【作者】 马凯；

【导师】 李凤兰； 高述民；

【作者基本信息】 北京林业大学 ， 植物学， 2004， 硕士

【摘要】 文冠果（Xanthoceras sorbifolia，Bunge．）是中国北方特有的木本油料植物，具有较高的药用价值。文冠果属杂性花，雄花的花药是可育的，两性花的花药是不育的。本论文通过双向电泳方法对文冠果不同时期两性花和雄花花药蛋白进行差异分析，并对差异蛋白在花器官中的存在和分布做了进一步研究，为深入研究雄性不育基因及表达调控奠定基础。 1．利用双向电泳技术对文冠果蕾期和开花期两性花花药和雄花花药的蛋白进行分离和差异分析，发现：在蕾期，两种类型花药蛋白无差异；至开花期，至少有4个蛋白(B_1-4)的表达存在差异。 2．利用制备型双向电泳技术分离、纯化、富集特异蛋白B₁（49.9kD），以该蛋白为抗原免疫新西兰兔，制备多克隆抗体，ELISA检测效价为500，Western Blot结果显示，该抗体具有较高的特异性。利用免疫组织化学分析方法，对文冠果不同时期两性花花药和雄花花药内B₁的存在和分布进行了研究，结果显示：B₁分布在花药的绒毡层中。推测B₁是一个影响绒毡层发育，影响雄蕊育性的重要蛋白。更多还原

【Abstract】 Xanthoceras sorbifolia Bunge. is the specific oil plant in north of China and has great officinal value. The flower of Xanthoceras sorbifolia Bunge. is polygamous. The anther of male flower is fertile and that of bisexual flower is sterile. In order to study male sterility-associated genes and their expressions and control, in this dissertation, the differences of anther’s proteins between the bisexual and male flower of Xanthoceras sorbifolia Bunge. were analyzed by two-dimensional electrophoresis. And the distribution of special protein was studied by immunohistochemical localization.1. The anther’s total proteins in the bisexual and male flower of Xanthoceras sorbifolia Bunge. from bud stage to florescence were analyzed by two-dimensional electrophoresis. The result showed: the proteins of two types of anther in bud stage had no difference, however there were at least 4 proteins (B1-4) expressed differently in florescence.2. The special protein B1 (49.9kD) was isolated, purified and gathered by preparative two-dimensional electrophoresis. And then it was used to immunize New-Zealand rabbit to prepare polyclonal antibody. The result of ELISA showed that the antibody titer reached 500. Western blotting assays showed that the antibody obtained was effective, specific and useful for further study. The immunohistochemical localization of B1 was carried out for the first time in two types of flowers of Xanthoceras sorbifolia Bunge.. The result showed that B1 distributed in tapetum. So it is speculated that B1 is an important protein related to the development of tapetum and male sterility.更多还原