Journal of Soils and Sediments

, Volume 17, Issue 8, pp 2156–2164 | Cite as

Changes in soil bacterial communities in an evergreen broad-leaved forest in east China following 4 years of nitrogen addition

  • Jun Cui
  • Jingjing Wang
  • Jun Xu
  • Chonghua Xu
  • Xiaoniu XuEmail author
Soils, Sec 5 • Soil and Landscape Ecology • Research Article



Evergreen broad-leaved forest ecosystems are common in east China, where they are both ecologically and economically important. However, nitrogen (N) addition over many years has had a detrimental effect on these ecosystems. The objective of this research was to evaluate the effect of 4 years of N addition on microbial communities in an evergreen broad-leaved forest in southern Anhui, China.

Materials and methods

Allochthonous N in the form of aqueous NH4NO3 and phosphorus (P) in the form of Ca(H2PO4)2·H2O were applied at three doses with a control (CK, stream water only without fertilizer): low-N (50 kg N ha−1 year−1), high-N (100 kg N ha−1 year−1) and high-N+P (100 kg N ha−1 year−1 + 50 kg P ha−1 year−1). Quantitative PCR analysis of microbial community size and Illumina platform-based sequencing analysis of the V3-V4 16S rRNA gene region were performed to characterize soil bacterial community abundance, structure, and diversity.

Results and discussion

Bacterial diversity was increased in low-N and high-N treatments and decreased in the high-N+P treatment, but α-diversity indices were not significantly affected by N additions. Proteobacteria, Acidobacteria, and Actinobacteria were the predominant phyla in all treatments, and the relative abundance of different genera varied among treatments. Only soil pH (P = 0.051) showed a weak correlation with the bacterial community in CK and low-N treatment.


The composition of the bacterial community and the abundance of different phyla were significantly altered by N addition. The results of the present study indicate that soil bacterial communities in subtropical evergreen broad-leaved forest are, to a certain extent, resilient to changes derived from N additions.


16S rRNA Bacterial community Evergreen broad-leaved forest Microbial diversity Nitrogen addition 



This work was supported by the National Science Foundation of China (NSFC, No. 31370626) and the National Basic Research Program of China (973 Program, No. 2010CB950602). We gratefully acknowledge field assistance from C. Zhang, X. Yu, D. Tian, L. Ke, Z. F. Wang, and W. Fan. In the laboratory, we gratefully acknowledge C. Shi and Y. L. Wang for assistance with molecular biology experiments.

Supplementary material

11368_2017_1671_MOESM1_ESM.docx (23 kb)
ESM 1 (DOCX 23 kb)


  1. Aber JD, Nadelhoffer KJ, Steudler P, Melillo JM (1989) Nitrogen saturation in northern forest ecosystems. Bioscience 39:378–386CrossRefGoogle Scholar
  2. Bardhan S, Jose S, Jenkins MA, Webster CR, Udawatta RP, Stehn SE (2012) Microbial community diversity and composition across a gradient of soil acidity in spruce–fir forests of the southern Appalachian Mountains. Appl Soil Ecol 61:60–68CrossRefGoogle Scholar
  3. Boxman AW, Blanck K, Brandrud TE (1998) Vegetation and soil biota response to experimentally changed nitrogen inputs in coniferous forest ecosystems of the NITREX project. Forest Ecol Manag 101:65–79CrossRefGoogle Scholar
  4. Brookes PC, Landman A (1985) Chloroform fumigation and the release of soil N: a rapid direct extraction method to measure microbial biomass in soil. Soil Biol Biochem 17:837–842Google Scholar
  5. Brons JK, van Elsas JD (2008) Analysis of bacterial communities in soil by use of denaturing gradient gel electrophoresis and clone libraries, as influenced by different reverse primers. Appl Environ Microbiol 74:2717–2727CrossRefGoogle Scholar
  6. Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Peña AG, Goodrich JK, Gordon JI (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7:335–336CrossRefGoogle Scholar
  7. Carson JK, Campbell L, Rooney D, Clipson N, Gleeson DB (2009) Minerals in soil select distinct bacterial communities in their microhabitats. FEMS Microbiol Ecol 67:381–388CrossRefGoogle Scholar
  8. Chao A (1984) Non-parametric estimation of the classes in a population. Scand J Stat 11:265–270Google Scholar
  9. Coleman DC, Whitman WB (2005) Linking species richness, biodiversity and ecosystem function in soil systems. Pedobiologia 49:479–497CrossRefGoogle Scholar
  10. Ding JL, Jiang X, Ma MC, Zhou BK, Guan DW, Zhao BS, Zhou J, Cao FM, Li L, Li J (2016) Effect of 35 years inorganic fertilizer and manure amendment on structure of bacterial and archaeal communities in black soil of northeast China. Appl Soil Ecol 105:187–195CrossRefGoogle Scholar
  11. Dion P (2008) Extreme views on prokaryote evolution. In: Dion P, Nautiyal CS (eds) Microbiology of extreme soils. Springer, Berlin, pp 45–70CrossRefGoogle Scholar
  12. Drenovsky RE, Vo D, Graham KJ, Scow KM (2004) Soil water content and organic carbon availability are major determinants of soil microbial community composition. Microb Ecol 48:424–430CrossRefGoogle Scholar
  13. Du E, Liu X (2014) High rates of wet nitrogen deposition in China: a synthesis. In: Sutton MA, Mason KE, Sheppard LJ, Sverdrup H, Haeuber R, Kevin HW (eds) Nitrogen deposition, critical loads and biodiversity. Proceedings of the International Nitrogen Initiative Workshop, linking experts of the Convention on Long-range Transboundary AirPollution and the Convention on Biological diversity. Springer, Netherlands, pp 48–56Google Scholar
  14. Dunbar J, Barns SM, Ticknor LO, Kuske CR (2002) Empirical and theoretical bacterial diversity in four Arizona soils. Appl Environ Microb 68:3035–3045CrossRefGoogle Scholar
  15. Fazi S, Amalfitano S, Pernthaler J, Puddu A (2005) Bacterial communities associated with benthic organic matter in headwater stream microhabitats. Environ Microbiol 7:1633–1640CrossRefGoogle Scholar
  16. Fierer N, Bradford MA, Jackson RB (2007) Toward an ecological classification of soil bacteria. Ecology 88:1354–1364CrossRefGoogle Scholar
  17. Fierer N, Lauber LC, Ramirez KS, Zaneveld J, Bradford MA, Knight R (2012) Comparative metagenomic, phylogenetic and physiological analyses of soil microbial communities across nitrogen gradients. ISME J 6:1007–1017CrossRefGoogle Scholar
  18. Frey SD, Ollinger S, Nadelhoffer K, Bowden R, Brzostek E, Burton A, Caldwell BA, Crow S, Goodale CL, Grandy AS, Finzi AC, Kramer MG, Lajtha K, LeMoine J, Martin M, McDowell WH, Minocha R, Sadowsky JJ, Templer PH, Wickings K (2014) Chronic nitrogen additions suppress decomposition and sequester soil carbon in temperate forests. Biogeochemistry 121:305–316CrossRefGoogle Scholar
  19. Galloway JN, Schlesinger WH, Levy H, Michaels A, Schnoor JL (1995) Nitrogen fixation: Anthropogenic enhancement-enviromental response. Global Biogeochem Cy 9:235–252CrossRefGoogle Scholar
  20. Galloway JN, Townsend AR, Erisman JW, Bekunda M, Cai Z, Freney JR, Martinelli LA, Seitzinger SP, Sutton MA (2008) Transformation of the nitrogen cycle: recent trends, questions, and potential solutions. Science 320:889–892CrossRefGoogle Scholar
  21. Hartman WH, Richardson CJ, Vilgalys R, Bruland GL (2008) Environmental and anthropogenic controls over bacterial communities in wetland soils. P Natl Acad Sci USA 105:17842CrossRefGoogle Scholar
  22. Holland EA, Braswell BH, Sulzman J, Lamarque JF (2005) Nitrogen deposition onto the United States and Western Europe: synthesis of observation and models. Ecol Appl 15:38–57CrossRefGoogle Scholar
  23. Huang W, Liu J, Wang YP, Zhou G, Han T, Li Y (2012) Increasing phosphorus limitation along three successional forests in southern China. Plant Soil 364:181–191CrossRefGoogle Scholar
  24. IPCC (2013) Climate change 2013: the physical science basis. In: Stocker TF, Qin D, Plattner G-K, Tignor M, Allen SK, Boschung J, Nauels A, Xia Y, Bex V, Midgley PM (eds) Contribution of Working Group I to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change. Cambridge University Press, CambridgeGoogle Scholar
  25. Jiang Y, Chen C, Xu Z, Liu Y (2011) Effects of single and mixed species forest ecosystems on diversity and function of soil microbial community in subtropical China. J Soils Sediments 12:228–240CrossRefGoogle Scholar
  26. Jones D, Willett V (2006) Experimental evaluation of methods to quantify dissolved organic nitrogen (DON) and dissolved organic carbon (DOC) in soil. Soil Biol Biochem 38:991–999CrossRefGoogle Scholar
  27. Lane DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. Wiley, Chichester, pp 115–175Google Scholar
  28. Li P, Han W, Zhang C, Tian D, Xu XN, Fang JY (2015) Nutrient resorption of Castanopsiseyrei varies at the defoliation peaks in spring and autumn in a subtropical forest, Anhui, China. Ecol Res 30:111–118CrossRefGoogle Scholar
  29. Lin YT, Jangid K, Whitman WB, Coleman DC, Chiu CY (2011) Soil bacterial communities in native and regenerated perhumid montane forests. Appl Soil Ecol 47:111–118CrossRefGoogle Scholar
  30. Lu XK, Mao QG, Gilliam F, Mo JM (2014) Nitrogen deposition contributes to soil acidification in tropical ecosystems. Glob Chang Biol 20:3790–3801CrossRefGoogle Scholar
  31. McCaig AE, Glover LA, Prosser JI (1999) Molecular analysis of bacterial community structure and diversity in unimproved and improved upland grass pastures. Appl Environ Microbiol 65:1721–1730Google Scholar
  32. Meng H, Li K, Nie M, Wan JR, Quan ZX, Fang CM, Chen JK, Gu JD, Li B (2013) Responses of bacterial and fungal communities to an elevation gradient in a subtropical montane forest of China. Appl Microbial Biot 97:2219–2230CrossRefGoogle Scholar
  33. Pei Z, Eichenberg D, Bruelheide H, Kröber W, Kühn P, Li Y, von Oheimb G, Purschke O, Scholten T, François Buscot F, Gutknecht JLM (2016) Soil and tree species traits both shape soil microbial communities during early growth of Chinese subtropical forests. Soil Biol Biochem 96:180–190CrossRefGoogle Scholar
  34. Philippot L, Andersson SGE, Battin TJ, Prosser JI, Schimel JP, Whitman WB, Hallin S (2010) The ecological coherence of high bacterial taxonomic ranks. Nat Rev Microbiol 8:523–529CrossRefGoogle Scholar
  35. Rousk J, Baath E, Brookes PC, Lauber CL, Lozupone C, Caporaso JG, Knight R, Fierer N (2010) Soil bacterial and fungal communities across a pH gradient in an arable soil. ISME J 4:1340–1351CrossRefGoogle Scholar
  36. Sessitsch A, Weilharter A, Gerzabek MH, Kirchmann H, Kandeler E (2001) Microbial population structures in soil particle size fractions of a long-term fertilizer field experiment. Appl Environ Microbiol 67:4215–4224CrossRefGoogle Scholar
  37. Song YC (1995) On the global position of the evergreen broad-leaved forests of China. In: Box EO, Peet RK, Masuzawa T, Yamada I, Fujiwara K, Maycock PF (eds) Vegetation science in forestry: global perspective based on Forest ecosystems of east and South-East Asia. Springer, Berlin, pp 69–84Google Scholar
  38. Sun H, Terhonen E, Koskinen K, Paulin L, Kasanen R, Asiegbu FO (2014) Bacterial diversity and community structure along different peat soils in boreal forest. Appl Soil Ecol 74:37–45CrossRefGoogle Scholar
  39. Tang J, Ding X, Wang L, Xu Q, Yang Z, Zhao J, Sun Q, Feng S, Zhang J (2012) Effects of wetland degradation on bacterial community in the Zoige Wetland of Qinghai-Tibetan Plateau (China). World J Microb Biot 28:649–657CrossRefGoogle Scholar
  40. Turlapati SA, Minocha R, Bhiravarasa PS, Tisa LS, Thomas WK, Minocha SC (2013) Chronic N-amended soils exhibit an altered bacterial community structure in Harvard Forest, MA, USA. FEMS Microbiol Ecol 83:478–493CrossRefGoogle Scholar
  41. Vance ED, Brookes PC, Jenkinson DS (1987) An extraction method for measuring soil microbial biomass C. Soil Biol Biochem 19:703–707Google Scholar
  42. Vitousek PM, Aber JD, Howarth RW, Likens GE, MAston PA, Schindler DW, Schlesinger WH, Tilman DG (1997) Human alteration of the global nitrogen cycle: sources and consequences. Ecol Appl 20:5–15CrossRefGoogle Scholar
  43. Wallenstein MD, McMahon S, Schimel J (2007) Bacterial and fungal community structure in Arctic tundra tussock and shrub soils. Microb Ecol 59:428–435CrossRefGoogle Scholar
  44. Wang X, Wang XL, Zhang WX, Shao YH, Zou XM, Liu T, Zhou LX, Wan SZ, Rao XQ, Li ZA, Fu SL (2016) Invariant community structure of soil bacteria in subtropical coniferous and broadleaved forests. Sci Rep 6:19071CrossRefGoogle Scholar
  45. Wu J, Joergensen RG, Pommerening B, Chaussod R, Brookes PC (1990) Measurement of soil microbial biomass C by fumigation-extraction-an automated procedure. Soil Biol Biochem 22:1167–1169CrossRefGoogle Scholar
  46. Wu YT, Gutknecht J, Nadrowski K, Geißler C, Kühn P, Scholten T, Both S, Erfmeier A, Böhnke M, Bruelheide H, Wubet T, Buscot F (2012) Relationships between soil microorganisms, plant communities, and soil characteristics in Chinese subtropical forests. Ecosystems 15:624–636CrossRefGoogle Scholar
  47. Yang J, Ke L, Cui J, Xu XN (2014) Responses of soil dissolved organic carbon and microbial biomass carbon to N and P addition in a subtropical evergreen broad-leaved forest. Chin J Soil Sci 45:902–909 (in Chinese with English abstract)Google Scholar
  48. Zeng J, Liu X, Song L, Lin X, Zhang H, Shen C, Chu H (2016) Nitrogen fertilization directly affects soil bacterial diversity andindirectly affects bacterial community composition. Soil Biol Biochem 92:41–49CrossRefGoogle Scholar
  49. Zhang C, Zhang L, Li P, Shi WT, Xu XN (2014a) Effect of elevated N deposition on litterfall productionand seasonality in a subtropical evergreen broad-leaved forest. Chin J Ecol 33:1205–1210 (in Chinese with English abstract)Google Scholar
  50. Zhang X, Wei H, Chen Q, Han X (2014b) The counteractive effects of nitrogen addition and watering on soil bacterial communities in asteppe ecosystem. Soil Biol Biochem 72:26–34CrossRefGoogle Scholar
  51. Zhou LL, Addo-Danso SD, Wu PF, Li SB, Zou XH, Zhang Y, Ma XQ (2016) Leaf resorption efficiency in relation to foliar and soil nutrient concentrations and stoichiometry of Cunninghamia lanceolata with stand development in southern China. J Soils Sediments 5:1448–1459CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Jun Cui
    • 1
    • 2
  • Jingjing Wang
    • 1
    • 2
  • Jun Xu
    • 1
  • Chonghua Xu
    • 1
  • Xiaoniu Xu
    • 1
    • 2
    Email author
  1. 1.School of Forestry and Landscape ArchitectureAnhui Agricultural UniversityHefeiChina
  2. 2.Collaborative Innovation Centre of Agro-forestry Industry in Dabieshan AreaHefeiChina

Personalised recommendations