双价抗真菌基因表达载体的构建及转基因西瓜的研究

Construction of a Plant Expression Vector Carrying Two Antifungal Genes and Its Transfer to Watermelon

摘要:

构建了同时含有番茄儿丁质酶基因（Chi3）和β-1，3-葡聚糖酶基因（Glu-Ac）的双价抗真菌基因植物表达载体，以西瓜（Citrullus lanatus）子叶块为外植体，采用根癌农杆菌（Agrobacterium tumefaciens）介导法，将Chi3和Glu-Ac同时导入西瓜栽培种“中育一号”，共获得46株抗性再生植株。经PCR、Southern blot和RT—PCR检测，表明外源基因己成功整合到西瓜基因组中，并在转录水平得到表达。利用尖孢镰刀菌西瓜专化型（Fusarium oxysporum）对转基因植株进行离体叶片抗病性检测，表明转基因植株对枯萎病的抗性均有不同程度的增强。

Abstract:

Using plant bivalent expression vector pCAMBIA1300 and pBI121 as basic components, a new binary vector harboring tomato chitinase gene Chi3 and tomato β-1,3-glucanase gene Glu-Ac, was constructed through sequential restriction digests and ligations recombination. The two genes driven by CaMV35s promoter were successfully transferred into watermelon cultivars “ZhongYuYiHao” via Agrobacterium-mediated transformation. Successful integration and expression of Chi3 and Glu-A c were confirmed by PCP,Southern blot analysis and RT-PCR. The reaction of detached leaves of transgenic plants to infection Fusarium oxysporum infection was examined. The results indicated that transgenic plants gained certain resistance.