引黄灌区不同种植年限紫花苜蓿土壤养分与细菌群落特征研究
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摘要
以宁夏引黄灌区种植年限为1～6年(2012-2017年)的紫花苜蓿地土壤为研究对象,采用16S rRNA扩增子测序技术对细菌群落组成、丰度和多样性等特征进行研究,分析了土壤养分与细菌种群多样性的相关关系。结果表明:不同种植年限苜蓿地土壤中共检测出细菌约40个门、78个纲、151个目、275个科和416个属;在门水平下细菌的优势门类为变形菌门、酸杆菌门和拟杆菌门,共占细菌总数的68%;随种植年限的增加,细菌菌群丰度和多样性变化表现为1 yr>5 yr>3 yr>4 yr>2 yr>6 yr;土壤pH、有机碳、全氮和碱解氮呈先降低后增大的变化趋势,且都在第5年时达到最大,与细菌群落组成变化规律基本保持一致。Spearman相关性分析结果表明,有机碳、全氮和碱解氮对细菌群落组成具有显著影响(P<0.05)。
The soil bacterial community composition, abundance and diversity were studied in alfalfa stands between 1-6 years of age(i.e. established 2012-2017), in the Ningxia Yellow River irrigation area. The determination of the soil bacterial community characteristics was achieved by DNA extraction followed by PCR amplification, and sequencing using the HiSeq platform. The 16 S rRNA amplifier sequencing technique was used in the study. The correlation between soil nutrients and bacterial population diversity was analyzed. The results indicate that there are about 40 phyla, 78 classes, 151 orders, 275 families and 416 genera of bacteria in the soils of the various alfalfa stands of different ages. At the phylum level, the dominant genera of bacteria detected were Proteobacteria, Acidobacteria and Bacteroidetes, accounting for 68% of the total bacterial population. The ranking of bacterial abundance and diversity with stand age was: 1 yr>5 yr>3 yr>4 yr>2 yr>6 yr. Soil pH, organic carbon, total nitrogen and alkali-hydrolysable nitrogen all decreased initially, and then increased with stand age, reaching their maxima at year 5. The result is basically consistent with findings of other studies of factors affecting bacterial community composition. Analysis using Spearman's correlation showed that organic carbon, total nitrogen and alkaline nitrogen were significantly(P<0.05) correlated with bacterial community composition. In general, the dominant taxa of soil bacterial communities present in the Yellow River irrigation area did not change with stand age, but population numbers and bacterial community diversity did vary from year to year, though not with any consistent trend over time. Soil organic carbon, soil alkali-hydrolysable nitrogen and soil total nitrogen were the main factors associated with changes in bacterial taxa present across years.
The soil bacterial community composition, abundance and diversity were studied in alfalfa stands between 1-6 years of age(i.e. established 2012-2017), in the Ningxia Yellow River irrigation area. The determination of the soil bacterial community characteristics was achieved by DNA extraction followed by PCR amplification, and sequencing using the HiSeq platform. The 16 S rRNA amplifier sequencing technique was used in the study. The correlation between soil nutrients and bacterial population diversity was analyzed. The results indicate that there are about 40 phyla, 78 classes, 151 orders, 275 families and 416 genera of bacteria in the soils of the various alfalfa stands of different ages. At the phylum level, the dominant genera of bacteria detected were Proteobacteria, Acidobacteria and Bacteroidetes, accounting for 68% of the total bacterial population. The ranking of bacterial abundance and diversity with stand age was: 1 yr>5 yr>3 yr>4 yr>2 yr>6 yr. Soil pH, organic carbon, total nitrogen and alkali-hydrolysable nitrogen all decreased initially, and then increased with stand age, reaching their maxima at year 5. The result is basically consistent with findings of other studies of factors affecting bacterial community composition. Analysis using Spearman's correlation showed that organic carbon, total nitrogen and alkaline nitrogen were significantly(P<0.05) correlated with bacterial community composition. In general, the dominant taxa of soil bacterial communities present in the Yellow River irrigation area did not change with stand age, but population numbers and bacterial community diversity did vary from year to year, though not with any consistent trend over time. Soil organic carbon, soil alkali-hydrolysable nitrogen and soil total nitrogen were the main factors associated with changes in bacterial taxa present across years.
引文
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[16] Huang Z,Liu Y,Cui Z,et al.Soil water storage deficit of alfalfa (Medicago sativa) grasslands along ages in arid area (China).Field Crops Research,2018,221:1-6.
[17] Bao S D.Soil agrochemical analysis.Beijing:China Agricultural Publishing House,2000:30-42.鲍士旦.土壤农化分析.北京:中国农业出版社,2000:30-42.
[18] Liu B R.Experimental technology of soil bacterial diversity based on PCR-DGGE and 16S rDNA.Research of Agricultural Science,2014,35(3):34-38.刘秉儒.基于PCR-DGGE和16S rDNA克隆技术研究土壤细菌多样性的实验技术.农业科学研究,2014,35(3):34-38.
[19] Jiao S,Liu Z,Liu Y,et al.Bacterial communities in oil contaminated soils:Biogeography and co-occurrence patterns.Soil Biology & Biochemistry,2016,(98):64-73.
[20] Qin J,Li Y,Cai Z,et al.A metagenome-wide association study of gut microbiota in type 2 diabetes.Nature,2012,490(7418):55-60.
[21] Haas B J.Chimeric 16S rRNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons.Genome Research,2011,21(3):494-504.
[22] Guo L L,Yin W L,Guo D L,et al.Variations of bacterial biodiversity in rhizosphere soils of oil tree peony cropping continuously for different years.Scientia Silvae Sinicae,2017,53(11):131-141.郭丽丽,尹伟伦,郭大龙,等.油用凤丹牡丹不同种植时间根际细菌群落多样性变化.林业科学,2017,53(11):131-141.
[23] Luo M,Han J,Jiang P A,et al.Diversity of culturable halophilic bacteria isolated from Lop Nur region in Xinjiang.Biodiversity Science,2009,17(3):288-295.罗明,韩剑,蒋平安,等.新疆罗布泊地区可培养嗜盐细菌多样性.生物多样性,2009,17(3):288-295.
[24] Bissett A,Richardson A E,Baker G,et al.Long-term land use effects on soil microbial community structure and function.Applied Soil Ecology,2011,(51):66-78.
[25] Lauber C L,Hamady M,Knight R,et al.Pyrosequencing-based assessment of soil pH as a predictor of soil bacterial community structure at the continental scale.Applied and Environmental Microbiology,2009,75(15):5111-5120.
[26] Chen H K.Soil microbiology.Shanghai:Shanghai Science and Technology Publishing House,1981.陈华葵.土壤微生物学.上海:上海科学技术出版社,1981.
[27] Li Y M,Hu J C,Wang S L,et al.Function and application of soil microorganisms in forest ecosystem.Chinese Journal of Applied Ecology,2004,(10):1943-1946.李延茂,胡江春,汪思龙,等.森林生态系统中土壤微生物的作用与应用.应用生态学报,2004,(10):1943-1946.
[28] Han Y F,Yi W H,Wang W B,et al.Soil bacteria diversity in continuous cropping poplar plantation by high throughput sequencing.Journal of Shandong University (Science Edition),2014,49(5):1-6.韩亚飞,伊文慧,王文波,等.基于高通量测序技术的连作杨树人工林土壤细菌多样性研.山东大学学报(理学版),2014,49(5):1-6.
[29] Nan L L,Shi S L,Yu J H.Soil microbial properties in alfalfa field with different growing years in aria desert oasis.Acta Agrestia Sinica,2016,24(5):975-980.南丽丽,师尚礼,郁继华.荒漠灌区不同种植年限苜蓿草地土壤微生物特性.草地学报,2016,24(5):975-980.
[30] Sui X,Zhang R T,Zhong H X,et al.Study on the bacterial diversity in the soil of launzhang wetland in Sanjiang Plain by high flux sequencing.Soils,2015,47(5):919-925.隋心,张荣涛,钟海秀,等.利用高通量测序对三江平原小叶章湿地土壤细菌多样性的研究.土壤,2015,47(5):919-925.
[31] Yang J,Zhou G Y,Tian Y Y,et al.Differential analysis of soil bacteria diversity in different mixed forests of Dalbergia odorifera.Acta Ecologica Sinica,2015,35(24):8117-8127.杨菁,周国英,田媛媛,等.降香黄檀不同混交林土壤细菌多样性差异分析.生态学报,2015,35(24):8117-8127.
[32] Liu J,Sui Y,Yu Z,et al.High throughput sequencing analysis of biogeographical distribution of bacterial communities in the black soils of northeast China.Soil Biology and Biochemistry,2014,70:113-122.
[33] Sul W J,Asuming-Brempong S,Wang Q,et al.Tropical agricultural land management influences on soil microbial communities through its effect on soil organic carbon.Soil Biology and Biochemistry,2013,65:33-38.
[34] Baker B J,Sheik C S,Taylor C A,et al.Community transcriptomic assembly reveals microbes that contribute to deep-sea carbon and nitrogen cycling.The International Society for Microbial Ecology Journal,2013,7(10):1962-1973.
[35] Heichel G H,Barnes D K,Vance Lucerne C P.Nitrogen fixation of alfalfa in the seeding year 1.Crop Science,1981,21(2):330-335.
[36] Yang Y H,Jiang P A.Studies on soil properties of with different cultivating ages.Journal of Soil and Water Conservation,2005,19(2):110-113.杨玉海,蒋平安.不同种植年限苜蓿地土壤理化特性研究.水土保持学报,2005,19(2):110-113.
[37] Liu E B.Experimental study on improving soil fertility of loess slope by planting alfalfa.Shaanxi Agricultural Science,2007,(4):60-62.刘恩斌.种植紫花苜蓿提高黄土坡地土壤肥力试验研究.陕西农业科学,2007,(4):60-62.
[2] Schnitzer S A,Klironomos J N,Hille R L J,et al.Soil microbes drive the classic plant diversity-productivity pattern.Ecology,2011,92(2):296-303.
[3] Schneider T,Keiblinger K M,Schmid E,et al.Who is who in litter decomposition?Metaproteomics reveals major microbial players and their biogeochemical functions.The International Society for Microbial Ecology Journal,2012,6(9):1749-1762.
[4] Meng F Q,Qiao Y H,Wu W L,et al.Environmental impacts and production performances of organic agriculture in China:A monetary valuation.Journal of Environmental Management,2017,188:49-57.
[5] Bardgett R D,van der Putten W H.Belowground biodiversity and ecosystem functioning.Nature,2014,515(7528):505-511.
[6] He Z,Van Nostrand J D,Deng Y,et al.Development and applications of functional gene microarrays in the analysis of the functional diversity,composition,and structure of microbial communities.Frontiers of Environmental Science & Engineering in China,2011,5(1):1-20.
[7] Yu P F,Wang Y,Ge Q Y.High-fluxed DNA sequencing technology and its application development.Journal of Nanjing Xiaozhuang University,2010,26(3):1-5.于聘飞,王英,葛芹玉.高通量DNA测序技术及其应用进展.南京晓庄学院学报,2010,26(3):1-5.
[8] Zhang X J,Chen X Q,Liu H B,et al.Preliminary study on nitrogen and phosphorus loss in Yellow River irrigated paddy field of Ningxia.Ecology and Environmental Sciences,2010,19(5):1202-1209.张学军,陈晓群,刘宏斌,等.宁夏引黄灌区稻田氮磷流失特征初探.生态环境学报,2010,19(5):1202-1209.
[9] Wang Y Q,Bai Y R,Zhan X L,et al.Spatial variability of soil nutrients at different sampling scales in farmland in the Yellow River irrigated area in Ningxia.Arid Zone Research,2014,31(2):209-215.王幼奇,白一茹,展秀丽,等.在不同尺度下宁夏引黄灌区农田土壤养分空间变异分析.干旱区研究,2014,31(2):209-215.
[10] Fu B Z,Gao X Q,Zhang R,et al.Comparison of different alfalfa cultivars in the Yellow River irrigation region of Ningxia.Journal of Northwest University of Agriculture and Forestry Science and Technology (Natural Science Edition),2017,45(2):71-78.伏兵哲,高雪芹,张蓉,等.宁夏引黄灌区不同紫花苜蓿品种比较.西北农林科技大学学报(自然科学版),2017,45(2):71-78.
[11] Yang H S,Cao M J,Fan F,et al.Effects of the number of growth years of alfalfa on the physical and chemical properties of soil.Chinese Journal of Grassland,2006,(6):29-32.杨恒山,曹敏建,范富,等.紫花苜蓿生长年限对土壤理化性状的影响.中国草地学报,2006,(6):29-32.
[12] Hu F C.Initial research report on soil fertility improvement by planting Medicago sativa.Pratacultural Science,2005,22(8):47-49.胡发成.种植苜蓿改良培肥地力的研究初报.草业科学,2005,22(8):47-49.
[13] Ren J Z.Main forage cultivation and seed production.Chengdu:Sichuan Science and Technology Press,1986:1-3.任继周.主要牧草栽培及种子生产.成都:四川科学技术出版社,1986:1-3.
[14] Zhang C X,Hao M D,Wang X G,et al.Study on soil nitrogen and fertility distribution characteristics in alfalfa field in gully region of the Loess Plateau.Acta Botanica Boreali-Occidentalia Sinica,2004,(6):1107-1111.张春霞,郝明德,王旭刚,等.黄土高原地区紫花苜蓿生长过程中土壤养分的变化规律.西北植物学报,2004,(6):1107-1111.
[15] Xu L J,Wang B,Xin X P.Soil nutrient and microbial characteristics associated alfalfa cultivated grassland.Acta Agrestia Sinica,2011,19(3):406-411.徐丽君,王波,辛晓平.紫花苜蓿人工草地土壤养分及土壤微生物特性.草地学报,2011,19(3):406-411.
[16] Huang Z,Liu Y,Cui Z,et al.Soil water storage deficit of alfalfa (Medicago sativa) grasslands along ages in arid area (China).Field Crops Research,2018,221:1-6.
[17] Bao S D.Soil agrochemical analysis.Beijing:China Agricultural Publishing House,2000:30-42.鲍士旦.土壤农化分析.北京:中国农业出版社,2000:30-42.
[18] Liu B R.Experimental technology of soil bacterial diversity based on PCR-DGGE and 16S rDNA.Research of Agricultural Science,2014,35(3):34-38.刘秉儒.基于PCR-DGGE和16S rDNA克隆技术研究土壤细菌多样性的实验技术.农业科学研究,2014,35(3):34-38.
[19] Jiao S,Liu Z,Liu Y,et al.Bacterial communities in oil contaminated soils:Biogeography and co-occurrence patterns.Soil Biology & Biochemistry,2016,(98):64-73.
[20] Qin J,Li Y,Cai Z,et al.A metagenome-wide association study of gut microbiota in type 2 diabetes.Nature,2012,490(7418):55-60.
[21] Haas B J.Chimeric 16S rRNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons.Genome Research,2011,21(3):494-504.
[22] Guo L L,Yin W L,Guo D L,et al.Variations of bacterial biodiversity in rhizosphere soils of oil tree peony cropping continuously for different years.Scientia Silvae Sinicae,2017,53(11):131-141.郭丽丽,尹伟伦,郭大龙,等.油用凤丹牡丹不同种植时间根际细菌群落多样性变化.林业科学,2017,53(11):131-141.
[23] Luo M,Han J,Jiang P A,et al.Diversity of culturable halophilic bacteria isolated from Lop Nur region in Xinjiang.Biodiversity Science,2009,17(3):288-295.罗明,韩剑,蒋平安,等.新疆罗布泊地区可培养嗜盐细菌多样性.生物多样性,2009,17(3):288-295.
[24] Bissett A,Richardson A E,Baker G,et al.Long-term land use effects on soil microbial community structure and function.Applied Soil Ecology,2011,(51):66-78.
[25] Lauber C L,Hamady M,Knight R,et al.Pyrosequencing-based assessment of soil pH as a predictor of soil bacterial community structure at the continental scale.Applied and Environmental Microbiology,2009,75(15):5111-5120.
[26] Chen H K.Soil microbiology.Shanghai:Shanghai Science and Technology Publishing House,1981.陈华葵.土壤微生物学.上海:上海科学技术出版社,1981.
[27] Li Y M,Hu J C,Wang S L,et al.Function and application of soil microorganisms in forest ecosystem.Chinese Journal of Applied Ecology,2004,(10):1943-1946.李延茂,胡江春,汪思龙,等.森林生态系统中土壤微生物的作用与应用.应用生态学报,2004,(10):1943-1946.
[28] Han Y F,Yi W H,Wang W B,et al.Soil bacteria diversity in continuous cropping poplar plantation by high throughput sequencing.Journal of Shandong University (Science Edition),2014,49(5):1-6.韩亚飞,伊文慧,王文波,等.基于高通量测序技术的连作杨树人工林土壤细菌多样性研.山东大学学报(理学版),2014,49(5):1-6.
[29] Nan L L,Shi S L,Yu J H.Soil microbial properties in alfalfa field with different growing years in aria desert oasis.Acta Agrestia Sinica,2016,24(5):975-980.南丽丽,师尚礼,郁继华.荒漠灌区不同种植年限苜蓿草地土壤微生物特性.草地学报,2016,24(5):975-980.
[30] Sui X,Zhang R T,Zhong H X,et al.Study on the bacterial diversity in the soil of launzhang wetland in Sanjiang Plain by high flux sequencing.Soils,2015,47(5):919-925.隋心,张荣涛,钟海秀,等.利用高通量测序对三江平原小叶章湿地土壤细菌多样性的研究.土壤,2015,47(5):919-925.
[31] Yang J,Zhou G Y,Tian Y Y,et al.Differential analysis of soil bacteria diversity in different mixed forests of Dalbergia odorifera.Acta Ecologica Sinica,2015,35(24):8117-8127.杨菁,周国英,田媛媛,等.降香黄檀不同混交林土壤细菌多样性差异分析.生态学报,2015,35(24):8117-8127.
[32] Liu J,Sui Y,Yu Z,et al.High throughput sequencing analysis of biogeographical distribution of bacterial communities in the black soils of northeast China.Soil Biology and Biochemistry,2014,70:113-122.
[33] Sul W J,Asuming-Brempong S,Wang Q,et al.Tropical agricultural land management influences on soil microbial communities through its effect on soil organic carbon.Soil Biology and Biochemistry,2013,65:33-38.
[34] Baker B J,Sheik C S,Taylor C A,et al.Community transcriptomic assembly reveals microbes that contribute to deep-sea carbon and nitrogen cycling.The International Society for Microbial Ecology Journal,2013,7(10):1962-1973.
[35] Heichel G H,Barnes D K,Vance Lucerne C P.Nitrogen fixation of alfalfa in the seeding year 1.Crop Science,1981,21(2):330-335.
[36] Yang Y H,Jiang P A.Studies on soil properties of with different cultivating ages.Journal of Soil and Water Conservation,2005,19(2):110-113.杨玉海,蒋平安.不同种植年限苜蓿地土壤理化特性研究.水土保持学报,2005,19(2):110-113.
[37] Liu E B.Experimental study on improving soil fertility of loess slope by planting alfalfa.Shaanxi Agricultural Science,2007,(4):60-62.刘恩斌.种植紫花苜蓿提高黄土坡地土壤肥力试验研究.陕西农业科学,2007,(4):60-62.