基于简化基因组测序技术的番茄雄性不育基因定位

发布时间：2018-11-23 19:08

【摘要】：【目的】获得番茄雄性不育基因的候选基因,提高番茄雄性不育基因定位的准确性,为该雄性不育基因的克隆及功能机制研究提供理论基础。【方法】以一个与苗期绿茎基因紧密连锁的花粉败育型材料为母本,以一个苗期紫茎可育系为父本,通过集群分离(BSA)法分别建立30个F2单株的不育和可育DNA池,基于简化基因组测序(SLAF-Seq)技术检测足够量的简化基因组扩增片段,通过关联分析,获得目标雄性不育基因的候选区域,并利用GO数据库对关联区域内的基因进行功能注释。【结果】通过SLAF测序共获得20.27 Mb的序列量;共获得103 250个SLAF标签,标签整体平均深度达160.39倍;完成了2个混合池间SLAF标签标记的多态性分析,共获得多态性标记6 892个;利用2个混合池进行关联分析,共得到8个与目标雄性不育基因相关的候选区域,区域平均大小为0.29Mb,区域内共有27个差异标记,146个候选基因。【结论】利用SLAF测序技术对番茄雄性不育基因进行了初步定位。
[Abstract]:[objective] to obtain candidate genes for tomato male sterility gene, and to improve the accuracy of location of male sterility gene in tomato. [methods] A pollen abortion type material closely linked to green stem gene at seedling stage was used as female parent and a purple stem fertile line at seedling stage as male parent. The sterile and fertile DNA pools of 30 F2 plants were established by cluster isolation (BSA) method. Sufficient simplified genomic amplification fragments were detected based on simplified genome sequencing (SLAF-Seq). Candidate regions of the target male sterility gene were obtained, and functional annotation of the genes in the associated region was carried out by using GO database. [results] 20.27 Mb sequence was obtained by SLAF sequencing. A total of 103,250 SLAF tags were obtained, with a total average depth of 160.39 times, and a total of 6892 polymorphic markers were obtained through the polymorphism analysis of SLAF tags between two mixed pools. Eight candidate regions associated with the target male sterility gene were obtained by association analysis with two mixed pools. The average size of the candidate regions was 0.29 Mb, and there were 27 differential markers in the region. [conclusion] SLAF sequencing was used to localize male sterile genes in tomato.
【作者单位】：新疆农业大学林学与园艺学院;新疆农业科学院园艺作物研究所;
【基金】：公益性行业(农业)科研专项(201303115) 大宗蔬菜产业技术体系乌鲁木齐试验站项目(CARS-25-G-51) 新疆维吾尔自治区园艺学重点学科基金项目(2016-10758-3)
【分类号】：S641.2

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