Na^+/H^+逆向转运蛋白基因PsnhaA的克隆及在大豆中的功能验证  

Cloning of Na^+/H^+ Antiporter Gene PsnhaA and Analysis on Function in Soybean

作　　者：罗茜[1] 张海玲[2] 徐香玲[1] 姚琳[1] 王全伟[1] 

机构地区：[1]哈尔滨师范大学生命科学与技术学院/黑龙江省分子细胞遗传与遗传育种重点实验室,黑龙江哈尔滨150025 [2]黑龙江省农业科学院,黑龙江哈尔滨150086

出　　处：《黑龙江农业科学》2015年第5期6-12,共7页Heilongjiang Agricultural Sciences

基　　金：黑龙江省自然科学基金重点资助项目(ZD201003);哈尔滨市科技创新人才研究专项资金资助项目(RC2013QN002103);黑龙江省教育厅科学技术研究资助项目(12511154);哈尔滨师范大学博士科研启动基金资助项目(KGB200903)

摘　　要：高盐对作物造成渗透胁迫和离子毒害,Na+/H+逆向转运蛋白是生物耐盐的关键因子,能够维持高盐胁迫下生物体的正常生长代谢。为促进作物的遗传转化,选育耐盐作物品种,从施氏假单胞菌(Pseudomonas stutzeri)中克隆质膜Na+/H+逆向转运蛋白基因PsnhaA,并构建到植物表达载体pBI121上,通过发根农杆菌介导的整株生长点注射法转化大豆。结果表明:PsnhaA基因已整合到转化大豆基因组中,并在转录水平获得表达。耐盐相关生理指标检测结果显示,盐胁迫后转基因植株的质膜相对电导率显著低于对照植株,叶绿素含量和脯氨酸含量显著高于对照植株。PsnhaA基因显著提高了大豆的耐盐水平,为农作物的耐盐性改良提供了优良的候选基因。In order to promote crop genetic transformation, breed salt-tolerant crop varieties, the osmotic stress and ion toxicity caused by high salt is one of the major abiotic stress factors that affect the crop growth and development. Na＋/H＋ antip0rter is the key factor in the salt-stress tolerance in organism. It can maintain normal growth and metabolism of organism under high salt stress. A plasma membrane Na＋/H＋ antiporter gene nhaA was cloned from Pseudornonas stutzeri,and the plant expression vector pBI121 was constructed. PsnhaA gene was transformed into soybean growing point via Agrobacterium- mediated transformation. The results showed that PsnhaA was integrated into the soybean genome, and could transcribed. Salt resistance analysis showed that the relative electronic conductivity of the transformed plants plasma membrane was significantly lower than that of the control under salt stress. While the content of chlorophyll and proline in the transformed plants were significantly higher than that in the controls. The expression of PsnhaA increased the salt stress tolerance of the transgenic soybean and provided excellent candidate genes for improving salt tolerance of crops.

关 键 词：NA+/H+逆向转运蛋白 克隆 大豆 遗传转化 

分 类 号：Q943.2[生物学—植物学] S565.1[农业科学—作物学]