多效唑对芒果园土壤细菌多样性的影响及PICRUSt基因功能预测分析
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摘要
为了探究多效唑对芒果园土壤微生物多样性和群落结构的影响,本研究以海南乐东某芒果园为研究对象,设置施用和不施用多效唑的土壤分别为处理组和对照组,通过Illumina Miseq测序平台对芒果园土壤进行16S rRNA高通量测序分析。2组样品通过高通量测序共得到3586个OTUs(operational taxonomic units),可注释到38个门、89个纲、195个目、378个科、673个属、1353个种。Alpha多样性分析表明,施用多效唑后,土壤细菌丰富度指数显著高于同时期未施用多效唑的土壤;但施药后土壤的细菌多样性显著降低。主成分分析表明,施用多效唑对土壤细菌的群落结构产生影响。与对照组相比,施用多效唑的土壤中变形菌门和浮霉菌门等含量显著增高,而放线菌门和厚壁菌门等含量显著降低。PICRUSt功能预测分析结果表明,芒果园土壤细菌主要涉及细胞生长与死亡、碳水化合物代谢、次生产物代谢的生物合成、氨基酸代谢等43个子功能,表现出功能上的丰富性。多效唑处理后,会降低土壤细菌的整体代谢能力。由此可见,施用多效唑会降低土壤细菌的多样性,改变土壤细菌的相对丰度,且对土壤细菌的功能会有一定的影响。
The Illumina Miseq 16 S rRNA high-throughput sequencing technology was used to study the bacterial community structure and diversity of two group soil samples with and without paclobutrazol treatment,respectively,from a mango orchard in Ledong,Hainan.A total of 3586 operational taxonomic units(OTUs) in two groups were obtained,which were classified as 38 phylums,89 classes,195 orders,378 families,673 genera and 1353 species.The results of the analysis of Alpha diversity showed that soil bacterial richness index was significantly higher in the paclobutrazo-treated soils,however compared with the soils without paclobutrazol treatment,the bacterial diversity was significantly decreased.The principal component analysis indicated that the community structure of soil bacteria was affected by paclobutrazol.The quantities of Proteobacteria and Planctomycetes were significantly increased in the paclobutrazol-treated soils,however,those of Actinobacteria and Firmicutes were significantly decreased.The PICRUSt analysis showed that the functional gene families were mainly related to 43 sub-functions including the cell growth and death,carbohydrate metabolism,biosynthesis of secondary metabolites,amino acid metabolism,and so on.The total metabolic capacity of soil bacteria could be reduced after treated with paclobutrazol.Therefore,the application of paclobutrazol can reduce the diversity of soil bacteria,and it also has some impacts on the relative abundance and functions of soil bacteria.
The Illumina Miseq 16 S rRNA high-throughput sequencing technology was used to study the bacterial community structure and diversity of two group soil samples with and without paclobutrazol treatment,respectively,from a mango orchard in Ledong,Hainan.A total of 3586 operational taxonomic units(OTUs) in two groups were obtained,which were classified as 38 phylums,89 classes,195 orders,378 families,673 genera and 1353 species.The results of the analysis of Alpha diversity showed that soil bacterial richness index was significantly higher in the paclobutrazo-treated soils,however compared with the soils without paclobutrazol treatment,the bacterial diversity was significantly decreased.The principal component analysis indicated that the community structure of soil bacteria was affected by paclobutrazol.The quantities of Proteobacteria and Planctomycetes were significantly increased in the paclobutrazol-treated soils,however,those of Actinobacteria and Firmicutes were significantly decreased.The PICRUSt analysis showed that the functional gene families were mainly related to 43 sub-functions including the cell growth and death,carbohydrate metabolism,biosynthesis of secondary metabolites,amino acid metabolism,and so on.The total metabolic capacity of soil bacteria could be reduced after treated with paclobutrazol.Therefore,the application of paclobutrazol can reduce the diversity of soil bacteria,and it also has some impacts on the relative abundance and functions of soil bacteria.
引文
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[12]袁志华,程波,常玉海,等.15%多效唑可湿性粉剂对土壤微生物多样性的影响研究[J].农业环境科学学报,2008(5):1848-1852.
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[14]Silva M S,Célia Maria,Vieira F,et al.Paclobutrazol effects on soil microorganisms[J].Applied Soil Ecology,2003,22(1):79-86.
[15]方治国,郝翠梅,姚文冲,等.空气微生物群落解析方法:从培养到非培养[J].生态学报,2016,36(14):4244-4253.
[16]王岩,沈锡权,吴祖芳,等.PCR-SSCP技术在微生物群落多态性分析中的应用进展[J].生物技术,2009,19(3):84-87.
[17]林生,庄家强,陈婷,等.不同年限茶树根际土壤微生物群落PLFA生物标记多样性分析[J].生态学杂志,2013,32(1):64-71.
[18]夏围围,贾仲君.高通量测序和DGGE分析土壤微生物群落的技术评价[J].微生物学报,2014,54(12):1489-1499.
[19]张丁予,章婷曦,王国祥.第二代测序技术的发展及应用[J].环境科学与技术,2016(9):96-102.
[20]Wu Y,Ke X,Hernández M,et al.Autotrophic growth of bacterial and archaeal ammonia oxidizers in freshwater sediment microcosms incubated at different temperatures[J].Applied and Environmental Microbiology,2013,79(9):3076-3084.
[21]Justyna P,Rebecchi A,Pisacane V,et al.Bacterial diversity in typical Italian salami at different ripening stages as revealed by high-throughput sequencing of 16S r RNA amplicons[J].Food Microbiology,2015,46:342-356.
[22]Langille M G I,Zaneveld J,Caporaso J G,et al.Predictive functional profiling of microbial communities using 16Sr RNA marker gene sequences[J].Nature Biotechnology,2013,31(9):814-821.
[23]Polónia A R M,Richard C D F,Coelho F J R d C,et al.Compositional analysis of archaeal communities in high and low microbial abundance sponges in the Misool coral reef system,Indonesia[J].Marine Biology Research,2018:1-14.
[24]董志颖,洪慢,胡晗静,等.过量氮输入对寡营养海水细菌群落代谢潜力的影响[J].环境科学学报,2018,38(2):457-466.
[25]张菲,田伟,孙峰,等.丹江口库区表层浮游细菌群落组成与PICRUSt功能预测分析[J].环境科学,2019(3):1-14.
[26]厉桂香,马克明.北京东灵山树线处土壤细菌的PICRUSt基因预测分析[J].生态学报,2018,38(6):2180-2186.
[27]孙峰,田伟,张菲,等.丹江口库区库滨带植被土壤细菌群落多样性及PICRUSt功能预测分析[J].环境科学,2019(1):1-13.
[28]程扬,刘子丹,沈启斌,等.秸秆生物炭施用对玉米根际和非根际土壤微生物群落结构的影响[J].生态环境学报,2018,27(10):1870-1877.
[29]鲍士旦.土壤农化分析[M].3版.北京:中国农业出版社,2000.
[30]Caporaso J G,Lauber C L,Walters W A,et al.Global patterns of 16S r RNA 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(Supplement 1):4516-4522.
[31]Waingwright M,吴文礼.农药对土壤微生物活性的影响[J].土壤学进展,1980(4):31-35.
[32]张娜娜,姜博,邢奕,等.有机磷农药污染土壤的微生物降解研究进展[J].土壤,2018,50(4):645-655.
[33]华健,陈曼佳,李芳柏,等.水稻土中五氯酚的降解转化动态及其对微生物群落的影[J].地球与环境,2018,46(3):225-230.
[34]刘倩倩.三种常用杀菌剂在麦田土壤中的消解动态及对土壤微生物的毒性评价[D].泰安:山东农业大学,2016.
[35]伍云丽.腈菌唑在茶园土壤中的消解动力学及其对土壤微生物活性影响的研究[D].杭州:浙江大学,2014.
[36]Kuo J,Wang Y W,Chen M,et al.The effect of paclobutrazol on soil bacterial composition across three consecutive flowering stages of mung bean[J].Folia Microbiologica,2019,64(2):197-205.
[37]Hua F,Tang F,Wei Z,et al.Persistence of repeated triadimefon application and its impact on soil microbial functional diversity[J].Journal of Environmental Science and Health Part B,2012,47(2):104-110.
[38]Jeffries T C,Rayu S,Nielsen U N,et al.Metagenomic functional potential predicts degradation rates of a model organophosphorus xenobiotic in pesticide contaminated soils[J].Frontiers in Microbiology,2018,9:147.
[39]Zafra G,Taylor T D,Absalón,et al.Comparative metagenomic analysis of PAH degradation in soil by a mixed microbial consortium[J].Journal of Hazardous Materials,2016,318:702-710.
[40]徐帅,林奕岑,周梦佳,等.基于高通量测定肉鸡回肠微生物多样性及PICRUSt基因预测分析[J].动物营养学报,2016,28(8):2581-2588.
[2]Vaz F L,Netto A M,Antonino A C D,et al.Modeling of the kinetics biodegradation of paclobutrazol in two soils of the semiarid northeast brazil[J].Quimica Nova,2012,35(1):77-81.
[3]孙定波.反季节芒果控梢催花技术[J].中国热带农业,2008(5):53-53.
[4]Kishore K,Singh H S,Kurian R M.Paclobutrazol use in perennial fruit crops and its residual effects:A review[J].Indian Journal of Agricultural Sciences,2015,85(7):863-872.
[5]Singh V K,Garg N,Bhirigwanshi S R.Effect of paclobutrazol doses on nutritional and microbiological properties of mango(Mangifera indica)orchard soils[J].Indian Journal of Agricultural Sciences,2005,75:738-739.
[6]Fierer N.Embracing the unknown:disentangling the complexities of the soil microbiome[J].Nature Reviews Microbiology,2017,15(10):579-590.
[7]Doran J W,Fraser D G,Culik M N,et al.Influence of alternative and conventional agricultural management on soil microbial processes and nitrogen availability[J].American Journal of Alternative Agriculture,1987,2(3):99-106.
[8]李旭东,徐莉娜,薛林贵.土壤微生物多样性研究的新进展[J].广州化工,2014(20):6-10.
[9]杨成德,龙瑞军,陈秀蓉,等.土壤微生物功能群及其研究进展[J].土壤通报,2008,39(2):421-425.
[10]许光辉,郑洪元.土壤微生物分析方法手册[M].北京:农业出版社,1986:102-110.
[11]赵秀芬,房增国,高祖明.多效唑对土壤微生物数量及部分酶活性的影响[J].南方农业学报,2005,36(3):234-235.
[12]袁志华,程波,常玉海,等.15%多效唑可湿性粉剂对土壤微生物多样性的影响研究[J].农业环境科学学报,2008(5):1848-1852.
[13]Gon?alves I C R,Araújo A S F,Carvalho E M S,et al.Effect of paclobutrazol on microbial biomass,respiration and cellulose decomposition in soil[J].European Journal of Soil Biology,2009,45(3):235-238.
[14]Silva M S,Célia Maria,Vieira F,et al.Paclobutrazol effects on soil microorganisms[J].Applied Soil Ecology,2003,22(1):79-86.
[15]方治国,郝翠梅,姚文冲,等.空气微生物群落解析方法:从培养到非培养[J].生态学报,2016,36(14):4244-4253.
[16]王岩,沈锡权,吴祖芳,等.PCR-SSCP技术在微生物群落多态性分析中的应用进展[J].生物技术,2009,19(3):84-87.
[17]林生,庄家强,陈婷,等.不同年限茶树根际土壤微生物群落PLFA生物标记多样性分析[J].生态学杂志,2013,32(1):64-71.
[18]夏围围,贾仲君.高通量测序和DGGE分析土壤微生物群落的技术评价[J].微生物学报,2014,54(12):1489-1499.
[19]张丁予,章婷曦,王国祥.第二代测序技术的发展及应用[J].环境科学与技术,2016(9):96-102.
[20]Wu Y,Ke X,Hernández M,et al.Autotrophic growth of bacterial and archaeal ammonia oxidizers in freshwater sediment microcosms incubated at different temperatures[J].Applied and Environmental Microbiology,2013,79(9):3076-3084.
[21]Justyna P,Rebecchi A,Pisacane V,et al.Bacterial diversity in typical Italian salami at different ripening stages as revealed by high-throughput sequencing of 16S r RNA amplicons[J].Food Microbiology,2015,46:342-356.
[22]Langille M G I,Zaneveld J,Caporaso J G,et al.Predictive functional profiling of microbial communities using 16Sr RNA marker gene sequences[J].Nature Biotechnology,2013,31(9):814-821.
[23]Polónia A R M,Richard C D F,Coelho F J R d C,et al.Compositional analysis of archaeal communities in high and low microbial abundance sponges in the Misool coral reef system,Indonesia[J].Marine Biology Research,2018:1-14.
[24]董志颖,洪慢,胡晗静,等.过量氮输入对寡营养海水细菌群落代谢潜力的影响[J].环境科学学报,2018,38(2):457-466.
[25]张菲,田伟,孙峰,等.丹江口库区表层浮游细菌群落组成与PICRUSt功能预测分析[J].环境科学,2019(3):1-14.
[26]厉桂香,马克明.北京东灵山树线处土壤细菌的PICRUSt基因预测分析[J].生态学报,2018,38(6):2180-2186.
[27]孙峰,田伟,张菲,等.丹江口库区库滨带植被土壤细菌群落多样性及PICRUSt功能预测分析[J].环境科学,2019(1):1-13.
[28]程扬,刘子丹,沈启斌,等.秸秆生物炭施用对玉米根际和非根际土壤微生物群落结构的影响[J].生态环境学报,2018,27(10):1870-1877.
[29]鲍士旦.土壤农化分析[M].3版.北京:中国农业出版社,2000.
[30]Caporaso J G,Lauber C L,Walters W A,et al.Global patterns of 16S r RNA 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(Supplement 1):4516-4522.
[31]Waingwright M,吴文礼.农药对土壤微生物活性的影响[J].土壤学进展,1980(4):31-35.
[32]张娜娜,姜博,邢奕,等.有机磷农药污染土壤的微生物降解研究进展[J].土壤,2018,50(4):645-655.
[33]华健,陈曼佳,李芳柏,等.水稻土中五氯酚的降解转化动态及其对微生物群落的影[J].地球与环境,2018,46(3):225-230.
[34]刘倩倩.三种常用杀菌剂在麦田土壤中的消解动态及对土壤微生物的毒性评价[D].泰安:山东农业大学,2016.
[35]伍云丽.腈菌唑在茶园土壤中的消解动力学及其对土壤微生物活性影响的研究[D].杭州:浙江大学,2014.
[36]Kuo J,Wang Y W,Chen M,et al.The effect of paclobutrazol on soil bacterial composition across three consecutive flowering stages of mung bean[J].Folia Microbiologica,2019,64(2):197-205.
[37]Hua F,Tang F,Wei Z,et al.Persistence of repeated triadimefon application and its impact on soil microbial functional diversity[J].Journal of Environmental Science and Health Part B,2012,47(2):104-110.
[38]Jeffries T C,Rayu S,Nielsen U N,et al.Metagenomic functional potential predicts degradation rates of a model organophosphorus xenobiotic in pesticide contaminated soils[J].Frontiers in Microbiology,2018,9:147.
[39]Zafra G,Taylor T D,Absalón,et al.Comparative metagenomic analysis of PAH degradation in soil by a mixed microbial consortium[J].Journal of Hazardous Materials,2016,318:702-710.
[40]徐帅,林奕岑,周梦佳,等.基于高通量测定肉鸡回肠微生物多样性及PICRUSt基因预测分析[J].动物营养学报,2016,28(8):2581-2588.
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