番茄抗细菌性斑点病的遗传特性和抗性基因的分离及鉴定植物病理学专业论文.docxVIP

华中农业人学硕士学位论文摘要 华中农业人学硕士学位论文 摘要 倍茄细菌性斑点病是一种影响番茄产量和品质的重要病害。发生危害遍及世界各 地。。近年来在我国北方普遍发生。研究表明推广抗性品种是防治该病最经济、最安全 而且最有效的措施。Manin等(1993)已经从抗性品种中克隆到了抗番茄细菌性斑点病基 研究的目标是： 麓 抗细菌性斑点病的遗传特性：(2)从我国番茄品种中获得抗 因Pto。在本研 我国尚未从番茄中克隆到该抗病基因，因此无法开展该基 因的研究工作。 开展了番茄抗细菌性斑点病的研究。f围绕着这一主题，7本 番茄细菌性斑点病基因PW的克隆。 试验中采用番茄抗病品种和感病品种，通过常规杂交及鉴定其子代的抗病性分离 情况，确定在抗性品种红珍珠和美味樱桃中该抗病性呈显性单基因遗传。 根据已报道P，o基因的同源序列设计引物，通过PCR扩增从lO个抗病番茄品种中 获得10个克隆。序列分析比较证明其中两个克隆(xx和BJX)和报道的Pm基因序列完 全一致，表明我们已经从中国的番茄品种中获得了含有该基因的克隆。克隆XX序列 编码区全长936核苷酸对，推导编码311个氨基酸，分子量为34 kDa的蛋白。其余几 个克隆与报道Pm基因序列的同源性在91％以上，因而为Pm基因同源序列。 构建了PfD基因的原核表达载体，转化大肠杆菌BL21(DE3DLysS菌株，成功地实 现了该基因的原核表达，结果产生出了表观分子量为31 kDa的特异性蛋白。 关键词：番茄细菌性斑点病，丁香假单胞杆菌番茄致病变种， 抗性遗传，基因克隆 Pto基因，原核表达 褥茄抗细菌性斑点病的遗传特性和抗性基粥的分离及鉴定Abstract 褥茄抗细菌性斑点病的遗传特性和抗性基粥的分离及鉴定 Abstract Tomato bacterial speck caused by Pseudomonas syringae pv．tomato(PST)is important disease for the production oftomato It has become problem in the north of China recent years．Previous studies showed that breeding for resistance was effective in the suppression of this disease．In order to exploit tomato resistance，a study was undertaken to determine the genetics of resistance to this bacterial disease in tomato varieties collected in China and gene cloning was used to confirm the existence ofthe resistance gene． Genetics of disease resistance was carried out by conventional crossings between resistant and susceptible varieties of tomato．Resistance in F1．F2 or BCl population was identified by inoculations with bacteria of PST．The resultS showed that the resistance in the two tomato varieties of Hong Zhen Zhu and Mei Wei Ying Tao，was controlled by single dormant gene． Two specific primers were synthesized based on the published sequence of the tomato gene Pto responsible for resistance to PST．Polymerase chain reaction was used to amply the resistant genes in the 1 0 resistant tomato varieties collected in China．The amplified DNA fragments were individually cloned into the plasmid vector in strain of Escherichia coli． Sequencings ofthe cloned