玉米早期发育阶段粒位效应的蛋白质组学分析  

Proteomics Analysis of Grain Position Effects During Early Developmental Stages of Maize

作　　者：于涛[1] 李耕[1] 刘鹏[1] 董树亭[1] 张吉旺[1] 赵斌[1] 柏晗[1] 

机构地区：[1]山东农业大学农学院/作物生物学国家重点实验室,山东泰安271018

出　　处：《中国农业科学》2016年第1期54-68,共15页Scientia Agricultura Sinica

基　　金：国家自然科学基金(31371576;31401339);国家"十二五"科技支撑计划(2011BAD16B14;2013BAD07B06-2);国家公益性行业(农业)科研专项经费项目(201203100;201203096);山东省高等学校科技计划项目(J14LF10);山东省农业重大应用技术创新课题

摘　　要：【目的】从蛋白质组学的角度研究早期发育阶段玉米上、中部籽粒差异表达的蛋白质,分析其功能,探明玉米粒位发育差异的分子机理。【方法】在大田条件下,以粒位效应显著的玉米品种登海661(DH661)为供试材料,90 000株/hm^2密度下种植,在开花期人工饱和授粉后0、3、6、12 d取果穗上部与中部籽粒。采用TCA-丙酮沉淀法提取籽粒总蛋白,双向电泳分离后获得蛋白质图谱。分别以0、3、6、12 d中部籽粒的凝胶作为参考胶,将上部籽粒凝胶与其进行比对,利用Image master 2D 7.0软件分析籽粒早期发育不同阶段上、中粒位蛋白质表达的差异。通过MALDI-TOF/TOF MS质谱分析及NCBI数据库搜索,对差异表达蛋白质进行鉴定并分析其涉及的生物学功能。【结果】较高密度种植后,果穗籽粒早期发育阶段共检测到超过1000个清晰蛋白质点。通过图像处理软件成对匹配分析,果穗上、中部籽粒早期发育阶段差异蛋白质点为66个,其中52个蛋白质点与NCBI数据库匹配,鉴定率为78.8%。差异蛋白质涉及籽粒呼吸与能量代谢(10个蛋白质点,19%)、胁迫与防御(9个蛋白质点,17%)、蛋白质代谢(9个蛋白质点,17%)、氮代谢(6个蛋白质点,11%)、细胞分化与增殖(5个蛋白质点,10%)、转录与翻译(5个蛋白质点,10%)、次生物质代谢(3个蛋白质点,6%)等功能范畴。对相关的差异蛋白质表达丰度分析,与中部籽粒相比,上部籽粒涉及细胞分化与增殖、呼吸与能量代谢的大部分蛋白质在一个或多个时间段内均显著下调,说明上部籽粒胚乳细胞增殖及呼吸能量代谢能力显著降低。同时,上部籽粒涉及胁迫与防御的多种抗氧化酶系、乙二醛酶1以及涉及蛋白质代谢的4个分子伴侣蛋白质在发育早期也处于低水平表达,说明上部籽粒应对逆境条件防御能力较弱且蛋白质结构不稳定。另外,与中部籽粒相比,上部籽粒氮代谢中丙氨酸转氨酶�【Objective】In order to understand the molecular mechanism of maize grain position effects, the function of differential expression proteins between upper and middle grains during the early developmental stages of maize were studied by using an approach of plant proteomics.【Method】Denghai661（DH661） with significant grain position effects was used as experimental material and planted at 90 000 plants/hm^2 in a field. Upper and middle grains were harvested after flowering artificialsaturation pollination at 0, 3, 6, and 12 d. The total proteins were extracted by the trichloroacetate（TCA）-acetone precipitation method, and the protein profiles were set up by two-dimensional gel electrophoresis（SDS-PAGE）. The upper grain gel was compared to middle grain gel as a reference at 0, 3, 6, 12 d, respectively. The differential expression proteins between the upper and middle grains were analyzed with Image master 2D 7.0. The functions of these differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry（MALDI-TOF/TOF） analysis and NCBI database searching.【Result】After high density planting, grain ears were detected at more than 1000 clear spots at early developmental stages. After a comparative proteomics analysis and MALDI-TOF/TOF mass spectrometry（MS）, 66 protein spots were found significantly differentially expressed between upper and middle grains during the early developmental stages, of which 52 protein spots were successfully matched in the NCBI database and the identification rate was about 78.8%. These proteins were involved in grain respiration and energy metabolism（10 spots, 19%）, stress and defense（9 spots, 17%）, protein metabolism（9 spots, 17%）, nitrogen metabolism（6 sports, 11%）, cell differentiation and proliferation（5 spots, 10%）, transcription and translation（5 spots, 10%）, secondary metabolism（3 spots, 6%）, and other function categories. Analyzing related proteins ex

关 键 词：玉米 粒位效应 早期发育 差异表达蛋白质 蛋白质功能 

分 类 号：S513[农业科学—作物学]