中国农业科学 ›› 2010, Vol. 43 ›› Issue (24): 5000-5008 .doi: 10.3864/j.issn.0578-1752.2010.24.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

玉米花粉与花丝早期互作的蛋白质组学分析

刘怀华,王莉雯,刘楠,刘旭,马侠,宁丽华,张华,崔德周,姜川,陈化榜

  

  1. (山东农业大学农学院/作物生物学国家重点实验室)
  • 收稿日期:2010-05-27 修回日期:2010-08-05 出版日期:2010-12-15 发布日期:2010-12-15
  • 通讯作者: 陈化榜

Proteomic Analyses of the Early Pollen-Silk Interaction in Maize

LIU Huai-hua, WANG Li-wen, LIU Nan, LIU Xu, MA Xia, NING Li-hua, ZHANG Hua, CUI De-zhou, JIANG Chuan, CHEN Hua-bang
  

  1. (山东农业大学农学院/作物生物学国家重点实验室)
  • Received:2010-05-27 Revised:2010-08-05 Online:2010-12-15 Published:2010-12-15
  • Contact: CHEN Hua-bang

摘要:

【目的】分离授粉早期玉米花丝的总蛋白质,为从蛋白质组角度揭示玉米花粉与花丝早期的相互作用奠定基础。【方法】以玉米(Zea mays L.)自交系Ga25为试材,采用三氯乙酸-丙酮法提取总蛋白质,运用双向电泳、质谱分析与检索技术,比较自交授粉后1 h和2 h的花丝蛋白质组的差异。【结果】与授粉1 h的蛋白质组相比,在授粉2 h后的蛋白质组中出现28个差异蛋白点,其中,6个蛋白质点特异表达,19个蛋白质点上调表达,3个蛋白质点下调表达;通过MALDI-TOF-MS质谱测序和MASCOT序列分析,注释了23个蛋白质点,其余5个为未知蛋白;功能分类表明,差异蛋白质分别参与细胞壁合成(21.4%)、防御/抗胁迫(17.9%)、细胞组织、蛋白命运、蛋白合成、转录等细胞过程。【结论】在玉米花粉与花丝早期的相互作用过程中蛋白质组有显著的变化,与授粉后1 h相比,授粉后2 h的蛋白质组中大部分差异蛋白呈上调趋势,并有特异蛋白质出现;分泌型过氧化物酶、膨胀素、果胶甲基化酶抑制蛋白、谷胱甘肽转移酶与未知蛋白4可能与玉米花粉和花丝的早期互作密切相关。

关键词: 玉米, 蛋白质组, 花粉与花丝早期互作, 双向电泳, 质谱分析

Abstract:

【Objective】 The purpose of this project was to study the early pollen-silk interaction in maize through proteomic analysis. 【Method】 Inbred line Ga25 was pollinated and total silk proteins were extracted after 1 h and 2 h pollination. Total proteins were extracted by TCA/acetone, separated by two-dimensional gel electrophoresis (2-DE) and analysed through MALDI-TOF-MS mass spectrometry. 【Result】 Compared with the pollen/silk proteome after 1 h pollination, the proteome after 2 h pollination contained 28 differentially expressed proteins, including 6 specifically expressed ones, 19 up-regulated and 3 down-regulated ones. And 23 of them were identified homologous to known proteins in various databases by MALDI-TOF-MS and MASCOT analysis. The predicted functions of the 23 proteins were related to cell wall biosynthesis (21.4%), defense/stress responses (17.9%), cellular organization, protein fate, protein synthesis and transcription. 【Conclusion】 The pollen/silk proteome profiles were different between the 1 h and 2 h pollination intervals. The majority of differentially expressed proteins were up-regulated; down-regulated and speciafically expressed proteins were also found in the 2 h pollination proteome profile. Secretory peroxidase, expansin, pectin methylesterase inhibitor, glutathione S-transferase and unknown protein 4 may play important roles in early pollen-silk interactions in maize.

Key words: maize (Zea mays L.), proteome profile, early pollen-silk interaction, 2-DE, MALDI-TOF-MS