表达细菌阿特拉津氯水解酶基因的转基因烟草对土壤中阿特拉津的生物降解  

Biodegradation of Atrazine in Soil by Transgenic Tobacco Plants Expressing Bacterial Atrazine Chlorohydrolase Gene

作　　者：斯华敏[1] 朱丽[1] 牟仁祥[2] 刘文真[1] 胡国成[1] 付亚萍[1] 陈铭学[2] 蔡宝立[3] 孙宗修[1] 

机构地区：[1]中国水稻研究所水稻生物学国家重点实验室,浙江杭州310006 [2]中国水稻研究所农业部稻米及制品质量监督及检验测试中心,浙江杭州310006 [3]南开大学微生物学系,天津300071

出　　处：《农业环境科学学报》2008年第2期596-601,共6页Journal of Agro-Environment Science

基　　金：浙江省自然科学基金(X306649)

摘　　要：植物修复(Phytoremediation)技术是消除或减少土壤环境中有机污染物的重要手段,本研究采用植物转基因技术对土壤中除草剂阿特拉津的降解进行了探索。通过农杆菌介导将阿特拉津氯水解酶基因AD1-atzA转入烟草中,获得了转基因植株。T1代植株在浇灌了20mg·L-1阿特拉津溶液的模拟污染土壤条件下生长45d,抗性植株的RT-PCR结果证实叶片中阿特拉津氯水解酶基因得到正常转录,液相色谱质谱分析在叶片中检出阿特拉津的水解产物羟基阿特拉津。结果表明,用转基因植物修复阿特拉津污染土壤是值得进一步探索的途径。Atrazine, the most widely used herbicide in the world, is one of the persistent toxic substances （PTS）. Atrazine chlorohydrolase, encoded by atrazine chlorohydrolase gene, can catalyze the conversion of toxic atrazine to nontoxic hydroxyatrazine. In our previous study, the bacterial atrazine chlorohydrolase gene （ADl-atzA） was cloned from A rthrobacter sp. strain AD1 and inserted into the expression vector p 1301-atzA. In order to explore the effect of transgenic tobacco plants expressing A Dl-atzA gene on biodegradation of atrazine, in this study, the AD1-atzA gene was introduced into tobacco by Agrobacterium tumefaciens mediated transformation and total of 102 plants were obtained. PCR analysis confirmed that the A D1-atzA gene was integrated into the genome of 74% of To plants. T1 transgenic plants were used for atrazine resistance experiment. The results showed that some of transgenic lines grew well on soil irrigated with 20 mg·L^-1 atrazine even treated for 45 days while the nontransgenic tobacco plants died completely under the same condition after 7 days. The survivable Tl transgenic tobacco plants were used for further analysis. RT-PCR analysis indicated that A Dl-atzA gene was expressed normally in transgenic plants and hydroxyatrazine was discovered in transgenic plants leaves by HPLC-MS method. However, neither RT-PCR band nor hydroxyatrazine was detected in wild-type tobacco leaves. These results proved that it was feasible to apply transgenic plants harbored A Dl-atzA gene to phytore-mediation of atrazine-contaminated soils.

关 键 词：阿特拉津 生物降解 阿特拉津氯水解酶基因 转基因烟草 植物修复 环境生物技术 

分 类 号：X53[环境科学与工程—环境工程]