| 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0.29 Mb，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。 |
| 关键词: 番茄 雄性不育 简化基因组测序 基因定位 |
| DOI： |
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| 基金项目: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） |
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| Mapping male-sterile gene in tomato by specific length amplified fragment sequencing |
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WANG Baike,LI Ning,TANG Yaping,et al
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| Abstract: |
| 【Objective】This study obtained candidates of male-sterile gene and improved the accuracy of locating male-sterile gene in tomato to provide theoretical basis for further cloning and revealing male sterilemechanism.【Method】A pollen abortion tomato line closely linked to a green stalk Marker at seedling stage was used as the female parent and a male fertility tomato line with purple stalk at seedling stage was used as the male parent.F2 generation male-sterile and male-fertility DNA pools were constructed with thirty individuals by BSA method.The specific length amplified fragment sequencing (SLAF-Seq) technology was used to detect sufficient fragments.Then,the candidate regions of sterile male gene were obtained by association analysis,and the function of candidate regions genes were annotated by the GO library.【Result】A total of 20.27 Mb reads data were obtained by SLAF-sequencing and 103 250 SLAF Markers were obtained by SLAF-exploiting with the average sequencing depth of 160.39 times.Then 6 892 polymorphism Markers were obtained between two mixed DNA pools by polymorphism analysis and eight candidate regions were determined.The average length of the eight candidate regions was 0.29 Mb,containing 27 polymorphic Markers and 146 candidate genes.【Conclusion】The male sterile gene was preliminarily located by SLAF-Seq. |
| Key words: tomato male sterile SLAF-Seq gene mapping |
