Difference in soil bacterial community composition depends on forest type rather than nitrogen and phosphorus additions in tropical montane rainforests

  • Pin LiEmail author
  • Congcong Shen
  • Lai Jiang
  • Zhaozhong Feng
  • Jingyun Fang
Original Paper


Rapid increase of nitrogen (N) deposition could alter nutrient availability, leading to changes in soil microbial processes and ecosystem carbon and nutrient cycling. However, the effects of N deposition on soil microbes remain elusive in the tropical rainforests in Asia. Here, we conducted a 3-year N addition experiment with four treatments (0, 20, 50, and 100 kg N ha−1 year−1) in a primary and secondary tropical montane forest in Hainan Island, China, to explore the effects of elevated N availability on soil microbial community composition. We also conducted a phosphorus (P) treatment (50 kg P ha−1 year−1) and a N + P treatment (50 kg N ha−1 year−1 + 50 kg P ha−1 year−1) to examine potential P limitation driven by N deposition in highly weathered tropical soils, using a bar-coded pyrosequencing technique. The composition of soil bacterial communities differed dramatically between the primary and secondary forests, but not significant dissimilarity among the fertilization treatments. The community composition, phylogenetic diversity and phylotype richness were significantly correlated with soil pH, total organic C (TOC), and total N (TN), respectively. There were significant differences between the primary and secondary forest in pH, TOC, and TN, but not among the fertilization treatments. These results suggest that differences in soil nutrient status between the primary and secondary forests due to different successional stages rather than chronic N fertilization may be the major factor affecting soil bacterial composition in the tropical montane rainforests.


Nitrogen and phosphorus addition Soil bacterial community Tropic Primary and secondary forest Pyrosequencing 



We thank Dr. Haiyan Chu for kindly allowing us to use the lab equipment, Dr. Yu Shi and Mr. Huaibo Sun for kindly providing us with experimental guidance. Thanks are also due to Dr. Xiaoting Xu and Dr. Haihua Shen for the assistance of sampling and analysis. We thank Alex Boon, PhD, from Liwen Bianji, Edanz Editing China (, for editing the English text of a draft of this manuscript.

Funding information

This study was financially supported by National Key Research and Development Program (2017YFC0503900).

Supplementary material

374_2019_1349_MOESM1_ESM.docx (365 kb)
ESM 1 (DOCX 364 kb)


  1. Bai ZZ, Yang G, Chen H, Zhu Q, Chen DX, Li YD, Wang X, Wu ZM, Zhou GY, Peng CH (2014) Nitrous oxide fluxes from three forest types of the tropical mountain rainforests on Hainan Island, China. Atmos Environ 92:469–477CrossRefGoogle Scholar
  2. Boot CM, Hall EK, Denef K, Baron JS (2016) Long-term reactive nitrogen loading alters soil carbon and microbial community properties in a subalpine forest ecosystem. Soil Biol Biochem 92:211–220CrossRefGoogle Scholar
  3. Camenzind T, Hempel S, Homeier J, Horn S, Velescu A, Wilcke W, Rillig MC (2014) Nitrogen and phosphorus additions impact arbuscular mycorrhizal abundance and molecular diversity in a tropical montane forest. Glob Chang Biol 20:3646–3659CrossRefGoogle Scholar
  4. Camenzind T, Homeier J, Dietrich K, Hempel S, Hertel D, Krohn A, Leuschner C, Oelmann Y, Olsson PA, Suárez JP, Rillig MC (2016) Opposing effects of nitrogen versus phosphorus additions on mycorrhizal fungal abundance along an elevational gradient in tropical montane forests. Soil Biol Biochem 94:37–47CrossRefGoogle Scholar
  5. Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Peña AG, Goodrich JK, Gordon JI, Huttley GA, Kelley ST, Knights D, Koenig JE, Ley RE, Lozupone CA, McDonald D, Muegge BD, Pirrung M, Reeder J, Sevinsky JR, Turnbaugh PJ, Walters WA, Widmann J, Yatsunenko T, Zaneveld J, Knight R (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7:335–336CrossRefGoogle Scholar
  6. Chen LY, Li P, Yang YH (2016a) Dynamic patterns of nitrogen: phosphorus ratios in forest soils of China under changing environment. J Geophys Res-Biogeo 121:2410–2421CrossRefGoogle Scholar
  7. Chen YL, Ding JZ, Peng YF, Li F, Yang GB, Liu L, Qin SQ, Fang K, Yang YH (2016b) Patterns and drivers of soil microbial communities in Tibetan alpine and global terrestrial ecosystems. J Biogeogr 43:2027–2039CrossRefGoogle Scholar
  8. Chu HY, Fierer N, Lauber CL, Caporaso JG, Knight R, Grogan P (2010) Soil bacterial diversity in the Arctic is not fundamentally different from that found in other biomes. Environ Microbiol 12:2998–3006CrossRefGoogle Scholar
  9. Cleveland CC, Liptzin D (2007) C:N:P stoichiometry in soil: is there a ‘Redfield ratio’ for the microbial biomass? Biogeochemistry 85:235–252CrossRefGoogle Scholar
  10. Contosta AR, Frey SD, Cooper AB (2015) Soil microbial communities vary as much over time as with chronic warming and nitrogen additions. Soil Biol Biochem 88:19–24CrossRefGoogle Scholar
  11. Cusack DF, Silver WL, Torn MS, Burton SD, Firestone MK (2011) Changes in microbial community characteristics and soil organic matter with nitrogen additions in two tropical forests. Ecology 92:621–632CrossRefGoogle Scholar
  12. Don A, Böhme IH, Dohrmann AB, Poeplau C, Tebbe CC (2017) Microbial community composition affects soil organic carbon turnover in mineral soils. Biol Fertil Soils 53:445–456CrossRefGoogle Scholar
  13. Du E, Zhou Z, Li P, Hu X, Ma Y, Wang W, Zheng C, Zhu J, He JS, Fang J (2013) NEECF: a project of nutrient enrichment experiments in China’s forests. J Plant Ecol 6:428–435CrossRefGoogle Scholar
  14. Faith DP (1992) Conservation evaluation and phylogenetic diversity. Biol Conserv 61:1–10CrossRefGoogle Scholar
  15. Fang JY, Li YD, Zhu B, Liu GH, Zhou GY (2004) Community structures and species richness in the montane rain forest of Jianfengling, Hainan Island, China. Biodivers Sci 12:29–43Google Scholar
  16. Fanin N, Hattenschwiler S, Schimann H, Fromin N (2015) Interactive effects of C, N and P fertilization on soil microbial community structure and function in an Amazonian rain forest. Funct Ecol 29:140–150CrossRefGoogle Scholar
  17. Fierer N, Jackson RB (2006) The diversity and biogeography of soil bacterial communities. Proc Natl Acad Sci USA 103:626–631CrossRefGoogle Scholar
  18. Galloway JN, Winiwarter W, Leip A, Leach AM, Bleeker A, Erisman JW (2014) Nitrogen footprints: past, present and future. Environ Res Lett 9:115003CrossRefGoogle Scholar
  19. Jiang L, Tian D, Ma SH, Zhou XL, Xu LC, Zhu JX, Jing X, Zheng CY, Shen HH, Zhou Z, Li YD, Zhu B, Fang JY (2017) The response of tree growth to nitrogen and phosphorus additions in a tropical montane rainforest. Sci Total Environ 618:1064–1070CrossRefGoogle Scholar
  20. Jing X, Chen X, Tang M, Ding ZJ, Jiang L, Li P, Ma SH, Tian D, Xu LC, Zhu JX, Ji CJ, Shen HH, Zheng CY, Fang JY, Zhu B (2017) Nitrogen deposition has minor effect on soil extracellular enzyme activities in six Chinese forests. Sci Total Environ 607–608:806–815CrossRefGoogle Scholar
  21. LeBauer DS, Treseder KK (2008) Nitrogen limitation of net primary productivity in terrestrial ecosystems is globally distributed. Ecology 89:371–379CrossRefGoogle Scholar
  22. Li WZ, Godzik A (2006) Cd-hit, a fast program for clustering and comparing large sets of protein or nucleotide sequences. Bioinformatics 22:1658–1659CrossRefGoogle Scholar
  23. Li P, Yang YH, Han WX, Fang JY (2014) Global patterns of soil microbial nitrogen and phosphorus stoichiometry in forest ecosystems. Glob Ecol Biogeogr 23:979–987CrossRefGoogle Scholar
  24. Li J, Li ZA, Wang FM, Zou B, Chen Y, Zhao J, Mo QF, Li YW, Li XB, Xia HP (2015) Effects of nitrogen and phosphorus addition on soil microbial community in a secondary tropical forest of China. Biol Fert Soils 51:207–215CrossRefGoogle Scholar
  25. Liu LL, Greaver TL (2010) A global perspective on belowground carbon dynamics under nitrogen enrichment. Ecol Lett 13:819–828CrossRefGoogle Scholar
  26. Liu L, Gundersen P, Zhang T, Mo JM (2012) Effects of phosphorus addition on soil microbial biomass and community composition in three forest types in tropical China. Soil Biol Biochem 44:31–38CrossRefGoogle Scholar
  27. Liu L, Zhang T, Gilliam FS, Gundersen P, Zhang W, Chen H, Mo JM (2013a) Interactive effects of nitrogen and phosphorus on soil microbial communities in a tropical forest. PLoS One 8:e61188. CrossRefGoogle Scholar
  28. Liu XJ, Zhang Y, Han WX, Tang A, Shen JL, Cui ZL, Vitousek P, Erisman JW, Goulding K, Christie P, Fangmeier A, Zhang FS (2013b) Enhanced nitrogen deposition over China. Nature 494:459–462CrossRefGoogle Scholar
  29. Liu L, Gundersen P, Zhang W, Zhang T, Chen H, Mo JM (2015) Effects of nitrogen and phosphorus additions on soil microbial biomass and community structure in two reforested tropical forests. Sci Rep 5:14378. CrossRefGoogle Scholar
  30. Lu XK, Mao QG, Gilliam FS, Luo YQ, Mo JM (2014) Nitrogen deposition contributes to soil acidification in tropical ecosystems. Glob Chang Biol 20:3790–3801CrossRefGoogle Scholar
  31. Lucas RW, Klaminder J, Futter MN, Bishop KH, Egnell G, Laudon H, Hogberg P (2011) A meta-analysis of the effects of nitrogen additions on base cations: implications for plants, soils, and streams. Forest Ecol Manag 262:95–104CrossRefGoogle Scholar
  32. Luyssaert S, Schulze ED, Borner A, Knohl A, Hessenmoller D, Law BE, Ciais P, Grace J (2008) Old-growth forests as global carbon sinks. Nature 455:213–215CrossRefGoogle Scholar
  33. Marklein AR, Houlton BZ (2012) Nitrogen inputs accelerate phosphorus cycling rates across a wide variety of terrestrial ecosystems. New Phytol 193:696–704Google Scholar
  34. Nelson DW, Sommers LE (1982) Total carbon, organic carbon, and organic matter. In: Page AL (ed) Methods of soil analysis. American Society of Agronomy, Madison, WI, pp 539–579Google Scholar
  35. Ngo KM, Turner BL, Muller-Landau HC, Davies SJ, Larjavaara M, Hassan NFN, Lum S (2013) Carbon stocks in primary and secondary tropical forests in Singapore. Forest Ecol Manag 296:81–89CrossRefGoogle Scholar
  36. Olsen SR, Cole CV, Watanabe FS, Dean LA (1954) Estimation of available phosphorus in soils by extraction with sodium bicarbonate. USDA circular 939. U.S. Govt. Printing Office, Washington, DCGoogle Scholar
  37. Pan Y, Birdsey RA, Fang J, Houghton R, Kauppi PE, Kurz WA, Phillips OL, Shvidenko A, Lewis SL, Canadell JG, Ciais P, Jackson RB, Pacala SW, McGuire AD, Piao S, Rautiainen A, Sitch S, Daniel H (2011) A large and persistent carbon sink in the world’s forests. Science 333:988–993CrossRefGoogle Scholar
  38. R Development Core Team (2015) R, a language and environment for statistical computing. Foundation for Statistical Computing, Vienna, AustriaGoogle Scholar
  39. Reay DS, Dentener F, Smith P, Grace J, Feely RA (2008) Global nitrogen deposition and carbon sinks. Nat Geosci 1:430–437CrossRefGoogle Scholar
  40. Reich PB, Oleksyn J (2004) Global patterns of plant leaf N and P in relation to temperature and latitude. PNAS 101:11001–11006CrossRefGoogle Scholar
  41. Schöler A, Jacquiod S, Vestergaard G, Schulz S, Schloter M (2017) Analysis of soil microbial communities based on amplicon sequencing of marker genes. Biol Fertil Soils 53:485–489CrossRefGoogle Scholar
  42. Shen C, Xiong J, Zhang H, Feng Y, Lin X, Li X, Liang W, Chu H (2013) Soil pH drives the spatial distribution of bacterial communities along elevation on Changbai Mountain. Soil Biol Biochem 57:204–211CrossRefGoogle Scholar
  43. Shen C, Ni Y, Liang W, Wang J, Chu H (2015) Distinct soil bacterial communities along a small-scale elevational gradient in alpine tundra. Front Microbiol 6:582.
  44. Shen C, Ge Y, Yang T, Chu HY (2017) Verrucomicrobial elevational distribution was strongly influenced by soil pH and carbon/nitrogen ratio. J Soils Sediments 17:2449–2456CrossRefGoogle Scholar
  45. Sommers LE, Nelson DW (1972) Determination of total phosphorus in soils: a rapid perchloric acid digestion procedure. Soil Sci Soc Am J 36:902–904CrossRefGoogle Scholar
  46. Stempfhuber B, Richter-Heitmann T, Bienek L, Schöning I, Schrumpf M, Friedrich M, Schulz S, Schloter M (2017) Soil pH and plant diversity drive co-occurrence patterns of ammonia and nitrite oxidizer in soils from forest ecosystems. Biol Fertil Soils 53:691–700CrossRefGoogle Scholar
  47. Strickland MS, Lauber C, Fierer N, Bradford MA (2009) Testing the functional significance of microbial community composition. Ecology 90:441–451CrossRefGoogle Scholar
  48. Terrer C, Vicca S, Stocker BD, Hungate BA, Phillips RP, Reich PB, Finzi AC, Prentice IC (2018) Ecosystem responses to elevated CO2 governed by plant–soil interactions and the cost of nitrogen acquisition. New Phytol 217:507–522CrossRefGoogle Scholar
  49. Tian D, Li P, Fang WJ, Xu J, Luo YK, Yan ZB, Zhu B, Wang JJ, Xu XN, Fang JY (2017) Growth responses of trees and understory plants to nitrogen fertilization in a subtropical forest in China. Biogeosciences 14:3461–3469CrossRefGoogle Scholar
  50. Turner BL, Wright SJ (2014) The response of microbial biomass and hydrolytic enzymes to a decade of nitrogen, phosphorus, and potassium addition in a lowland tropical rain forest. Biogeochemistry 117:115–130CrossRefGoogle Scholar
  51. van der Heijden MGA, Bardgett RD, Straalen NMV (2008) The unseen majority: soil microbes as drivers of plant diversity and productivity in terrestrial ecosystems. Ecol Lett 11:296–310CrossRefGoogle Scholar
  52. Vestergaard G, Schulz S, Schöler A, Schloter M (2017) Making big data smart—how to use metagenomics to understand soil quality. Biol Fert Soils 53:479–484CrossRefGoogle Scholar
  53. Vitousek PM, Porder S, Houlton BZ, Chadwick OA (2010) Terrestrial phosphorus limitation: mechanisms, implications, and nitrogen-phosphorus interactions. Ecol Appl 20:5–15CrossRefGoogle Scholar
  54. Weintraub SR, Wieder WR, Cleveland CC, Townsend AR (2013) Organic matter inputs shift soil enzyme activity and allocation patterns in a wet tropical forest. Biogeochemistry 114:313–326CrossRefGoogle Scholar
  55. Wright SJ (2005) Tropical forests in a changing environment. Trends Ecol Evol 20:553–560CrossRefGoogle Scholar
  56. Xiong J, Liu Y, Lin X, Zhang H, Zeng J, Hou J, Yang Y, Yao T, Knight R, Chu H (2012) Geographic distance and pH drive bacterial distribution in alkaline lake sediments across Tibetan Plateau. Environ Microbiol 14:2457–2466CrossRefGoogle Scholar
  57. Yang Y, Li P, Ding J, Zhao X, Ma W, Ji C, Fang J (2014) Increased topsoil carbon stock across China’s forests. Glob Chang Biol 20:2687–2696CrossRefGoogle Scholar
  58. Zak DR, Pregitzer KS, King JS, Holmes WE (2000) Elevated atmospheric CO2 fine roots and the response of soil microorganisms: a review and hypothesis. New Phytol 147:201–222CrossRefGoogle Scholar
  59. Zeng J, Liu XJ, Song L, Lin XG, Zhang HY, Shen CC, Chu HY (2016) Nitrogen fertilization directly affects soil bacterial diversity and indirectly affects bacterial community composition. Soil Biol Biochem 92:41–49CrossRefGoogle Scholar
  60. Zhou Z, Jiang L, Du EZ, Hu HF, Li YD, Chen DX, Fang JY (2013) Temperature and substrate availability regulate soil respiration in the tropical mountain rainforests, Hainan Island, China. J Plant Ecol 6:325–334CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Urban and Regional Ecology, Research Center for Eco-Environmental SciencesChinese Academy of SciencesBeijingChina
  2. 2.College of Resources and EnvironmentUniversity of Chinese Academy of SciencesBeijingChina
  3. 3.Department of Ecology, College of Urban and Environmental Sciences, and Key Laboratory for Earth Surface Processes of the Ministry of EducationPeking UniversityBeijingChina
  4. 4.Collaborative Innovation Center of Atmospheric Environment and Equipment Technology, School of Environmental Science and EngineeringNanjing University of Information Science & TechnologyNanjingChina

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