土壤细菌群落对湿热灭菌蚕沙还田的响应
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摘要
【目的】本文旨在揭示无外源微生物干扰下,土壤微生物群落对蚕沙中养分物质的利用状况,探索蚕沙无害化还田后的生态效应。【方法】通过向土壤中添加湿热灭菌蚕沙的方式来模拟无害蚕沙还田,并采用高通量测序技术测定土壤细菌群落结构及多样性,以揭示土壤微生物对于湿热灭菌蚕沙还田的快速响应状况。【结果】湿热灭菌蚕沙还田对土壤细菌结构无显著的影响(P> 0. 05),但减少了土壤细菌的种类,其Chao I指数、Shannon多样性指数和Simpson多样性指数显著降低(P <0. 05)。较对照而言,与纤维素降解相关的厚壁菌门、放线菌门和变形菌门细菌的相对丰度在湿热灭菌蚕沙还田条件下会显著增加,特别是一些链霉菌属、芽孢杆菌目、梭菌科、毛螺菌科、短波单胞杆菌属、固氮螺旋菌属和假黄单胞菌属的细菌类群。【结论】土壤细菌群落的结构和功能相对稳定,湿热灭菌蚕沙还田会快速激活与纤维素代谢相关的细菌类群,并通过增加相对丰度的方式促进土壤自我修复。
【Objective】This paper aimed to reveal the utilization of soil microbial communities to nutrient material from silkworm excrement without the interference of exogenous microorganisms and explore the ecological efficiency of its harmless returning to the field. 【Method】The behaviours of adding moist-heat sterilization silkworm excrement was used to simulate sterile silkworm returning,the high-throughput sequencing was used to determine the community structure and diversity of soil microbial,and the collaborative response of soil microbes to silkworm would be revealed. 【Result】The adding moist-heat sterilization silkworm excrement had no significant effect on bacterial structure of soil( P> 0. 05),however,the species number of soil bacteria was reduced,and Chao I index,the Simpson diversity index and the Shannon diversity index were significantly reduced( P < 0. 05). Compared with the control,the relative abundance of the cellulolytic bacteria of firmicutes,proteobacteria and actinomycetes was increased significantly under the condition of moist-heat sterilization silkworm excrement returning,especially some bacterial species of Streptomyces,Bacillales,Clostridiaceae,Lachnospiraceae,Brevundimonas,Azospirillum and Pseudoxanthomonas.【Conclusion】The soil bacterial community structure and function was relatively stable,and the cellulose-degrading bacteria could be activated quickly after adding moist-heat sterilization silkworm excrement to soil and promoted soil self-healing by increasing their relative abundance.
【Objective】This paper aimed to reveal the utilization of soil microbial communities to nutrient material from silkworm excrement without the interference of exogenous microorganisms and explore the ecological efficiency of its harmless returning to the field. 【Method】The behaviours of adding moist-heat sterilization silkworm excrement was used to simulate sterile silkworm returning,the high-throughput sequencing was used to determine the community structure and diversity of soil microbial,and the collaborative response of soil microbes to silkworm would be revealed. 【Result】The adding moist-heat sterilization silkworm excrement had no significant effect on bacterial structure of soil( P> 0. 05),however,the species number of soil bacteria was reduced,and Chao I index,the Simpson diversity index and the Shannon diversity index were significantly reduced( P < 0. 05). Compared with the control,the relative abundance of the cellulolytic bacteria of firmicutes,proteobacteria and actinomycetes was increased significantly under the condition of moist-heat sterilization silkworm excrement returning,especially some bacterial species of Streptomyces,Bacillales,Clostridiaceae,Lachnospiraceae,Brevundimonas,Azospirillum and Pseudoxanthomonas.【Conclusion】The soil bacterial community structure and function was relatively stable,and the cellulose-degrading bacteria could be activated quickly after adding moist-heat sterilization silkworm excrement to soil and promoted soil self-healing by increasing their relative abundance.
引文
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[23]刘欣. Paenibacillus sp. E18双功能木聚糖酶基因克隆及性质研究[D].北京:中国农业科学院硕士学位论文,2009.
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[25]Sadhu S,Maiti T K. Cellulase production by bacteria:a review[J]. British Microbiology Research Journal,2013,3(3):235-258.
[26]刘超.一株降解纤维素的链霉菌的分离纯化及其纤维素降解特性研究[D].东北师范大学硕士学位论文,2012.
[27]王佳佳,奚永兰,常志州,等.麦秸不同部位生物降解速率差异[J].农业资源与环境学报,2015,32(1):74-80.
[28]吴翔,甘炳成,黄忠乾,等. 2株降解纤维素放线菌的鉴定[J].西南农业学报,2014,27(1):117-120.
[29]罗立津,万立,陈宏,等.耐低温木质纤维素降解菌群的富集培养及其种群结构分析[J].农业生物技术学报,2015,23(6):727-737.
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[32]侯丽媛,江经纬,蒋建东,等.假黄单胞菌株J1的筛选及木质纤维素降解基因的生物信息学分析[J].南京农业大学学报,2016,39(4):573-581.
[33]Kumar M,Revathi K,Khanna S. Biodegradation of cellulosic andlignocellulosic waste by Pseudoxanthomonas sp R-28[J]. Carbohy-drate Polymers,2015,134:761-766.
[34]Du R,Yan J B,Li S Z,et al. Cellulosic ethanol production by nat-ural bacterial consortia is enhanced by Pseudoxanthomonas taiwanen-sis[J]. Biotechnology for Biofuels,2015,8(1):10.
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[36]王行.暖化湿地土—水界面碳磷耦合迁移特征及微生物生态学响应—微宇宙研究[D].杭州:浙江大学博士学位论文,2014.
[37]余倩,殷强仲,赵丹燕.成都市主要河流水中噬菌蛭弧菌的调查[J].现代预防医学,1994,21(3):174-176.
[38]Yair S,Yaacov D,Susan K J,et al. Small eats big:ecology and di-versity of Bdellovibrio and like organisms,and their dynamics inpredator-prey interactions[J]. Agronomic,2003,23(5-6):275-284.
[39]秦生巨.噬菌蛭弧菌及其微生物生态制剂在水生动物养殖上的应用[J].水产科技情报,2007,34(1):29-32.
[40]周玖英,陈孝进,帅荣,等.星形诺卡菌引起脑脓肿1例[J].实验与检验医学,2011,29(4):458-458.
[41]苑美玉. 1例星形诺卡菌引起肺内感染的病例分析[J].国际检验医学杂志,2014,35(20):2858-2859.
[42]Conville P S,Witebsky F G. Organisms designated as Nocardia as-teroides drug pattern type VI are members of the species Nocardiacyriacigeorgica[J]. Journal of Clinical Microbiology,2007,45(7):2257-2259.
[43]Nawrath T,Mgode G F,Weetjens B,et al. The volatiles of patho-genic and nonpathogenic mycobacteria and related bacteria[J]. Beil-stein Journal of Organic Chemistry,2012,8(1):290-299.
[44]Saubolle M A,Sussland D. Nocardiosis:review of clinical and la-boratory experience[J]. Journal of Clinical Microbiology,2003,41(10):4497.
[2]王谢,张建华,姚莉,等.蚕沙组分的计量分析及开发应用概况[J].蚕业科学,2016,42(5):918-925.
[3]吴耀峰.合理处理蚕沙防止病原扩散[J].北方蚕业,1985(3):12.
[4]赵德胜,韩文清,张明斗.培养基灭菌温度与时间的选择[J].价值工程,2010,29(5):205-206.
[5]陈晓萍.蚕沙堆肥及其堆肥产品的生物学效应研究[D].南昌:江西大学硕士学位论文,2011.
[6]蒲洁.农牧交错带植被恢复的土壤微生物响应[D].杨凌:西北农林科技大学硕士学位论文,2015.
[7]Caporaso J G,Lauber C L,Walters W A,et al. Ultra-high-through-put microbial community analysis on the Illumina Hi Seq and Mi Seqplatforms[J]. ISME Journal,2012,6(8):1621-1624.
[8]陶莲,刁其玉.青贮发酵对玉米秸秆品质及菌群构成的影响[J].动物营养学报,2016,28(1):198-207.
[9]Magoˇc T,Salzberg S L. FLASH:fast length adjustment of shortreads to improve genome assemblies[J]. Bioinformatics,2011,27(21):2957-2963.
[10]Caporaso J G,Kuczynski J,Stombaugh J,et al. QIIME allows a-nalysis of high-throughput community sequencing data[J]. NaturalMethods,2010,7(5):335-336.
[11]Bokulich N A,Sathish S,Faith J J,et al. Quality-filtering vastlyimproves diversity estimates from Illumina amplicon sequencing[J].Natural Methods,2012,10(1):57-59.
[12]Sickle J V. Using mean similarity dendrograms to evaluate classifi-cations[J]. Journal of Agricultural,Biological,and EnvironmentalStatistics,1997,2(4):370-388.
[13]Underwood M A,Arriola J,Gerber C W,et al. Bifidobacteriumlongum subsp. infantis in experimental necrotizing enterocolitis:al-terations in inflammation,innate immune response,and the microbi-ota[J]. Pediatric Research,2014,76(4):326.
[14]马海霞.堆肥生境微生物区系动态变化和优势丝状真菌胞外酶系功能分析[D].济南:山东大学硕士学位论文,2015.
[15]李彦红.牲畜粪便暗发酵产氢-产甲烷过程中参数优化及其微生物群落特性[D].郑州:郑州大学硕士学位论文,2016.
[16]张家齐.园林废弃物堆腐微生物过程及纤维素降解菌筛选研究[D].北京:中国林业科学研究院硕士学位论文,2012.
[17]肖文萍.藏猪肠道微生物多样性的研究[D].杨凌:西北农林科技大学硕士学位论文,2012.
[18]周帅,王晨阳,范朝刚.变性梯度凝胶电泳研究脓毒症大鼠肠道菌群变化[J].肠外与肠内营养,2014,21(4):236-240.
[19]杨梅梅.中药复方与益生素对热应激下肉鸡肠道菌群及生长的影响[D].广州:华南农业大学硕士学位论文,2016.
[20]高凤芹.柳枝稷发酵产甲烷过程中纤维素降解机制及微生物多样性变化[D].北京:中国农业大学博士学位论文,2015.
[21]鲜文东,明红,李文均.高温放线菌科最新研究进展[J].微生物学报,2015,55(1):1-11.
[22]李康宁.浑善达克沙地生物土壤结皮中降解纤维素菌的分离、鉴定及降解活性分析[D].呼和浩特:内蒙古农业大学硕士学位论文,2010.
[23]刘欣. Paenibacillus sp. E18双功能木聚糖酶基因克隆及性质研究[D].北京:中国农业科学院硕士学位论文,2009.
[24]霍云龙. Paenibacillus cookii GX-4的β-葡萄糖苷酶基因的克隆、表达、酶学性质研究与分子改造[D].南宁:广西大学硕士学位论文,2013.
[25]Sadhu S,Maiti T K. Cellulase production by bacteria:a review[J]. British Microbiology Research Journal,2013,3(3):235-258.
[26]刘超.一株降解纤维素的链霉菌的分离纯化及其纤维素降解特性研究[D].东北师范大学硕士学位论文,2012.
[27]王佳佳,奚永兰,常志州,等.麦秸不同部位生物降解速率差异[J].农业资源与环境学报,2015,32(1):74-80.
[28]吴翔,甘炳成,黄忠乾,等. 2株降解纤维素放线菌的鉴定[J].西南农业学报,2014,27(1):117-120.
[29]罗立津,万立,陈宏,等.耐低温木质纤维素降解菌群的富集培养及其种群结构分析[J].农业生物技术学报,2015,23(6):727-737.
[30]蒲一涛,钟毅沪,周万龙.固氮菌和纤维素分解菌的混合培养及其对生活垃圾降解的影响[J].环境科学与技术,1999(1):15-18.
[31]许第发.蚯蚓和微生物的协同作用对有机质降解和肥效的影响[D].杭州:浙江大学硕士学位论文,2005.
[32]侯丽媛,江经纬,蒋建东,等.假黄单胞菌株J1的筛选及木质纤维素降解基因的生物信息学分析[J].南京农业大学学报,2016,39(4):573-581.
[33]Kumar M,Revathi K,Khanna S. Biodegradation of cellulosic andlignocellulosic waste by Pseudoxanthomonas sp R-28[J]. Carbohy-drate Polymers,2015,134:761-766.
[34]Du R,Yan J B,Li S Z,et al. Cellulosic ethanol production by nat-ural bacterial consortia is enhanced by Pseudoxanthomonas taiwanen-sis[J]. Biotechnology for Biofuels,2015,8(1):10.
[35]麻攀,谭成玉,白雪芳,等.几丁质、壳聚糖及其降解产物对巨噬细胞的作用[J].细胞与分子免疫学杂志,2008,24(10):1027-1028.
[36]王行.暖化湿地土—水界面碳磷耦合迁移特征及微生物生态学响应—微宇宙研究[D].杭州:浙江大学博士学位论文,2014.
[37]余倩,殷强仲,赵丹燕.成都市主要河流水中噬菌蛭弧菌的调查[J].现代预防医学,1994,21(3):174-176.
[38]Yair S,Yaacov D,Susan K J,et al. Small eats big:ecology and di-versity of Bdellovibrio and like organisms,and their dynamics inpredator-prey interactions[J]. Agronomic,2003,23(5-6):275-284.
[39]秦生巨.噬菌蛭弧菌及其微生物生态制剂在水生动物养殖上的应用[J].水产科技情报,2007,34(1):29-32.
[40]周玖英,陈孝进,帅荣,等.星形诺卡菌引起脑脓肿1例[J].实验与检验医学,2011,29(4):458-458.
[41]苑美玉. 1例星形诺卡菌引起肺内感染的病例分析[J].国际检验医学杂志,2014,35(20):2858-2859.
[42]Conville P S,Witebsky F G. Organisms designated as Nocardia as-teroides drug pattern type VI are members of the species Nocardiacyriacigeorgica[J]. Journal of Clinical Microbiology,2007,45(7):2257-2259.
[43]Nawrath T,Mgode G F,Weetjens B,et al. The volatiles of patho-genic and nonpathogenic mycobacteria and related bacteria[J]. Beil-stein Journal of Organic Chemistry,2012,8(1):290-299.
[44]Saubolle M A,Sussland D. Nocardiosis:review of clinical and la-boratory experience[J]. Journal of Clinical Microbiology,2003,41(10):4497.