云杉木质素高降解的菌株筛选
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摘要
以新疆南山、库尔德宁等地区感染白腐病的雪岭云杉为材料,从中分离得到4株具有木质素降解能力的真菌,并确定其分类学地位,初步研究菌株对木质素的降解能力。通过含有0.04%愈创木酚的马铃薯培养基进行初筛,含有1%的苯胺蓝的马铃薯培养基进行复筛,扫描电镜观察法及称重法对木质素降解能力进行初步测定,得到2-A、1-G、2-B、3-E四种菌,对木质素降解率分别为5.82%、6.23%、3.80%、5.14%。18S r DNA序列分析确定2-A在分类学上属于葡萄座腔菌目,葡萄座腔菌科,微壳色单隔孢属(Microdiplodia sp.); 1-G为三隔镰孢菌(Fusarium tricinctum),镰孢菌属(Fusarium)。
Collecting rotting wood of spruce in Xinjiang,such as Nam Shan,Aletai Kuerdening,for research meteaial. Lignin degrading fungi from infected spruce were extracted and separated,its taxonomic status were determined,and the lignin degradation rate were tested. Using potato medium containing 0.04% guaiacol for screening,containing 1% aniline blue for rescreening,then four lignin degrading ability of the fungi were dicovered by electronic microscopy and weighing method. The resulting lignin degradation rate of 2-A,1-G,2-B,3-E were 5. 82%,6. 23%,3. 80%,5. 14%,respectively. Using the measured 18 S r DNA sequence characterization,2-A in the taxonomy belongs to Botryosphaeriales,Botryosphaeriaceae,Microdiplodia sp.; 1-G belongs to the Fusarium sp.
Collecting rotting wood of spruce in Xinjiang,such as Nam Shan,Aletai Kuerdening,for research meteaial. Lignin degrading fungi from infected spruce were extracted and separated,its taxonomic status were determined,and the lignin degradation rate were tested. Using potato medium containing 0.04% guaiacol for screening,containing 1% aniline blue for rescreening,then four lignin degrading ability of the fungi were dicovered by electronic microscopy and weighing method. The resulting lignin degradation rate of 2-A,1-G,2-B,3-E were 5. 82%,6. 23%,3. 80%,5. 14%,respectively. Using the measured 18 S r DNA sequence characterization,2-A in the taxonomy belongs to Botryosphaeriales,Botryosphaeriaceae,Microdiplodia sp.; 1-G belongs to the Fusarium sp.
引文
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[20]任丹,张波,张小平,等.川楝内生真菌的遗传及PKS、NRPS基因的多样性[J].中草药,2014,45(10):1 461-1 467.
[21] HIGUCHI T. Lignin biochemistry:biosynthesis and biodegradation[J]. Wood Science and Technology,1990,24(1):23-63.
[2] HORN S J,VAAJE-KOLSTAD G,WESTERENG B,et al. Novel enzymes for the degradation of cellulose[J]. Biotechnology for Biofuels,2012,5(1):45.
[3] ARANTES V,MILAGRES A M F,FILLEY T R,et al. Lignocellulosic polysaccharides and lignin degradation by wood decay fungi:the relevance of nonenzymatic fenton-based reactions[J]. Journal of Industrial Microbiology&Biotechnology,2011,38(4):541-555.
[4] MARTONE P T,ESTEVEZ J M,LU F C,et al. Discovery of lignin in seaweed reveals convergent evolution of cell-wall architecture[J]. Current Biology,2009,19(2):169-175.
[5] HARBORNE J B. Methods in plant biochemistry. Volume 1:plant phenolics[M]. London:Academic Press,1989.
[6]庄雪峰,陈礼辉,林金国,等.杉木心材6种溶剂提取物的抗菌活性[J].森林与环境学报,2016,36(2):231-235.
[7]吴柳萍,纪志荣,连素兰,等.福建省森林病虫害发生风险的概率分析[J].森林与环境学报,2017,37(1):95-101.
[8] REID I D. Biodegradation of lignin[J]. Canadian Journal of Botany,1995,73(S1):1 011-1 018.
[9] JAFARI N,SOUDI M R,KASRA-KERMANSHAHI R. Biodegradation perspectives of azo dyes by yeasts[J]. Microbiology,2014,83(5):484-497.
[10] VARMAN A M,HE L,FOLLENFANT R,et al. Decoding how a soil bacterium extracts building blocks and metabolic energy from ligninolysis provides road map for lignin valorization[J]. Proceedings of the National Academy of Sciences of the United States of America,2016,113(40):E5802-E5811.
[11] HATAKKA A. Lignin-modifying enzymes from selected white-rot fungi:production and role from in lignin degradation[J].FEMS Microbiology Reviews,1994,13(2/3):125-135.
[12] TAMURA K,NEI M,KUMAR S. Prospects for inferring very large phylogenies by using the neighbor-joining method[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(30):11 030-11 035.
[13] TAMURA K,PETERSON D,PETERSON N,et al. MEGA5:molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance, and maximum parsimony methods[J]. Molecular Biology and Evolution, 2011,28(10):2 731-2 739.
[14]张杰.白腐真菌的筛选、诱变及其对秸秆降解能力的研究[D].扬州:扬州大学,2010.
[15] ROTHSCHILD N,NOVOTNY'C,AEK V,et al. Ligninolytic enzymes of the fungus Irpex lacteus(Polyporus tulipiferae):isolation and characterization of lignin peroxidase[J]. Enzyme and Microbial Technology,2002,31(5):627-633.
[16] EGGERT C,TEMP U,ERIKSSON K E L. The ligninolytic system of the white rot fungus Pycnoporus cinnabarinus:purification and characterization of the laccase[J]. Applied and Environmental Microbiology,1996,62(4):1 151-1 158.
[17] PALMIERI G,GIARDINA P,BIANCO C,et al. Copper induction of laccase isoenzymes in the ligninolytic fungus Pleurotus ostreatus[J]. Applied and Environmental Microbiology,2000,66(3):920-924.
[18] REGALADO V,PERESTELO F,RODRGUEZ A,et al. Activated oxygen species and two extracellular enzymes:laccase and aryl-alcohol oxidase, novel for the lignin-degrading fungus Fusarium proliferatum[J]. Applied Microbiology and Biotechnology,1999,51(3):388-390.
[19] ARUN A,RAJA P P,ARTHI R,et al. Polycyclic aromatic hydrocarbons(PAHs)biodegradation by basidiomycetes fungi,Pseudomonas isolate,and their cocultures:comparative in vivo and in silico approach[J]. Applied Biochemistry and Biotechnology,2008,151(2/3):132-142.
[20]任丹,张波,张小平,等.川楝内生真菌的遗传及PKS、NRPS基因的多样性[J].中草药,2014,45(10):1 461-1 467.
[21] HIGUCHI T. Lignin biochemistry:biosynthesis and biodegradation[J]. Wood Science and Technology,1990,24(1):23-63.
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