摘要
利用基因组重测序技术进行基因定位目前已在水稻功能基因组学研究中得到广泛应用。其中,Mutmap+技术因无需进行杂交操作,且无需背景亲本的基因组信息,具有更广阔的应用前景。然而,目前Mutmap+技术筛选候选突变位点的方法依赖于复杂的数据计算,要求研究者具有较高的生物信息学知识。根据Mutmap+的实验原理,设计一种简单的筛选候选突变位点的方法。该方法对混池测序的表型池与非表型池的突变指数分别加以限定,即表型池突变指数等于1,非表型池突变指数小于0.5。对测序所得突变位点进行简单排序,即可得到候选突变位点。运用该筛选方法,从6个水稻不育突变体成功克隆突变基因,并对其中1个突变体进行了细胞学表型的验证。该方法可简化Mutmap+技术的数据分析流程,便于将Mutmap+技术更好地运用到水稻功能基因组学研究中。
Genome re-sequencing technology is widely used in rice gene mapping and functional genomics studies. Mutmap+, a versatile method based on genome re-sequencing, has a broader application prospects because of its independence of hybridization and genomic sequence of parental variety. However, current procedure to screen candidate mutation sites of Mutmap+depends on complex data calculation, which requires a high level of bioinformatics knowledge. In this study, we designed a simple method to screen mutation sites in Mutmap+. This method defines the mutation index of phenotype pool and non-phenotype pool. The mutation index of phenotype pool is equal to 1, and the mutation index of non-phenotype pool is less than 0.5. By simply sorting the mutation sites obtained from sequencing, candidate mutation sites can be obtained. Using this screening method, we successfully cloned mutated genes from six rice sterile mutants and verified the cytological phenotype of one of the mutants. This method can simplify the data analysis process of Mutmap+technology, so as to better apply it to the research of rice functional genomics.
作者
王延妍
李颖颖
王子瑞
毛馨晨
唐家琪
于恒秀
张超
WANG Yanyan;LI Yingying;WANG Zirui;MAO Xinchen;TANG Jiaqi;YU Hengxiu;ZHANG Chao(Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Key Laboratory of Plant Functional Genomics of the Ministry of Education/College of Agriculture,Yangzhou University,Yangzhou 225009,China)
出处
《扬州大学学报（农业与生命科学版）》
CAS
北大核心
2024年第1期27-32,共6页
Journal of Yangzhou University：Agricultural and Life Science Edition
基金
江苏省自然科学基金资助项目(BK20200951)。
作者简介
王延妍(2002-),女,江苏镇江人,扬州大学学生,主要从事水稻育性调控基因的克隆研究;通信作者:张超,扬州大学讲师、博士,主要从事水稻功能基因组学研究,E-mail:chaozhang@yzu.edu.cn。
