转录因子DREB1A基因和Bar基因双价植物表达载体的构建及对马铃薯遗传转化的研究

doi:10.11686/cyxb20140312

摘要/Abstract

摘要： 马铃薯在生长发育过程中受各种生物和非生物胁迫的影响,干旱是其中最常见和危害最严重的非生物胁迫因素之一,常常导致单产不高,总产不稳,严重影响着马铃薯产业的发展。本研究以改善马铃薯抗旱性为目的,采用PCR方法从拟南芥中克隆了转录因子DREB1A基因和诱导型启动子rd29A。利用DNA重组技术成功构建了诱导型启动子rd29A驱动转录因子DREB1A基因和CaMV35S启动子驱动Bar基因的双价植物表达载体pBI121-rd29-BDR,并通过农杆菌介导法对马铃薯进行遗传转化,PPT筛选得到22株抗性苗,PCR和RT-PCR检测证明DREB1A基因已整合到陇薯10号马铃薯基因组中并在转基因植株中转录表达,有望提高转基因马铃薯的抗旱性。目前,作者正在进行转基因马铃薯的抗旱性分析研究。

Abstract: Potato growth and productivity has been extremely affected by drought-stress. It is important to improve drought tolerance of potato to increase yield under stress conditions. To improve drought resistance of potato, the drought response transcription factor DREBIA gene and rd29A promoter were isolated from Arabidopsis thaliana by PCR. Using recombinant DNA technology, the bivalent plant expression vector pBI121-rd29-BDR, which contains two expression cassettes, rd29A-DREB1A-NOS and CaMV35S-Bar-NOS, was successfully constructed. Potato stems were used for the transformation. Excised stems (0.5-1.0 cm) were infected with Agrobacterium LBA4404/pBI121-rd29-BDR (OD₆₀₀=0.5-0.8) and grown on a co-cultivation medium for 3 days in darkness, then placed on callus induction medium plates containing 2 mg/L PPT and 200 mg/L cefotaxime for callus formation. Twenty two resistant seedlings were obtained. The PCR and RT-PCR testing showed that DREB1A was not only integrated into the genome of the potato Longshu 10, but was also expressed in transgenic plants. To verify drought resistance of transgenic potato, analyses were carried out.

中图分类号:

S532.03

贾小霞,齐恩芳,王一航,文国宏,龚成文,王红梅,李建武,马胜,胡新元. 转录因子DREB1A基因和Bar基因双价植物表达载体的构建及对马铃薯遗传转化的研究[J]. 草业学报, 2014, 23(3): 110-117.

JIA Xiao-xia,QI En-fang,WANG Yi-hang,WEN Guo-hong,GONG Cheng-wen,WANG Hong-mei,LI Jian-wu,MA Sheng,HU Xin-yuan. Construction of a bivalent plant expression vector of DREB1A and Bar genes and studies of genetic transformation of potato[J]. Acta Prataculturae Sinica, 2014, 23(3): 110-117.

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