红麻线粒体基因atp9克隆及不育细胞质分子标签的利用  

Cloning mitochondrial gene atp9 and utilization molecular marker associated with male sterile cytoplasm in kenaf

作　　者：赵艳红[1,2] 廖小芳[1] 赵洪涛[2] 黄其椿[2] 唐兴富[2] 李初英[2] 周瑞阳[1] 

机构地区：[1]广西大学农学院,南宁530005 [2]广西农业科学院经济作物研究所,南宁530007

出　　处：《南方农业学报》2015年第6期964-970,共7页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(31460366);国家麻类产业技术体系建设专项项目(CARS-19-S14);广西农业科学院科技发展基金重点项目(2015JZ09);广西农业科学院基本科研业务专项项目(桂农科2014YQ09)

摘　　要：【目的】快速克隆红麻线粒体atp9基因编码区(CDS)序列并分析其系统进化关系,为atp9基因功能验证奠定基础;利用基于atp9基因开发的不育细胞质分子标签MM556检测红麻种质资源的细胞质育性,为鉴定红麻不育细胞质种质资源提供参考。【方法】以红麻细胞质雄性不育系P3A及保持系P3B为材料,采用同源克隆方法克隆线粒体基因atp9的CDS区并进行序列比对和进化分析。利用分子标签MM556对84份未知细胞质育性的红麻种质资源进行不育细胞质材料筛选,并利用6个金钱吊芙蓉品种进行验证。【结果】红麻细胞质雄性不育系与保持系的CDS区碱基均为339 bp,两者间存在4个碱基的差异,推导编码112个氨基酸。红麻atp9基因与葡萄的亲缘关系最近,其节点支持率为72,其次为十字花科的油菜和拟南芥,与韭菜亲缘关系最远。MM556分子标签鉴定结果表明,84份红麻种质资源中,UG93-2ms-1、UG93-2ms-2、UG93-2ms-3、UG93-2ms-4、UG93-2-22、KN250、KN250-1、KN142、ZB90和09-7等10份材料具有不育细胞质,其中UG93-2ms-1、UG93-2ms-2、UG93-2ms-3和UG93-2ms-4均为红麻野败型UG93雄性不育突变体的衍生株系。经对金钱吊芙蓉品种的分子检测和田间育性验证,发现MM556分子标签同样适用于金钱吊芙蓉材料育性的辨别。【结论】克隆获得的红麻atp9基因编码区序列及筛选获得的不育细胞质种质资源可用于进一步研究红麻细胞质雄性不育分子机理;基于atp9基因开发的分子标签MM556可用于红麻不育细胞质的鉴定。【Objective】The present study was conducted to clone the coding sequence（CDS） region of mitochondrial atp9 gene of kenaf and analyze its evaluation relationships with other crops in order to lay the foundation for atp9 gene function verification. A molecular marker MM556 associated with male sterile cytoplasm, which was developed based on gene atp9, was used for detecting the fertility of kenaf germplasm resources in order to provide references for detecting kenaf male sterile cytoplasm germplasm resources. 【Method】The CDS region was cloned from kenaf cytoplasmic male sterility（CMS） line P3 A and its maintainer line P3 B by using homological cloning method, followed by sequence alignment between P3 A and P3 B. The phylogenetic tree of gene atp9 was analyzed amongst different crops. Meanwhile, a specific molecular marker MM556 was used for screening male sterile cytoplasm materials from 84 accessions of kenaf germplasm resources with unknown cytoplasmic fertility. 【Result 】The CDS region of atp9 was 339 bp in length for P3 A and P3 B,and only four base pairs＇ difference was found in P3 A and P3 B. CDS region was contained 112 deduced amino acids.Phylogenetic analysis results showed that, atp9 gene of kenaf had the closest relationship with grape being 72 of node approval rating, followed by rape and Arabidopsis, and the farthest one was leek. Ten varieties viz., UG93-2ms-1, UG93-2ms-2, UG93-2ms-3, UG93-2ms-4, UG93-2-22, KN250, KN250-1, KN142, ZB90 and 09-7 containing male sterile cytoplasm were identified from 84 germplasm resources of kenaf by using molecular marker MM556; out of which, 4 varieties viz., UG93-2ms-1, UG93-2ms-2, UG93-2ms-3, UG93-2ms-4 were derivative strain from wild-abortive type mutation of kenaf variety UG93. The results of molecular detection and field fertility verification of Hibisius radjatus showed that, molecular marker MM556 was also suitable for fertility validation of tetraploid Hibisius radjatus as well. 【Conclusion】The obtained CDS region of atp9 gene and scr

关 键 词：红麻 不育细胞质 atp9 分子标签 育性检测 

分 类 号：S563.5[农业科学—作物学]