高通量测序解析微生物肥料对红壤有机农田土壤微生物群落的影响
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摘要
高通量测序技术可用于精确分析土壤微生物群落,从微生物群落结构和多样性的角度阐释微生物肥料对有机农田根区土壤微生物群落的影响。在红壤有机农田轮作种植条件下,施用微生物肥料后利用Illumina MiSeq高通量测序技术结合相关生物信息学分析土壤细菌和真菌的多样性指数及群落结构。结果表明,从6个有机农田根区土壤样本中获得7 729个细菌分类操作单元(operational taxonomic units,简称OTU)和3 271个真菌OTU,细菌和真菌文库测序覆盖率均在99%以上。微生物肥料会显著降低土壤细菌和真菌种群多样性,且可在一定程度上降低细菌群落丰富度,显著降低真菌群落丰富度;并减少根区土壤特有细菌和真菌物种数量。放线菌门、变形菌门和酸杆菌门是优势细菌,子囊菌门是优势真菌;微生物肥料会提高放线菌门、变形菌门、厚壁菌门和担子菌门的相对丰度,分别比对照组提高29. 46%、9. 17%、129. 33%、165. 73%;但会降低酸杆菌门、绿弯菌门、子囊菌门、接合菌门的相对丰度,分别比对照组降低30. 14%、33. 50%、17. 27%、86. 33%。因此,施用微生物肥料可改变红壤有机农田根区土壤细菌和真菌的丰富度和多样性,有助于控制作物病害发生。
引文
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[29]韩哲,刘守伟,潘凯,等.不同栽培模式对黄瓜根际土壤酶活性及细菌群落结构的影响[J].植物营养与肥料学报,2012,18(4):922-931.
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[31]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.
[32]Di Pietro A,Gut-Rella M,Pachlatko J P,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.
[2]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]国家认证认可监督管理委员会.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,2017:1-3.
[4]Bokulich N A,Mills D A.Improved selection of internal transcribed spacer-specific primers enables quantitative,ultra-highthroughput profiling of fungal communities[J].Applied and Environmental Microbiology,2013,79(8):2519-2526.
[5]Caporaso J G,Lauber C L,Walters W A,et al.Ultra-highthroughput microbial community analysis on the Illumina Hi Seq and Mi Seq platforms[J].The ISME Journal,2012,6(8):1621-1624.
[6]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研究进展[J].中国农学通报,2014,30(15):256-260.
[7]鲁如坤.土壤农业化学分析方法[M].北京:中国农业科学技术出版社,2000:107-109.
[8]Xu N,Tan G C,Wang H Y,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]Magoc T,Salzberg S L.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics,2011,27(21):2957-2963.
[10]Bolger A M,Lohse M,Usadel B.Trimmomatic:a flexible trimmer for Illumina sequence data[J].Bioinformatics,2014,30(15):2114-2120.
[11]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.
[12]Wang Q,Garrity G M,Tiedje J M,et al.Naive bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy[J].Applied and Environmental Microbiology,2007,73(16):5261-5267.
[13]Pitta D W,Parmar N,Patel A K,et al.Bacterial diversity dynamics associated with different diets and different primer pairs in the rumen of Kankrej cattle[J].PLo S One,2014,9(11):e111710.
[14]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.
[15]Chen H,Boutros P C.VennDiagram:a package for the generation of highly-customizable Venn and Euler diagrams in R[J].BMCBioinformatics,2011,12:35.
[16]杜思瑶,于淼,刘芳华,等.设施种植模式对土壤细菌多样性及群落结构的影响[J].中国生态农业学报,2017,25(11):1615-1625.
[17]Zhao S,Liu D Y,Ling N,et al.Bio-organic fertilizer application significantly reduces the Fusarium oxysporum population and alters the composition of fungi communities of watermelon Fusarium wilt rhizosphere soil[J].Biology and Fertility of Soils,2014,50(5):765-774.
[18]杨亚东,王志敏,曾昭海.长期施肥和灌溉对土壤细菌数量、多样性和群落结构的影响[J].中国农业科学,2018,51(2):290-301.
[19]Liu J J,Sui Y E,Yu Z H,et al.High throughput sequencing analysis of biogeographical distribution of bacterial communities in the black soils of northeast China[J].Soil Biology and Biochemistry,2014,70:113-122.
[20]陈丹梅,段玉琪,杨宇虹,等.长期施肥对植烟土壤养分及微生物群落结构的影响[J].中国农业科学,2014,47(17):3424-3433.
[21]Caporaso J G,Kuczynski J,Stombaugh J,et al.QIIME allows analysis of high-throughput community sequencing data[J].Nature Methods,2010,7(5):335-336.
[22]吴江利,罗学刚,李宝强,等.微生物菌肥作用下荒漠土壤微生物群落结构和功能研究[J].中国农学通报,2015,31(9):216-223.
[23]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.
[24]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.
[25]Luo J,Ran W,Hu J,et al.Application of Bio-Organic fertilizer significantly affected fungal diversity of soils[J].Soil Science Society of America Journal,2010,74(6):2039-2048.
[26]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.
[27]Jiang Z Q,Guo Y H,Li S M,et al.Evaluation of biocontrol efficiencies of different Bacillus preparations and different field application methods against Phytophthora bight of bell pepper[J].Biological Control,2006,36(2):216-223.
[28]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.
[29]韩哲,刘守伟,潘凯,等.不同栽培模式对黄瓜根际土壤酶活性及细菌群落结构的影响[J].植物营养与肥料学报,2012,18(4):922-931.
[30]姜飞,刘业霞,艾希珍,等.嫁接辣椒根际土壤微生物及酶活性与根腐病抗性的关系[J].中国农业科学,2010,43(16):3367-3374.
[31]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.
[32]Di Pietro A,Gut-Rella M,Pachlatko J P,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.