论文部分内容阅读
从整体水平阐明西瓜对西瓜枯萎病菌1号生理小种的抗病分子机制和抗病相关基因的表达特征,以高抗枯萎病菌1号生理小种的西瓜品种“卡红(Calhoun Gray)”为试材,接种西瓜枯萎病菌和蒸馏水的根尖组织作为测验方(Tester)和驱动方(Driver),构建西瓜枯萎病菌胁迫的SSH-c DNA正向文库。利用反向Northern斑点杂交技术对文库中克隆进行杂交筛选。随机测序300个阳性克隆,序列比对分析,利用RT-PCR技术分析抗病相关基因的表达特性。259条EST成功测序,167条与已知基因具有较高的同源性,占全部ESTs的65.5%,其中与抗病和防卫相关的有64条23种,占24.7%;卡红对枯萎病菌1号生理小种的抗性相关基因主要涉及抗病信号传导、抗病防卫、转录因子、次生代谢合成和细胞保护等方面;Aquaporin和Peroxidase基因在接种后表达量均增加。Calhoun Gray对枯萎病菌侵染作出的反应是全方位多方面的,抗病相关基因主要集中在系统获得性抗性反应中,获得了一些Calhoun Gray与野生西瓜PI296341在与枯萎病生理小种1互作中差异表达的基因,为深入研究西瓜与枯萎病菌互作的分子机制以及关键基因的功能分析奠定基础。
From the overall level of watermelon Fusarium oxysporum fumigatus No. 1 physiological races resistant molecular mechanism and resistance-related gene expression characteristics of high resistance to Fusarium wilt No. 1 physiological races of watermelon varieties "Calhoun Gray As the test material, the root tip tissues of Fusarium oxysporum f. Sp. And distilled water were inoculated as Tester and Driver to construct the SSH-c DNA positive library of Fusarium oxysporum f. Hybridization screening of clones in the library was performed using reverse Northern blot hybridization. Randomly sequenced 300 positive clones, sequence alignment analysis, the use of RT-PCR analysis of resistance-related genes expression characteristics. 259 ESTs were successfully sequenced and 167 had high homology with known genes, accounting for 65.5% of all ESTs, of which 64 were resistant to disease and defense, accounting for 24.7%, accounting for 24.7% The resistance-related genes of Physiological Clone 1 mainly involved in resistance signal transduction, disease resistance and defense, transcription factor, secondary metabolism and cytoprotection. The expression of Aquaporin and Peroxidase genes increased after inoculation. The response of Calhoun Gray to Fusarium wilt was comprehensive and varied. The genes related to disease resistance were mainly concentrated in the system-acquired resistance. Some Calhoun Gray and wild watermelon PI296341 The differentially expressed genes laid the foundation for the further study on the molecular mechanism of interaction between watermelon and Fusarium wilt and the functional analysis of key genes.