一种腐熟菌剂对粪肥腐熟过程中菌群结构及其肥效的影响
|
摘要
【目的】研究添加一种自制的腐熟菌剂,对牛粪、羊粪、鸡粪三种粪肥发酵腐熟后菌群结构和数量及其肥效的影响。【方法】采用高通量测序技术及RT-PCR技术,对三种腐熟后的粪肥细菌菌群结构和丰度进行分析;采用国标化学法分析有机肥主要理化指标;采用番茄育苗试验评价苗期肥效。【结果】在牛、羊、鸡粪中添加菌剂处理的样本中厚壁菌门(Firmicutes)和变形菌门(Proteobacteria)均有较高水平的聚类,菌群丰富度低且优势菌群种类单一,其中,在牛粪样品NA中乳酸菌属(Lactobacillu)OTUs占样品总OTUs的44%;育苗后穴盘样品优势菌群丰富度明显增加,其中丛毛单胞菌属(Comamonas)、短波单胞菌属(Brevundmonas)、变形杆菌(Dyadobacter)、鞘氨醇单胞菌(Sphingomonas)和假单胞菌(Pseudpmonas)均表现较高的丰度,而未加菌剂处理的腐熟样品和穴盘样品N5和N5P菌群变化不明显。甲基营养型芽孢杆菌(Bacillus methylotrophicus)和类球红细菌(Rhodobacter sphaeroides)在腐熟粪肥样本NA和JA中均有较高的拷贝数。添加菌剂处理粪肥能明显促进肥料酸化,使其pH在4.78～5.90,有机质含量提升到56%以上,全氮大于5%;番茄育苗试验中,牛粪NA、羊粪YA和鸡粪JA处理平均株高分别比对照组增高了47.16%、46.28%和47.65%。【结论】该腐熟菌剂能有效增加粪肥中功能菌群数量、优化优势菌群结构、并促进肥料酸化、显著提高肥料有机质和全氮含量,腐熟后的粪肥对番茄苗期生长肥效显著。
【Objective】 To research the effect of self-developed decomposing bacteria agent on the bacterial community and manurial efficiency in manure.【Method】High-flux sequencing and RT-PCR were used to investigate the microflora in three kinds of manure. The main physicochemical indexes of manure were analyzed by international standard chemical method, and tomatoes seedling raising test was used to evaluate the fertilizer efficiency.【Result】Firmicutes and Proteobacteria dominate at the end of fermentation in three kinds of manure. There were low diversity and few dominant communities of bacteria. The OTUs of Lactobacillu had 44% proportion of total OTUs. The bacterial diversity was abundant in sample undergoing tomatoes seedling raising including Comamonas, Brevundmonas, Dyadobacter, Sphingomonas and Pseudpmonas, but there was no obvious change in untreated sample N5 and N5 P. The results of RT-PCR showed that Bacillus methylotrophicus and Rhodobacter sphaeroides had large quantities in NA and JA. The manure could be obviously acidized and keep its pH Value between 4.78-5.90. The organic content increased to more than 56% and total nitrogen content was more than 5%. Through tomato seedling raising test average heights of tomato plants were 47.16%, 46.28% and 47.65%, respectively higher than those of the control group.【Conclusion】The decomposing bacteria agent can increase dominant microbiota in manure and accelerate the acidification of fertilizer dramatically. The fertilizer has a positive function on the growth of tomato plants in the seedling stage.
【Objective】 To research the effect of self-developed decomposing bacteria agent on the bacterial community and manurial efficiency in manure.【Method】High-flux sequencing and RT-PCR were used to investigate the microflora in three kinds of manure. The main physicochemical indexes of manure were analyzed by international standard chemical method, and tomatoes seedling raising test was used to evaluate the fertilizer efficiency.【Result】Firmicutes and Proteobacteria dominate at the end of fermentation in three kinds of manure. There were low diversity and few dominant communities of bacteria. The OTUs of Lactobacillu had 44% proportion of total OTUs. The bacterial diversity was abundant in sample undergoing tomatoes seedling raising including Comamonas, Brevundmonas, Dyadobacter, Sphingomonas and Pseudpmonas, but there was no obvious change in untreated sample N5 and N5 P. The results of RT-PCR showed that Bacillus methylotrophicus and Rhodobacter sphaeroides had large quantities in NA and JA. The manure could be obviously acidized and keep its pH Value between 4.78-5.90. The organic content increased to more than 56% and total nitrogen content was more than 5%. Through tomato seedling raising test average heights of tomato plants were 47.16%, 46.28% and 47.65%, respectively higher than those of the control group.【Conclusion】The decomposing bacteria agent can increase dominant microbiota in manure and accelerate the acidification of fertilizer dramatically. The fertilizer has a positive function on the growth of tomato plants in the seedling stage.
引文
[1]李振玉,等.有机肥和生物菌剂在我国农业生产中的作用及地位[M].北京:中国农业科学技术出版社,2017.LI Zhen-yu,et al.(2017).Role and status of organic fertilizer and biological bacterium agent in agricultural production [M].Beijing:China Agricultural Science and Technology Press.(in Chinese)
[2] 李博文,等.微生物肥料研发与应用[M].北京:中国农业出版社,2016.LI Bo-wen,et al.(2016).Research and application of microbial fertilizer [M].Beijing:China Agriculture Press.(in Chinese)
[3] 沈德龙,曹凤明,李力.我国生物有机肥的发展现状与展望[J].中国土壤与肥料,2007,(6):1-5.SHEN De-long,CAO Feng-ming,LI Li.(2007).Development status and prospect of microbial organic fertilizer in China [J].Soil and Fertilizer in China,(6):1-5.(in Chinese)
[4] 张余莽,周海军,张景野,等.生物有机肥的研究进展[J].吉林农业科学,2010,35(3):37-40.ZHANG Yu- mang,ZHOU Hai-jun,ZHANG Jing- ye,et al.(2010).Progress of Studies of Bio-organic Fertilize [J].Jilin Agricultural Sciences,35(3):37-40.(in Chinese)
[5] Ling,N.,Xue,C.,Huang,Q.,Yang,X.,Xu,Y.,& Shen,Q.(2010).Development of a mode of application of bioorganic fertilizer for improving the biocontrol efficacy tofusariumwilt.BioControl (Dordrecht),55(5):673-683.
[6] 田伟.牛粪高温堆肥过程中的物质变化、微生物多样性以及腐熟度评价研究 [D].南京:南京农业大学,2012.TIAN Wei,(2012).Study of substance change,microbialdiversity and maturity assessment during thermophilic composting of dairy manure [D].PhD Dissertation.Nanjing Agricultural University,Nanjing.(in Chinese)
[7] 李杰,郁继华,冯致,等.不同微生物菌剂对牛粪好氧堆肥的影响[J],干旱区资源与环境,2014,28(2):109-113.LI Jie,YU Ji-hua,FENG Zhi,et al.(2014).Effect of different microbial agents on cow manure in aerophile composting [J].Journal of Arid Land Resources and Environment,28(2):109-113.(in Chinese)
[8] 李杰.不同微生物菌剂对牛粪和玉米秸秆高温腐熟的影响[D].兰州:甘肃农业大学,2013.LI Jie.(2013).Effect of different microbial agents on cow manure and corn stalk in composting [D].Master Dissertation.Gansu Agricultural University,Lanzhou.(in Chinese)
[9] 李敬波.牛粪堆肥过程中优势菌群变化及复合菌剂制备[D].大连:大连理工大学,2014.LI Jing-bo,(2014).Changes of dominant microbiota during cattle manure composting and preparation of a microbial inoculant for composting [D].Master Dissertation.Dalian University of Technology,Daliang.(in Chinese)
[10] 李鸣雷,商鸿生,谷洁,等.促进农业废弃物腐解的复合微生物菌剂的筛选[J].西北农业学报,2005,14(2):101-104.LI Ming- lei,SHANG Hong -sheng,GU Jie,et al.(2005 ).Screening of the composite microorganism for promoting the decomposition of agriculture residue [J].Journal of Northwest Agriculture,14(2):101-104.(in Chinese)
[11] Liu,J.,Yang,H.,Zhao,M.,& Zhang,X.H.(2014).Spatial distribution patterns of benthic microbial communities along the pearl estuary,china.Systematic and Applied Microbiology,37(8):578-589.
[12] Miao,Y.,Liao,R.,Zhang,X.X.,Wang,Y.,Wang,Z.,& Shi,P.,et al.(2015).Metagenomic insights into cr (vi) effect on microbial communities and functional genes of an expanded granular sludge bed reactor treating high-nitrate wastewater.Water Research:43-52.
[13] 刘雅慈,何泽,张胜,等.油气田土壤甲烷氧化菌实时荧光定量PCR检测技术的建立与应用[J].微生物学通报,2014,41( 6) :1 071- 1 081.LIU Ya-ci,HE Ze,ZHANG Sheng,et al.(2014).Developmentand application of a fluorescent quantitative real-time PCR technique for detection of methane-oxidizing bacteria in oil and gas field soil [J].Microbiology China,41(6) :1,071-1,081.(in Chinese)
[14] 袁亚男,刘文忠.实时荧光定量PCR 技术的类型、特点与应用[J].中国畜牧兽医,2008,(3) :27 - 30.YUAN Ya -nan,LIU Wen-zhong.( 2008).The types,characteristics and applications of real-time fluoescent quantitative PCR techniques[J].China Animal Husbandry and Veterinary Medicine,(3) :27- 30.(in Chinese)
[15] 陈竞,冯蕾,杨新平,等.羊粪沼液微生物总DNA提取方法优化比较研究[J].新疆农业科学,2013,50(12):2 280-2 285.CHENG Jing,FENG Lei,YANG Xin-ping,et al.(2013).The Optimization Extraction Methods of Total Microbial DNA of Sheep Dung Biogas Slurry [J].Xinjiang Agricultural Sciences,50(12):2,280-2,285 .(in Chinese)
[16] 杨井泉,张云峰,高磊,等.2株兼具除臭功能的纤维素降解细菌的分离鉴定[J].家蓄生态学报,2018,39(9):46-51.YANG Jing-quan,ZHANG Yun-feng,GAO Lei.(2018).Isolation and Identification of two strains of cellulose-decomposing Bacterium with the function of deodorization [J].Journal of Domestic Animal Ecology,39(9):46-51.(in Chinese)
[17] 李世贵,王飞,顾金刚,等.微生物菌剂在农业废弃物堆肥腐熟过程中的应用及其田间试验效果[J].微生物学杂志,2011,31 (6):62-65.LI Shi-gui,WANG Fei,GU Jin-gang,et al.(2011).Application of microbial agent in the compost decomposition process of agricultural junks and its effect in the field experiment [J].Journal of Microbiology,31 (6):62-65.(in Chinese)
[18] 王钧漫.不同农田土壤微生物群落对石油污染及种植刺槐的响应[D].杨凌:西北农林科技大学,2016.WANG Jun-man.(2016).The Response of Microbial Community to Petroleun Pollution and Planting Robinia Pseudoacacia in Different Agricultural Soils [D].Master Dissertation.Northwestern A & F University,Yangling.(in Chinese).
[19] 程红胜,张玉华,向欣,等.不同菌剂对猪粪发酵中腐植酸含量的影响.[J]南方农业学报,2013,44(12):2 018-2 022.CHEN Hong-sheng,ZHANG Yu-hua,XIANG Xin,et al.(2013).Effects of microbial inoculum on humic acid content of fermented pig manure [J].Journal of Southern Agriculture,44(12):2,018-2,022.(in Chinese)
[20] Juteau,P.,Rho,D.,Larocque,R.,& Leduy,A.(1999).Analysis of the relative abundance of different types of bacteria capable of toluene degradation in a compost biofilter.Applied Microbiology and Biotechnology,52(6):863-868.
[2] 李博文,等.微生物肥料研发与应用[M].北京:中国农业出版社,2016.LI Bo-wen,et al.(2016).Research and application of microbial fertilizer [M].Beijing:China Agriculture Press.(in Chinese)
[3] 沈德龙,曹凤明,李力.我国生物有机肥的发展现状与展望[J].中国土壤与肥料,2007,(6):1-5.SHEN De-long,CAO Feng-ming,LI Li.(2007).Development status and prospect of microbial organic fertilizer in China [J].Soil and Fertilizer in China,(6):1-5.(in Chinese)
[4] 张余莽,周海军,张景野,等.生物有机肥的研究进展[J].吉林农业科学,2010,35(3):37-40.ZHANG Yu- mang,ZHOU Hai-jun,ZHANG Jing- ye,et al.(2010).Progress of Studies of Bio-organic Fertilize [J].Jilin Agricultural Sciences,35(3):37-40.(in Chinese)
[5] Ling,N.,Xue,C.,Huang,Q.,Yang,X.,Xu,Y.,& Shen,Q.(2010).Development of a mode of application of bioorganic fertilizer for improving the biocontrol efficacy tofusariumwilt.BioControl (Dordrecht),55(5):673-683.
[6] 田伟.牛粪高温堆肥过程中的物质变化、微生物多样性以及腐熟度评价研究 [D].南京:南京农业大学,2012.TIAN Wei,(2012).Study of substance change,microbialdiversity and maturity assessment during thermophilic composting of dairy manure [D].PhD Dissertation.Nanjing Agricultural University,Nanjing.(in Chinese)
[7] 李杰,郁继华,冯致,等.不同微生物菌剂对牛粪好氧堆肥的影响[J],干旱区资源与环境,2014,28(2):109-113.LI Jie,YU Ji-hua,FENG Zhi,et al.(2014).Effect of different microbial agents on cow manure in aerophile composting [J].Journal of Arid Land Resources and Environment,28(2):109-113.(in Chinese)
[8] 李杰.不同微生物菌剂对牛粪和玉米秸秆高温腐熟的影响[D].兰州:甘肃农业大学,2013.LI Jie.(2013).Effect of different microbial agents on cow manure and corn stalk in composting [D].Master Dissertation.Gansu Agricultural University,Lanzhou.(in Chinese)
[9] 李敬波.牛粪堆肥过程中优势菌群变化及复合菌剂制备[D].大连:大连理工大学,2014.LI Jing-bo,(2014).Changes of dominant microbiota during cattle manure composting and preparation of a microbial inoculant for composting [D].Master Dissertation.Dalian University of Technology,Daliang.(in Chinese)
[10] 李鸣雷,商鸿生,谷洁,等.促进农业废弃物腐解的复合微生物菌剂的筛选[J].西北农业学报,2005,14(2):101-104.LI Ming- lei,SHANG Hong -sheng,GU Jie,et al.(2005 ).Screening of the composite microorganism for promoting the decomposition of agriculture residue [J].Journal of Northwest Agriculture,14(2):101-104.(in Chinese)
[11] Liu,J.,Yang,H.,Zhao,M.,& Zhang,X.H.(2014).Spatial distribution patterns of benthic microbial communities along the pearl estuary,china.Systematic and Applied Microbiology,37(8):578-589.
[12] Miao,Y.,Liao,R.,Zhang,X.X.,Wang,Y.,Wang,Z.,& Shi,P.,et al.(2015).Metagenomic insights into cr (vi) effect on microbial communities and functional genes of an expanded granular sludge bed reactor treating high-nitrate wastewater.Water Research:43-52.
[13] 刘雅慈,何泽,张胜,等.油气田土壤甲烷氧化菌实时荧光定量PCR检测技术的建立与应用[J].微生物学通报,2014,41( 6) :1 071- 1 081.LIU Ya-ci,HE Ze,ZHANG Sheng,et al.(2014).Developmentand application of a fluorescent quantitative real-time PCR technique for detection of methane-oxidizing bacteria in oil and gas field soil [J].Microbiology China,41(6) :1,071-1,081.(in Chinese)
[14] 袁亚男,刘文忠.实时荧光定量PCR 技术的类型、特点与应用[J].中国畜牧兽医,2008,(3) :27 - 30.YUAN Ya -nan,LIU Wen-zhong.( 2008).The types,characteristics and applications of real-time fluoescent quantitative PCR techniques[J].China Animal Husbandry and Veterinary Medicine,(3) :27- 30.(in Chinese)
[15] 陈竞,冯蕾,杨新平,等.羊粪沼液微生物总DNA提取方法优化比较研究[J].新疆农业科学,2013,50(12):2 280-2 285.CHENG Jing,FENG Lei,YANG Xin-ping,et al.(2013).The Optimization Extraction Methods of Total Microbial DNA of Sheep Dung Biogas Slurry [J].Xinjiang Agricultural Sciences,50(12):2,280-2,285 .(in Chinese)
[16] 杨井泉,张云峰,高磊,等.2株兼具除臭功能的纤维素降解细菌的分离鉴定[J].家蓄生态学报,2018,39(9):46-51.YANG Jing-quan,ZHANG Yun-feng,GAO Lei.(2018).Isolation and Identification of two strains of cellulose-decomposing Bacterium with the function of deodorization [J].Journal of Domestic Animal Ecology,39(9):46-51.(in Chinese)
[17] 李世贵,王飞,顾金刚,等.微生物菌剂在农业废弃物堆肥腐熟过程中的应用及其田间试验效果[J].微生物学杂志,2011,31 (6):62-65.LI Shi-gui,WANG Fei,GU Jin-gang,et al.(2011).Application of microbial agent in the compost decomposition process of agricultural junks and its effect in the field experiment [J].Journal of Microbiology,31 (6):62-65.(in Chinese)
[18] 王钧漫.不同农田土壤微生物群落对石油污染及种植刺槐的响应[D].杨凌:西北农林科技大学,2016.WANG Jun-man.(2016).The Response of Microbial Community to Petroleun Pollution and Planting Robinia Pseudoacacia in Different Agricultural Soils [D].Master Dissertation.Northwestern A & F University,Yangling.(in Chinese).
[19] 程红胜,张玉华,向欣,等.不同菌剂对猪粪发酵中腐植酸含量的影响.[J]南方农业学报,2013,44(12):2 018-2 022.CHEN Hong-sheng,ZHANG Yu-hua,XIANG Xin,et al.(2013).Effects of microbial inoculum on humic acid content of fermented pig manure [J].Journal of Southern Agriculture,44(12):2,018-2,022.(in Chinese)
[20] Juteau,P.,Rho,D.,Larocque,R.,& Leduy,A.(1999).Analysis of the relative abundance of different types of bacteria capable of toluene degradation in a compost biofilter.Applied Microbiology and Biotechnology,52(6):863-868.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |