铅锌矿区耐砷细菌的分离、鉴定及性质研究

doi:10.13560/j.cnki.biotech.bull.1985.2016-0983

摘要/Abstract

摘要： 旨在从湖南康家湾铅锌矿区的重金属污染土壤样品中筛选出耐高浓度砷的细菌菌株。用稀释涂布法分离耐砷细菌;根据16S rRNA基因系统发育分析鉴定分离得到的砷高耐受性菌株并检测菌株内含有的砷耐受性相关基因;用砷钼蓝法测定耐砷细菌的砷氧化还原能力;并通过吲哚乙酸（IAA）定量实验检测优势菌株产IAA的能力。结果显示,从土壤样品中共分离出152株耐砷细菌,其中6株细菌对As⁵⁺和As³⁺的耐受性值分别高达800 mmol/L和20 mmol/L;并且这6株耐砷细菌分属于5个不同的属：假单胞菌属、苍白杆菌属、芽孢杆菌属、威廉氏菌属和节细菌属;菌株Tw31、Tw133、Sw149和Tw222中存在砷还原酶基因arsC,Bw218和Tw222中存在砷离子外排基因arsB/ACR3（2）;在72 h之内,菌株Tw133和Tw222的As³⁺氧化率（约17 %）和As⁵⁺还原率（约35 %）均高于其他菌株;尤其是菌株Tw133在144 h具有48.66 %的As⁵⁺还原率;且这两株菌分别能产生42.86 μg/mL和24.36 μg/mL的IAA。筛选出的Tw133和Tw222菌株在砷耐受性、砷氧还能力和产IAA能力等方面展现出了较明显的优势,为深入研究细菌的砷耐受性机制提供了实验材料。

关键词: 耐砷细菌, 耐砷基因, 砷氧化还原, IAA

Abstract: The aim of this study is to isolate highly arsenic-tolerant bacterial strains from heavy metal contaminated soils from the Kangjiawan lead-zinc mine,Hunan Province. Serial dilution and plating method was used to isolate arsenic-tolerant bacteria,and the strains were identified by 16S rRNA sequence analysis. Then the detection of arsenic tolerance-related genes was carried out. The arsenic molybdenum blue experiment was conducted to evaluate arsenic oxidation-reduction ability of arsenic-tolerant bacteria. Furthermore,the indole acetic acid（IAA）produced by the dominant bacterial strains was quantitatively determined. A total of 152 arsenic-tolerant bacterial strains were isolated from soils samples,and six strains were resistant to As⁵⁺ and As³⁺ up to 800 mmol/L and 20 mmol/L,respectively. The six arsenic-tolerant bacterial strains belonged to five different genera,Pseudomonas,Ochrobactrum,Bacillus,Williamsia,and Arthrobacter. In arsenic tolerance-related gene detection experiments,arsenate reductase gene arsCexisted in strains Tw31,Tw133,Sw149,and Tw222,and strains Bw218 and Tw222 contained arsenic ion efflux permease gene arsB/ACR3（2）. Within 72 h,the As³⁺ oxidation rate and As⁵⁺reduction rate of strains Tw133 and Tw222 were higher than those of other strains,which were about 17 % and 35 %,respectively. In particular,strain Tw133 had an As⁵⁺reduction rate of 48.66 % at 144 h. Moreover,the synthetic quantities of IAA in the two strains were 42.86 μg/mL and 24.36 μg/mL,respectively. The isolated strains Tw133 and Tw222 showed obvious advantages in terms of arsenic tolerance,arsenic oxidation-reduction capacity and IAA production,providing experimental materials for the further study of arsenic-tolerant mechanism in bacteria.

Key words: arsenic-tolerant bacteria, arsenic-tolerant gene, arsenic oxidation-reduction, IAA?

吴丹，张志鹏，马玉超. 铅锌矿区耐砷细菌的分离、鉴定及性质研究[J]. 生物技术通报, 2017, 33(5): 210-218.

WU Dan ZHANG Zhi-peng MA Yu-chao. Isolation,Identification and Characterization of Arsenic-tolerant Bacteria from Lead-zinc Mine Tailing[J]. Biotechnology Bulletin, 2017, 33(5): 210-218.

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