羽衣甘蓝胁迫应答基因BoRS1转化烟草的初步研究  

The Study of Transformation of Tobacco with the Stress Responsible Gene BoRS1 from Brassica oleracea var. acephala

作　　者：唐东芹[1] 钱虹妹[2] 赵凌侠[2] 唐克轩[2] 黄丹枫[1] 

机构地区：[1]上海交通大学农业与生物学院,上海201101 [2]上海交通大学复旦交大诺丁汉植物生物技术研发中心,上海2000301

出　　处：《生物工程学报》2005年第3期489-492,共4页Chinese Journal of Biotechnology

摘　　要：将克隆于羽衣甘蓝的胁迫应答基因BoRS1连入中间载体p35S 2 30 0 ::gus ::noster相应位点,成功地构建了含BoRS1基因的植物双元表达载体p35S 2 30 0 ::BoRS1::noster,并通过农杆菌介导法对烟草进行了遗传转化。PCR检测结果表明目的基因BoRS1已成功地导入并整合到烟草基因组中。RT PCR分析显示,在不同的转基因烟草植株中BoRS1表达量存在差异。转BoRS1烟草的耐干性和甘露醇胁迫研究表明,BoRS1基因的表达对提高植物抗干旱胁迫能力有一定的作用。Transgenic tobacco plants expressing a stress responsive gene BoRS1, isolated from Brassica oleracea var. acephala, under the control of the 35S promoter of the Cauliflower mosaic virus were produced. Some plants were further used to test the effect of high level BoRS1 expression on drought stress resistance. The presence of transgene in putative transgenic plants was confirmed by PCR analysis. Thirty-six among 130 transformants showed amplification of predicted fragment of BoRS1 while no amplification was observed in the control. Some transgenic lines confirmed by PCR analysis were analyzed through semi-quantitative one-step RT-PCR for the expression of BoRS1 gene. Amplification of 1.4 kb cDNA product revealed transcription of BoRS1 gene. Meanwhile, differential intensity of the cDNA band indicated variable expression levels of the transgene among different transformed lines. The water loss of detached leaves from the transgenic plants was slower than that of the control. Transgenic tobaccos and the non-transgenic controls were used for further drought stress experiments by using different concentration of mannitol. The transformants showed higher tolerance to drought stress than non-transgenic plants and different transgenic lines exhibited different tolerance during drought stress. These results showed that the BoRS1 gene probably play role in enhancing the ability to drought stress.

关 键 词：BoRS1基因 烟草 干旱胁迫 

分 类 号：S336[农业科学—作物遗传育种]