RNA干涉水稻端粒酶再生植株的初步研究

Preliminary study on the effect of RNA interference telomerase on rice

依据水稻端粒酶基因的相关生物学信息,构建了含有水稻端粒酶序列RNA干扰结构的植物表达载体pC am 23A-1-2,并利用农杆菌介导法将目的基因导入到粳稻品种日本晴中,获得了78个独立转化子,共165棵转基因植株。通过PCR和Southern杂交分析,证明RNA干扰结构已整合进水稻基因组中;应用染色体末端限制性片段分析法,显示在转基因水稻中端粒DNA序列长度有逐代缩短的趋势,初步证明这些转基因水稻中端粒酶亚基已失活,但是T0和T1代转基因水稻植株的主要农艺性状没有发生明显变化。

According to the biological information of rice telomerase, the vector pCam23A-1-2 containing RNAi structure of telomerase was constructed and the target gene was transferred into the japonica rice cultivar, Nipponbare mediated with Agrobacterium. Many transgenic plants were obtained. PCR and Southern blotting analysis indicated that the T-DNA had been integrated into the genome of the transgenic rice plants. Terminal restriction fragments （TRF） analysis showed that the length of telomere DNA was getting short gradually in T0 and T1 transgenie rice plants. However, no significant variance was detected in the main agronomical characters of the transgenic rice both in To and Tt generation.