作物学报2011，Vol.37Issue(7)：1134-1143,10.DOI:10.3724/SP.J.1006.2011.0113
小麦生理型和遗传型雄性不育系及其保持系小花完整叶绿体蛋白质组分比较研究
Comparison of Floret Intact Chloroplast Proteome in Male Sterile LineInduced by CHA-SQ-1, Cytoplasmic-nuclear Sterile and Its Normal Fertile Lines in Wheat
摘要
Abstract
We extracted floret chloroplast proteins from the genetic male sterile line ms(S)-1376, the physiological male sterile line ms(A)-1376 induced by chemical hybridizing agent SQ-1, and their counterpart maintainer line (A)-1376 (normal fertility). These proteins were separated via 2-D electrophoresis with immobilized pH 4-7 gradients as the first dimension and SDS-PAGE as the second one. We established a set of technique available for intact chloroplast isolation and 2-DE of proteins in wheat floret and compared the effect of intact chloroplast purification using two methods. Based on the 2-DE technique, we explored the difference of the floret chloroplast proteins among the three materials tested in their early uninucleate anther stage. The result showed that we obtained high purity of intact chloroplast using discontinuous sucrose density gradient centrifugation with three-step gradient densities of 30%, 45%, and 60%. Proteins were extracted from chloroplast by TCA-acetone and analyzed by PDQuest software, we got as much as 239 protein spots within pH 4-7 and with molecular weight of 14.4-66.2 kD on each 2-DE gel. We analyzed six differential expressed proteins by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF)mass spectrometry (MS) and bioinformatical database, and found they were acyl-CoA dehydrogenase domain protein, calmodulin-binding protein phosphatase, multiple catalytic peptidase, heat-shock protein 60, light receptor protein 2 and a protein of unknown function. These proteins are involved in series of physiological reactions such as metabolism of energy substances, chloroplast defendance. Chloroplasts signal transduction and plant growth, the differential expression of those proteins among the three materials tested is likely related to the male sterility in wheat.关键词
小麦/遗传型和生理型雄性不育/完整叶绿体蛋白质组/差异表达/2D-PAGE/MALDI-TOF-MSKey words
Wheat/ Genetic and physiological male sterilities/ Intact chloroplast proteins/ Differential expression/ 2-dimensional electrophoresis/ MALDI-TOF-MS引用本文复制引用
李莉 ,王书平 ,张改生 ,王亮明 ,宋瑜龙 ,张龙雨 ,牛娜 ,马守才..小麦生理型和遗传型雄性不育系及其保持系小花完整叶绿体蛋白质组分比较研究[J].作物学报,2011,37(7):1134-1143,10.基金项目
本研究由国家高技术研究发展计划(863计划)重大专项(2009AA101102),国家自然科学基金项目(31071477),高等学校博士学科点专项科研基金(20090204110024),陕西省“13115”科技创新工程重大科技专项(2010ZDKG-68)和西北农林科技大学拔尖人才支持计划项目资助. （863计划）
