一株木质素降解菌的筛选鉴定及其在堆肥中的应用
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摘要
为提高农林废弃物堆肥中木质素的降解效率,促进堆肥腐熟,提高堆肥品质,研究筛选了一株高效木质素降解菌株,并应用于猪粪与枯枝落叶混合堆肥。研究结果表明,筛选菌株YZC3对木质素的降解率达到79.2%,可同时产生木质素过氧化物酶、锰过氧化物酶和漆酶,其中锰过氧化物酶活性最强,为16.4 U/mL。经形态学和ITS分子鉴定,确定YZC3其为粉红粘帚霉(Clonostachys rosea strain)。将其应用于堆肥中,可延长高温持续天数6d和二次发酵天数3 d,堆肥结束时T值降为0.59,而对照堆肥为0.86,显著促进了堆肥腐熟。YZC3还有助于提高堆肥产品中腐植酸含量,减少堆肥中全氮的损失,是一种极具应用价值的堆肥菌剂。
In order to improve the degradation efficiency of lignin in agricultural and forestry waste compost,promote compost maturity and improve compost quality,a strain of high-efficiency lignin degradation strain was studied and applied to pig manure and litter mixed compost.The results showed that the degradation rate of lignin by screening strain YZC3 reached 79.2%,which could produce lignin peroxidase,manganese peroxidase and laccase simultaneously.The highest manganese peroxidase activity was 16.4 U/mL.Based on morphology and ITS molecular identification,YZC3 was identified as Clonostachys rosea strain.Applying it to compost can extend the high temperature lasting days for 6 d and the second fermentation days for 3 d.At the end of composting,the T value is reduced to 0.59 and the control compost is 0.86,which significantly promotes compost maturity.YZC3 also helps to increase the humic acid content in compost products and reduce the loss of TN in compost.It is a highly useful composting agent.
In order to improve the degradation efficiency of lignin in agricultural and forestry waste compost,promote compost maturity and improve compost quality,a strain of high-efficiency lignin degradation strain was studied and applied to pig manure and litter mixed compost.The results showed that the degradation rate of lignin by screening strain YZC3 reached 79.2%,which could produce lignin peroxidase,manganese peroxidase and laccase simultaneously.The highest manganese peroxidase activity was 16.4 U/mL.Based on morphology and ITS molecular identification,YZC3 was identified as Clonostachys rosea strain.Applying it to compost can extend the high temperature lasting days for 6 d and the second fermentation days for 3 d.At the end of composting,the T value is reduced to 0.59 and the control compost is 0.86,which significantly promotes compost maturity.YZC3 also helps to increase the humic acid content in compost products and reduce the loss of TN in compost.It is a highly useful composting agent.
引文
[1]Himmel M E,Ding S Y,Johnson D K,et al.Biomass recalcitrance:engineering plants and enzymes for biofuels production[J].Science,2007,315(5813):804-807.
[2]王庆福,黄清铧,梁磊,等.灵芝木质素降解酶研究及其潜在应用进展[J].热带作物学报,2015,36(7):1361-1367.
[3]徐衣显,刘晓,王伟.我国生物质废物污染现状与资源化发展趋势[J].再生利用,2008,1(5):31-34.
[4]路瑶,魏贤勇,宗志敏,等.木质素的结构研究与应用[J].化学进展,2013,25(05):838-858.
[5]武志远.木质素生物合成途径中关键酶基因的分子特征[J].农村科学实验,2017,(1):89-90.
[6]黄映芳,蔡明亮,吴敏雅.造纸黑液中木质素降解的研究现状与进展[J].纸和造纸,2018,(1):12-17.
[7]Moubarik A,Grimi N,Boussetta N,et al.Isolation and characterization of lignin from Moroccan sugar cane bagasse:Production of ligninphenol-formaldehyde wood adhesive[J].Industrial Crops and Products,2013,45:296-302.
[8]彭晓光,杨林娥,张磊.生物法降解秸秆木质素研究进展[J].现代农业科技,2010,(1):18-20.
[9]刘杰凤,马超,王春,等.海洋微生物纤维素酶及半纤维素酶基因克隆与表达研究进展[J].生物技术通报,2012,(6):36-42.
[10]冯波,林元山,胡超,等.一株木质素降解菌的筛选、鉴定及其漆酶发酵条件的优化[J].湖南师范大学(自然科学学报),2015,(2):12-16,73.
[11]钱静亚,张正沛,季蓉蓉,等.3株真菌固态发酵产木质素降解酶的研究[J].江苏农业科学,2017,45(5):277-280.
[12]马英辉,李利军,卢美欢,等.木质素降解放线菌的鉴定及产酶特性研究[J].环境污染与防治,2018,40(8):902-912.
[13]张鹏飞,李素艳,余克非,等.木质素降解细菌的筛选及园林废弃物降解研究[J].安徽农业大学学报,2018,45(4):676-681.
[14]陈莹,张俊涛,阮琳.园林废弃物堆肥中木质素降解菌的鉴定及其降解能力研究[J].华南农业大学学报,2014,(6):94-98.
[15]王宜磊,朱陶,邓振旭.愈创木酚法快速筛选漆酶产生菌[J].生物技术,2007,17(4):40-42.
[16]蔡磊,尹峻峰,杨丽萍,等.几种简便的木质素降解真菌定性筛选方法[J].微生物学通报,2002,29(1):67-69.
[17]倪启亮,王敦球,戚拓业.木质素降解酶的酶活测定方法探讨[J].广西农学报,2008,23(2):54-57.
[18]Faison B D,Kirk T K.Factors involved in the regulation of a ligninase activity in phanerochaete chrysosporium[J].Applied and Environmental Microbiology,1985,49:299-304.
[19]杨胜.饲料分析及饲料质量检测技术(第一版)[M].北京:北京农业大学出版社,1999.58-63.
[20]吕伟成,张绍升.香蕉枯萎病菌1号和4号生理小种rDNA-ITS的比较[J].福建农林大学学报(自然科学版),2009,38(3):231-233.
[21]司梦莹,师德强,戴友芝,等.高效木质素降解菌的筛选及降解性能研究[J].广西科学,2015,(1):48-52,59.
[22]刘月,许修宏,徐杰,等.功能菌剂对堆肥中木质纤维素降解的影响[J].中国土壤与肥料,2014,(4):81-86.
[23]宋彩红,贾璇,李鸣晓,等.沼渣与畜禽粪便混合堆肥发酵效果的综合评价[J].农业工程学报,2013,29(24):227-232.
[24]Morel T L,Colin F,Germon J C,et al.Methods for the evaluation of the maturity of municipal refuse compost//Gasser J KR.Composting of agricultural and other wastes[C]//London:Elsevier Appled Science publishers,1985.56-72.
[25]Plaza C,Nogales R,Senesi N,et al.Organic matter humification by vermicomposting of cattle manure alone and mixed with two-phase olive pomace[J].Bioresource Technology,2008,99(11):5085-5089.
[26]Zhang L,Sun X Y.Changes in physical,chemical,and microbiological properties during the two-stage co-composting of green waste with spent mushroom compost and biochar[J].Bioresource Technology,2014,171:274-284.
[27]Lopez M J,Vargas-García M C,Suárez-Estrella F,et al.Biodelignific ation and humi fication of horticultural plant residues by fungi[J].International Biodeteri oration&Biodegradation,2006,57:24-30.
[28]郁红艳.农业废物堆肥化中木质素降解及其微生物特性研究[D].长沙:湖南大学,2007.120-127.
[29]张力,喜霞,大勇.白腐真菌木质素降解酶系研究进展[J].吉林畜牧兽医,2009,30(2):9-12.
[30]鲍文英,江经纬,周云,等.一株木质纤维素降解菌的筛选及其全基因组分析[J].微生物学报,2016,56(5):765-777.
[31]Ambert-Balay K,Dougherty M,Tien M.Reactivity of manganese peroxidase:site-directed mutagenesis of residues in proximity to the porphyrin ring[J].Archives of Biochemistry and Biophysics,2000,382(1):89-94.
[32]杨暖,张妙直,宋洪英.木质素过氧化物酶应用研究进展[J].现代农业科学,2009,16(4):19-20.
[33]赖超凤,李爽,彭丽丽,等.漆酶及其在有机合成中应用的研究进展[J].化工进展,2010,29(7):1300-1308.
[34]Tuomela M,Vikman M,Hatakka A,et al.Biodegradation of lignin in a compost environment:a review[J].Bioresource Technology,2000,72(2):169-183.
[35]Sivasubramanian S,Namasivayam S K R.Phenol degradation studies using microbial consortium isolated from environmental sources[J].Journal of Environmental Chemical Engineering,2015,3(1):243-252.
[36]Carvalho A L D A,Rezende L C D,Costa L B,et al.Optimizing the mass production of Clonostachys rosea by liquid-state fermentation[J].Biological Control,2018,118:16-25.
[36]Zhang L,Yang J,Niu Q,et al.Investigation on the infection mechanism of the fungus Clonostachys rosea against nematodes using the green fluorescent protein[J].Applied Microbiology&Biotechnology,2008,78(6):983-990.
[38]Anwar W,Ali S,Nawaz K,et al.Entomopathogenic fungus Clonostachysrosea as a biocontrol agent against whitefly(Bemisia tabaci)[J].Biocontrol Science and Technology,2018,28(8):750-760.
[39]赵士振.粉红粘帚霉防治果蔬灰霉病的研究[D].武汉:华中农业大学,2015.
[40]李丹阳,邓启得,雍明丽,等.稻曲病菌菌核降解微生物的筛选与作用机制分析[J].中国生物防治学报,2016,32(2):258-264.
[41]王淑芳,马桂珍,李世东,等.应用实时荧光定量技术评价粉红粘帚霉对水稻纹枯病的田间防治效果[J].植物病理学报,2014,(4):422-427.
[42]王瑞虎.粉红粘帚霉诱导番茄果实对灰霉病的非寄主抗性研究[D].哈尔滨:东北农业大学,2013.
[43]李书强,李林会,沈江洁,等.生防菌对黄瓜枯萎病防效及其对黄瓜诱导抗性测定[J].河北科技师范学院学报,2017,(1):53-58.
[44]高航.粉红粘帚霉对四种林果枝干病原菌的生防作用研究[D].泰安:山东农业大学,2016.
[45]Sun Z B,Zhang J,Sun M H,et al.Identification of genes related to chlamydospore formation in Clonostachys rosea 67-1.[J].Microbiologyopen,2018:e00624.
[2]王庆福,黄清铧,梁磊,等.灵芝木质素降解酶研究及其潜在应用进展[J].热带作物学报,2015,36(7):1361-1367.
[3]徐衣显,刘晓,王伟.我国生物质废物污染现状与资源化发展趋势[J].再生利用,2008,1(5):31-34.
[4]路瑶,魏贤勇,宗志敏,等.木质素的结构研究与应用[J].化学进展,2013,25(05):838-858.
[5]武志远.木质素生物合成途径中关键酶基因的分子特征[J].农村科学实验,2017,(1):89-90.
[6]黄映芳,蔡明亮,吴敏雅.造纸黑液中木质素降解的研究现状与进展[J].纸和造纸,2018,(1):12-17.
[7]Moubarik A,Grimi N,Boussetta N,et al.Isolation and characterization of lignin from Moroccan sugar cane bagasse:Production of ligninphenol-formaldehyde wood adhesive[J].Industrial Crops and Products,2013,45:296-302.
[8]彭晓光,杨林娥,张磊.生物法降解秸秆木质素研究进展[J].现代农业科技,2010,(1):18-20.
[9]刘杰凤,马超,王春,等.海洋微生物纤维素酶及半纤维素酶基因克隆与表达研究进展[J].生物技术通报,2012,(6):36-42.
[10]冯波,林元山,胡超,等.一株木质素降解菌的筛选、鉴定及其漆酶发酵条件的优化[J].湖南师范大学(自然科学学报),2015,(2):12-16,73.
[11]钱静亚,张正沛,季蓉蓉,等.3株真菌固态发酵产木质素降解酶的研究[J].江苏农业科学,2017,45(5):277-280.
[12]马英辉,李利军,卢美欢,等.木质素降解放线菌的鉴定及产酶特性研究[J].环境污染与防治,2018,40(8):902-912.
[13]张鹏飞,李素艳,余克非,等.木质素降解细菌的筛选及园林废弃物降解研究[J].安徽农业大学学报,2018,45(4):676-681.
[14]陈莹,张俊涛,阮琳.园林废弃物堆肥中木质素降解菌的鉴定及其降解能力研究[J].华南农业大学学报,2014,(6):94-98.
[15]王宜磊,朱陶,邓振旭.愈创木酚法快速筛选漆酶产生菌[J].生物技术,2007,17(4):40-42.
[16]蔡磊,尹峻峰,杨丽萍,等.几种简便的木质素降解真菌定性筛选方法[J].微生物学通报,2002,29(1):67-69.
[17]倪启亮,王敦球,戚拓业.木质素降解酶的酶活测定方法探讨[J].广西农学报,2008,23(2):54-57.
[18]Faison B D,Kirk T K.Factors involved in the regulation of a ligninase activity in phanerochaete chrysosporium[J].Applied and Environmental Microbiology,1985,49:299-304.
[19]杨胜.饲料分析及饲料质量检测技术(第一版)[M].北京:北京农业大学出版社,1999.58-63.
[20]吕伟成,张绍升.香蕉枯萎病菌1号和4号生理小种rDNA-ITS的比较[J].福建农林大学学报(自然科学版),2009,38(3):231-233.
[21]司梦莹,师德强,戴友芝,等.高效木质素降解菌的筛选及降解性能研究[J].广西科学,2015,(1):48-52,59.
[22]刘月,许修宏,徐杰,等.功能菌剂对堆肥中木质纤维素降解的影响[J].中国土壤与肥料,2014,(4):81-86.
[23]宋彩红,贾璇,李鸣晓,等.沼渣与畜禽粪便混合堆肥发酵效果的综合评价[J].农业工程学报,2013,29(24):227-232.
[24]Morel T L,Colin F,Germon J C,et al.Methods for the evaluation of the maturity of municipal refuse compost//Gasser J KR.Composting of agricultural and other wastes[C]//London:Elsevier Appled Science publishers,1985.56-72.
[25]Plaza C,Nogales R,Senesi N,et al.Organic matter humification by vermicomposting of cattle manure alone and mixed with two-phase olive pomace[J].Bioresource Technology,2008,99(11):5085-5089.
[26]Zhang L,Sun X Y.Changes in physical,chemical,and microbiological properties during the two-stage co-composting of green waste with spent mushroom compost and biochar[J].Bioresource Technology,2014,171:274-284.
[27]Lopez M J,Vargas-García M C,Suárez-Estrella F,et al.Biodelignific ation and humi fication of horticultural plant residues by fungi[J].International Biodeteri oration&Biodegradation,2006,57:24-30.
[28]郁红艳.农业废物堆肥化中木质素降解及其微生物特性研究[D].长沙:湖南大学,2007.120-127.
[29]张力,喜霞,大勇.白腐真菌木质素降解酶系研究进展[J].吉林畜牧兽医,2009,30(2):9-12.
[30]鲍文英,江经纬,周云,等.一株木质纤维素降解菌的筛选及其全基因组分析[J].微生物学报,2016,56(5):765-777.
[31]Ambert-Balay K,Dougherty M,Tien M.Reactivity of manganese peroxidase:site-directed mutagenesis of residues in proximity to the porphyrin ring[J].Archives of Biochemistry and Biophysics,2000,382(1):89-94.
[32]杨暖,张妙直,宋洪英.木质素过氧化物酶应用研究进展[J].现代农业科学,2009,16(4):19-20.
[33]赖超凤,李爽,彭丽丽,等.漆酶及其在有机合成中应用的研究进展[J].化工进展,2010,29(7):1300-1308.
[34]Tuomela M,Vikman M,Hatakka A,et al.Biodegradation of lignin in a compost environment:a review[J].Bioresource Technology,2000,72(2):169-183.
[35]Sivasubramanian S,Namasivayam S K R.Phenol degradation studies using microbial consortium isolated from environmental sources[J].Journal of Environmental Chemical Engineering,2015,3(1):243-252.
[36]Carvalho A L D A,Rezende L C D,Costa L B,et al.Optimizing the mass production of Clonostachys rosea by liquid-state fermentation[J].Biological Control,2018,118:16-25.
[36]Zhang L,Yang J,Niu Q,et al.Investigation on the infection mechanism of the fungus Clonostachys rosea against nematodes using the green fluorescent protein[J].Applied Microbiology&Biotechnology,2008,78(6):983-990.
[38]Anwar W,Ali S,Nawaz K,et al.Entomopathogenic fungus Clonostachysrosea as a biocontrol agent against whitefly(Bemisia tabaci)[J].Biocontrol Science and Technology,2018,28(8):750-760.
[39]赵士振.粉红粘帚霉防治果蔬灰霉病的研究[D].武汉:华中农业大学,2015.
[40]李丹阳,邓启得,雍明丽,等.稻曲病菌菌核降解微生物的筛选与作用机制分析[J].中国生物防治学报,2016,32(2):258-264.
[41]王淑芳,马桂珍,李世东,等.应用实时荧光定量技术评价粉红粘帚霉对水稻纹枯病的田间防治效果[J].植物病理学报,2014,(4):422-427.
[42]王瑞虎.粉红粘帚霉诱导番茄果实对灰霉病的非寄主抗性研究[D].哈尔滨:东北农业大学,2013.
[43]李书强,李林会,沈江洁,等.生防菌对黄瓜枯萎病防效及其对黄瓜诱导抗性测定[J].河北科技师范学院学报,2017,(1):53-58.
[44]高航.粉红粘帚霉对四种林果枝干病原菌的生防作用研究[D].泰安:山东农业大学,2016.
[45]Sun Z B,Zhang J,Sun M H,et al.Identification of genes related to chlamydospore formation in Clonostachys rosea 67-1.[J].Microbiologyopen,2018:e00624.
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