天麻与蜜环菌相互作用中差异表达蛋白质的研究
其他题名Differentially expressed proteins in the interaction between Gastrodia elata and Armillaria mellea
张晓勤
学位类型硕士
导师曾英
2003
学位授予单位中国科学院昆明植物研究所
学位授予地点中国科学院昆明植物研究所
学位专业植物学
关键词蛋白质组学 植物蛋白质组 天麻 蜜环菌 双向电泳 图像分析 质谱鉴定
摘要本论文分两部分,第一部分为综述。通过查阅多篇文献详述了当前蛋白质组学研究技术方法进展,以及植物蛋白质组学方面的应用进展,包括在植物组织器官与亚细胞蛋白质组的研究、植物遗传学与生理学方面的研究及蛋白质组信息学的进展,并展望了植物蛋白质组学的研究趋势。第二部分为实验论文。兰科植物天麻的生长依赖于蜜环菌的侵染,只有当天麻的营养茎与蜜环菌建立营养关系,才能形成次生球茎并生长发育,当年生次生球茎不被蜜环菌侵染,前人已分离出天麻抗真菌蛋白(GAFP)并研究了该蛋白在天麻防御过程中的作用。但是在两者复杂的关系中可能还有其它的表达产物扮演重要的角色,通过蛋白质组分析可进一步理解两者间相互作用的分子机理。采用蛋白质组学研究的基本技术一双向聚丙烯酰胺凝胶电泳(2D PAGE),对天麻的营养茎和次生茎以及蜜环菌的蛋白质进行分离。通过改善制备样品的方法和优化电泳方式及参数,得到了比较理想的双向电泳图谱,用Melanie图像分析软件对天麻营养球茎与次生球茎的固相pH梯度(IPG)双向电泳图像进行初步的比较,营养球茎和次生球茎的蛋白点数分别约为889和901个,两者约52%的蛋白点相匹配。图谱上有一些点不能匹配,而一些匹配的点在表达量上也存在很明显的差异,这些蛋白点被称为差异表达的蛋白质。对蜜环菌双向电泳图谱分析,检测出蛋白点数约为926个,天麻营养茎和蜜环菌图像比较基本上找不到可以匹配的蛋白点。对部分在次生球茎中增量表达的蛋白点用质谱(MALDI-TOF-MS)进行肽质量指纹谱(PMF)鉴定,并在数据库中进一步检索同源性,因当前数据库中植物种类及条目有限,检索到的蛋白同源性不是很高,随着各种数据库的充实,相信这些“未知”蛋白会成为“已知”,同时这些蛋白质重要的生理功能也将被揭晓。
其他摘要This thesis included two parts. The first part was the review about the progress of proteomics and plant proteome, focusing on technological aspects of proteome analysis including sample preparation, protein identification and protein-protein interactions, and recent advances in plant proteomics such as organ and subcellular proteome, applications to genetic and physiological studies as well as proteomic information were also presented. The second part was the experiment. The growth of Gasti'odia elata depended on the invading by Armillaria mellea, and Gastrodia antifungal protein (GAFP) was separated and studied which maybe played an important role in the symbiosis. But other proteins which haven't been investigated probably participated in the symbiosis. So proteomics was adopted in this thesis to further study. Proteins extracted from nutritive and secondary corms of Gastrodia elata and Armillaria mellea were separated by two dimensional polyacrylamide gel electrophoresis (2D PAGE), followed by Melanie image software analysis. Parameters for sample preparation and running electrophoresis were critically investigated. Approximately 900 protein spots were detected in each 2D PAGE gel, and 52% spots were matched well between nutritive and secondary conns, but there were no matched spots between nutritive corm and Armillaria mellea. Some increased displayed proteins from secondary corms of Gastrodia elata in comparison with nutritive corms were identified by MALDI-TOF-MS, followed by databases homologous searches. With the insufficiency of current databases, the proteins were still in ambiguous attributes.
页数56
语种中文
文献类型学位论文
条目标识符http://ir.kib.ac.cn/handle/151853/646
专题昆明植物所硕博研究生毕业学位论文
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张晓勤. 天麻与蜜环菌相互作用中差异表达蛋白质的研究[D]. 中国科学院昆明植物研究所. 中国科学院昆明植物研究所,2003.
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