摘要
阿特拉津氯水解酶(AtzA)能有效催化有毒的阿特拉津脱氯生成无毒的羟基阿特拉津。本文将来自假单胞菌ADP的atzA-ADP和来自节杆菌AD1的atzA-NK分别插入Ti载体pBin438,构建了atzA植物表达载体pBin438-atzA-ADP和pBin438-atzA-NK。并通过农杆菌介导法将其转入烟草,经基因组PCR筛选及RT-PCR分析,获得16株转atzA-ADP烟草株系和11株转atzA-NK烟草株系。在含150mgL-1阿特拉津的1/2MS培养基上生长50d时,株系401的降解能力最高,达79.26%,远高于野生型烟草的0.47%,说明转基因烟草可用于建立阿特拉津残留环境的植物修复系统。
Atrazine （2-chloro-4-ethylamino-6-isopropylamno-1,3,5-triazine） is a wildly used herbicide in the world for controlling broad-leaf weeds in corn and sugarcane. However, atrazine is degraded slowly in the environment with an average half-life ranging from 4 to 57 weeks, and has been detected in soil as well as in ground and surface water in several countries. Atrazine chlorohydrolase （AtzA） catalyses the hydrolytic dechlorination and detoxification of atrazine to hydroxyatrazine, a non-herbicidal and non-toxic substance and is more strongly sorbed to soil than atrazine. In the present paper, atzAs were amplified from pMD4 plasmid （containing atzA-ADP） and pET21b-atzA-NK plasmid （containing atzA-NK）, and inserted into plant expression plasmid pBin438. The recombinant plasmid was successfully introduced into tobacco leaf disks by Agrobacterium-mediated transformation and 136 plantlets were regenerated from selected medium contained 20 mg L^-1 kanamycin and 150 μg mL^-1 atrazine, and then were subjected to analyses of PCR and RT-PCR. The PCR using primers atzA-7/atzA-2, atzA-5/atzA-8, and atzA-7/atzA-8, which are specific for atzA, revealed that 42 of 72 （58.3%） pBin438-atzA-ADP and 38 of 64 （59.4%） of pBin438-atzA-NK regenerated plants were transgenic, respectively. Further RT-PCR indicated that 38.1% （16 of 42） of atzA-ADP and 28.9% （11 of 38） of atzA-NK, expressed AtzA in transgenic tobacco plants, respectively. After 50 days＇ growth on 1/2MS medium contained 150 mg L^-1 atrazine, the transgenic plants degraded a large amount of atrazine presented in the medium with degrading as high as 79.26% in line 401, whereas 0.47% in the wild type. These results indicated that the transgenic tobacco plants can catalyze atrazine efficiently.
出处
《作物学报》
CAS
CSCD
北大核心
2008年第5期783-789,共7页
Acta Agronomica Sinica
基金
国家自然科学基金项目(30671183)
关键词
阿特拉津氯水解酶
转基因
农杆菌介导法
烟草
阿特拉津降解能力
Atrazine chlorohydrolase （AtzA）
Transgenic
Agrobacterium-mediate transformation
Tobacco
Phytoremediation capability of atrazine
作者简介
王绘砖（1981-），女，博士研究生，主要从事分子遗传学研究。
通讯作者（Corresponding author）：陈德富（1965-），男，博士，教授，博士生导师。E-mail：chendefu@nankai．edu．cn
