枯草芽胞杆菌菌肥对有机冬瓜根区土壤微生态的影响
|
摘要
【背景】微生物肥料已广泛应用于我国有机作物的种植,其对有机种植土壤微生态的影响尚需科学评测。【目的】高通量测序技术可用于精确分析土壤微生物群落,从细菌、真菌群落结构和多样性的角度阐释枯草芽胞杆菌菌肥对有机农田根区土壤微生物群落的影响。【方法】在有机农田轮作种植条件下,施用枯草芽胞杆菌菌肥后提取冬瓜根区土壤基因组DNA,通过PCR扩增建立文库,利用IlluminaMiSeq高通量测序技术,并结合相关生物信息学方法分析土壤细菌16SrRNA基因V3-V4区和真菌ITS1区的多样性指数及群落结构;测定根区土壤化学性质及酶活性,分析有机冬瓜果实品质,并作相关分析。【结果】从6个有机冬瓜根区土壤样本中获得14199个细菌操作分类单元(OTU)和3378个真菌OTU,细菌和真菌文库测序覆盖率分别在98%、99%以上。枯草芽胞杆菌菌肥会在一定程度上提高土壤细菌种群多样性而降低真菌种群多样性,丰富了细菌群落结构,但显著降低了真菌群落丰富度(P<0.05);并减少了根区土壤特有细菌和真菌物种。变形菌门、厚壁菌门和放线菌门是优势细菌,子囊菌门是优势真菌;枯草芽胞杆菌菌肥会提高绿弯菌门和子囊菌门的相对丰度,比例分别为46.23%、10.01%;降低变形菌门和担子菌门的相对丰度,比例分别为11.14%、74.72%。枯草芽胞杆菌菌肥显著降低了土壤pH,显著提高了有机冬瓜果实总氨基酸、可溶性固形物等营养成分含量(P<0.05)。【结论】施用枯草芽胞杆菌菌肥改变有机冬瓜根区土壤细菌和真菌的丰富度和多样性,降低了土壤pH,提高了有机冬瓜果实品质。
[Background] Microbial fertilizers have been widely used in the cultivation of organic crops in China, and their effects on the micro-ecology in soils of organic farm still needs scientific evaluation.[Objective] The high-throughput sequencing technology can be used to analyze soil microbial community accurately. We explained effects of Bacillus subtilis microbial fertilizer on organic farm root-zone soil bacterial and fungal community in the view of structure and diversity. [Methods] Under organic and rotate farming conditions, soil genomic DNA was extracted after application of Bacillus subtilis microbial fertilizer. And PCR amplification was made to establish libraries. Then, the 16 S rRNA genes V3-V4 regions of soil bacterium and fungal ITS1 regions were sequenced by Illumina high-throughput sequencing technology on MiSeq platform, and related biological analysis was conducted to explain the changes of soil bacterial and fungal diversities and structures. In addition, soil chemical properties and enzyme activities, nutritional quality of organic Chinese watermelon were determined. [Results] A total of 14 199 bacterial operational taxonomic units(OTUs) and 3 378 fungal OTUs were obtained from 6 organic Chinese watermelon root-zone soil samples. The sequencing coverage of bacterial and fungal libraries were above 98% and 99%, respectively. Bacillus subtilis microbial fertilizer would increase the soil bacterial diversity while reduce the fungal diversity to a certain degree, and increased the abundance of bacteria, but it reduced the abundance of fungi significantly(P<0.05), and reduced the number of bacterial and fungal unique OTU. Proteobacteria, Firmicutes and Actinobacteria were the dominant bacteria, and Ascomycota was dominant fungi. It increased the relative abundance of Chloroflexi and Ascomycota by46.23% and 10.01%, respectively. And, it reduced the relative abundance of Proteobacteria and Basidiomycota by 11.14% and 74.72%, respectively. It significantly reduced the pH of soil(P<0.05).Wherein, it significantly increased the content of total amino acids and soluble solid in organic Chinese watermelon(P<0.05). [Conclusion] The abundance and diversity of soil bacteria and fungi were changed with Bacillus subtilis microbial fertilizer applied. And soil pH was reduced, nutritional quality of organic Chinese watermelon was increased also.
[Background] Microbial fertilizers have been widely used in the cultivation of organic crops in China, and their effects on the micro-ecology in soils of organic farm still needs scientific evaluation.[Objective] The high-throughput sequencing technology can be used to analyze soil microbial community accurately. We explained effects of Bacillus subtilis microbial fertilizer on organic farm root-zone soil bacterial and fungal community in the view of structure and diversity. [Methods] Under organic and rotate farming conditions, soil genomic DNA was extracted after application of Bacillus subtilis microbial fertilizer. And PCR amplification was made to establish libraries. Then, the 16 S rRNA genes V3-V4 regions of soil bacterium and fungal ITS1 regions were sequenced by Illumina high-throughput sequencing technology on MiSeq platform, and related biological analysis was conducted to explain the changes of soil bacterial and fungal diversities and structures. In addition, soil chemical properties and enzyme activities, nutritional quality of organic Chinese watermelon were determined. [Results] A total of 14 199 bacterial operational taxonomic units(OTUs) and 3 378 fungal OTUs were obtained from 6 organic Chinese watermelon root-zone soil samples. The sequencing coverage of bacterial and fungal libraries were above 98% and 99%, respectively. Bacillus subtilis microbial fertilizer would increase the soil bacterial diversity while reduce the fungal diversity to a certain degree, and increased the abundance of bacteria, but it reduced the abundance of fungi significantly(P<0.05), and reduced the number of bacterial and fungal unique OTU. Proteobacteria, Firmicutes and Actinobacteria were the dominant bacteria, and Ascomycota was dominant fungi. It increased the relative abundance of Chloroflexi and Ascomycota by46.23% and 10.01%, respectively. And, it reduced the relative abundance of Proteobacteria and Basidiomycota by 11.14% and 74.72%, respectively. It significantly reduced the pH of soil(P<0.05).Wherein, it significantly increased the content of total amino acids and soluble solid in organic Chinese watermelon(P<0.05). [Conclusion] The abundance and diversity of soil bacteria and fungi were changed with Bacillus subtilis microbial fertilizer applied. And soil pH was reduced, nutritional quality of organic Chinese watermelon was increased also.
引文
[1]Shen DL,Li J,Jiang X,et al.GB 20287-2006 Microbial inoculants in agriculture[S].Beijing:Standards Press of China,2006(in Chinese)沈德龙,李俊,姜昕,等.GB 20287-2006农用微生物菌剂[S].北京:中国标准出版社,2006
[2]Xu JG,Sun T,Li S.Application of microbial fertilizers in agricultural production of China[J].Crops,2016(1):1-6(in Chinese)许景钢,孙涛,李嵩.我国微生物肥料的研发及其在农业生产中的应用[J].作物杂志,2016(1):1-6
[3]Certification and Accreditation Administration of the People’s Republic of China and China Agricultural University.China Organic Product Certification and Organic Industry Development(2016)[M].Beijing:China Zhijian Publishing House,Standards Press of China,2017:1-3(in Chinese)国家认证认可监督管理委员会,中国农业大学.中国有机产品认证与有机产业发展(2016)[M].北京:中国质检出版社、中国标准出版社,2017:1-3
[4]Bokulich AN,Mills DA.Improved selection of internal transcribed spACEr-specific primers enables quantitative,ultra-high-throughput profiling of fungal communities[J].Applied and Environmental Microbiology,2013,79(8):2519-2526
[5]Caporaso JG,Lauber CL,Walters WA,et al.Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms[J].The ISME Journal,2012,6(8):1621-1624
[6]Lou J,Liu Y,Li Y.Review of high-throughput sequencing techniques in studies of soil microbial diversity[J].Chinese Agricultural Science Bulletin,2014,30(15):256-260(in Chinese)楼骏,柳勇,李延.高通量测序技术在土壤微生物多样性研究中的应用[J].中国农学通报,2014,30(15):256-260
[7]Lu RK.The Analysis Methods of Soils and Agrochemistry[M].Beijing:China Agricultural Science and Technology Press,2000:107-109(in Chinese)鲁如坤.土壤农业化学分析方法[M].北京:中国农业科技出版社,2000:107-109
[8]Xu N,Tan GC,Wang HY,et al.Effect of biochar additions to soil on nitrogen leaching,microbial biomass and bacterial community structure[J].European Journal of Soil Biology,2016,74:1-8
[9]The State Food and Drug Administration of the People’s Republic of China,The National Health and Family Planning Commission of the People’s Republic of China.GB 5009.124-2016 National food safety standard:determination of amino acids in foods[S].Beijing:China Standards Press,2016(in Chinese)中华人民共和国国家食品药品监督管理总局,中华人民共和国国家卫生和计划生育委员会.GB 5009.124-2016食品安全国家标准食品中氨基酸的测定[S].北京:中国标准出版社,2016
[10]The Ministry of Agriculture of the People’s Republic of China.NY/T 2637-2014 Determination of soluble solids content of fruits and vegetables:refractometer method[S].Beijing:China Standards Press,2014(in Chinese)中华人民共和国农业部.NY/T 2637-2014水果和蔬菜可溶性固形物含量的测定折射仪法[S].北京:中国标准出版社,2014
[11]The Ministry of Agriculture of the People’s Republic of China.NY/T 1278-2007 Determination of soluble sugar in vegetables and products shaffer-somogyi[S].Beijing:China Standards Press,2007(in Chinese)中华人民共和国农业部.NY/T 1278-2007蔬菜及其制品中可溶性糖的测定铜还原碘量法[S].北京:中国标准出版社,2007
[12]The State Administration of Quality Supervision,Inspection and Quarantine of the People’s Republic of China.GB/T 12456-2008Determination of total acid in foods[S].Beijing:China Standards Press,2008(in Chinese)中华人民共和国国家质量监督检验检疫总局.GB/T 12456-2008食品中总酸的测定[S].北京:中国标准出版社,2008
[13]The Ministry of Health of the People’s Republic of China.GB5009.5-2016 National food safety standard:determination of protein in foods[S].Beijing:China Standards Press,2016(in Chinese)中华人民共和国卫生部.GB 5009.5-2016食品安全国家标准食品中蛋白质的测定[S].北京:中国标准出版社,2016
[14]The Ministry of Health of the People’s Republic of China.GB/T5009.10-2003 Determination of crude fiber in vegetable foods[S].Beijing:China Standards Press,2003(in Chinese)中华人民共和国卫生部.GB/T 5009.10-2003植物类食品中粗纤维的测定[S].北京:中国标准出版社,2003
[15]The Ministry of Health of the People’s Republic of China.GB5009.86-2016 National food safety standards:Determination of reductive-form ascorbic acid in foods[S].Beijing:China Standards Press,2016(in Chinese)中华人民共和国卫生部.GB 5009.86-2016食品安全国家标准食品中抗坏血酸的测定[S].北京:中国标准出版社,2016
[16]The Ministry of Health of the People’s Republic of China.GB5009.33-2016 National food safety standard determination of nitrite and nitrate in foods[S].Beijing:China Standards Press,2016(in Chinese)中华人民共和国卫生部.GB 5009.33-2016食品安全国家标准食品中亚硝酸盐与硝酸盐的测定[S].北京:中国标准出版社,2016
[17]Mago?T,Salzberg SL.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics,2011,27(21):2957-2963
[18]Bolger AM,Lohse M,Usadel B.Trimmomatic:a flexible trimmer for Illumina sequence data[J].Bioinformatics,2014,30(15):2114-2120
[19]Quast C,Pruesse E,Yilmaz P,et al.The SILVA ribosomal RNAgene database project:improved data processing and web-based tools[J].Nucleic Acids Research,2013,41(D1):D590-D596
[20]Wang Q,Garrity GM,Tiedje JM,et al.Na?ve Bayesian classifier for rapid assignment of r RNA sequences into the new bacterial taxonomy[J].Applied and Environmental Microbiology,2007,73(16):5261-5267
[21]Pitta DW,Parmar N,Patel AK,et al.Bacterial diversity dynamics associated with different diets and different primer pairs in the rumen of Kankrej cattle[J].PLoS One,2014,9(11):e111710
[22]He Z,Zhao TT,Xing ZL,et al.Analysis of bacterial community composition in landfill cover soil[J].China Environmental Science,2015,35(12):3744-3753(in Chinese)何芝,赵天涛,邢志林,等.典型生活垃圾填埋场覆盖土微生物群落分析[J].中国环境科学,2015,35(12):3744-3753
[23]Chen HB,Boutros PC.VennDiagram:a package for the generation of highly-customizable Venn and Euler diagrams in R[J].BMC Bioinformatics,2011,12(1):35
[24]Gu X,Sun Q,Wang R,et al.Microbial fertilizer and organic fertilizer on soil improvement effect of plastic shed watermelon[J].Agricultural Research in the Arid Areas,2017,35(3):219-225(in Chinese)顾欣,孙权,王锐,等.菌肥与有机肥配施对拱棚西瓜土壤的改良效果[J].干旱地区农业研究,2017,35(3):219-225
[25]Schloss PD,Handelsman J.Status of the microbial census[J].Microbiology and Molecular Biology Reviews,2004,68(4):686-691
[26]Roesch LF,Fulthorpe RR,Riva A,et al.Pyrosequencing enumerates and contrasts soil microbial diversity[J].The ISMEJournal,2007,1(4):283-290
[27]Caporaso JG,Kuczynski J,Stombaugh J,et al.QIIME allows analysis of high-throughput community sequencing data[J].Nature Methods,2010,7(5):335-336
[28]You S,Zhang LM,Ji SG,et al.Impact of biocontrol agent Bacillus subtilis on bacterial communities in tobacco rhizospheric soil[J].Chinese Journal of Applied Ecology,2014,25(11):3323-3330(in Chinese)游偲,张立猛,计思贵,等.枯草芽孢杆菌菌剂对烟草根际土壤细菌群落的影响[J].应用生态学报,2014,25(11):3323-3330
[29]Kang J,Zhang SY,Han T,et al.Research on rhizosphere soil microbial diversity of two typical kinds of disease in yam[J].Biotechnology Bulletin,2017,33(7):107-113(in Chinese)康捷,章淑艳,韩韬,等.两种麻山药典型病害根际土壤微生物多样性的研究[J].生物技术通报,2017,33(7):107-113
[30]Dawar S,Wahab S,Tariq M,et al.Application of Bacillus species in the control of root rot diseases of crop plants[J].Archives of Phytopathology and Plant Protection,2010,43(4):412-418
[31]Jiang ZQ,Guo YH,Li SM,et al.Evaluation of biocontrol efficiency of different Bacillus preparations and field application methods against Phytophthora blight of bell pepper[J].Biological Control,2006,36(2):216-223
[32]Peng Z,Wang CJ,Chen QH,et al.Effects of bio-fertilizer“Ningdun”on controlling Phytophthora sojae and promoting soybean growth[J].Acta Agricultural Shanghai,2014,30(6):95-98(in chinese)彭震,王春娟,陈庆河,等.生物肥料“宁盾”对大豆疫霉病的防效及对毛豆的促生作用[J].上海农业学报,2014,30(6):95-98
[33]Jones RT,Robeson MS,Lauber CL,et al.A comprehensive survey of soil acidobacterial diversity using pyrosequencing and clone library analyses[J].The ISME Journal,2009,3(4):442-453
[34]Tian D,Ma X,Li YE,et al.Research on soil bacteria under the impact of sealed CO2 leakage by high-throughput sequencing technology[J].Environmental Science,2013,34(10):4096-4104(in Chinese)田地,马欣,李玉娥,等.利用高通量测序对封存CO2泄漏情景下土壤细菌的研究[J].环境科学,2013,34(10):4096-4104
[35]Liu J.Effects of soil microbial communities from cucumber rotation and continuous monocropping systems on cucumber seeding growth[D].Harbin:Master’s Thesis of Northeast Agricultural University,2015(in Chinese)刘杰.连作及轮作土壤微生物菌群对黄瓜生长的影响[D].哈尔滨:东北农业大学硕士学位论文,2015
[36]Li FH,Li M,Liu JQ,et al.Effect of biochar on fungal abundance of rhizosphere soil and cucumber root growth in greenhouse[J].Transactions of the Chinese Society for Agricultural Machinery,2017,48(4):265-270,341(in Chinese)李发虎,李明,刘金泉,等.生物炭对温室黄瓜根际土壤真菌丰度和根系生长的影响[J].农业机械学报,2017,48(4):265-270,341
[37]Di Pietro A,Gut-Rella M,Pachiatko JP,et al.Role of antibiotics produced by Chaetomium globosum in biocontrol of Pythium ultimum,a causal agent of damping-off[J].Phytopathology,1992,82(2):131-135
[38]Zhao ZT.Phylogenetic and comparative genomic analysis of fungal gene families related to fungal development and pathogenecity[D].Yangling:Doctoral Dissertation of Northwest Agriculture and Forestry University,2014(in Chinese)赵中涛.真菌生长发育和致病相关基因家族的比较和进化基因组学研究[D].杨凌:西北农林科技大学博士学位论文,2014
[39]Xing WF,Yu ZY,Chen LJ,et al.Control effect of biological fertilizer“Ningdun”on Fusarium oxysporum f.sp.melonis[J].Jiangsu Agricultural Sciences,2014,42(3):78-81(in Chinese)邢卫峰,于侦云,陈刘军,等.生物肥料“宁盾”对甜瓜枯萎病的防治效果[J].江苏农业科学,2014,42(3):78-81
[40]Fierer N,Bradford MA,Jackson RB.Toward an ecological classification of soil bacteria[J].Ecology,2007,88(6):1354-1364
[41]Xu YC,Shen QR,Ran W.Effect of zero-tillage and application of manure on soil microbial biomass C,N and P after sixteen years of cropping[J].Acta Pedologica Sinica,2002,39(1):89-96(in Chinese)徐阳春,沈其荣,冉炜.长期免耕与施用有机肥对土壤微生物生物量碳、氮、磷的影响[J].土壤学报,2002,39(1):89-96
[42]Ning CC,Wang JW,Cai KZ.The effects of organic fertilizers on soil fertility and soil environmental quality:A review[J].Ecology and Environmental Sciences,2016,25(1):175-181(in Chinese)宁川川,王建武,蔡昆争.有机肥对土壤肥力和土壤环境质量的影响研究进展[J].生态环境学报,2016,25(1):175-181
[43]Shan D,He JL,Xing ED,et al.Effects of microbial fertilizer on microorganism and soil enzyme activity in coal mine dump of typical steppe[J].Bulletin of Soil and Water Conservation,2017,37(3):81-85(in Chinese)珊丹,何京丽,邢恩德,等.微生物菌肥对草原矿区排土场土壤微生物与土壤酶活性的影响[J].水土保持通报,2017,37(3):81-85
[44]Sun DL,Jiang X,Wu QL,et al.Intragenomic heterogeneity of16S r RNA genes causes overestimation of Prokaryotic diversity[J].Applied and Environmental Microbiology,2013,79(19):5962-5969
[45]Zhou NY.The relationship between soil bacterial community and application of chemical fertilizers and straw returning[J].Microbiology China,2015,42(10):2048(in Chinese)周宁一.土壤细菌群落与施用化肥及秸秆还田的关系[J].微生物学通报,2015,42(10):2048
[46]Zhou SS,Xu J,Zhu H,et al.Gut microbiota-involved mechanisms in enhancing systemic exposure of ginsenosides by coexisting polysaccharides in ginseng decoction[J].Scientific Reports,2016,6:22474
[2]Xu JG,Sun T,Li S.Application of microbial fertilizers in agricultural production of China[J].Crops,2016(1):1-6(in Chinese)许景钢,孙涛,李嵩.我国微生物肥料的研发及其在农业生产中的应用[J].作物杂志,2016(1):1-6
[3]Certification and Accreditation Administration of the People’s Republic of China and China Agricultural University.China Organic Product Certification and Organic Industry Development(2016)[M].Beijing:China Zhijian Publishing House,Standards Press of China,2017:1-3(in Chinese)国家认证认可监督管理委员会,中国农业大学.中国有机产品认证与有机产业发展(2016)[M].北京:中国质检出版社、中国标准出版社,2017:1-3
[4]Bokulich AN,Mills DA.Improved selection of internal transcribed spACEr-specific primers enables quantitative,ultra-high-throughput profiling of fungal communities[J].Applied and Environmental Microbiology,2013,79(8):2519-2526
[5]Caporaso JG,Lauber CL,Walters WA,et al.Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms[J].The ISME Journal,2012,6(8):1621-1624
[6]Lou J,Liu Y,Li Y.Review of high-throughput sequencing techniques in studies of soil microbial diversity[J].Chinese Agricultural Science Bulletin,2014,30(15):256-260(in Chinese)楼骏,柳勇,李延.高通量测序技术在土壤微生物多样性研究中的应用[J].中国农学通报,2014,30(15):256-260
[7]Lu RK.The Analysis Methods of Soils and Agrochemistry[M].Beijing:China Agricultural Science and Technology Press,2000:107-109(in Chinese)鲁如坤.土壤农业化学分析方法[M].北京:中国农业科技出版社,2000:107-109
[8]Xu N,Tan GC,Wang HY,et al.Effect of biochar additions to soil on nitrogen leaching,microbial biomass and bacterial community structure[J].European Journal of Soil Biology,2016,74:1-8
[9]The State Food and Drug Administration of the People’s Republic of China,The National Health and Family Planning Commission of the People’s Republic of China.GB 5009.124-2016 National food safety standard:determination of amino acids in foods[S].Beijing:China Standards Press,2016(in Chinese)中华人民共和国国家食品药品监督管理总局,中华人民共和国国家卫生和计划生育委员会.GB 5009.124-2016食品安全国家标准食品中氨基酸的测定[S].北京:中国标准出版社,2016
[10]The Ministry of Agriculture of the People’s Republic of China.NY/T 2637-2014 Determination of soluble solids content of fruits and vegetables:refractometer method[S].Beijing:China Standards Press,2014(in Chinese)中华人民共和国农业部.NY/T 2637-2014水果和蔬菜可溶性固形物含量的测定折射仪法[S].北京:中国标准出版社,2014
[11]The Ministry of Agriculture of the People’s Republic of China.NY/T 1278-2007 Determination of soluble sugar in vegetables and products shaffer-somogyi[S].Beijing:China Standards Press,2007(in Chinese)中华人民共和国农业部.NY/T 1278-2007蔬菜及其制品中可溶性糖的测定铜还原碘量法[S].北京:中国标准出版社,2007
[12]The State Administration of Quality Supervision,Inspection and Quarantine of the People’s Republic of China.GB/T 12456-2008Determination of total acid in foods[S].Beijing:China Standards Press,2008(in Chinese)中华人民共和国国家质量监督检验检疫总局.GB/T 12456-2008食品中总酸的测定[S].北京:中国标准出版社,2008
[13]The Ministry of Health of the People’s Republic of China.GB5009.5-2016 National food safety standard:determination of protein in foods[S].Beijing:China Standards Press,2016(in Chinese)中华人民共和国卫生部.GB 5009.5-2016食品安全国家标准食品中蛋白质的测定[S].北京:中国标准出版社,2016
[14]The Ministry of Health of the People’s Republic of China.GB/T5009.10-2003 Determination of crude fiber in vegetable foods[S].Beijing:China Standards Press,2003(in Chinese)中华人民共和国卫生部.GB/T 5009.10-2003植物类食品中粗纤维的测定[S].北京:中国标准出版社,2003
[15]The Ministry of Health of the People’s Republic of China.GB5009.86-2016 National food safety standards:Determination of reductive-form ascorbic acid in foods[S].Beijing:China Standards Press,2016(in Chinese)中华人民共和国卫生部.GB 5009.86-2016食品安全国家标准食品中抗坏血酸的测定[S].北京:中国标准出版社,2016
[16]The Ministry of Health of the People’s Republic of China.GB5009.33-2016 National food safety standard determination of nitrite and nitrate in foods[S].Beijing:China Standards Press,2016(in Chinese)中华人民共和国卫生部.GB 5009.33-2016食品安全国家标准食品中亚硝酸盐与硝酸盐的测定[S].北京:中国标准出版社,2016
[17]Mago?T,Salzberg SL.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics,2011,27(21):2957-2963
[18]Bolger AM,Lohse M,Usadel B.Trimmomatic:a flexible trimmer for Illumina sequence data[J].Bioinformatics,2014,30(15):2114-2120
[19]Quast C,Pruesse E,Yilmaz P,et al.The SILVA ribosomal RNAgene database project:improved data processing and web-based tools[J].Nucleic Acids Research,2013,41(D1):D590-D596
[20]Wang Q,Garrity GM,Tiedje JM,et al.Na?ve Bayesian classifier for rapid assignment of r RNA sequences into the new bacterial taxonomy[J].Applied and Environmental Microbiology,2007,73(16):5261-5267
[21]Pitta DW,Parmar N,Patel AK,et al.Bacterial diversity dynamics associated with different diets and different primer pairs in the rumen of Kankrej cattle[J].PLoS One,2014,9(11):e111710
[22]He Z,Zhao TT,Xing ZL,et al.Analysis of bacterial community composition in landfill cover soil[J].China Environmental Science,2015,35(12):3744-3753(in Chinese)何芝,赵天涛,邢志林,等.典型生活垃圾填埋场覆盖土微生物群落分析[J].中国环境科学,2015,35(12):3744-3753
[23]Chen HB,Boutros PC.VennDiagram:a package for the generation of highly-customizable Venn and Euler diagrams in R[J].BMC Bioinformatics,2011,12(1):35
[24]Gu X,Sun Q,Wang R,et al.Microbial fertilizer and organic fertilizer on soil improvement effect of plastic shed watermelon[J].Agricultural Research in the Arid Areas,2017,35(3):219-225(in Chinese)顾欣,孙权,王锐,等.菌肥与有机肥配施对拱棚西瓜土壤的改良效果[J].干旱地区农业研究,2017,35(3):219-225
[25]Schloss PD,Handelsman J.Status of the microbial census[J].Microbiology and Molecular Biology Reviews,2004,68(4):686-691
[26]Roesch LF,Fulthorpe RR,Riva A,et al.Pyrosequencing enumerates and contrasts soil microbial diversity[J].The ISMEJournal,2007,1(4):283-290
[27]Caporaso JG,Kuczynski J,Stombaugh J,et al.QIIME allows analysis of high-throughput community sequencing data[J].Nature Methods,2010,7(5):335-336
[28]You S,Zhang LM,Ji SG,et al.Impact of biocontrol agent Bacillus subtilis on bacterial communities in tobacco rhizospheric soil[J].Chinese Journal of Applied Ecology,2014,25(11):3323-3330(in Chinese)游偲,张立猛,计思贵,等.枯草芽孢杆菌菌剂对烟草根际土壤细菌群落的影响[J].应用生态学报,2014,25(11):3323-3330
[29]Kang J,Zhang SY,Han T,et al.Research on rhizosphere soil microbial diversity of two typical kinds of disease in yam[J].Biotechnology Bulletin,2017,33(7):107-113(in Chinese)康捷,章淑艳,韩韬,等.两种麻山药典型病害根际土壤微生物多样性的研究[J].生物技术通报,2017,33(7):107-113
[30]Dawar S,Wahab S,Tariq M,et al.Application of Bacillus species in the control of root rot diseases of crop plants[J].Archives of Phytopathology and Plant Protection,2010,43(4):412-418
[31]Jiang ZQ,Guo YH,Li SM,et al.Evaluation of biocontrol efficiency of different Bacillus preparations and field application methods against Phytophthora blight of bell pepper[J].Biological Control,2006,36(2):216-223
[32]Peng Z,Wang CJ,Chen QH,et al.Effects of bio-fertilizer“Ningdun”on controlling Phytophthora sojae and promoting soybean growth[J].Acta Agricultural Shanghai,2014,30(6):95-98(in chinese)彭震,王春娟,陈庆河,等.生物肥料“宁盾”对大豆疫霉病的防效及对毛豆的促生作用[J].上海农业学报,2014,30(6):95-98
[33]Jones RT,Robeson MS,Lauber CL,et al.A comprehensive survey of soil acidobacterial diversity using pyrosequencing and clone library analyses[J].The ISME Journal,2009,3(4):442-453
[34]Tian D,Ma X,Li YE,et al.Research on soil bacteria under the impact of sealed CO2 leakage by high-throughput sequencing technology[J].Environmental Science,2013,34(10):4096-4104(in Chinese)田地,马欣,李玉娥,等.利用高通量测序对封存CO2泄漏情景下土壤细菌的研究[J].环境科学,2013,34(10):4096-4104
[35]Liu J.Effects of soil microbial communities from cucumber rotation and continuous monocropping systems on cucumber seeding growth[D].Harbin:Master’s Thesis of Northeast Agricultural University,2015(in Chinese)刘杰.连作及轮作土壤微生物菌群对黄瓜生长的影响[D].哈尔滨:东北农业大学硕士学位论文,2015
[36]Li FH,Li M,Liu JQ,et al.Effect of biochar on fungal abundance of rhizosphere soil and cucumber root growth in greenhouse[J].Transactions of the Chinese Society for Agricultural Machinery,2017,48(4):265-270,341(in Chinese)李发虎,李明,刘金泉,等.生物炭对温室黄瓜根际土壤真菌丰度和根系生长的影响[J].农业机械学报,2017,48(4):265-270,341
[37]Di Pietro A,Gut-Rella M,Pachiatko JP,et al.Role of antibiotics produced by Chaetomium globosum in biocontrol of Pythium ultimum,a causal agent of damping-off[J].Phytopathology,1992,82(2):131-135
[38]Zhao ZT.Phylogenetic and comparative genomic analysis of fungal gene families related to fungal development and pathogenecity[D].Yangling:Doctoral Dissertation of Northwest Agriculture and Forestry University,2014(in Chinese)赵中涛.真菌生长发育和致病相关基因家族的比较和进化基因组学研究[D].杨凌:西北农林科技大学博士学位论文,2014
[39]Xing WF,Yu ZY,Chen LJ,et al.Control effect of biological fertilizer“Ningdun”on Fusarium oxysporum f.sp.melonis[J].Jiangsu Agricultural Sciences,2014,42(3):78-81(in Chinese)邢卫峰,于侦云,陈刘军,等.生物肥料“宁盾”对甜瓜枯萎病的防治效果[J].江苏农业科学,2014,42(3):78-81
[40]Fierer N,Bradford MA,Jackson RB.Toward an ecological classification of soil bacteria[J].Ecology,2007,88(6):1354-1364
[41]Xu YC,Shen QR,Ran W.Effect of zero-tillage and application of manure on soil microbial biomass C,N and P after sixteen years of cropping[J].Acta Pedologica Sinica,2002,39(1):89-96(in Chinese)徐阳春,沈其荣,冉炜.长期免耕与施用有机肥对土壤微生物生物量碳、氮、磷的影响[J].土壤学报,2002,39(1):89-96
[42]Ning CC,Wang JW,Cai KZ.The effects of organic fertilizers on soil fertility and soil environmental quality:A review[J].Ecology and Environmental Sciences,2016,25(1):175-181(in Chinese)宁川川,王建武,蔡昆争.有机肥对土壤肥力和土壤环境质量的影响研究进展[J].生态环境学报,2016,25(1):175-181
[43]Shan D,He JL,Xing ED,et al.Effects of microbial fertilizer on microorganism and soil enzyme activity in coal mine dump of typical steppe[J].Bulletin of Soil and Water Conservation,2017,37(3):81-85(in Chinese)珊丹,何京丽,邢恩德,等.微生物菌肥对草原矿区排土场土壤微生物与土壤酶活性的影响[J].水土保持通报,2017,37(3):81-85
[44]Sun DL,Jiang X,Wu QL,et al.Intragenomic heterogeneity of16S r RNA genes causes overestimation of Prokaryotic diversity[J].Applied and Environmental Microbiology,2013,79(19):5962-5969
[45]Zhou NY.The relationship between soil bacterial community and application of chemical fertilizers and straw returning[J].Microbiology China,2015,42(10):2048(in Chinese)周宁一.土壤细菌群落与施用化肥及秸秆还田的关系[J].微生物学通报,2015,42(10):2048
[46]Zhou SS,Xu J,Zhu H,et al.Gut microbiota-involved mechanisms in enhancing systemic exposure of ginsenosides by coexisting polysaccharides in ginseng decoction[J].Scientific Reports,2016,6:22474