【作者】 郭晓瑞；

【导师】 祖元刚；

【作者基本信息】 东北林业大学 ， 植物学， 2004， 硕士

【摘要】 本文主要运用蛋白质组学的技术和方法，研究在高温胁迫过程中，外源ABA对长春花幼苗耐热性提高方面的影响及其蛋白质组学基础，揭示了ABA作为重要植物抗逆调节物在提高植物耐热性方面具有重要的意义。 外源ABA处理能维持SOD和POD的活性，有效降低膜脂过氧化作用；叶绿素含量测定结果表明，外施ABA预处理可以缓解热胁迫对长春花幼苗叶绿素a、b的破坏。 确立了长春花幼苗叶片总蛋白的最佳提取方案和用于双向凝胶电泳的蛋白定量方法。在此基础上，运用双向凝胶电泳技术对长春花幼苗叶片热胁迫过程中蛋白表达情况进行分析，并对其中32个表达差异明显的蛋白点进行肽质量指纹谱鉴定，有10个蛋白点的鉴定结果较好，其中3个为小分子量热激蛋白：HSP33，HSP18.2和HSP20.0。通过对该3个蛋白点在不同凝胶上的表达情况进行分析，发现HSP33和HSP18.2在热激处理条件下被监测到，HSP20.0在外施ABA后进行热激处理过程中被监测到，进一步分析发现HSP18.2和HSP20.0均为组成型表达，且在热胁迫条件下HSP18.2的表达量先升高后明显随热胁迫时间的延长而下降，而外施ABA后HSP20.0的表达量随热胁迫时间的延长逐渐升高。更多还原

【Abstract】 The unpretreated and ABA pretreated Cantharanthus roseus saplings were exposed to high temperature stress. The physiological indexes correlative to thermo-tolerance were studied in the laboratory, and the expression of total proteins were also detected in the study. Through these we attempted to probe into the mechanism of ABA on Cantharanthus roseus saplings during the heat stress. The result as follows:The ABA pretreated can maintain the activity of SOD and POD, reduce the excessive oxidation of membrane lipid effectively and maintain the stability of membrane structure, also can release the heat injury of chlorophyll a and chlorophyll b.The optimal program for total proteins extraction of Cantharanthus roseus saplings’ leaves were selected and the quantification method for proteins from 2-DE were established. The protein expression was analyzed and 32 differentially expressed points were identified through PMF. 10 points were identified better, and 3 points were small heat shock proteins. They were HSP33, HSP18.2, HSP20.0 respectively. Having analyzed the expression of the three points in different gels, we found HSP33 and HSP18.2 could be detected under heat stress without ABA pretreated. while HSP20.0 could be detected under the heat stress with ABA pretreated .The further analyze made it clear that HSP18.2 and HSP20.0 were both constitutively expressed. During the heat stress, the expression of HSP18.2 was up-regulated at the beginning, and decreased markedly with the prolongation of heat stress time. On the contrary, the expression of HSP20.0 rose with the elongation of heat stress time.更多还原