一株菊酯类农药降解菌的分离鉴定及其降解酶基因的克隆  

Isolation,identification and degrading gene cloning of a pyrethroids-degrading bacterium

作　　者：张松柏[1,2] 张德咏[1,2] 刘勇[1,2] 罗香文 成飞雪 罗源华 

机构地区：[1]中南大学研究生院隆平分院,长沙410125 [2]湖南省植物保护研究所,长沙410125

出　　处：《微生物学报》2009年第11期1520-1526,共7页Acta Microbiologica Sinica

基　　金：国家"863计划"(2006AA10Z401);国家"十一五"科技支撑计划项目(2006BAD17B08;2006BAD08A08)~~

摘　　要：【目的】筛选分离高效降解菊酯类农药的光合细菌,研究其降解特性,并对该菌株中降解酶基因进行克隆与初步分析。【方法】根据分离菌株的细胞形态结构、活细胞光吸收特征、生理生化特征及其16SrDNA序列系统发育分析鉴定降解菌,气相色谱法测定该菌株降解菊酯类农药的能力,PCR方法克隆降解酶基因。【结果】菌株PSB07-21属红假单胞菌属(Rhodopseudomonas sp.),其降解最佳条件为3000lx、35℃、pH7,在此条件下培养15d对600mg/L甲氰菊酯、氯氰菊酯、联苯菊酯降解率分别为66.63,43.25,50.18%。经序列测定与分析表明,克隆到1个329bp的ORF,该ORF预测的编码蛋白与2OG-Fe(Ⅱ)oxygenase的同源性为37.0%。添加低浓度Fe(Ⅱ)对该菌株降解菊酯类农药有一定促进作用。【结论】PSB07-21能有效的降解多种菊酯类农药,该研究为菊酯类农药生物修复提供理论依据。[Objective]This study aimed to isolate, identify and clone degrading gene of a synthetic pyrethroids degrading bacterium. [ Methods] A photosynthetic bacterial strain PSB07-21 capable of degrading several synthetic pyrethroids efficiently was isolated by an enrichment culture. PSB07-21 was identified based on its morphology, physiology and phylogenetic analysis of 16S rDNA sequence. The degradation ability of this strain was evaluated with gas chromatography. The degrading gene was cloned with PCR. [Results] PSB07-21 was closely related to Rhodopseudomonas sp. The optimum condition of degrading synthetic pyrethroidss was at 35℃, pH 7 and 3000 lx. PSB07-21 could degrade fenpropathrin, cypermethrin and bipthenthrin by 66.63%, 43.25% and 50.18% in a concentration of 600 mg/L at day 15, respectively. We cloned a putative gene which was 326bplong with 37.0% identical to 2OG-Fe（ Ⅱ） oxygenase gene. When compensating low concentration Fe（ Ⅱ ） in PSB medium with synthetic pyrethroids, the degradation efficiency of PSB07-21 was enhanced. [ Conclusion] The strain has the potential application to synthetic pyrethroids bioremediation.

关 键 词：红假单胞菌PSB07-21 菊酯类农药 降解酶基因 克隆 

分 类 号：X172[环境科学与工程—环境科学]