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OsbHLH120是一个参与非生物胁迫响应基因,与水稻的抗旱性密切相关。本研究根据RNAi表达载体设计原则,从水稻日本晴基因组DNA扩增OsbHLH120基因片段,将长度400 bp干扰目的片段通过正反两个方向与载体p TCK303连接。从pTCK303载体的启动子、终止子和Rice Itron结构中设计两对检测引物,直接将检测引物的PCR产物测序验证载体是否构建成功。通过农杆菌介导法将构建的OsbHLH120RNAi干扰表达载体导入水稻品种日本晴,获得18株转基因阳性苗。本研究结果将有利于进一步研究OsbHLH120在水稻抗旱过程中的作用。
OsbHLH120 is a gene involved in abiotic stress response and is closely related to drought resistance in rice. In this study, OsbHLH120 gene fragment was amplified from Nipponbare genomic DNA according to the design principle of RNAi expression vector and the fragment of 400 bp in length was ligated to vector pTCK303 in both forward and reverse directions. Two pairs of detection primers were designed from the promoter, terminator and Rice Itron structure of pTCK303 vector, and the PCR products of the detection primer were directly sequenced to verify whether the vector was successfully constructed. The constructed OsbHLH120 RNA interference expression vector was introduced into the rice variety Nipponbare through agrobacterium-mediated method, and 18 transgenic positive plants were obtained. The results of this study will be helpful to further study the role of OsbHLH120 in drought resistance of rice.