白菜型冬油菜温敏不育系PK3-12S育性转换的差异蛋白质组学分析  

Differential proteomics analysis of fertility transformation of the winter rape thermo-sensitive sterile line PK3-12S(Brassica rapa L.)

作　　者：米文博 方园[1] 刘自刚[1] 徐春梅 刘高阳 邹娅 徐明霞 郑国强 曹小东 方新玲 MI Wen-Bo;FANG Yuan;LIU Zi-Gang;XU Chun-Mei;LIU Gao-Yang;ZOU Ya;XU Ming-Xia;ZHENG Guo-Qiang;CAO Xiao-Dong;FANG Xin-Ling(Gansu Provincial Key Laboratory of Arid Land Crop Sciences/Key Laboratory of Crop Genetics Improvement and Germplasm Enhancement of Gansu Province/Gansu Research Center of Rapeseed Engineering and Technology/College of Agronomy,Gansu Agricultural University,Lanzhou 730070,Gansu,China)

机构地区：[1]甘肃省干旱生境作物生物学重点实验室/甘肃省作物遗传改良与种质创新重点实验室/甘肃省油菜工程与技术研究中心/甘肃农业大学农学院,甘肃兰州730070

出　　处：《作物学报》2020年第10期1507-1516,共10页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31660404);国家重点基础研究发展计划项目(2018YFD0100502-2);甘肃省高校科研成果转化培育项目(2018D-13);国家现代农业产业技术体系建设专项(CARS-13);甘肃省科技重大专项项目(17ZD2NA016-4)资助。

摘　　要：为揭示温敏不育系PK3-12S育性转换机制,本研究以白菜型冬油菜温敏不育系PK3-12花药为材料,采用2-DE和LC-MS/MS质谱鉴定等差异蛋白组学方法,分离鉴定了PK3-12在不育/可育条件下花药差异表达蛋白质,并对差异表达蛋白进行了生物信息学分析;进而采用RT-PCR检测了PK3-12在不育/可育条件下花蕾发育进程中差异蛋白编码基因表达量变化。结果表明,高温不育条件下,PK3-12花药形态瘪小,药室有少量败育花粉,育性转换受1对隐性基因控制,表达变化量在2倍以上差异蛋白质点31个,其中增量表达蛋白质点6个,减量表达蛋白质点11个,表达完全抑制蛋白点12个,不育花药特异表达蛋白点2个。质谱鉴定出15个差异蛋白质,参与信号转导通路、二羧基乙醛酸代谢、糖酵解代谢、次生合成代谢、氨基酸生物合成、分支酸生物合成、碳代谢途径等细胞过程。Rubisco亚基连接蛋白编码基因BrrbcL开放读码框(open reading frame,ORF)长度为1095 bp,编码364个氨基酸;与可育花蕾相比,发育进程中不育花蕾BrrbcL基因、膜联蛋白基因(ANN)、BetVI过敏原家族基因(BetVI)表达明显下调,表明上述基因可能参与了温敏不育系PK3-12S育性的转换。To reveal the fertility switching mechanism of temperature-sensitive sterility line PK3-12S(Brassica rapa L.),the differentially expressed proteins were isolated and identified using anthers of PK3-12S in sterile/fertile conditions by 2-DE and LC-MS/MS mass spectrometry.The expression level variations of differentially expressed genes were examined by RT-PCR in PK3-12 flower buds during sterility/fertile development.The result showed that the sterile anther size of PK3-12 was small with a little abortive pollen in the anther room under high temperature.The trait of fertility transformation was controlled by a pair of recessive alleles.There were 31 differentially expressed proteins with more than two times of the expression level,including six protein spots with increasing expression,11 protein spots with reduced expression,12 protein spots with complete inhibition,and two protein spots with induced expressed.Fifteen differentially expressed proteins involved in the cellular processes such as signal transduction pathways,glyoxylate and dicarboxylate metabolism,glycolysis gluconeogenesis,biosynthesis of secondary metabolites,biontheses of amino acids,chorismate biosynthesis,and carbon metabolism pathways were identified by mass spectrometry.The BrrbcL gene,encoding a Rubisco subunit-binding accessory protein,had an open reading frame(ORF)in length of 1095 bp encoded 364 amino acids.Compared with fertile anthers,the expression level of BrrbcL gene,annexin gene(ANN)and BetVI allergen family gene(BetVI)was significantly down-regulated during sterile anthers development,which indicated that these genes maybe participate in the fertility transformation of the thermo-sensitive sterile line PK3-12S.

关 键 词：白菜型冬油菜 温敏不育 差异蛋白组学 基因表达 

分 类 号：S565.4[农业科学—作物学]