中国农业科学Issue(8):1550-1557,8.
黄瓜成熟瓜网纹基因H遗传定位及候选基因分析
Molecular Mapping and Candidate Gene Analysis for Heavy Netting Gene (H) of Mature Fruit of Cucumber (Cucumis sativus L.)
摘要
Abstract
Objective]Heavy netting of mature fruit is one of the important phenotypes of cucumber. Molecular mapping and candidate gene analysis for this trait will provide a theoretical basis for improvement of fruit traits. It also can lay a good foundation for fine mapping and gene cloning. [Method] Cucumber inbred lines PI205996(P1) without heavy netting mature fruit and PI263079 (P2) with heavy netting mature fruit were used as the experiment materials for genetic analysis and gene mapping in this study. Bulked segregation analysis (BSA) was performed in the F2 population with 230 individuals using 2 112 SSR markers. The sequence and re-sequencing information of 9 930 and 115 core germplasms were used to develop new SSR markers in the primary mapping region of the heavy netting gene by Primer 6.0. JoinMap 4.0 and MapInspect softwares were employed to construct a SSR linkage map for the gene. Bioinformatics analysis was conducted to detect candidate genes. [Result] Genetic analysis showed that a single dominant nuclear gene (H) controlled the heavy netting of mature fruit trait in PI263079. There were 255 of 2 112 SSR primers showed polymorphism in the parent lines and the polymorphic rate was 12.1%. Using the 255 polymorphic markers, the DNA of seven individuals with heavy netting fruit and seven individuals without heavy netting fruit was analyzed. Nine SSR markers on cucumber chromosome 5 were identified to be linked with the H gene. Flanking markers SSR13006 and CSWCT-17 were 3.6 and 8.2 cM away from the H gene, respectively. A total of 97 pairs of new SSR primers were developed based on the sequence information in the primary mapping region of the H gene. And only four polymorphic markers were detected between the parent lines with the polymorphic rate of 4.1%. Finally, 13 SSR markers were identified to be linked with the H gene after analysis of the F2 mapping population using the four new developed molecular markers. Flanking markers SSR13006 and SSR-90 were 3.6 and 1.7 cM away from the H gene, respectively. The physical distance between SSR13006 and SSRH-90 was 297.7 kb containing 29 predicted genes. Csa5G591790 was speculated as a possible candidate gene.[Conclusion]The heavy netting of mature fruit trait of PI263079 was controlled by one dominant nuclear gene (H). It was located on the Chr.5 of cucumber delimited in a physical distance of 297.7 Kb. The results in this study will be of great benefit to fine mapping and gene cloning for the H gene and MAS of heavy netting of mature fruit.关键词
黄瓜/成熟瓜网纹/SSR标记/基因定位/候选基因Key words
Cucumis sativus L./heavy netting of mature fruit/SSR marker/gene mapping/candidate gene引用本文复制引用
王敏,顾兴芳,苗晗,刘书林,王烨,Todd C.Wehner,张圣平..黄瓜成熟瓜网纹基因H遗传定位及候选基因分析[J].中国农业科学,2014,(8):1550-1557,8.基金项目
国家“863”计划项目(2012AA100105)、国家现代农业产业技术体系建设专项资金(CARS-25)、农业部园艺作物生物学与种质创制重点实验室资助项目 ()