玉米诱变与原自交系株高变化的蛋白质组学及差异基因的功能分析  

Function analysis of proteomic and differential genes of plant height between mutagenic line and control inbred of maize

作　　者：吕二锁[1] 逯晓萍[1] 王树彦[1] 薛春雷 韩平安[1] 董婧[1] 任锐 李美娜 

机构地区：[1]内蒙古农业大学农学院,内蒙古呼和浩特010019 [2]呼和浩特市合创农业科技研究中心,内蒙古呼和浩特011500 [3]呼和浩特市种子管理站,内蒙古呼和浩特010020

出　　处：《西北农林科技大学学报（自然科学版）》2015年第6期88-98,共11页Journal of Northwest A&F University(Natural Science Edition)

基　　金：内蒙古自治区科学技术项目(20131410);内蒙古呼和浩特科学技术项目(2013-重-计-1)

摘　　要：【目的】研究玉米经EMS处理后诱变系与原自交系的蛋白表达机制,为从蛋白质组学角度揭示诱变后选育材料在株高发生变化及生物产量明显提高等方面的分子理论奠定基础。【方法】以玉米(Zea mays L.)优良自交系AMD16和诱变后选育材料为试材,采用双向电泳、质谱分析以及检索技术,比较选育材料与原自交系叶片蛋白质组的差异,并对其候选基因进行分离、克隆,构建植物表达载体p3300-ZmMDH6,最后进行拟南芥的遗传转化与鉴定。【结果】EMS诱变后选育材料的蛋白质组中出现21个差异蛋白质点,其中5个蛋白质点在选育材料中特异表达,1个蛋白质点下调表达,7个蛋白质点上调表达,有2个差异蛋白质点在原自交系中表达,而未在选育材料中表达。通过MALDI-TOF-MS质谱测序和MASCOT序列分析,鉴定出了15个差异表达蛋白质点,其功能涉及生物细胞代谢/能量代谢、防御/抗胁迫、细胞蛋白合成和叶绿素合成等细胞过程。用RT-PCR方法克隆了其候选基因ZmMDH6的编码区cDNA,长度1 296bp,由432个氨基酸残基组成,分子质量46.80ku,等电点5.79;并构建植物表达载体进而转化拟南芥,经检测证明已获得T1代转基因植株。【结论】玉米诱变系与原自交系在蛋白丰度上存在明显的差异;差异表达蛋白涉及到各个生长发育过程,可能与玉米生物产量的提高和株高的改变有密切关系。【Objective】This paper studied the expression differences of mutagenic line and inbred line of maize through proteomic analysis to provide reference for improving plant height and biomass of mutagenic maize line.【Method】The proteomic differences of mutagenesis and inbred line AMD16 were compared using two-dimensional gel electrophoresis(2-DE),MALDI-TOF-MS mass spectrometry and retrieval technology,and the candidate genes were separated and cloned to construct plant expression vector p3300-ZmMDH6 before being introduced into Arabidopsis and identified.【Result】The proteome of mutagenicline contained 21 differentially expressed protein spots,including 5specifically expressed spots,7up-regulated spots and 1down-regulated spots.Meanwhile there were two differences expressed in inbred line AMD16,but not expressed in the breeding materials.15 of them were identified as homologous to known proteins in the databases by MALDI-TOF-MS and MASCOT analysis,and its functions involved in biological cell metabolism/energy metabolism,defense/stress responses,protein synthesis in cells,and chlorophyll synthesis.The complete coding region cDNA of ZmMDH6 was amplified by reverse transcription polymerase chain reaction(RT-PCR).The genes had the length of 1 296 bp and constituted 432 amino acid residues with molecular of 46.80 ku and pI of 5.79.The plant expression vector for the gene was constructed and introduced into Arabidopsis.The T1 transgenic plants were identified by PCR.【Conclusion】Protein abundance was significantly different between mutagenic line and inbred line of maize.The different expression of protein involved in all growth and development processes,indicating that it related to biomass and plant height.

关 键 词：玉米 蛋白质组 苹果酸脱氢酶 基因克隆 基因表达 

分 类 号：S[农业科学]