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细菌乙酰胆碱氧化酶基因(codA)在烟草的表达与抗盐能力的分析
Expression of Choline Oxidase Gene (codA) Enhances Salt Tolerance of the Tobacco
【摘要】 将土壤细菌 (A .globiformis)的乙酰胆碱氧化酶 (COD)基因 (codA)通过农杆菌介导转入到烟草中 ,应用抗性筛选获得了抗性植株。PCR检测结果表明 :codA已整合到抗性植株染色体中 ;Western印迹鉴定及金标免疫分子定位的结果表明 :乙酰胆碱氧化酶基因 (codA)在转基因烟草中得到表达 ,表达产物COD定位在叶绿体中。通过对转基因植株的抗盐能力分析 ,结果表明转基因植株比对照植株具有更高的抗盐性 ,其中幼小植株 (1.0~ 1.5cm)可在 40 0mmol/LNaCl的培养基上存活 30天以上 ,并获得具有一定抗盐性状的转基因植株 (T4 40 0 ) ,能在30 0mmol/LNaCl浓度下较好生长 ;较大植株 (6~ 8cm)能在 40 0mmol/LNaCl浓度下较好生长
【Abstract】 The codA gene for choline oxidase, which converts choline into betaine. This enzyme, cloned from a soil bacterium Arthrobactor globiformis, has been transferred and expressed in tobacco by the Agrobatcterium method transformation through the binary plasmid pGAH/codA. The pGAH/codA carried with Km R and Hyg R, and coding sequence for a transit peptide from Rubisco small subunit gene (rbcS) was inserted between 35 S promoter and codA, so that the COD could be introduced into the chloroplast by this transit peptide. The transformed plants were screened on the medium containing the Km and Hyg. PCR, Western and gold immunolocalization tests showed that the codA gene has integrated into the tobacco DNA genome and its protein was expressed and the mature peptide has gone into the chloroplasts by the transit peptide. The results of salttolerance measuring for transgenic plants showed that the transgenic plants were more tolerance to the salt than the control plants. The young transgenic plants (1.0—1.5 cm) could survive at 400 mmol/L NaCl MS medium for more than 30 days. From them the higher tolerance plant T4 400 was obtained, which could grow at the 300 mmol/L NaCl MS medium well. The transgenic plants (6—8 cm) could grow normally at the 400 mmol/L NaCl MS medium while wild plants failed to do so. So the transferred plant with codA enhanced its tolerance to the salt stress.
【Key words】 tobacco; codA gene; transgenic plant; expression; salt resistance;
- 【文献出处】 生物化学与生物物理学报 ,Acta Biochimica Et Biophysica Sinica , 编辑部邮箱 ,2001年05期
- 【分类号】Q786
- 【被引频次】41
- 【下载频次】231