Lü Xin, Peng Xia-Wei, Hu Qing, Ma An-Zhou, Jiang Ze-Ping, Wei Yuan
College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China.
Huan Jing Ke Xue. 2013 Jan;34(1):263-70.
Naturally occurring bacteria isolates capable of metabolizing pesticides have received considerable attention because they offer the possibility of both environmentally friendly and in situ remediation. The effect of herbicide quizalofop-p-ethyl on bacterial community in soil was analyzed using the technique of PCR-DGGE for isolating strains biodegrading quizalofop-p-ethyl. Results indicated that the soil bacterial community structures significantly changed after adding quizalofop-p-ethyl. The bacterial diversity of soil showed an increasing-decreasing-increasing trend. The largest changes occurred in the 9th day and then became stabilized. According to the sequencing results of bands in DGGE profiles, it inferred that members of bacterial genera Pseudomonas, Massilia and Burkholderia had tolerance to quizalofop-p-ethyl, and the potential for degradation. These microbial groups could be used to isolate and screen as the indigenous microbial resources to reduce pesticide residues. Digoxigenin-labeled probes had been synthesized based on the sequencing results of bands in the DGGE profiles, and three bacterial strains capable of biodegrading quizalofop-p-ethyl were isolated from soil by colony in situ hybridization technique. The strain named L1 was able to utilize quizalofop-p-ethyl as the sole source of carbon. The strain was identified as Pseudomonas sp., based on the phylogenetic analysis of 16S rRNA. The degrading ability of strain L1 in minimal medium with quizalofop-p-ethyl was investigated by HPLC. The quizalofop-p-ethyl content decreased by almost 50% after 7 days, and the biomass of strain L1 increased while the content of quizalofop-p-ethyl was decreased. This confirmed that the strain L1 had the capacity of degradation. This result provided a basis for future research on degradation mechanism and functional genes.
能够代谢农药的天然细菌分离株受到了广泛关注,因为它们提供了环境友好型原位修复的可能性。采用PCR-DGGE技术分析了除草剂精喹禾灵对土壤细菌群落的影响,以分离降解精喹禾灵的菌株。结果表明,添加精喹禾灵后土壤细菌群落结构发生了显著变化。土壤细菌多样性呈先增加后减少再增加的趋势。最大的变化发生在第9天,然后趋于稳定。根据DGGE图谱中条带的测序结果,推断假单胞菌属、马赛菌属和伯克霍尔德菌属的成员对精喹禾灵具有耐受性和降解潜力。这些微生物类群可作为本地微生物资源进行分离筛选,以减少农药残留。根据DGGE图谱中条带的测序结果合成了地高辛标记探针,并通过菌落原位杂交技术从土壤中分离出三株能够降解精喹禾灵的细菌菌株。命名为L1的菌株能够利用精喹禾灵作为唯一碳源。基于16S rRNA的系统发育分析,该菌株被鉴定为假单胞菌属。通过高效液相色谱法研究了菌株L1在含有精喹禾灵的基本培养基中的降解能力。7天后,精喹禾灵含量下降了近50%,随着精喹禾灵含量的下降,菌株L1的生物量增加。这证实了菌株L1具有降解能力。该结果为今后降解机理和功能基因的研究提供了依据。