Pi-ta^+和Bchi基因双价表达载体构建及水稻的遗传转化

Construction and Genetic Transformation of Pi-ta^+ and Bchi Genes Co-expressing Vector in Rice

稻瘟病是水稻上最重要的病害之一。本研究所用Pi-ta+基因是一个来自景洪直立型紫秆普通野生稻的基因,该基因属于稀有的抗稻瘟病Pi-ta+等位基因,具有抗稻瘟病的能力;几丁质酶是一种以几丁质为底物的水解酶,它可以降解病原真菌细胞壁中的几丁质,破坏细胞新物质的沉积使病原体死亡。为获得抗稻瘟病的水稻新材料和进一步培育聚合多个抗稻瘟病基因的转基因水稻新品种,本研究采用一种新的方法构建了Pi-ta+和Bchi的双价表达载体pCAMBIA1300-Pi-ta+-Bchi,通过农杆菌介导法转入栽培稻品种云资粳41中,经过一定浓度的甘露糖筛选培养,分化得到的再生苗经氯酚红显色法和PCR检测,获得11棵转基因阳性植株,离体检测证明外源基因的导入增强了转基因水稻的稻瘟病抗性。

Rice blast is one of the most serious diseases in the rice.Pi-ta＋ gene has the ability to blast resistance.The Pi-ta＋ gene has been proved to be a rare existing Pi-ta＋ allele from Jinghong erect type of common wild rice（Oryza rufipogon Griff） in this study.Chitinase can degrade the chitin in pathogenic fungal cell wall and destroy the deposition of new cell material causing pathogen death.The gene encoding Chitinase can help to control rice blast disease.In order to obtain new materials of rice resistant to blast disease,aggregating more blast resistance genes in genetically modified rice varieties is necessary.In this study,a new method was used to build a co-expressing vector pCAMBIA1300-Pi-ta＋-Bchi which was transformed to calli of rice cultivar ＇Yunzijing 41＇ via Agrobacterium tumefaciens system.Eleven transgenic plants were got after screening with a certain concentration of mannose,through CPR assaying and PCR analysing.In vitro testing proved that transgenic plants enhanced the resistance to Piricularia oryzae Cav.