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Inheritance of resistance in lettuce to Plasmopara lactucae-radicis and expression of the beet curly top virus coat protein gene in transgenic tobacco.

机译:生菜中抗疟原虫的抗性遗传和甜菜卷曲顶病毒外壳蛋白基因在转基因烟草中的表达。

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摘要

The research presented in this dissertation involves the study of two systems in order to analyze and develop resistance in plants to pathogens. The first study considered the interaction between lettuce (Lactuca sativa L.) and Plasmopara lactucae-radicis, a recently described casual agent of downy mildew. This unique fungus is the only known casual agent of downy mildew that is restricted to the roots of its host. Thirty-eight lettuce cultivars were screened for resistance to P. lactucae-radicis. Two-wk-old lettuce plants grown hydroponically were challenged with this fungus and evaluated 2 and 3 wk after inoculation for resistance. Root necrosis and production of sporangia on roots was considered a susceptible reaction. Five cultivars were determined to be resistant to this fungus. Resistant cultivars, however, were colonized by the fungus but did not support the production of sporangia on roots. Data from F2 and F3 progenies demonstrated that resistance was conferred by a single recessive allele (plr). Fungal infection of susceptible plants resulted in significant decreases in fresh root and shoot weights and leaf number compared to decreases associated with infection of resistant plants. RFLP probes have been identified for mapping the plr gene. The second topic of this dissertation dealt with the development of transgenic tobacco plants that express a chimeric gene that consisted of the 35-S promoter and the coat protein gene of Beet Curly Top Virus, a member of the geminivirus group. Expression of viral coat protein genes in plants has resulted in resistance to the virus from which the coat protein gene was obtained. This type of "coat protein-mediated" resistance has not been demonstrated for any geminivirus. Tobacco leaf discs were inoculated with an Agrobacterium tumefaciens line that contained the chimeric gene. Three transgenic lines were determined by Southern and Northern analysis, and ELISA, to express the chimeric gene and produce coat protein. Trials to determine the resistance of these plant lines to BCTV are pending federal approval.
机译:本文的研究涉及两个系统的研究,以分析和发展植物对病原体的抗性。第一项研究考虑了莴苣(Lactuca sativa L.)与最近描述的霜霉病临时病原体Plasmopara lactucae-radicis之间的相互作用。这种独特的真菌是唯一已知的霜霉病临时病原体,其仅限于寄主的根部。筛选了38个莴苣品种对乳酸假单胞菌的抗性。用这种真菌攻击以水培方式生长的两周龄的莴苣植物,并在接种后2周和3周评估其抗性。根部坏死和根部孢子囊的产生被认为是易感反应。确定了五个品种对该真菌具​​有抗性。但是,抗性品种被真菌定殖,但不支持根部产生孢子囊。来自F2和F3后代的数据表明,抗性由单个隐性等位基因(plr)赋予。与抗性植物感染相关的减少相比,易感植物的真菌感染导致鲜根和苗重以及叶数的显着减少。已经确定了RFLP探针可定位plr基因。本论文的第二个主题涉及表达嵌合基因的转基因烟草植物的开发,该嵌合基因由35-S启动子和甜菜卷曲顶病毒(双生病毒组的成员)的外壳蛋白基因组成。病毒外壳蛋白基因在植物中的表达导致了对获得外壳蛋白基因的病毒的抗性。这种“外壳蛋白介导的”抗药性尚未在任何双生病毒中得到证实。烟草叶盘用含有嵌合基因的根癌农杆菌系接种。通过Southern和Northern分析以及ELISA确定三个转基因品系,以表达嵌合基因并产生外壳蛋白。确定这些植物系对BCTV抗性的试验尚待联邦政府批准。

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