摘要
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。
Resistance system induced by Fusarium oxysporum f. sp. niveurn in watermelon was described from the whole level, and expression characteristics of the related genes were analyzed. ＂Calhoun Gray＂, a watermelon variety with high resistance to Fusarium wilt physiological race I was used as the test material, and a cDNA library was constructed by suppression subtractive hybridization （SSH） using root tip tissues sam- pled at 0~6~12,24,36,48 and 72 h after inoculation with race 1 ofF. oxysporum f. sp. niveurn or distilled wa- ter. The induced clones were verified by reverse Northern dot-blot, and 300 positive clones were randomly se- quenced, and the expression of two important genes was studied by RT-PCR. A total of 259 differentially ex- pressed sequences were screened out, and 167 of them were quite similar to the known genes, accounting for 65.5%; Among of them, 64 genes are related to disease resistance and defense, accounting for 24.7%. Blast results showed that the resistance related genes of Calhoun Gray included signal transduction, disease defense, transcription factor, secondary metabolism synthesis, plant cell protection, etc. Further analysis indicated that expression of aquaporin and peroxidase genes were both increased after inoculation with Fusarium wilt physio-logical race 1. The interaction between Calhoun Gray and F. oxysporum involves many aspects, which is similar to that of PI296341. The resistance related genes are mainly those participating in systemic acquired resistance reaction, and genes exclusively expressed in Calhoun Gray but not in PI296341 were gained. These results make a good foundation for further exploration of molecular mechanism between watermelon and F. oxysporum and identification of the key resistant genes.
出处
《植物病理学报》
CAS
CSCD
北大核心
2015年第1期22-32,共11页
Acta Phytopathologica Sinica
基金
广西自然科学基金重点项目(2010GXNSFD013031)
第46批教育部留学回国人员科研启动基金,自治区主席科技资金(1324102)
