农杆菌介导的水稻双基因共转化初步研究

利用农杆菌介导法将分别位于不同质粒上的gus基因和nptⅡ基因按照1︰1和1︰3的浸染浓度混合共转化粳稻品种鄂宜105和中花12种子胚诱导的愈伤组织，结果从43棵经潮霉素反复筛选的再生植株中获得21棵共转化植株。经PCR分析表明2个基因的共转化率分别为47.37%和60.00%。Southern Blot和Western Blot检测发现gus基因已经整合到水稻基因组中并且表达。遗传分析证实外源基因在T1中大多以孟德尔方式进行遗传。分析了影响转化率的几个因素如浸染菌量比例和品种之间的关系以期进一步提高共转化的效率。

A Preliminary Study on Agrobacterium Tumefaciens-mediated Co-transformation of Double Genes in Rice

Gus and nptⅡ genes were constructed separately with their respective vectors and co-transformed into mature seed-derived calli of Japanica rice cultivars Eyi105 and Zhonghua 12 with a strain dosage ratio of 1:1 and 1:3 respectively through Agrobacterium tumefaciens-mediated transformation. 21 transgenic plants contained gus gene were gained from 43 regeneration plants. The results of PCR showed that co-transformation efficiency of these two genes were 47.37% and 60.00%, respectively. The results of Southern Blot and Western Blot analysis showed that Gus has been integrated into rice genome. Genetic analysis confirmed Mendelian segregation was popular among T1 plants in progeny. In this study, some key problems were discussed such as the relation of transformation rate and strain dosage percent, some way will be help to support the mutualism effect.