棉花CMS-D2和CMS-D8不育系线粒体全基因组比较分析  

Comparative analysis of whole mitochondrial genomes between CMS-D2 and CMS-D8 sterile lines in cotton

作　　者：葛李爽 冯娟娟 张梦[1] 郭立平[1] 戚廷香[1] 张学贤[1] 李永旗[1] 唐会妮[1] 乔秀琴[1] 邢朝柱[1] 吴建勇[1] Ge Lishuang;Feng Juanjuan;Zhang Meng;Guo Liping;Qi Tingxiang;Zhang Xuexian;Li Yongqi;Tang Huini;Qiao Xiuqin;Xing Chaozhu;Wu Jianyong(Institute of Cotton Research of Chinese Academy of Agricultural Sciences/National Key Laboratory of Cotton Bio-breeding and Integrated Utilization/Key Laboratory for Cotton Genetic Improvement,Ministry of Agriculture and Rural Affairs,Anyang,Henan 455000,China)

机构地区：[1]中国农业科学院棉花研究所/棉花生物育种与综合利用全国重点实验室/农业农村部棉花生物学与遗传育种重点实验室,河南安阳455000

出　　处：《棉花学报》2024年第1期28-38,共11页Cotton Science

基　　金：河南省自然科学基金面上项目“orf610a调控棉花细胞质雄性不育的分子机制研究”(232300421168)。

摘　　要：【目的】探究哈克尼西棉不育胞质(CMS-D2)和三裂棉不育胞质(CMS-D8)的不育系线粒体基因组之间的序列结构差异,为筛选鉴定不育相关基因奠定基础。【方法】根据D2A和D8A这2个不育系的线粒体基因组测序组装结果,使用Synteny and Rearrangement Identifier(Sy RI)软件鉴定结构变异,用Plotsr可视化分析包含共线性及非共线性区域的重组变异位点。以D8A线粒体基因组注释结果为参考,用D2A线粒体基因组注释的开放阅读框(open reading frame,ORF)编码的氨基酸序列进行tblastn比对,筛选出D2A线粒体基因组中特有的ORF,并进行聚合酶链式反应(polymerase chain reaction,PCR)验证、相对表达量分析和生物信息学分析。【结果】D2A和D8A这2个不育系线粒体基因组之间存在2个部分重叠且相邻的倒易位区域。在D2A中发现17个特异的ORF,PCR验证了D2A中存在的6个特异ORF(orf114e、orf121b-1、orf121b-2、orf138b-2、orf186a-2和orf317a-2)。3~4 mm花蕾中orf121b-1和orf121b-2的相对表达量较高。orf114e、orf186a-2和orf317a-2具有典型的跨膜结构域和嵌合基因结构,符合不育基因的部分特征。【结论】D2A和D8A线粒体基因组间存在2个相邻的倒易位区域。D2A中存在6个特异的ORF,其中orf114e、orf186a-2和orf317a-2可能与棉花CMS-D2孢子体败育有关。[Objective]This research aims to investigate the sequence structural differences between the mitochondrial genomes of Gossypium harknessii sterile cytoplasm(CMS-D2)and G.trilobum sterile cytoplasm(CMS-D8),which can establish a foundation for the screening and identification of sterility-related genes.[Methods]Based on the sequencing and assembled results of the mitochondrial genome of D2A and D8A,the Synteny and Rearrangement Identifier(SyRI)software was used to identify structural variations,and recombination variation sites containing collinear and non-collinear regions were visualized with Plotsr.The annotation results of the D8A mitochondrial genome were utilized as a reference,and the amino acid sequences encoded by the open reading frame(ORF)annotated in the D2A mitochondrial genome were used for tblastn comparison to screen out the unique ORF in the D2A mitochondrial genome.Subsequently,polymerase chain reaction(PCR)verification,relative expression level analysis,and bioinformatics analysis were performed.[Results]The mitochondrial genomes of the sterile lines D2A and D8A had two partially overlapped and adjacent inverted translocation regions.Seventeen specific ORF were found in D2A mitochondrial genome,and PCR verified the presence of 6 specific ORF(orf114e,orf121b-1,orf121b-2,orf138b-2,orf186a-2,and orf317a-2)in D2A.The relative expression levels of orf121b-1 and orf121b-2 were higher in 3-4 mm flower buds.orf114e,orf186a-2,and orf317a-2 have typical transmembrane domains and chimeric gene structures,which are consistent with some characteristics of sterility genes.[Conclusion]Two adjacent inverted translocation regions were identified between the mitochondrial genomes of D2A and D8A.There are six specific ORF in D2A,among which orf114e,orf186a-2,and orf317a-2 may be potentially related to sporophytic abortion of CMS-D2.

关 键 词：棉花 细胞质雄性不育 线粒体基因组 孢子体不育 配子体不育 开放阅读框 

分 类 号：S562[农业科学—作物学]