生物炭不同施用量对烟草根际土壤微生物多样性的影响
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摘要
为研究生物炭不同施用量对烟草根际土壤微生物多样性的影响。以烤烟品种云烟87根际土壤为研究对象,对不施用生物炭的对照处理,以及3个生物炭施用量递增处理的根际土壤ITS2区和16S r DNA-V4区进行高通量测序,对下机数据进行生物信息学分析,得到了各处理根际土壤微生物的OTU丰度、分布、α多样性、群落种类组成及丰度信息,并对群落组成及丰度分别进行了PCA聚类分析及UPGMA聚类分析。结果表明,在50~150 g/棵的施用范围内,增加生物炭的施用量提高了根际土壤细菌种类的多样性和分布的均匀程度,真菌则相反;施用生物炭50,100,150 g/棵处理后,变形菌门细菌的丰度相比不施用生物炭的对照稍有降低,分别降低了4.1%,2.7%,0.7%;酸杆菌门细菌的丰度相比对照分别增加了10.4%,8.1%,7.7%,蓝菌门细菌的丰度相比对照分别降低了11.4%,11.4%,11.2%,放线菌门细菌的丰度均低于对照,各处理间的芽单胞菌门细菌丰度差异甚微。施用生物炭50 g/棵处理后,接合菌门真菌丰度比对照(50.92%)下降了12.38%,之后随着生物炭施用量的提高,接合菌门真菌丰度逐渐上升至53.68%;与之相反,施用生物炭50 g/棵处理后,子囊菌门真菌丰度比对照(30.63%)升高了10.15%,之后随着生物炭施用量的提高,子囊菌门真菌丰度逐渐下降至29.11%;施用生物炭50 g/棵处理后,担子菌门和壶菌门真菌丰度基本不变,之后随着生物炭施用量的提高,担子菌门真菌丰度呈现出先上升后降低的趋势;各处理对球囊菌门真菌丰度影响不大。研究生物炭对烟草根际微生态的影响方式以及作用规律,可为生物炭在烟田的应用提供理论依据。
In order to study the influence of different application amounts of charcoal on the microbial diversity in tobacco rhizosphere soil,with the rhizosphere soil of flue-cured tobacco variety Yunyan 87 as the research object,we compared the situation by applying different amounts of charcoal,and conducted high-throughput sequencing of rhizosphere soil ITS2 region and 16 S r DNA-V4 region with progressive treatment with 3 application amounts of charcoal; through bioinformatics analysis of data,we obtained the OTU abundance,distribution,α diversity,community species composition and abundance information of microorganisms in rhizosphere soil when different treatment methods were used,and conducted PCA clustering analysis and UPGMA clustering analysis of the com-munity composition and abundance. The results showed that within the application scope of 50-150 g/tree,by increasing the application amounts of charcoal,it could increase the variety diversity and distribution uniformity of bacteria in rhizosphere soil,while it would reduce the variety diversity and distribution uniformity of fungus; compared to the control group where no charcoal was applied,after applying the charcoal with the amounts of 50,100,150 g/tree,the abundances of Proteobacteria bacteria had declined by 4. 1%,2. 7% and 0. 7% respectively; the abundances of Acidobacteria bacteria increased by 10. 4%,8. 1% and 7. 7% respectively; the abundances of Actinobacteria bacteria were all lower than the abundance of control group; the difference in abundance of Gemmatimonadetes bacteria between various groups was very small. After applying the charcoal with the amount of 50 g/tree,the abundance of Zygomycota fungus had declined by 12. 38% compared to the control group( 50. 92%),and then,with the increased of the application amount of charcoal,the abundance of Zygomycota fungus gradually increased to 53. 68%; on the contrary,after applying the charcoal with the amount of 50 g/tree,the abundance of Ascomycota fungus had increased by 10. 15% compared to the control group( 30. 63%),and then,with the increased of the application amount of charcoal,the abundance of Ascomycota fungus gradually declined to 29. 11%;after applying the charcoal with the amount of 50 g/tree,the abundances of Basidiomycota and Chytridiomycota funguses almost had no change,and then,with the increased of the application amount of charcoal,the abundances of Basidiomycota and Chytridiomycota funguses presented the trend of increased first,and then declined; various treatment methods did not have significant influence on the abundance of Glomeromycota fungus. By studying the influencing method and action rules of charcoal application on the micro-ecology of tobacco rhizosphere,it can provide theoretical basis to the application of charcoal in tobacco field.
In order to study the influence of different application amounts of charcoal on the microbial diversity in tobacco rhizosphere soil,with the rhizosphere soil of flue-cured tobacco variety Yunyan 87 as the research object,we compared the situation by applying different amounts of charcoal,and conducted high-throughput sequencing of rhizosphere soil ITS2 region and 16 S r DNA-V4 region with progressive treatment with 3 application amounts of charcoal; through bioinformatics analysis of data,we obtained the OTU abundance,distribution,α diversity,community species composition and abundance information of microorganisms in rhizosphere soil when different treatment methods were used,and conducted PCA clustering analysis and UPGMA clustering analysis of the com-munity composition and abundance. The results showed that within the application scope of 50-150 g/tree,by increasing the application amounts of charcoal,it could increase the variety diversity and distribution uniformity of bacteria in rhizosphere soil,while it would reduce the variety diversity and distribution uniformity of fungus; compared to the control group where no charcoal was applied,after applying the charcoal with the amounts of 50,100,150 g/tree,the abundances of Proteobacteria bacteria had declined by 4. 1%,2. 7% and 0. 7% respectively; the abundances of Acidobacteria bacteria increased by 10. 4%,8. 1% and 7. 7% respectively; the abundances of Actinobacteria bacteria were all lower than the abundance of control group; the difference in abundance of Gemmatimonadetes bacteria between various groups was very small. After applying the charcoal with the amount of 50 g/tree,the abundance of Zygomycota fungus had declined by 12. 38% compared to the control group( 50. 92%),and then,with the increased of the application amount of charcoal,the abundance of Zygomycota fungus gradually increased to 53. 68%; on the contrary,after applying the charcoal with the amount of 50 g/tree,the abundance of Ascomycota fungus had increased by 10. 15% compared to the control group( 30. 63%),and then,with the increased of the application amount of charcoal,the abundance of Ascomycota fungus gradually declined to 29. 11%;after applying the charcoal with the amount of 50 g/tree,the abundances of Basidiomycota and Chytridiomycota funguses almost had no change,and then,with the increased of the application amount of charcoal,the abundances of Basidiomycota and Chytridiomycota funguses presented the trend of increased first,and then declined; various treatment methods did not have significant influence on the abundance of Glomeromycota fungus. By studying the influencing method and action rules of charcoal application on the micro-ecology of tobacco rhizosphere,it can provide theoretical basis to the application of charcoal in tobacco field.
引文
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[11]赵秋芳,马海洋,王辉,等.生物炭对香草兰生长及根际土壤微生物的影响[J].湖北农业科学,2015,54(22):5647-5651.
[12]刘领,王艳芳,宋久洋,等.生物炭与氮肥减量配施对烤烟生长及土壤酶活性的影响[J].河南农业科学,2016,45(2):62-66.
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[21]Van Zwieten L,Kimber S,Morris S,et al.effects of biochar from slow pyrolysis of papermill waste on agronomic performance and soil fertility[J].Plant and Soil,2010,327(1/2):235-246.
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[25]Yamato M,Okimori Y,Wibowo I F,et al.Effects of the application of charred bark of Acacia mangium on the yield of maize,cowpea and peanut,and soil chemical properties in South Sumatra,Indonesia[J].Soil Science&Plant Nutrition,2006,52(4):489-495.
[26]Li X,Zhang H,Wu M,et al.Effect of methamidophos on soil fungi community in microcosms by plate count,DGGE and clone library analysis[J].Journal of Environmental Sciences-China,2008,20(5):619-625.
[27]Grossman J M,O'neill B E,Tsai S M,et al.Amazonian anthrosols support similar microbial communities that differ distinctly from those extant in adjacent,unmodified soils of the same mineralogy[J].Microbial Ecology,2010,60(1):192-205.
[28]陈泽斌,李冰,王定康,等.薄荷内生细菌的多样性及组成分析[J].浙江农业学报,2016,28(1):56-63.
[29]赵帅,周娜,赵振勇,等.基于高通量测序分析盐角草根部内生细菌多样性及动态规律[J].微生物学报,2016,56(6):1000-1008.
[30]徐慧敏,闫海,马松,等.鞘氨醇单胞菌USTB-05对微囊藻毒素的生物降解[J].中国环境科学,2014,34(5):1316-1321.
[2]赵殿峰,徐静,罗璇,等.生物炭对土壤养分、烤烟生长以及烟叶化学成分的影响[J].西北农业学报,2014,23(3):85-92.
[3]陈尧,郑华,石俊雄,等.施用化肥和菜籽粕对烤烟根际微生物的影响[J].土壤学报,2012,49(1):198-203.
[4]宋久洋,刘领,陈明灿,等.生物质炭施用对烤烟生长及光合特性的影响[J].河南科技大学学报:自然科学版,2014,35(4):68-72.
[5]李航,董涛,王明元.生物炭对香蕉苗根际土壤微生物群落与代谢活性的影响[J].微生物学杂志,2016,36(1):42-48.
[6]韩光明,孟军,曹婷,等.生物炭对菠菜根际微生物及土壤理化性质的影响[J].沈阳农业大学学报,2012,43(5):515-520.
[7]孙大荃,孟军,张伟明,等.生物炭对棕壤大豆根际微生物的影响[J].沈阳农业大学学报,2011,42(5):521-526.
[8]毛家伟,张锦中,张翔,等.豫东烟区生物炭对烤烟生长发育及经济性状的影响[J].安徽农业科学,2013,41(35):13516-13517.
[9]叶协锋,李志鹏,于晓娜,等.生物炭用量对植烟土壤碳库及烤后烟叶质量的影响[J].中国烟草学报,2015,21(5):33-41.
[10]湛方栋,陆引罡,关国经,等.烤烟根际微生物群落结构及其动态变化的研究[J].土壤学报,2005,42(3):488-494.
[11]赵秋芳,马海洋,王辉,等.生物炭对香草兰生长及根际土壤微生物的影响[J].湖北农业科学,2015,54(22):5647-5651.
[12]刘领,王艳芳,宋久洋,等.生物炭与氮肥减量配施对烤烟生长及土壤酶活性的影响[J].河南农业科学,2016,45(2):62-66.
[13]Caporaso J G,Lauber C L,Walters W A,et al.Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample[J].Proceedings of the National Academy of Sciences of the United States of America,2011,108(S):4516-4522.
[14]Hess M,Sczyrba A,Egan R,et al.Metagenomic discovery of biomass-degrading genes and genomes from cow rumen[J].Science,2011,331(616):463-467.
[15]Edgar R C,Haas B J,Clemente J C,et al.UCHIME improves sensitivity and speed of chimera detection[J].Bioinformatics,2011,27(16):2194-2200.
[16]Edgar R C.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.
[17]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.
[18]Desantis T Z,Hugenholtz P,Larsen N,et al.Greengenes,a chimera-checked 16S rRNA gene database and workbench compatible with ARB[J].Applied and Environmental Microbiology,2006,72(7):5069-5072.
[19]Gulino L M,Ouwerkerk D,Kang A Y,et al.Shedding light on the microbial community of the macropod foregut using 454-amplicon pyrosequencing[J].PLo S One,2013,8(4):e61463.
[20]Graber E R,Meller Harel Y,Kolton M,et al.Biochar impact on development and productivity of pepper and tomato grown in fertigated soilless media[J].Plant and Soil,2010,337(1/2):481-496.
[21]Van Zwieten L,Kimber S,Morris S,et al.effects of biochar from slow pyrolysis of papermill waste on agronomic performance and soil fertility[J].Plant and Soil,2010,327(1/2):235-246.
[22]Steinbeiss S,Gleixner G,Antonietti M.Effect of biochar amendment on soil carbon balance and soil microbial activity[J].Soil Biology and Biochemistry,2009,41(6):1301-1310.
[23]Matthias C R,Marcel W,Mohamed S,et al.Material derived from hydrothermal carbonization:effects on plant growth and Arbuscular mycorrhiza[J].Applied Soil Ecology,2010,45(3):238-242.
[24]Rondon M A,Lehmann J,Ramírez J,et al.Biological nitrogen fixation by common beans(Phaseolus vulgaris L.)increases with bio-char additions[J].Biology and Fertility of Soils,2007,43(6):699-708.
[25]Yamato M,Okimori Y,Wibowo I F,et al.Effects of the application of charred bark of Acacia mangium on the yield of maize,cowpea and peanut,and soil chemical properties in South Sumatra,Indonesia[J].Soil Science&Plant Nutrition,2006,52(4):489-495.
[26]Li X,Zhang H,Wu M,et al.Effect of methamidophos on soil fungi community in microcosms by plate count,DGGE and clone library analysis[J].Journal of Environmental Sciences-China,2008,20(5):619-625.
[27]Grossman J M,O'neill B E,Tsai S M,et al.Amazonian anthrosols support similar microbial communities that differ distinctly from those extant in adjacent,unmodified soils of the same mineralogy[J].Microbial Ecology,2010,60(1):192-205.
[28]陈泽斌,李冰,王定康,等.薄荷内生细菌的多样性及组成分析[J].浙江农业学报,2016,28(1):56-63.
[29]赵帅,周娜,赵振勇,等.基于高通量测序分析盐角草根部内生细菌多样性及动态规律[J].微生物学报,2016,56(6):1000-1008.
[30]徐慧敏,闫海,马松,等.鞘氨醇单胞菌USTB-05对微囊藻毒素的生物降解[J].中国环境科学,2014,34(5):1316-1321.