水稻OsCERK2转基因后代的鉴定与分析

Identification and Analysis of OsCERK2 Transgenic Progeny of Rice

为了研究水稻Os CERK2基因的功能,构建了Os CERK2基因过表达和RNAi载体,利用农杆菌介导的遗传转化获得转基因植株,经多代筛选和分子检测,获得了4个超量表达的过表达纯系和2个转录水平下降的RNAi纯系。转基因植株经病原微生物细胞壁成分喷雾处理,采用半定量RT-PCR方法分析转基因植株中病程相关(PR)基因的表达情况,结果发现,诱导后过表达植株中PR基因表达增强,而RNAi植株中表达下降;抗病性鉴定结果表明,过表达转基因植株对白叶枯菌致病小种PXO99的抗性增强,而RNAi植株与野生型差异不显著。结果表明,过量表达Os CERK2基因可能是水稻抗病的有效途径。

To study the biological function of OsCERK2 gene, we constructed the OsCERK2 gene overexpression, and RNA interference vectors by modifying pCAMBIA1300 and pTCK303 as framework plasmid, and then trans- ferred the vectors respectively into the rice through Agrobacterium-mediated genetic transformation. Molecular detection produced four overexpressing homozygous lines and two RNAi homozygous lines with reduced transcrip- tional level. The transgenic plants of the OE lines and RNAi lines were sprayed with peptidoglycan prepared from the cell wall of both race PXO99 （Xoo- R6） of Xanthomonas oryzae pv. oryzae （PGNXoo） and the strain JB01- 25 of X. oryzae pv. oryzicola （PGNXoc） and ChitinMo from the cell wall of Magnaporthe oryzae, with water as control. The expression of pathogenesis related （PR） genes were determined by using the semi-quantitative RT- PCR. The results showed that the plants of overexpressing lines improved the expression of their PR genes while the plants of RNAi expressing line reduced the expression of the PR genes, as compared with the nontransgenic rice. Furthermore, the plants of overexpressing transgenic lines exhibited high tolerance to PXO99, but there were not significantly different between the RNAi plants and the wild type. These results indicated that OsCERK2 overexpressing gene may be an effective way to disease tolerance of rice.