2 (Sonneveld 2006): GCC ATT GTT GCA CAA ATT GA
Results Control Cultivars Determined References Summit S1S2 S1S2 Colney S5S6 S5S6 Inge S4S9 S4S9 Schneider S3S12 S3S12 Cryall-seeding S2S7 S2S7 Sumburst S3 S3S4' Noble - S6S13 Cristobalina - S3S6
Results for our ten varieties Cultivars Crystal Champaing Ronde grosse Gemelle Summerland Carly O'Lory Determined S3S5 S3S6 S2S9 S5S17 S1S9 7-91C - Big Lory - Mister Early - Riaño - Walter C -
Markers for Fruit size
Markers for fruit size Colors indicate whether the haplotype is associated with large, small, or presumed small fruit size 190-237 190-255 190-225 190-257 192-250 202-250 192-223 192-225 204-235 190--223 190-235 204-223 204-255 LARGE PRESUMED SMALL SMALL NO EFFECT
Conclusions The goals of the STSM were achieved A set of markers for fingerprinting Markers for hybrid selection Proposal Consensus a set of markers for fingerprinting Protocol for S-alleles MAS
Acknowledgements THANKS FOR YOUR ATTENTION
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