Scientia Agricultura Sinica ›› 2018, Vol. 51 ›› Issue (13): 2622-2630.doi: 10.3864/j.issn.0578-1752.2018.13.017

• RESEARCH NOTES • Previous Articles    

Development and Application of SSR New Molecular Marker for Seedless Traits in Grape

MA YaRu1, FENG JianCan2, LIU ChongHuai1, FAN XiuCai1, SUN HaiSheng1, JIANG JianFu1, ZHANG Ying1   

  1. 1Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou 450009; 2College of Horticulture, Henan Agricultural University, Zhengzhou 450002
  • Received:2017-11-20 Online:2018-07-01 Published:2018-07-01

Abstract: 【Objective】The objective of this study is to develop a new SSR molecular marker for seedless traits in grape, and to identify the early seedless traits of grape hybrids, and to lay the foundation for molecular marker assisted selection breeding. 【Method】RAD-seq was carried out in 133 grape materials including the female parent ‘Red Globe’, the male parent ‘Centennial Seedless’ and their F1 hybrids, and the sequencing results were compared to the reference genome. SAMtools software was used to generate CaSFS software required for pileup and GLF files, and the effective SNP sets between parents were obtained by filtration. A window sliding method was used to determine the genotype of each window, and selected one window per 15 SNP and slided one SNP at a time. The genotype of each individual was obtained and the bin generated. Linkage analysis was carried out based on Joinmap software for generated bin, and genetic linkage map was constructed between ‘Red Globe’ and ‘Centennial Seedless’. The QTL loci was mapped using WindowQTLCartographer2.0 software. 35 SSR molecular markers consistent with the SSR characteristic sequence were identified by Perl programming language in the QTL mapping interval, and then 35 pairs of SSR molecular marker primers were designed. The molecular marker was screened in the differences with the seedless traits between parents by HRM technology, and the accuracy of seedless typing, the rate of seedless detection and the rate of seedless holding were detected in 131 F1 hybrid population and natural population including 65 grape varieties.【Result】On the genetic linkage map between ‘Red Globe’ and ‘Centennial Seedless’, the seedless traits of grape were mapped on chromosome 18, and the mapping interval was 26 835 846-26 960 426, and the contribution rate to seedless was 77.9%, and the LOD threshold was 26.3. The molecular marker VvSD10 was screened in the differences with the seedless traits between parents, and there was a 111 bp loci in seedless traits of grape. Identification of seedless traits in grape F1 hybrid population and natural population by HRM technology using 111 bp loci on VvSD10 marker, the results indicated that in F1 hybrid population and natural population, the correct typing rate of molecular marker VvSD10 was 97% and 94%, respectively, and the rate of seedless detection of both populations was 56%, the rate of seedless holding was 77% and 85%, respectively. 【Conclusion】Seedless typing in grape by Molecular marker VvSD10 could provide high accurate information in both genetic and natural populations and laid the foundation for molecular marker assisted breeding program in seedless grape.

Key words: Vitis vinifera L., seedless, QTL mapping, SSR molecular marker, assisted breeding

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