一株土壤高温纤维素降解菌的筛选及鉴定
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摘要
为开发纤维素降解菌剂,分别采用富集培养法、刚果红纤维素培养基法以及滤纸崩解试验,从长沙周边稻田土壤中分离高温纤维素降解菌。结果表明:通过筛选共获得降解菌34株,其中降解菌Z3的降解效果较好,透明圈直径与菌落直径的比值为2.69,显著大于2;该菌株能在10 d内将滤纸完全降解;根据形态特征观察、生理生化特征、16S rDNA序列测定以及系统发育分析,鉴定菌株Z3为枯草芽孢杆菌(Bacillus subtilis)。该菌株环境适应性好,分解纤维素能力强,具有潜在的应用价值。
In order to develop cellulose-degrading microbial inoculum,we isolated high-temperature resistant cellulose-degrading bacteria from paddyfield soil around Changsha by enrichment culture method,cellulose-congo red medium method,and filter paper disintegration test.The results show that 34 strains are obtained,among which,strain Z3 has good degradation effect with the diameter ratio of transparent circle to colony of 2.69,significantly greater than 2.Moreover,strain Z3 can completely degrade filter paper within 10 d.Furthermore,strain Z3 is identified as Bacillus subtilis by morphologic characterization,physiological and biochemical charactoristics,16 S rDNA sequencing,and phylogenic analysis.Strain Z3 has high adaptability to environment and good cellulose-degrading ability,which has potential application value.
In order to develop cellulose-degrading microbial inoculum,we isolated high-temperature resistant cellulose-degrading bacteria from paddyfield soil around Changsha by enrichment culture method,cellulose-congo red medium method,and filter paper disintegration test.The results show that 34 strains are obtained,among which,strain Z3 has good degradation effect with the diameter ratio of transparent circle to colony of 2.69,significantly greater than 2.Moreover,strain Z3 can completely degrade filter paper within 10 d.Furthermore,strain Z3 is identified as Bacillus subtilis by morphologic characterization,physiological and biochemical charactoristics,16 S rDNA sequencing,and phylogenic analysis.Strain Z3 has high adaptability to environment and good cellulose-degrading ability,which has potential application value.
引文
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[16] 田敏,张庆华,毛忠贵,等.一株高温兼性纤维素降解细菌的分离鉴定及特性分析[J].安全与环境学报,2013,13(2):31-36.TIAN M,ZHANG Q H,MAO Z G,et al.Isolation and identification of a thermophilic cellulolytic facultative anaerobic bacterium and analysis of its properties[J].Journal of Safety and Environment,2013,13(2):31-36.
[2] 刘振东,李贵春,杨晓梅,等.我国农业废弃物资源化利用现状与发展趋势分析[J].安徽农业科学,2012,40(26):13068-13070.LIU Z D,LI G C,YANG X M,et al.Status and development trend of resource utilization ways of agricultural residues in China[J].Journal of Anhui Agricultural Sciences,2012,40(26):13068-13070.
[3] 彭靖.对我国农业废弃物资源化利用的思考[J].生态环境学报,2009,18(2):794-798.PENG J.Review and dicussion on utilization of agricultural waste resources in China[J].Ecology and Environmnet,2009,18(2):794-798.
[4] 朱能武.堆肥微生物学研究现状与发展前景[J].氨基酸和生物资源,2005,27(4):40-44.ZHU N W.Microbiology of composting:a review and prospect[J].Amino Acids & Biotic Resources,2005,27(4):40-44.
[5] TENGERDY R P,SZAKACS G.Bioconversion of lignocellulose in solid substrate fermentation[J].Biochemical Engineering Journal,2003,13(2):169-179.
[6] 黄得扬,陆文静,王洪涛,等.高效纤维素分解菌在蔬菜-花卉秸秆联合好氧堆肥中的应用[J].环境科学,2004,25(2):145-149.HUANG D Y,LU W J,WANG H T,et al.Application of high-efficient cellulose utilization microorganisms in co-composting of vegetable wastes and flower stalk[J].Chinese Journal of Environmental Science,2004,25(2):145-149.
[7] HAKI G D,RAKSHIT S K.Developments in industrially important thermostable enzymes:a review[J].Bioresource Technology,2003,89(1):17-34.
[8] 周德庆.微生物学实验手册[M].上海:上海科学技术出版社,1986.
[9] 东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:162.
[10] 常慧萍,张红,杨雪,等.高温纤维素降解细菌的筛选与鉴定[J].河南教育学院学报(自然科学版),2012,21(3):5-7.CHANG H P,ZHANG H,YANG X,et al.Screening and identification of high-temperature resistant cellulose-decomposing bacteria[J].Journal of Henan Education Institute(Natural Science Edition),2012,21(3):5-7.
[11] 王海峰,包木太,韩红,等.一株枯草芽孢杆菌分离鉴定及其降解稠油特性[J].深圳大学学报(理工版),2009,26(3):221-227.WANG H F,BAO M T,HAN H,et al.Isolation and identification of a Bacillus subtilis and its characterization in degrading heavy oil [J].Journal of Shenzhen University(Science & Engineering),2009,26(3):221-227.
[12] 田伟,王磊,李刚,等.高温纤维素降解菌的筛选及内切酶基因的克隆表达[J].农业环境科学学报,2014,33(4):788-793.TIAN W,WANG L,LI G,et al.Isolation of thermophilic cellulose-decomposing bacteria and cloning and expression of an endoglucanase gene[J].Journal of Agro-Environment Science,2014,33(4):788-793.
[13] 张楠,杨兴明,徐阳春,等.高温纤维素降解菌的筛选和酶活性测定及鉴定[J].南京农业大学学报,2010,33(3):82-87.ZHANG N,YANG X M,XU Y C,et al.Screening,cellulase activity measurement and identification of cellulose-decomposing bacteria[J].Journal of Nanjing Agricultural University,2010,33(3):82-87.
[14] 易旻,杨玉婷,李梦霖,等.一株耐高温纤维素降解菌的分离筛选及鉴定[J].吉首大学学报(自然科学版),2017,38(4):72-77.YI M,YANG Y T,LI M L,et al.Screening and identification of a high-temperature resistant cellulose-degrading bacterium[J].Journal of Jishou University(Natural Science Edition),2017,38(4):72-77.
[15] 周望平,肖兵南,邱美珍,等.牛粪中高温纤维素降解菌的性能研究[J].纤维素科学与技术,2009,17(3):23-27.ZHOU W P,XIAO B N,QIU M Z,et al.Study on performance of the pyro-cellulose degradation bacterium from cattle dung[J].Journal of Cellulose Science and Technology,2009,17(3):23-27.
[16] 田敏,张庆华,毛忠贵,等.一株高温兼性纤维素降解细菌的分离鉴定及特性分析[J].安全与环境学报,2013,13(2):31-36.TIAN M,ZHANG Q H,MAO Z G,et al.Isolation and identification of a thermophilic cellulolytic facultative anaerobic bacterium and analysis of its properties[J].Journal of Safety and Environment,2013,13(2):31-36.