山药根际土壤微生物16S rRNA多样性及影响因素
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摘要
通过采集山药连作2年的植株根际土壤及根茎,采用高通量测序技术研究根际土壤细菌多样性,分析7个样本中菌群的组成、丰度、α多样性、β多样性、菌群差异性。采用典型相关分析(Canonical Correlation Analysis,CCA)分析优势菌群与土壤化学因子、根茎糖类物质的关系,探讨根际土壤中细菌群落与土壤化学指标、根茎糖类物质的相关性。结果表明:山药根际土壤细菌优势群落为变形菌、绿弯菌、酸杆菌、放线菌;土壤pH是影响细菌群落结构及分布的重要因子;酸杆菌和硝化螺旋菌与土壤中蔗糖酶、脲酶活性及总有机碳、碱解氮呈正相关,奇古菌和绿弯菌与土壤总有机碳、碱解氮呈正相关,土壤中碳、氮养分又与山药根茎中可溶性糖、粗多糖及淀粉呈正相关关系,说明这些微生态环境因子可能对山药根茎糖类物质积累有促进作用。
【Objective】Chinese yam(Dioscorea opposita) is a parannial herbal liane popular as vegetable in this country. Its tuber is consumed as both food and medicinal herb for its functions of antioxidation, anti-aging, immunity regulation, blood sugar reduction, etc. Soil ecoenvironmental factors and soil nutrient status are major factors affecting growth and quality of the plant. Soil microbes, in particular, participate in recycling of carbon, nitrogen and some other nutrient elements in the soil, flowing of energy, formation of soil humus, shaping and amelioration of soil structure. This study aims to explore relationships of the soil bacterial community in the rhizosphere of yam with soil nutrients and contents of saccharides in the tuber, and to elucidate major soil ecological factors affecting nutrient accumulation in yam tuber, in an attempt to provide certain reference for development of yam-specfic bio-manure and assessment of soil environment for cultivation of the plant.【Method】Seven samples of rhizosphere soil and rhizomes of 2-year old Chinese yam plants were collected for analysis of composition, abundance, Alpha diversity, Beta diversity and bacteria diversity of soil microbes with the aid of the high-throughput sequencing technology, and for analysis of relationships dominant soil bacterial groups and soil microbial community in rhizosphere with soil chemical factors and saccharides in root with the canonical correlation analysis method.【Result】Results show that Shannon and Chaol indices were significantly and negatively related to soil phosphatese activity; proteobacteria, chloroflexi, acidobacteria and actinobacteria were the dominant bacterial groups in the rhizosphere of Chinese yam; soil pH was an important factor affecting structure and distribution of the soil bacterial community; acidobacteria and nitrifying spirulina was related to sucrase and urease activities in soil; archaea and chloroflexi was positively related to soil organic carbon and nitrogen; soil carbon, nitrogen nutrient was positively related to soluble sugar, crude polysaccharides and starch in the root of yam. Chloroflexi and Thaumarchaeota, dominant soil bacterial groups were positively related to total organic carbon, total nitrogen, AK, AP and phosphatase activity in rhizosphere soil, and to content of starch and crude polysaccharide(spearman correlation index is 0.54, P<0.01) in tuber, too, which indicate that the above-mentioned chemical properties plays important roles in growth of the plant; Acidobacteria was negatively related to soil pH, AP and catalase and positively to total organic carbon, AK and invertase activity in soil, and negatively related to crude polysaccharide in tuber, too; Nitrospirae and Actinobacteria was positively related to activity of urease and catalase, and negatively to phosphatase and sucrase; Proteobacteria and Nitrospirae was positively related to soluble sugar and negatively related to crude polysaccharide.【Conclusion】All the findings demonstrate that the dominant bacterial groups in the rhizosphere of Chinese yam are proteobacteria, chlorobacteria, acidobacteria and actinomycetes; Soil pH is an important factor affecting structure of the soil bacterial community.Acidobacteri and nitrospira may improve carbon and nitrogen levels in the soil through stimulating the activity of sucrase and urease in soil; archaea and chlorobacteria promotes TOC and AN recycling in rhizosphere soil and increase TOC and AN nutrient supply in the soil, which in turn promotes accumulation of soluble sugar, crude polysaccharides and starch in rhizome of yam.
【Objective】Chinese yam(Dioscorea opposita) is a parannial herbal liane popular as vegetable in this country. Its tuber is consumed as both food and medicinal herb for its functions of antioxidation, anti-aging, immunity regulation, blood sugar reduction, etc. Soil ecoenvironmental factors and soil nutrient status are major factors affecting growth and quality of the plant. Soil microbes, in particular, participate in recycling of carbon, nitrogen and some other nutrient elements in the soil, flowing of energy, formation of soil humus, shaping and amelioration of soil structure. This study aims to explore relationships of the soil bacterial community in the rhizosphere of yam with soil nutrients and contents of saccharides in the tuber, and to elucidate major soil ecological factors affecting nutrient accumulation in yam tuber, in an attempt to provide certain reference for development of yam-specfic bio-manure and assessment of soil environment for cultivation of the plant.【Method】Seven samples of rhizosphere soil and rhizomes of 2-year old Chinese yam plants were collected for analysis of composition, abundance, Alpha diversity, Beta diversity and bacteria diversity of soil microbes with the aid of the high-throughput sequencing technology, and for analysis of relationships dominant soil bacterial groups and soil microbial community in rhizosphere with soil chemical factors and saccharides in root with the canonical correlation analysis method.【Result】Results show that Shannon and Chaol indices were significantly and negatively related to soil phosphatese activity; proteobacteria, chloroflexi, acidobacteria and actinobacteria were the dominant bacterial groups in the rhizosphere of Chinese yam; soil pH was an important factor affecting structure and distribution of the soil bacterial community; acidobacteria and nitrifying spirulina was related to sucrase and urease activities in soil; archaea and chloroflexi was positively related to soil organic carbon and nitrogen; soil carbon, nitrogen nutrient was positively related to soluble sugar, crude polysaccharides and starch in the root of yam. Chloroflexi and Thaumarchaeota, dominant soil bacterial groups were positively related to total organic carbon, total nitrogen, AK, AP and phosphatase activity in rhizosphere soil, and to content of starch and crude polysaccharide(spearman correlation index is 0.54, P<0.01) in tuber, too, which indicate that the above-mentioned chemical properties plays important roles in growth of the plant; Acidobacteria was negatively related to soil pH, AP and catalase and positively to total organic carbon, AK and invertase activity in soil, and negatively related to crude polysaccharide in tuber, too; Nitrospirae and Actinobacteria was positively related to activity of urease and catalase, and negatively to phosphatase and sucrase; Proteobacteria and Nitrospirae was positively related to soluble sugar and negatively related to crude polysaccharide.【Conclusion】All the findings demonstrate that the dominant bacterial groups in the rhizosphere of Chinese yam are proteobacteria, chlorobacteria, acidobacteria and actinomycetes; Soil pH is an important factor affecting structure of the soil bacterial community.Acidobacteri and nitrospira may improve carbon and nitrogen levels in the soil through stimulating the activity of sucrase and urease in soil; archaea and chlorobacteria promotes TOC and AN recycling in rhizosphere soil and increase TOC and AN nutrient supply in the soil, which in turn promotes accumulation of soluble sugar, crude polysaccharides and starch in rhizome of yam.
引文
[1]邵礼梅，许世伟.山药化学成分及现代药理研究进展.中医药学报，2017,45(2):125—127Shao L M,Xu S W. Advances in chemical constituents and modern pharmacology of Chinese yam(In Chinese). Acta Chinese Medicine and Pharmacology,2017,45(2):125—127
[2]李光宇，吴次芳.土壤微生物研究在农田质量评价中的应用.土壤学报，2018,55(3):543—556Li G Y,Wu C F. Application of soil microbial studies to farmland quality evaluation(In Chinese). Acta Pedologica Sinica,2018,55(3):543—556
[3] Liu Z,Liu G,Fu B,et al. Relationship between plant species diversity and soil microbial functional diversity along a longitudinal gradient in temperate grasslands of Hulunbeir,Inner Mongolia,China. Ecological Research,2008,23(3):511—518
[4]孙凯宁，杨宁，王克安，等.山药连作对土壤微生物群落及土壤酶活性的影响.水土保持研究，2015,22(6):95—98Sun K N,Yang N,Wang K A,et al. Effect of Chinese Yam continuous cropping on the soil microbial community and soil enzyme activity(In Chinese). Research of Soil&Water Conservation,2015,22(6):95—98
[5]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.微生物学报，2016,56(6):1000—1008Zhao S,Zhou N,Zhao Z Y,et al. Endophytic bacterial diversity and dynamics in root of Salicornia europaea estimated via high throughput sequencing(In Chinese).Acta Microbiologica Sinica,2016,56(6):1000—1008.
[6]戴雅婷，闫志坚，解继红，等.基于高通量测序的两种植被恢复类型根际土壤细菌多样性研究.土壤学报，2017,54(3):735—748Dai Y T,Yan Z J,Xie J H,et al. Soil bacteria diversity in rhizosphere under two types of vegetation restoration based on high throughput sequencing(In Chinese). Acta Pedologica Sinica,2017,54(3):735—748
[7]李岩，杨晓东，秦璐，等.两种盐生植物根际土壤细菌多样性和群落结构.生态学报，2018,38(9):3118—3131Li Y,Yang X D,Qin L,et al. The bacterial diversity and community structures in rhizosphere soil of two halophytes,Lycium ruthenicum and Kalidium capsicum(In Chinese).Acta Ecologica Sinica,2018,38(9):3118—3131
[8]张红霞，魏方方，王家顺，等.安顺山药根茎与土壤中矿质元素相关性分析.西南大学学报（自然科学版），2017,39(5):31—36Zhang H X,Wei F F,Wang J S,et al. Correlation analysis of mineral elements in yam tuber and in its environmental soil in Anshun of Guizhou Province(In Chinese). Journal of Southwest University(Natural Science Edition),2017,39(5):31—36
[9]鲍士旦.土壤农化分析.北京：中国农业出版社，2000Bao S D. Soil andagricultural chemistry analysis(In Chinese). Beijing:China Agriculture Press,2000
[10]关松荫.土壤酶及其研究法.北京：农业出版社，1986Gua S Y. Soil enzyme andits research methods(In Chinese). Beijing:China Agriculture Press,1986
[11]张志良，瞿伟菁.植物生理学实验指导.北京：高等教育出版社，2003Zhang Z L,Qu W J. Experimental instruction in plant physiology(In Chinese). Beijing:Higher Education Press,2003
[12]张意静.食品分析技术.北京：中国轻工业出版社，2001Zhang Y J. Analysis of food technology(In Chinese).Beijing:China Light Industry Press,2001
[13]杨秉珣，刘泉，董廷旭.川西北不同沙化程度草地土壤细菌群落特征.水土保持研究，2018,25(6):45—52Yang B X,Liu Q,Dong T X. Soil bacterial communities of grasslands with different deserti?cation degrees in northwest Sichuan(In Chinese). Research of Soil and Water Conservation,2018,25(6):45—52
[14]于海玲.施氮量对土壤微生物群落组成特征的影响研究.长春：吉林农业大学，2017Yu H L. Effect of nitrogen application on composition character of soil microbial communities(In Chinese).Changchun:Jilin Agricultural University,2017
[15]金奇.鄱阳湖典型湿地土壤微生物群落特征及对环境变化的响应研究.南昌：江西师范大学，2017Jin Q. Characteristics of soil microbial community in the Poyang Lake wetland and its response to the environmental change(In Chinese). Nanchang:Jiangxi Normal University,2017
[16]王佩雯，朱金峰，陈征，等.高通量测序技术下连作植烟土壤细菌群落与土壤环境因子的耦合分析.农业生物技术学报，2016,24(11):1754—1763Wang P W,Zhu J F,Chen Z,et al. Coupling analysis based on high throughput sequencing technology of soil bacterial community and soil environmental factors in continuous cropping tobacco soil(In Chinese). Journal of Agricultural Biotechnology,2016,24(11):1754—1763
[17]王文波，王延平，王华田，等.杨树人工林连作与轮作对土壤氮素细菌类群和氮素代谢的影响.林业科学，2016,52(5):45—54Wang W B,Wang Y P,Wang H T,et al. Effect of different continuous cropping and rotation of poplar plantation on soil nitrogen bacteria community characteristics and nitrogen metabolism(In Chinese). Journal of Forest Sciences,2016,52(5):45—54
[18]张晶，林先贵，尹睿.参与土壤氮素循环的微生物功能基因多样性研究进展.中国生态农业学报，2009,17(5):1029—1034Zhang J,Lin X G,Yin R. Advances in functional gene diversity of microorganism in relation to soil nitrogen cycling(In Chinese). Chinese Journal of Ecological Agriculture,2009,17(5):1029—1034
[19]朱平，陈仁升，宋耀选，等.祁连山中部4种典型植被类型土壤细菌群落结构差异.生态学报，2017,37(10):3505—3514Zhu P,Chen R S,Song Y X,et al. Soil bacterial community composition and diversity of four representative vegetation types in the middle section of the Qilian Mountains,China(In Chinese). Acta Ecologica Sinica,2017,37(10):3505—3514
[20]刘兴，王世杰，刘秀明，等.贵州喀斯特地区土壤细菌群落结构特征及变化.地球与环境，2015,43(5):490—497Liu X,Wang S J,Liu X M,et al. Compositional characteristics and variations of soil microbial community in Karst Area of Puding County,Guizhou Province,China(In Chinese). Earth and Environment,2015,43(5):490—497
[21]王鹏，陈波，张华.基于高通量测序的鄱阳湖典型湿地土壤细菌群落特征分析.生态学报，2017,37(5):1650—1658Wang P,Chen B,Zhang H. High throughput sequencing analysis of bacterial communities in soils of a typical Poyang Lake wetland(In Chinese). Acta Ecologica Sinica,2017,37(5):1650—1658
[22]赵丰云，杨湘，董明明，等.加气灌溉改善干旱区葡萄根际土壤化学特性及细菌群落结构.农业工程学报，2017,33(22):119—126Zhao F Y,Yang X,Dong M M,et al. Aeration irrigation improving grape rhizosphere soil chemical properties and bacterial community structure in arid area(In Chinese).Transactions of the Chinese Society of Agricultural Engineering,2017,33(22):119—126
[23]张丽梅，贺纪正.一个新的古菌类群——奇古菌门（Thaumarchaeota）.微生物学报，2012,52(4):411—421Zhang L M,He J Z. A novel archaeal phylum:Thaumarchaeota—A review. Acta Microbiologica Sinica,2012,52(4):411—421
[24]侯雪燕.土壤pH对硝化作用和氨氧化微生物群落结构的影响.重庆：西南大学，2014Hou X Y. Effects of soil pH on nitri?cation and community structure of ammonia-oxidizing microorganisms(In Chinese). Chongqing:Southwest University,2014
[2]李光宇，吴次芳.土壤微生物研究在农田质量评价中的应用.土壤学报，2018,55(3):543—556Li G Y,Wu C F. Application of soil microbial studies to farmland quality evaluation(In Chinese). Acta Pedologica Sinica,2018,55(3):543—556
[3] Liu Z,Liu G,Fu B,et al. Relationship between plant species diversity and soil microbial functional diversity along a longitudinal gradient in temperate grasslands of Hulunbeir,Inner Mongolia,China. Ecological Research,2008,23(3):511—518
[4]孙凯宁，杨宁，王克安，等.山药连作对土壤微生物群落及土壤酶活性的影响.水土保持研究，2015,22(6):95—98Sun K N,Yang N,Wang K A,et al. Effect of Chinese Yam continuous cropping on the soil microbial community and soil enzyme activity(In Chinese). Research of Soil&Water Conservation,2015,22(6):95—98
[5]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.微生物学报，2016,56(6):1000—1008Zhao S,Zhou N,Zhao Z Y,et al. Endophytic bacterial diversity and dynamics in root of Salicornia europaea estimated via high throughput sequencing(In Chinese).Acta Microbiologica Sinica,2016,56(6):1000—1008.
[6]戴雅婷，闫志坚，解继红，等.基于高通量测序的两种植被恢复类型根际土壤细菌多样性研究.土壤学报，2017,54(3):735—748Dai Y T,Yan Z J,Xie J H,et al. Soil bacteria diversity in rhizosphere under two types of vegetation restoration based on high throughput sequencing(In Chinese). Acta Pedologica Sinica,2017,54(3):735—748
[7]李岩，杨晓东，秦璐，等.两种盐生植物根际土壤细菌多样性和群落结构.生态学报，2018,38(9):3118—3131Li Y,Yang X D,Qin L,et al. The bacterial diversity and community structures in rhizosphere soil of two halophytes,Lycium ruthenicum and Kalidium capsicum(In Chinese).Acta Ecologica Sinica,2018,38(9):3118—3131
[8]张红霞，魏方方，王家顺，等.安顺山药根茎与土壤中矿质元素相关性分析.西南大学学报（自然科学版），2017,39(5):31—36Zhang H X,Wei F F,Wang J S,et al. Correlation analysis of mineral elements in yam tuber and in its environmental soil in Anshun of Guizhou Province(In Chinese). Journal of Southwest University(Natural Science Edition),2017,39(5):31—36
[9]鲍士旦.土壤农化分析.北京：中国农业出版社，2000Bao S D. Soil andagricultural chemistry analysis(In Chinese). Beijing:China Agriculture Press,2000
[10]关松荫.土壤酶及其研究法.北京：农业出版社，1986Gua S Y. Soil enzyme andits research methods(In Chinese). Beijing:China Agriculture Press,1986
[11]张志良，瞿伟菁.植物生理学实验指导.北京：高等教育出版社，2003Zhang Z L,Qu W J. Experimental instruction in plant physiology(In Chinese). Beijing:Higher Education Press,2003
[12]张意静.食品分析技术.北京：中国轻工业出版社，2001Zhang Y J. Analysis of food technology(In Chinese).Beijing:China Light Industry Press,2001
[13]杨秉珣，刘泉，董廷旭.川西北不同沙化程度草地土壤细菌群落特征.水土保持研究，2018,25(6):45—52Yang B X,Liu Q,Dong T X. Soil bacterial communities of grasslands with different deserti?cation degrees in northwest Sichuan(In Chinese). Research of Soil and Water Conservation,2018,25(6):45—52
[14]于海玲.施氮量对土壤微生物群落组成特征的影响研究.长春：吉林农业大学，2017Yu H L. Effect of nitrogen application on composition character of soil microbial communities(In Chinese).Changchun:Jilin Agricultural University,2017
[15]金奇.鄱阳湖典型湿地土壤微生物群落特征及对环境变化的响应研究.南昌：江西师范大学，2017Jin Q. Characteristics of soil microbial community in the Poyang Lake wetland and its response to the environmental change(In Chinese). Nanchang:Jiangxi Normal University,2017
[16]王佩雯，朱金峰，陈征，等.高通量测序技术下连作植烟土壤细菌群落与土壤环境因子的耦合分析.农业生物技术学报，2016,24(11):1754—1763Wang P W,Zhu J F,Chen Z,et al. Coupling analysis based on high throughput sequencing technology of soil bacterial community and soil environmental factors in continuous cropping tobacco soil(In Chinese). Journal of Agricultural Biotechnology,2016,24(11):1754—1763
[17]王文波，王延平，王华田，等.杨树人工林连作与轮作对土壤氮素细菌类群和氮素代谢的影响.林业科学，2016,52(5):45—54Wang W B,Wang Y P,Wang H T,et al. Effect of different continuous cropping and rotation of poplar plantation on soil nitrogen bacteria community characteristics and nitrogen metabolism(In Chinese). Journal of Forest Sciences,2016,52(5):45—54
[18]张晶，林先贵，尹睿.参与土壤氮素循环的微生物功能基因多样性研究进展.中国生态农业学报，2009,17(5):1029—1034Zhang J,Lin X G,Yin R. Advances in functional gene diversity of microorganism in relation to soil nitrogen cycling(In Chinese). Chinese Journal of Ecological Agriculture,2009,17(5):1029—1034
[19]朱平，陈仁升，宋耀选，等.祁连山中部4种典型植被类型土壤细菌群落结构差异.生态学报，2017,37(10):3505—3514Zhu P,Chen R S,Song Y X,et al. Soil bacterial community composition and diversity of four representative vegetation types in the middle section of the Qilian Mountains,China(In Chinese). Acta Ecologica Sinica,2017,37(10):3505—3514
[20]刘兴，王世杰，刘秀明，等.贵州喀斯特地区土壤细菌群落结构特征及变化.地球与环境，2015,43(5):490—497Liu X,Wang S J,Liu X M,et al. Compositional characteristics and variations of soil microbial community in Karst Area of Puding County,Guizhou Province,China(In Chinese). Earth and Environment,2015,43(5):490—497
[21]王鹏，陈波，张华.基于高通量测序的鄱阳湖典型湿地土壤细菌群落特征分析.生态学报，2017,37(5):1650—1658Wang P,Chen B,Zhang H. High throughput sequencing analysis of bacterial communities in soils of a typical Poyang Lake wetland(In Chinese). Acta Ecologica Sinica,2017,37(5):1650—1658
[22]赵丰云，杨湘，董明明，等.加气灌溉改善干旱区葡萄根际土壤化学特性及细菌群落结构.农业工程学报，2017,33(22):119—126Zhao F Y,Yang X,Dong M M,et al. Aeration irrigation improving grape rhizosphere soil chemical properties and bacterial community structure in arid area(In Chinese).Transactions of the Chinese Society of Agricultural Engineering,2017,33(22):119—126
[23]张丽梅，贺纪正.一个新的古菌类群——奇古菌门（Thaumarchaeota）.微生物学报，2012,52(4):411—421Zhang L M,He J Z. A novel archaeal phylum:Thaumarchaeota—A review. Acta Microbiologica Sinica,2012,52(4):411—421
[24]侯雪燕.土壤pH对硝化作用和氨氧化微生物群落结构的影响.重庆：西南大学，2014Hou X Y. Effects of soil pH on nitri?cation and community structure of ammonia-oxidizing microorganisms(In Chinese). Chongqing:Southwest University,2014
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