OsbHLH1基因过表达载体构建及转化水稻研究

摘要

OsbHLH1基因编码bHLH类转录因子,与水稻耐寒性相关.以pCAMBIA3300为母体,利用玉米泛素(Ubiquitin)启动子构建了能使OsbHLH1基因过表达且含有Bar基因的植物表达载体p3300-Ubi-Ω- OsbHLH1.利用基因枪法将其转化到粳稻品种东稻3号中,获得再生苗47株.选取其中9株进行PCR、Southern检测,结果表明目的基因已经整合到水稻基因组中;草铵膦叶片涂布试验结果表明,Bar基因在水稻植株中正常表达;低温处理后,除45号植株外,其余8株光合速率的变化率显著低于非转基因对照,初步证明OsbHLH1基因在水稻中过表达显著提高了水稻的耐低温能力.%OsbHLH1 gene which codes one of transcription factors of the bHLH family,is involved in cold tolerance. An over-expression vector including OsbHLH1 gene and Bar gene was constructed based on pCAMBIA3300. The target genes were transformed into Dongdao number 3 by particle bombardment. Forty seven plantlets were regenerated.Nine of them were analyzed by PCR and Southern blotting, and the results showed that the target genes had been integrated into the rice genome. The spreading of glufosinate on the leaves indicated that the Bar gene had been expressed normally in rice. Under low temperature treatment, the changes of net photosynthetic rate of the regenerated plantlets, except plantlet number 45, were significantly lower than that of check, which showed that over-expression of OsbHLH1 gene could be able to enhance the cold tolerance of rice.