一株纤维素分解菌的鉴定及对两种草坪草凋落物分解活性的研究

草业学报 ›› 2011, Vol. 20 ›› Issue (6): 170-179.

一株纤维素分解菌的鉴定及对两种草坪草凋落物分解活性的研究

芦光新^1,2,陈秀蓉^1*,杨成德¹,薛莉¹, 刘雯¹,卞静¹

1.甘肃农业大学草业学院草业生态系统教育部重点实验室中-美草地畜牧业可持续发展研究中心,甘肃兰州 730070;
2.青海大学农牧学院草业科学系,青海西宁 810016

收稿日期:2011-06-23 出版日期:2011-06-25 发布日期:2011-12-20
通讯作者: E-mail:chenxiurong@gsau.edu.cn
作者简介:芦光新(1974-),男,青海湟中人,教授,在读博士。E-mail:lugx74@qq.com
基金资助:
国家自然科学基金项目(30471232)资助。

Identification of cellulose decomposing fungi strain F₁ and decomposition activity to two kinds of lawn grass litter

LU Guang-xin^1,2, CHEN Xiu-rong¹, YANG Cheng-de¹, XUE Li¹, LIU Wen¹, BIAN Jing¹

1.Pratacultural College, Gansu Agricultural University, Key Laboratory of Grassland Ecosystem Ministry of Education, Sino-U.S.Centers for Grazingland Ecosystem Sustainability, Agricultural University, Lanzhou 730070, China;
2.Department of Grassland Science, Agriculture and Animal Husbandry College, Qinghai University, Xining 810016, China

Received:2011-06-23 Online:2011-06-25 Published:2011-12-20

摘要/Abstract

摘要： 对1株分离筛选自东祁连山高寒草地土壤真菌菌株F₁的分类学地位及对多年生黑麦草和白三叶凋落物的分解活性进行了研究。采用形态学和rDNA-ITS分子生物学相结合的方法,对菌株F₁的分类学地位进行了确定,初步鉴定为Microdochium bolleyi;应用摇瓶液体发酵法,通过接种和不接种处理,分别以2种草坪草凋落物为底物,于25℃,150 r/min震荡培养42 d,测定了2种草坪草凋落物诱导真菌分泌木质纤维素酶活力动态变化和草坪草凋落物细胞壁物质(CWM)的降解率。结果表明,1)接种处理后,2种草坪草细胞壁非水溶性物质(ICWM)的量显著减少(P<0.05);2)漆酶出现最早,其次是纤维素酶和木聚糖酶,邻苯二酚氧化酶和愈创木酚氧化酶出现最晚;3)与白三叶凋落物为底物的发酵体系相比,黑麦草凋落物ICWM和CWM降解率高,且差异显著(P<0.05);4)除漆酶之外,以黑麦草凋落物诱导菌株F₁分泌的滤纸酶、羧甲基纤维素酶、木聚糖酶、邻苯二酚氧化酶和愈创木酚氧化酶活力比白三叶诱导相应的酶活力高(P<0.05)。

Abstract: A fungal isolate (strain F₁) was isolated from east part of grassland soil in Qilian alpine. Taxonomic status and decomposition activity to two kinds of lawn grass litter were studied in this study. Combinated morphology with rDNA-ITS molecular biology methods, the fungus was Microdochium bolleyi. The dynamics of lignocellulolytic enzymes secreted by M. bolleyi and cell wall material(CWM) decomposition of perennial ryegrass (Lolium perenne) and white clover (Trifolium repens) litter were investigated for 42 d at 25℃ on a rotary shaker at 150 r/min in flask liquid fermentation method. 1) Insoluble cell wall material(ICWM) of two kinds of lawn grass litter significantly reduced after inoculation strain F₁ (P<0.05); 2) When two kinds of lawn grass litter acts as substrate respectively, the laccase secreted by strain F₁ appears first, followed by the cellulase enzyme and xylanase, catechol oxidase and guaiacol oxidase occurs at the latest; 3) Perennial ryegrass compared with white clover, the insoluble cell wall material (ICWM) and cell wall material (CWM) degradation rate of perennial ryegrass litter was higher (P<0.05); 4) The FPAase, CMCase, xylanase, pycocatechol oxidase and guaiacol oxidase activity except for the laccase secreted by M. bolleyi in perennial ryegrass litter were higher than those in white clover litter (P<0.05).

中图分类号:

芦光新,陈秀蓉,杨成德,薛莉, 刘雯,卞静. 一株纤维素分解菌的鉴定及对两种草坪草凋落物分解活性的研究[J]. 草业学报, 2011, 20(6): 170-179.

LU Guang-xin, CHEN Xiu-rong, YANG Cheng-de, XUE Li, LIU Wen, BIAN Jing. Identification of cellulose decomposing fungi strain F₁ and decomposition activity to two kinds of lawn grass litter[J]. Acta Prataculturae Sinica, 2011, 20(6): 170-179.

参考文献

[1] Lyndl R, Paul J W, Vanzyl W H, et al. Micribial cellulose utilization: Fundamentals and biotechnology[J]. Microbiology and Molecular Biology Reviews, 2002, 66(3): 506-577.
[2] Hammel K E. Fungal degradation of lignin[A]. In: Cadisch G, Giller K E. Plant Litter Quality and Decomposition[C]. Wallingford, UK: Cab International, 1997: 33-45.
[3] Nowak J, Nowak D, Chevallier P. Analysis of composite structure and primordial wood remains in petrified wood[J]. Applied Spectroscopy, 2007, 61(8): 889-895.
[4] Hankin L, Anagnostakis S L. Solid medium containing carboxymethyl cellulose to detect CX cellulase activity of microorganisms[J]. Journal of General Microbiology, 1977, 98: 109-105.
[5] Smith J L, Paul E A. The Significance of Soil microbial Biomass Estimations[M]. New York: Marcel Dekker, 1990: 357-398.
[6] 章家恩, 刘文高. 微生物资源的开发利用与农业可持续发展[J]. 土壤与环境, 2001, 10(2): 154-157.
[7] Odum E P. Fundamentals of Ecology (3rd edition)[M]. Philadelphia: W. B. Saunders Company, 1971: 574.
[8] Burns R G. Enzyme activity in oil: some theoretical and practical consider-actions[A]. In: Burns R G. Soil Enzymes[C]. London, UK: Academic Press, 1978: 295-340.
[9] Kirk T K. Degradation of lignin[A].In: Gibson, David T.Microbial Degradation of Organic Compounds[M]. Newyork: Marcel Dekker, 1984: 399-437.
[10] Falcon M A, Rodriguez A, Carnicero A, et al. Isolation of microorganisms with lignin transformation potential from soil of tenerife island[J]. Soil Biology and Biochemistry, 1995, 27: 121-126.
[11] Sugden A M. Ecology: Diversity and ecosystem resilience[J]. Science, 2000, 290: 233-235.
[12] Casey H, Alexander L, Maren N, et al. A constant flux of diverse thermophilic bacteria into the cold arctic seabed[J]. Science, 2009, 325: 1541-1544.
[13] 王国荣, 陈秀蓉, 张俊忠, 等. 东祁连山高寒灌丛草地土壤微生物生理功能群的动态分布研究[J]. 草业学报, 2011, 20(2): 31-38.
[14] 张俊忠, 陈秀蓉, 杨成德, 等. 东祁连山高寒草地土壤可培养真菌多样性分析[J]. 草业学报, 2010, 19(2): 124-132.
[15] 陈秀蓉, 南志标. 细菌多样性及其在农业生态系统中的作用[J]. 草业科学, 2002, 19(9): 34-38.
[16] 芦光新, 陈秀蓉, 杨成德, 等. 纤维素分解功能菌及其在草业系统界面中的利用潜势[J]. 草业科学, 2011, 28(6): 1156-1161.
[17] 方中达. 植病研究方法[M]. 北京: 科学出版社, 1996: 122-154.
[18] Thompson J D, Gibson T J, Plewniak F, et al. The clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Research, 1997, 25: 4876-4882.
[19] Tamura K, Dudley J, Nei M, et al. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0[J]. Molecular Biology and Evolution, 2007, 24: 1596-1599.
[20] Larkin M A, Blackshields G, Brown N P, et al. Clustal W and clustal X version 2.0[J]. Bioinformatics, 2007, 23(21): 2947-2948.
[21] Miller G L. Use of dinitrosalicylic acid reagent for determination of reducing sugars[J]. Analytical Chemistry, 1959, 31: 426-428.
[22] 唐启元, 冯明光. 实用统计分析及其DPS数据处理系统[M]. 北京: 科学出版社, 2002.
[23] 王宜磊, 邓振旭. 糙皮侧耳多糖分解酶和木质素酶活性研究[J]. 食用菌, 1998, 40(5): 7-8.
[24] 王宜磊, 邓振旭, 赵良田. 彩绒革盖菌愈创木酚氧化酶活性研究[J]. 植物学通报, 2000, 17(5): 462-465.
[25] 郝杰杰, 宋福强, 田兴军, 等. 几株半知菌对马尾松落叶的分解——木质纤维素酶的活性动力学[J]. 林业科学, 2006, 42(11): 69-75.
[26] Lee J. Biological conversion of lignocellulosic biomass to ethanol[J]. Journal of Biotechnology, 1997, 56: 1-24.
[27] Takashima S, Iikura H, Nakamura A, et al. Over production of recombinant Trichoderma reesei cellulases by Aspergillus oryzae and their enzymatic properties[J]. Biotechnology Letter, 1998, 65: 163-171.
[28] Boominathan K, Reedy C A. Fungal degradation of lignin[A]. In: Arora D S, Elander R P, Mukerji K G, et al. Handbook of Applied Mycology[M]. New York: Marcel Dekker, 1992: 763-782.
[29] Thurston C F. The structure and function of fungal laccase[J]. Microbiology, 1994, 140: 19-26.
[30] Wong Y, Yu J. Laccase-catalysed decolorization of synthetic dyes[J]. Water Research, 1999, 33: 3512-3520.