黑暗条件下木薯叶绿体比较蛋白质组学初步研究  

A Primary Study of Comparative Proteomics of Chloroplast in Cassava under Dark Condition

作　　者：徐兵强[1,2] 贺庭琪[1,2] 王力敏[1,2] 王丹[2] 孙勇[2] 王旭初[2] 郭安平[2] 

机构地区：[1]海南大学农学院,海南海口570228 [2]中国热带农业科学院热带生物技术研究所,海南海口571101

出　　处：《江西农业大学学报》2015年第2期269-277,共9页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家重点基础研究发展计划(973计划)项目(2010CB12660-2;5);海南省自然科学基金项目(314118)

摘　　要：以"华南8号"木薯叶绿体为研究材料,采用改进酚抽法提取总蛋白,通过一维和二维电泳,经Image Master分析,得到差异蛋白点进行质谱鉴定。结果表明:改进酚抽法可以从木薯叶绿体中提取高质量总蛋白;获得分离性好、分辨率及重复性较高的双向电泳图谱;比较黑暗条件下1,3,5 d木薯叶绿体的蛋白质表达谱,发现186个差异表达蛋白点;经MALDI-TOF MS质谱鉴定,获得28个差异蛋白,这些蛋白主要参与光合作用、碳固定、能量代谢、脯氨酸代谢、胁迫防御等代谢途径。本研究初步阐述了木薯叶绿体在黑暗条件下的蛋白质组变化,在蛋白质水平上鉴定出部分蛋白质的表达可能受光调控。The chloroplast of the cassava cuhivar ＇South China 8＇ （SC8） was used to extract the chloro- plast protein with an improved phenol （BPP） method, and these proteins expressed profiles under dark condition were determined with 1-DE and 2-DE.The obtained typical 2-DE gels were analyzed by ImageMaster software to determine the differential expressed protein spots, and some of them were manually excised for protein in-gel digestion and subsequently identified by MALDI-TOF MS. The main results included：high quality proteins were obtained from chloroplast in cassava using phenol method ; high resolution of 2-DE reference maps with good reproducibility were generated for proteomics;the comparison of the 2-DE gels for chloroplast of the 1,3,5 day under dark treatment found 186 differential protein spots ; twenty-eight proteins were finally identified by MALDI-TOF MS, which were involved in photosynthesis, carbon fixation, energy metabolism, proline metabolism, stress defense, and so on.This study represents a comprehensive dynamic protein profile and light- regulated network of SC8 cassava chloroplast under dark treatment.

关 键 词：木薯叶绿体 暗处理 比较蛋白质组学 质谱 

分 类 号：S533[农业科学—作物学]