【作者】 肖勇；

【导师】 孟金陵；

【作者基本信息】 华中农业大学 ， 发育生物学， 2006， 硕士

【摘要】 菌核病(stem rot)是一种世界性病害，该病害严重影响油菜产量，在长江流域尤为严重，而且目前尚未发现对菌核病完全免疫的材料，所以寻找抗菌核病机理和培育抗病品种是油菜研究中急需解决的问题之一。 本文主要分为两个部分：第一部分工作，对芯片结果进行Northern验证，筛选与菌核病抗性相关的基因。第二部分工作：对实验室自行合成的新型甘蓝型油菜以及收集的常规品种种质资源进行菌核病抗性鉴定，筛选高抗油菜种质资源，为提高甘蓝型油菜抗菌核病性打下基础。 第一部分工作 刘仁虎博士利用拟南芥的cDNA芯片(包含9120个EST，大约6200个基因)研究了油菜受核盘菌和草酸胁迫下，局部组织基因的表达变化情况。本实验在此研究结果基础上，结合赵建伟(教授)利用F2-3所定位的菌核病抗性相关QTL，通过油菜-拟南芥比较作图，我们拟定了13个与菌核病抗性相关的候选基因。对部分基因Norhtern验证，其Nothern杂交的结果与芯片结果不一致。 对抗病材料宁RS-1和感病材料恢5200的活体叶片和离体叶片进行核盘菌的接种实验，实验结果显示：抗病亲本宁RS-1叶片上椭圆面积与感病亲本恢5200叶片上的椭圆面积没有显著差异。 第二部分工作 本实验的主要目的是对实验室收集和通过远缘杂交合成的材料进行菌核病抗性评估，筛选对菌核病具有较高抗(耐)性的材料。研究内容可以分为以下几个部分。 1．对新型甘蓝型油菜合成途径Ⅰ(如图5)中所需的109个埃芥(Brassica carinata)亲本株系进行菌核病抗性鉴定，选取中双9号为抗性对照，两次重复实验结果显示：三个埃芥亲本株系的菌核病抗性显著强于对照中双9号。 2．对新型甘蓝型油菜合成途径Ⅰ(如图5)中所需的埃芥亲本衍生的5倍体和6倍体材料进行菌核病抗性鉴定，五次重复实验结果显示：一个6倍体单株的菌核病抗性显著强于对照中双9号。取该抗性单株的花粉，观察细胞核内染色体数目，结果显示：其细胞内染色体数目为40条。所以该材料并非整6倍体。 3．对实验室自行合成的新型甘蓝型油菜和收集的常规品种种质资源的离体叶片进行核盘菌的接种实验。结果显示：有5个新型甘蓝型油菜叶片上的病斑椭圆面积略小于对照中双9号，没达到统计学上的显著水平。更多还原

【Abstract】 The paper is separated into two parts. In the first work, my work focus on making use of Northern blot to validate the result of microarray. In the second work, my works focus on screening out resistant cultivars, and contributes to the resistant breeding purpose of rapeseed.Part ⅠThe result of Doctor Liu’s microarray combined with the QTL related to resistance to Sclerotinia sclerotiorum was analyzed. Through this combinated analysis, we screened 13 candidate genes that may be correlated to resistant response to Sclerotinia sclerotiorum. The expressions of these genes were validated by Northern blot. The results of the experiment show that the candidate genes’ expression appeared no singinificant change between resistance materials and susceptible materials. Parents , NRS-1 and H5200, are inoculated to assess the level of resistance to Sclerotinia sclerotiorum. The experiment shows that the lesion areas between resistant parent NRS-1 and susceptible parent H5200 have no significant different.Part ⅡThe aim of this part of the experiments is to assess the resistance to Sclerotinia sclerotiorum for a large number of materials, inclunding new-type Brassica napus and general cultivars. Screen out many materials that show good resistance to Sclerotinia sclerotiorum. The content of the research could be separated into four parts as follows.1. Evaluate 109 Brassica carinata cultivars’resistance to Sclerotinia sclerotiorum. Zhongshuang 9 is select as a control. Zhongshuang 9 is a resistanct material. The results of the experiment show three Brassica carinata cultivars are significantly more resistance than Zhongshuang 92. Resistance to Sclerotinia sclerotiorum of some materials is evaluated, including Hexaploids and pentaploids derivate from the 110 Brassica carinata. The results of the experiments identified one Hexaploid which is significant more resistant than Zhongshuang 9.3. Some new-type Brassica napus and general cultivars are inoculated to assess their更多还原