作 者 ：曹云英,段骅,王志琴,刘立军,杨建昌*
期 刊 ：生态学报 2010年 30卷 22期 页码：6009~6018
Keywords:high temperature, rice, heat-tolerant cultivar, leaf, protein expression,
摘 要 ：为明确高温对耐热性不同水稻品种叶片蛋白质表达的影响，以耐热性不同的2个籼稻品种双桂1号（不耐热）和黄华占（耐热）为材料, 分别于苗期、减数分裂期及抽穗 (始穗后0—10 d)和灌浆早期(始穗后11—20 d)进行高温处理，之后取材并采用双向凝胶电泳技术研究高温对不同水稻品种叶片蛋白质表达的影响。结果表明，高温胁迫导致叶片中蛋白质的变化呈4种状况：新蛋白质的产生，一些蛋白质表达量上调，一些蛋白质的表达被抑制，一些蛋白质表达量下调。蛋白质表达变化在两品种以及4个处理时期的表现不同，总体表现为在热敏感品种中表达谱发生变化的蛋白质总数高于耐热品种。质谱分析表明，差异蛋白质主要涉及光合作用和信号转导，该类蛋白质在热敏感品种中表现为不表达或表达量下降，而在耐热品种则表现为有新诱导的蛋白质的产生或表达量上调，表明参与光合作用和信号转导的蛋白质在水稻耐热机制中发挥了重要作用。
Abstract:The global temperature has been continuously increasing in recent decades due to the explosion of population, the development of industries with emission of greenhouse gas, and excessive deforestation. High temperature in summer occurs more frequently and keeps longer in many regions of all over the world, leading to adverse effects on crop growth and development. Rice (Oryza sativa L.) is one of the most important staple food crops in China. The decrease in grain yield due to high temperature stress during the growth period has become a serious problem in rice production, especially in the Yangtze-Huaihe River Valley. Although effects of high temperatures on grain yield and quality have been studied, the mechanism involved in the injury of high temperature to rice growth is not understood. As proteomic approach is a new technique in plant stress physiology in recent years, this study investigated whether and how high temperature stress regulates protein expression in rice leaves. Two indica rice cultivars differing in heat tolerance, Shuanggui 1 (heat-sensitive) and Huanghuazhan (heat-tolerant), were pot-grown at Yangzhou University farm, Yanghzou, Jiangsu Province, China, and treated with high temperature stress at the stages of seedlings (during the 4th leaf, 38℃(day)/30℃(night)), meiosis (from 14 days before heading to the initial of heading, 35℃/25.6℃), heading (0-10 d after heading, 33.4℃/20.9℃) and early grain filling (11-20 d after heading, 33.4℃/20.9℃). The temperatures for controls were 25℃/20℃ at the seedling stage, 32.7℃/25.6℃ at meiosis, 29.9℃/20.9℃ at the heading, and 29.9℃/20.9℃ at the early grain filling stage. The protein expression in the leaf was observed by using two-dimensional gel electrophoresis with immobilized pH 4-7 gradients as the first dimension and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as the second dimension. The results showed that the changes in protein expressions under high temperature stress exhibited four statuses: some proteins were induced, some proteins were up-regulated, some proteins were inhibited, and some proteins were down-regulated. Protein expressions varied with the treatment stages and the two cultivars. Among the four treatment stages, protein expressions induced by high temperature stress were the most at the early grain filling stage, followed by the seedling stage, the third at meiosis, and the least at the heading stage. Protein expressions were more in the heat-sensitive cultivar than in the heat-tolerant cultivar. The mass spectrosmetric analysis indicated that the differences in protein expressions were mainly those involved in photosynthesis and signal transduction, such as Os01g0791600, Os12g0291400, ankyrin-like protein, ribulose-1,5-bisphosphate carboxylase/oxygenase (large subunit), Hypothetical protein OsI_22016, and Rubisco activase small isoform precursor. These proteins were inhibited or down-regulated in the heat-sensitive cultivar Shuanggui 1, but were induced or up-regulated in the heat-tolerant cultivar Huanghuazhan. The results suggest that proteins involved in photosynthesis and signal transduction play a major role in heat tolerance of rice. The possible regulatory mechanism of these proteins in rice leaves in response to high temperature was discussed.
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