小麦BNS雄性不育系及其转换系花药差异蛋白鉴定与分析

doi:10.3724/SP.J.1006.2011.01540

摘要/Abstract

摘要： BNS是一个新发现的温敏核型小麦雄性不育系, 它有良好的不育性和转换性, 被认为在小麦杂种优势利用上有重要价值。为探讨BNS的不育机制, 以不育系及其转换系的单核早期、单核晚期到二核期花药为材料, 用2-DE和MALDI-TOF-MS方法分离鉴定2个系的差异蛋白。结果发现, 在转换系中, ATP合酶α和β亚基、胞质苹果酸脱氢酶、线粒体醛脱氢酶亚基、Rubisco亚基等呼吸、光合能量代谢蛋白表达丰富, 但在不育系中这些蛋白缺失或下调。在不育系中还分离出山梨醇脱氢酶、组蛋白H2B.2、Harpin诱导子1、延伸因子TU等非小麦正常代谢蛋白。根据这些差异蛋白的功能, 推测ATP合酶α、β亚基蛋白最可能是BNS不育的源头蛋白, 它的表达可能受其上游的温度感应子调控。BNS的温度感应子隐性突变后转录启动温度阈值上调, 在较低温下, α、β亚基基因不表达或表达量小, 导致ATP合酶数量减少, 继而影响ATP合成。ATP数量减少, 使小孢子ATP供应短缺, 进一步影响到小孢子的发育, 使代谢异常, 花粉败育。

关键词: BNS小麦雄性不育, 花药差异蛋白, 双向凝胶电泳, MALDI-TOF-MS, ATP合酶

Abstract: BNS is a new type of thermo-sensitive nuclear male-sterile lines in wheat (Triticum aestivum L.). The mechanism of itssterility was studied using two-dimensional electrophoresis and MALDI-TOF-MS methods. Anthers of the sterile line (SL) and the conversion line (CL, fertile pollen rate higher than 50%) were obtained from plants with a series seeding dates, and two set of samples were used to compare the differentially expressed proteins which was extracted from the anthers at early stage of uninucleate and from late stage of uninucleate to binucleate stage of pollen development. The abundant expressions of proteins involved in respiratory and photosynthetic energy metabolism were identified in the CL, such as ATP synthase α and β subunit, NAD (P) Rossmann fold binding protein, cytoplasmic malate dehydrogenase, mitochondrial aldehyde dehydrogenase subunit, and Rubisco subunit protein. However, they were absent or down-regulated in the SL. Besides, a few abnormal proteins were detected in the SL, including sorbitol dehydrogenase, maturase K, histone H2B.2, Harpin-induced 1, and elongation factor TU. According to the functions of these differentially expressed proteins, we inferred that ATP synthase α and β subunits were most likely the source protein for the sterility of BNS. Their expressions are regulated by a temperature sensor that is located at upstream of the subunit gene operon in BNS. Mutation of the temperature sensor is determined by the temperature higher than a threshold, which may startup the transcriptions of α and β subunit genes. When the temperature is lower than the threshold, the expressions of α and β subunits are in very low levels, which affect the assembly of ATP synthase, and result in deficiency of ATP supply for the growth of microspore. This effect can be amplified in downstream physiological functions, such as development of respiratory and photosynthetic enzymes, which finally causes abnormal metabolism and pollen sterility.

Key words: Wheat BNS male-sterility, Anther differentially expressed proteins, Two-dimensional gel electrophoresis, MALDI-TOF-MS, ATP synthase

李友勇, 茹振钢, 苏晴, 付庆云. 小麦BNS雄性不育系及其转换系花药差异蛋白鉴定与分析[J]. 作物学报, 2011, 37(09): 1540-1550.

LI You-Yong, RU Zhen-Gang, SU Qing, FU Qing-Yun. Identification and Analysis of Differentially Expressed Proteins of BNS Male Sterile Line and Its Conversion Line of Wheat[J]. Acta Agron Sin, 2011, 37(09): 1540-1550.

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