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目的对采自河南省临颍县麦田的土壤样品进行苯磺隆耐受菌的筛选和初步鉴定。方法利用选择性培养基,经富集培养,从长期施用苯磺隆的麦田土壤中筛选苯磺隆耐受菌。采用基于16S rRNA基因序列的系统发育分析和生物学特性研究,对菌株进行初步分类鉴定。结果共分离到耐受苯磺隆的纯菌12株,其中一株LYBRD3-3对苯磺隆耐受能力最强,苯磺隆耐受浓度为150μg/L。经过对该菌形态观察、生理生化特征及基于16S rRNA基因序列的系统发育分析表明,LYBRD3-3与无色菌属的Achromobacter insolitus LMG 6003T(AY170847)序列具有最高的相似性,同源性为99.58%,初步鉴定该菌为无色菌属菌。结论分离自麦田土壤的苯磺隆耐受菌为进一步筛选苯磺隆降解功能菌株提供菌种资源,进而为污染土壤的原位生物修复提供基础研究。
Objective To screen and preliminary identify the soil samples from wheat field in Linying County of Henan Province. Methods The selective culture medium was used to screen for the tolerance of tribenuron-methyl-resistant bacteria in the soil of long-term application of tribenuron-methyl in enrichment culture. Based on the phylogenetic analysis and biological characteristics of 16S rRNA gene sequences, the strains were initially classified and identified. Results A total of 12 isolates were tolerated from tribenuron-methyl. One strain of LYBRD3-3 was the strongest tolerant to tribenuron-methyl and the tolerable tribenuron-methyl was 150μg / L. The morphological, physiological and biochemical characteristics of the strain and phylogenetic analysis based on 16S rRNA gene sequence showed that LYBRD3-3 had the highest similarity with Achromobacter insolitus LMG 6003T (AY170847) of Achromobacter with a homology of 99.58 %, Preliminary identification of the bacteria is colorless bacteria. CONCLUSIONS: The isolates of tribenuron-methyl (MTB) -resistant bacteria from the soil of wheat field provide the resources of bacteria for further screening of functional strains degraded by tribenuron-methyl and provide the basic research for the in situ bioremediation of contaminated soils.