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园艺学报 ›› 2017, Vol. 44 ›› Issue (2): 215-222.doi: 10.16420/j.issn.0513-353x.2016-0519

• 研究论文 • 上一篇    下一篇

苹果抗炭疽菌叶枯病基因SNPInDel标记的HRM筛选

刘源霞1,兰进好2,柏素花3,孙晓红3,刘春晓1,张玉刚1,戴洪义1,*   

  1. 1青岛农业大学园艺学院,青岛市园艺植物遗传改良与育种重点实验室,山东青岛 266109;2青岛农业大学农学与植物保护学院,山东青岛 266109;3青岛农业大学生命科学学院,山东青岛 266109
  • 出版日期:2017-02-25 发布日期:2017-02-25
  • 基金资助:

    国家自然科学基金项目(31372032);国家现代农业产业技术体系建设专项资金项目(CARS-28-01-07);山东省良种产业化工程项目(620902);“泰山学者”建设工程和青岛市民生科技计划项目(15-9-2-99-nsh,14-2-3-38-nsh);‘十二五’国家科技支撑计划项目(2013BAD02B01);山东省优秀中青年科学家科研奖励基金项目(BS2014SW027)

Screening of SNP and InDel Markers to Glomerella Leaf Spot Resistance Gene Locus in Apple Using HRM Technology

LIU Yuanxia1,LAN Jinhao2,BAI Suhua3,SUN Xiaohong3,LIU Chunxiao1,ZHANG Yugang1,and DAI Hongyi1,*   

  1. 1Qingdao Key Laboratory of Genetic Improvement and Breeding in Horticultural PlantsCollege of HorticultureQingdao Agricultural UniversityQingdaoShandong 266109China2College of Agronomy and Crop ProtectionQingdao Agricultural UniversityQingdaoShandong 266109China3College of Life SciencesQingdao Agricultural UniversityQingdao,Shandong 266109China
  • Online:2017-02-25 Published:2017-02-25

摘要:

以207株‘金冠’与‘富士’苹果的F1杂交分离群体实生树为试材,挑选抗炭疽菌叶枯病基因Rgls位点区域内的,第15条染色体上,位于SSR标记S0304673和S0405127之间500 kb物理距离内,由全基因组重测序技术所获得的与抗该病相关的10个SNP及10个InDel标记,通过高分辨率熔解曲线(High Resolution Melting,HRM)技术筛选出与基因Rgls位点紧密连锁的2个SNP及4个InDel标记。通过对重组个体的基因型及表型分析,发现标记InDel4227和InDel4254表现出与Rgls基因位点共分离,将基因Rgls精细定位于标记InDel4199 和SNP4299之间100 kb的物理距离内。对该基因位点两侧4.1 ~ 4.3 Mb距离内的基因进行统计分析,表明在该区段内存在40个有功能注释的基因,涉及到细胞组成、分子功能和生物过程方面共19个GO分类。

关键词: 苹果, 高分辨率熔解曲线, 炭疽菌叶枯病, SNP标记, InDel标记, 精细定位

Abstract:

In present study,10 SNP and 10 InDel markers associated with Glomerella leaf spot(GLS) resistance in the region ofRgls,on apple linkage group(LG)15,in a region of 500 kb between S0304673 and S0405127 markers,obtained by using the whole genome sequencing technology were tested in the population of 207 F1 individuals derived from a cross between‘Golden Delicious’and‘Fuji’apples by High Resolution Melting(HRM). Two SNP and 4 InDel markers tightly linked to the Rgls locus were screened. Through analyzing the genotypes and phenotypes of the recombinant individuals,the Rglslocus was fine mapped within a distance of 100 kb between InDel4199 and SNP4299 markers on the‘Golden Delicious’genome sequence and was co-segregated with InDel4227 and InDel4254 markers. Forty genes were predicted within the region of 4.1–4.3 Mb flanked the Rglslocus,including cellular component,molecular function and biological process in a total of 19 GO classifications.

Key words: apple, HRM, Glomerella leaf spot, SNP marker, InDel marker, fine mapping

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