凉水保护区土壤产类漆酶-多铜氧化酶细菌群落结构  

The community structure of laccase-like multicopper oxidase-producing bacteria in soil of Liangshui Nature Reserve

作　　者：赵丹[1,2] 谷惠琦[1] 崔岱宗[1] 范晓旭[1,2] 张曦[1] 赵敏[1] 

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040 [2]黑龙江大学生命科学学院微生物省高校重点实验室,哈尔滨150080

出　　处：《生态学报》2012年第13期4062-4070,共9页Acta Ecologica Sinica

基　　金：世界自然基金(WWF;CN0078.01);国家自然科学基金(30170775;30671702);国家支撑项目(2008BADA1B01-10)共同资助

摘　　要：在凉水国家级自然保护区3种主要林型红松(Pinus koraiensis)、白桦(Betula platyphylla)及云杉(Picea dietrich)林采集林下土壤样品,以铜离子作为筛选剂处理后,结合平板分离法与基于16S rDNA V3区片段的变性梯度凝胶电泳(DenaturingGradient Gel Electrophoresis,DGGE)技术,调查了土壤样品中产类漆酶-多铜氧化酶(laccase-like multicopper oxidase,LMCO)细菌的群落结构。这是研究产类漆酶-多铜氧化酶细菌在环境中存在的种、属及分布的新尝试。平板分离获得10株细菌均为芽孢杆菌属(Bacillus sp.),其中梭状芽孢杆菌(Bacillus fusiformis)未见相关报道。通过DGGE图谱分析可知,产类漆酶-多铜氧化酶细菌在研究地不同林型土壤中的群落结构无明显差异,在红松林土壤中多样性最为丰富。DGGE条带测序结果表明,取样地土壤中产类漆酶细菌主要为罗尔斯顿菌属(Ralstonia sp.)、肠杆菌属(Enterobacter sp.)、芽孢杆菌属和一些未培养细菌。Abstract： Laccases catalyze the oxidation of various aromatics, particularly phenolic and amine substrates, making them valuable in industrial applications. Laccases also play an important role in soil organic matter （SOM） turnover processes and the global carbon cycle due to their involvement in the synthesis and degradation of lignin as well as transformation of lignified substrates and humic substances. Laccases belong to the protein family of muhicopper oxidases characterized by copper atoms in the active center. Laccases or laccase-like multicopper oxidases （LMCO） have been extensively studied especially in fungi. Recently, increasing evidence points to a wide occurrence of LMCO in bacteria. As bacterial communities are known to decompose pollutants and municipal wastes involving large quantities of phenolic substances and organic matter, it can be deduced that bacterial LMCO might also participate in lignin degradation and SOM cycling. Copper atoms not only constitute the catalytic center of laccases but can also confer copper resistance to bacteria. Consequently, copper ions have a well known use in isolating LMCO-producing bacteria. To gain deeper insights into theLiangshui nature reserve, Heilongjiang Province. Soil samples were collected in three patterns of forests： Pinus koraiensis, Betula platyphyl,!a and Picea dietrich. Soil samples were enriched in M9 basal medium and then screened in LB medium with gradient Cu2.. Ten bacteria strains were isolated and identified based on traditional plating methods of bacteria separation. The results shawed that they all belonged to Bacillus sp. , falling into four different species： Bacillus pumilus, Bacillus subtilis, Bacillus fusiformis and Bacillus amyloliquefaciens, among which no laccase-relative results have been reported on Bacillus fusiformis. To investigate the structural variation of LMCO-producing bacteria, 16S-rDNA-V3-fragment-based DGGE and clone sequence BLAST technologies were applied. The data based on DGGE plot indicated there was

关 键 词：漆酶 类漆酶-多铜氧化酶 细菌 DGGE 

分 类 号：S154.3[农业科学—土壤学]