Development of markerassisted breeding strategies for strawberry The example of the everbearing trait (or perpetual flowering trait) Béatrice Denoyes, Amélia Gaston Philippe Chartier, Justine Perrotte Final EUBerry Meeting, Skierniewice 13th 16th October, 2014, Poland
Strawberry development cycle in France June bearing varieties Ever bearing varieties Nov. Dec. January Febuary March April May June July August Vegetative growth Florale initiation Runnering Flowering Fruit production Production
Perpetual flowering : a major QTL, FaPFRU Capitola CF1116 FaPFRU Quantitative notations for PF and RU - 2 groups of genotypes for flowering : PF and SF - continuous distribution for runnering 1 major QTL fot both traits, RU and PF Antagonism between the two traits in the population. Quantitative perpetual flowering suggesting other genetic controls Gaston et al., JXB, 2013
LGIV-Fb1 0 Adding markers in the genomic region of interest for improving the use of markers of MAS tgat134 LOD 0.0 F-LGIVb v007224 LOD 5.4 caaa410 9.5 13.5 21.6 25.6 32.1 gatg165 EMFv007 cttg228 tcaa260 ccaa410 22.9 31.3 39.1 45.6 46.2 54.5 65.8 70.7 74.2 81.8 85.8 92.2 ah05268 u1h5251r u1h5249c tgat134 k103124 k103120 c014138 caaa410 gatg165 EMFv007 cttg228 tcaa260 ccaa410 63.7 catc139 123.4 catc139 LGIV-Fb2 0 8.7 11.8 gatt284 gata295 UDF008 165.7 173.1 186.5 190.6 192.6 195.7 198.6 Bx089196 Bx083206 gatt284 Bx064216 Bx250222Bx250218 gata295 UDF008
Same introgression in different cultivars Capitola F-LGIV-b 0.0 5.3 9.7 14.0 18.2 25.5 29.5 36.0 tgat134r caaa410r gatg165r Bx048345 v007190c cttg228c tcaa260c ccaa410c 67.1 catc139c Mara des Bois F-LGIV-b 101.1 103.6 108.7 110.2 110.7 114.4 118.1 120.2 123.3 126.2 Bx089196 Bx083206 Bx005330 Bx215131 Bx052225 Bx059350 Bx056206 gatt284c Bx064216 Bx250222 Bx250218 gata295c u008123c 4.2 7.4 1.2 Bx089196 Bx083206 Bx058248 Bx063240 Bx063232 Bx059350 Bx059238 Bx056206 Bx064216 Bx064222
The use of developed markers in different segregating populations Genotypes of the parents Polyacrylamide gel Presence of markers in PF Different situations for OF genotypes: Presence or absence of markers Bx089 Bx083 Bx005 Phenotype (PF or OF) Génotypes Perpetual flowering Capitola 1 1 1 PF 1 1 1 1 1 1 Mara des Bois 1 1 1 PF 1 DD (c) 1 1 1 DD Parent_PF1 1 1 1 PF NA 0 1 1 1 1 Parent_PF2 1 1 1 PF NA 0 1 1 1 1 Once flowering CF1116 0 0 0 OF 0 0 0 0 0 0 Pajaro 0 0 0 OF 0 0 0 0 0 0 Parent_OF1 1 1 1 OF NA 0 1 1 1 1 Parent_OF2 0 0 0 OF NA 0 0 0 0 0 Parent_OF3 0 0 0 OF NA 0 0 0 0 0 Parent_OF4 1 1 1 OF NA 0 0 0 0 1 DD, double dose. Bx215 Bx052 Bx059 Bx056 Bx064 Bx250
The use of developed markers in different segregating populations Results in samples of segregating populations Population Parent 1 Parent 2 N ind. Bx064 Bx089 Both Markers present & Rem. Both markers absent and NOT Rem. Population1 Parent_OF1 Parent_PF1 10 6 6 5/7 0/3 Population2 Parent_OF1 Parent_PF2 10 5 5 4/4 0/6 Population3 Parent_PF1 Parent_OF2 10 9 7 3/4 4/6 Population4 Parent_PF2 Parent_OF2 10 9 9 7/7 2/3 Population5 Parent_OF3 Parent_PF1 10 9 8 1/1 7/9 Population6 Parent_OF4 Parent_PF1 10 8 6 3/3 2/7 Population7 Parent_OF4 Parent_PF2 9 6 6 0/2 6/7 Population8 Capitola CF1116 218 110 100 95/97 90/90 The markers can be used only when they are present in the PF parents and absent in the OF parent (Populations 3, 4 and 5)
Summary of results obtained in genetic resources RECOMBINANTS Distribution of phenotypes Markers Bx056-Bx005 total N PF N OF N uncertain Nb of genotypes 1-1 35 17 10 8 Nb of genotypes 0-0 23 0 18 5 Nb of genotypes 1-0 7 0 6 1 Nb of genotypes 0-1 9 3 3 3 Nb of genotypes? 7 Total 81 PF: Perpetual Flowering or everbearing / OF: Once flowering or June bearing The presence of markers in OF genotypes can be explained by pedigree. Old genotypes have had a double recombination around the trait Name Bx005 PF or OF Bx056 Ciflorette (Mamie x Earlyglow) 1 OF 1 Earlyglow 1 OF 1 Mamie 1 OF 1 In conclusion: The presence of markers in OF genotypes is not rare. Therefore, parents have to be genotyped before the use of markers in MAS for the everbearing trait.
What we can gain by using MAS for the everbearing trait? MAS: Screening on S0: MAS can enable to produce more seedlings and evaluate just the right combination. Exemple: introduction of a qualitative traits present in a everbearer variety (taste) for a june bearer project: Make more crosses (1000 seedlings) and keep just the 0-0 types: 2 weeks lab work to increase of 50% the probability of getting the right phenotype Screening for S1: to avoid false positive for everbearers on second year: flowering observations alone on seedlings S0 indicating a possible everbearer phenotype is sometime not confirmed on the next year on the propagated plant leading to a wrong cultivation system: -> wrong use of limited number of plots (cost effective). 50 plots available: 100 candidates S1 to analyse. Cost of 2 analysis < cost of 1 plot The gain of the probability increase of success is difficult to evaluate The loss due to a junebearer cultivar taking the room space of a everbearer cultivar in a everbearer cultivation system trial is in the 100 s MAS will be more efficient on several traits: everbearing + fruit quality + disease tolerance
THANK YOU FOR YOUR ATTENTION Final EUBerry Meeting, Skierniewice 13th 16th October, 2014, Poland