秸秆降解菌的筛选及其纤维素降解性能的研究
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摘要
通过秸秆粉平板、刚果红染色法以及小麦秸秆降解率来筛选高效秸秆降解菌,并测定菌株的胞外纤维素酶活来研究其秸秆纤维素的降解能力。结果表明,在所筛选的4株真菌中,菌株z-5的秸秆降解率最高,为47%;其次是菌株4-1、3-1和2-2,分别为45%、32%和31%,滤纸腐解实验结果与秸秆降解试验结果相似。4株秸秆降解菌均具有外切葡聚糖酶、内切葡聚糖酶和β-葡萄糖苷酶等3种纤维素酶,但变化趋势不同。菌株z-5和菌株4-1的三种酶活呈现出较好的协同性,菌株2-2和菌株3-1协同性较弱。通过秸秆粉平板,秸秆降解率测定以及滤纸孔洞法可以更准确快速地得到目的菌株;纤维素酶在秸秆降解中起着主导作用。
High-efficiency straw-degrading bacteria were screened by straw powder plate,Congo red staining method and degradation rate of wheat straw. The extracellular cellulase activities of the strains were determined to study the degradation ability of the straw cellulose. The results showed that among the 4 strains of fungi screened,the strain z-5 had the highest degradation rate of 47%,and that of strain 4-1,3-1 and 2-2 were 45%,32% and 31%,respectively. The filter paper-decomposing test results were similar to the straw degradation test results.Four strains all had exo-1,4-β-D-glucanase,endo-1,4-β-D-glucanase,and β-glucosidase,but their degradation trends were different. The three enzyme activities of strain z-5 and strain 4-1 showed better synergy,and that of strain 2-2 and strain 3-1 were less synergistic. In sum,the target strain can be obtained more accurately and quickly through the straw powder plate test,straw degradation rate determination and filter paper hole method,and the cellulase plays a leading role in straw degradation.
High-efficiency straw-degrading bacteria were screened by straw powder plate,Congo red staining method and degradation rate of wheat straw. The extracellular cellulase activities of the strains were determined to study the degradation ability of the straw cellulose. The results showed that among the 4 strains of fungi screened,the strain z-5 had the highest degradation rate of 47%,and that of strain 4-1,3-1 and 2-2 were 45%,32% and 31%,respectively. The filter paper-decomposing test results were similar to the straw degradation test results.Four strains all had exo-1,4-β-D-glucanase,endo-1,4-β-D-glucanase,and β-glucosidase,but their degradation trends were different. The three enzyme activities of strain z-5 and strain 4-1 showed better synergy,and that of strain 2-2 and strain 3-1 were less synergistic. In sum,the target strain can be obtained more accurately and quickly through the straw powder plate test,straw degradation rate determination and filter paper hole method,and the cellulase plays a leading role in straw degradation.
引文
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[13]王海滨,韩立荣,冯俊涛,等.高效纤维素降解菌的筛选及复合菌系的构建[J].农业生物技术学报, 2015, 23(4):421-431.
[14]王贤丰,单洪伟,张家松,等.从海水环境分离筛选甘蔗渣纤维素降解菌[J].微生物学通报, 2015, 42(6):981-989.
[15]孙江慧.几株食用菌对秸秆木质纤维素降解能力的研究[D].南京:南京农业大学, 2012.
[16]全国饲料工业标准化技术委员会. NY/Y9122004饲料添加剂纤维素酶活力的测定[S].北京:中华人民共和国农业部,2005.
[17]王洪媛,范丙全.三株高效秸秆纤维素降解真菌的筛选及其降解效果[J].微生物学报, 2010, 50(7):870-875.
[18]王海滨,韩立荣,冯俊涛,等.高效纤维素降解菌的筛选及复合菌系的构建[J].农业生物技术学报, 2015, 23(4):421-431.
[2]钟杭,朱海平,黄锦法.稻麦秸秆全量还田对作物产量和土壤的影响[J].浙江农业学报, 2002, 14(6):344-347.
[3]黄婷苗,郑险峰,侯仰毅,等.秸秆还田对冬小麦产量和氮、磷、钾吸收利用的影响[J].植物营养与肥料学报, 2015, 21(4):853-863.
[4]赵蒙蒙,姜曼,周祚万.几种农作物秸秆的成分分析[J].材料导报, 2011, 25(16):122-125.
[5]邱雁临.纤维素酶的研究和应用前景[J].粮食与饲料工业,2001(8):30-31.
[6]Tomme P, Van TH, Pettersson G, et al. Studies of the cellulolytic system of Trichoderma reesei QM 9414. Analysis of domain function in two cellobiohydrolases by limited proteolysis[J]. European Journal of Biochemistry, 1988, 170(3):575-581.
[7]Singh R, Varma AJ, Laxman RS, et al. Hydrolysis of cellulose derived from steam exploded bagasse by Penicillium cellulases:comparison with commercial cellulose[J]. Bioresour Technol,2009, 100(24):6679-6681.
[8]Okunowo WO, Gbenle GO, Osuntoki AA, et al. Production of cellulolytic and xylanolytic enzymes by a phytopathogenic Myrothecium roridum and some avirulent fungal isolates from water hyacinth[J]. African Journal of Biotechnology, 2010, 9(15):1074-1078.
[9]GeeraertsHAM,VandammeEJ.Cellulolyticpropertiesof Chaetomium crispatum[J]. Journal of Chemical Technology&Biotechnology Biotechnology, 2010, 33(2):107-113.
[10]刘晓梅,邹亚杰,胡清秀,等.菌渣纤维素降解菌的筛选与鉴定[J].农业环境科学学报, 2015, 34(7):1384-1391.
[11]沈萍.微生物学实验[M].第3版.北京:高等教育出版社,1999.
[12]吴文韬,鞠美庭,刘金鹏,等.一株纤维素降解菌的分离、鉴定及对玉米秸秆的降解特性[J].微生物学通报, 2013, 40(4):712-719.
[13]王海滨,韩立荣,冯俊涛,等.高效纤维素降解菌的筛选及复合菌系的构建[J].农业生物技术学报, 2015, 23(4):421-431.
[14]王贤丰,单洪伟,张家松,等.从海水环境分离筛选甘蔗渣纤维素降解菌[J].微生物学通报, 2015, 42(6):981-989.
[15]孙江慧.几株食用菌对秸秆木质纤维素降解能力的研究[D].南京:南京农业大学, 2012.
[16]全国饲料工业标准化技术委员会. NY/Y9122004饲料添加剂纤维素酶活力的测定[S].北京:中华人民共和国农业部,2005.
[17]王洪媛,范丙全.三株高效秸秆纤维素降解真菌的筛选及其降解效果[J].微生物学报, 2010, 50(7):870-875.
[18]王海滨,韩立荣,冯俊涛,等.高效纤维素降解菌的筛选及复合菌系的构建[J].农业生物技术学报, 2015, 23(4):421-431.
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