Enzyme Activities and Microbial Communities in Subtropical Forest Soil Aggregates to Ammonium and Nitrate-Nitrogen Additions

  • 1. Northeast Normal University, College of Geographic Science, Changchun 130024, China;
    2. Key Laboratory of Ecosystem Network Observation and Modeling, Institute of Geographic Sciences and Natural Resources Research, Chinese Academy of Sciences, Beijing 100101, China;
    3. College of Resources and Environment, University of Chinese Academy of Sciences, Beijing 100190, China

Received date: 2017-02-02

  Revised date: 2017-03-28

  Online published: 2017-05-20

Supported by

National Natural Science Foundation of China (41571251, 41571130043); Technology Innovation Program of Chinese Academy of Sciences (201604).


A laboratory incubation experiment was established to examine the impacts of nitrate and ammonium nitrogen additions on soil microbial attributes of a subtropical Pinus elliottii forest ecosystem in southern China. Soils were subjected to three different treatments: the control with no nitrogen addition (CK), the ammonium nitrogen addition (NH4+-N), and the nitrate nitrogen addition (NO3--N). Samples from bulk and two different size fractions (macroaggregate (>250 μm) and microaggregate (53-250 μm)) were analyzed for soil properties, enzyme activities and microbial communities on day 7 and 15 of the incubation. Our study demonstrated that NH4+-N had a greater influence on soil microbial activities than NO3--N. NH4+-N additions resulted in significant increases in β-1,4-glucosidase (βG) and β-1,4-N-acetyl glucosaminidase (NAG) enzyme activities in bulk, macroaggregate and microaggregate soils after 7 and 15 days incubation. NO3--N additions only significantly increased in βG and NAG enzyme activities in bulk, macroaggregate soils after 7 and 15 days incubation, but not in microaggregate. All NH4+-N and NO3--N additions resulted in significant increases in gram-positive bacterial PLFAs in microaggregates. Only a significant correlation between soil nutrient contents and enzyme activities in macroaggregates was founded, which suggests that the soil aggregation structure played an important role in the determining enzyme activities.

Cite this article

WEI Yan, WANG Zhongqiang, ZHANG Xinyu, YANG Hao, LIU Xiyu, LIU Wenjing . Enzyme Activities and Microbial Communities in Subtropical Forest Soil Aggregates to Ammonium and Nitrate-Nitrogen Additions[J]. Journal of Resources and Ecology, 2017 , 8(3) : 258 -267 . DOI: 10.5814/j.issn.1674-764x.2017.03.006


[1] Allison S D, Czimczik C I, Treseder K K. 2008. Microbial activity and soil respiration under nitrogen addition in Alaskan boreal forest. Global Change Biol, 14(5): 1156-1168.
[2] Allison S D, Vitousek P M. 2005. Responses of extracellular enzymes to simple and complex nutrient inputs. Soil Biol Biochem, 37(5): 937-944.
[3] An S S, Mentler A, Mayer H, et al. 2010. Soil aggregation, aggregate stability, organic carbon and nitrogen in different soil aggregate fractions under forest and shrub vegetation on the Loess Plateau, China. Catena, 81(3): 226-233.
[4] Bååth E, Anderson T H. 2003. Comparison of soil fungal/bacterial ratios in a pH gradient using physiological and PLFA-based techniques. Soil Biol Biochem, 35(7): 955-963.
[5] Bergstrom D W, Monreal C, King D J. 1998. Sensitivity of soil enzyme activities to conservation practices. Soil Sci Soc Am J, 62(5): 1286- 1295.
[6] Bettina H M S, Karsten K, Sabine B, et al. 2011. Microbial immobilization and mineralization of dissolved organic nitrogen from foerst floors. Soil Biol Biochem, 43(8): 1742-1745.
[7] Burns R G, DeForest J L, Marxsen J, et al. 2013. Soil enzymes in a changing environment: Current knowledge and future directions. Soil Biol Biochem, 58(2): 216-234.
[8] Burns R G, Dick R P. 2002. Enzymes in the environment: activity, ecology, and applications. CRC Press, New York.
[9] Calderon F J, Jackson L E, Scow K M, et al. 2001. Short-term dynamics of nitrogen, microbial activity, and phospholipid fatty acids after tillage. Soil Sci Soc Am J, 65(1): 118-126.
[10] Carreiro M M, Sinsabaugh R L, Repert D A, et al. 2000. Microbial enzyme shifts explain litter decay responses to simulated nitrogen deposition. Ecology, 81(9): 2359-2365.
[11] Carson J K, Gonzalez-Quiñones V, Murphy D V, et al. 2010. Low pore connectivity increases bacterial diversity in soil. Appl Environ Microb, 76(12): 3936-3942.
[12] Cusack D F, Silver W L, Torn M S, et al. 2011. Changes in microbial community characteristics and soil organic matter with nitrogen additions in two tropical forests. Ecology, 92(3): 621-632.
[13] Cusack D F, Silver W L, Torn M S, et al. 2011. Effects of nitrogen additions on above-and belowground carbon dynamics in two tropical forests. Biogeochemistry, 104(1): 203-225.
[14] Cusack D F, Torn M S, McDowell W H, et al. 2010. The response of heterotrophic activity and carbon cycling to nitrogen additions and warming in two tropical soils. Global Change Biol, 16(9): 2555-2572.
[15] DeForest J L, Zak D R, Pregitzer K S, et al. 2004. Atmospheric nitrate deposition, microbial community composition, and enzyme activity in northern hardwood forests. Soil Sci Soc Am J, 68(1): 132-138.
[16] Drenovsky R E, Elliott G N, Graham K J, et al. 2004. Comparison of phospholipid fatty acid (PLFA) and total soil fatty acid methyl esters (TSFAME) for characterizing soil microbial communities. Soil Biol Biochem, 36(11): 1793-1800.
[17] Drenovsky R E , Vo D, Graham K J, et al. 2004. Soil water content and organic carbon availability are major determinants of soil microbial community composition. Microbial Ecol, 48(3): 424-430.
[18] Ekenler M, Tabatabai M. 2002. β-Glucosaminidase activity of soils: effect of cropping systems and its relationship to nitrogen mineralization. Biol Fert Soils, 36(5): 367-376.
[19] Elliott E T. 1986. Aggregate structure and carbon, nitrogen, and phosphorus in native and cultivated soils. Soil Sci Soc Am J, 50(3): 627-633.
[20] Evans C D, Goodale C L, Caporn S J, et al. 2008. Does elevated nitrogen deposition or ecosystem recovery from acidification drive increased dissolved organic carbon loss from upland soil? A review of evidence from field nitrogen addition experiments. Biogeochemistry, 91(1): 13-35.
[21] Fansler S J, Smith J L, Bolton J H, et al. 2005. Distribution of two C cycle enzymes in soil aggregates of a prairie chronosequence. Biol Fert Soils, 42(1): 17-23.
[22] Fierer N, Jackson R B. 2006. The diversity and biogeography of soil bacterial communities. P Natl Acad Sci USA, 103(3): 626-63.
[23] Frey S D, Knorr M, Parrent J L, et al. 2004. Chronic nitrogen enrichment affects the structure and function of the soil microbial community in temperate hardwood and pine forests. Forest Ecol Manag, 196(1): 159-171.
[24] Frostegård Å, Bååth E. 1996. The use of phospholipid fatty acid analysis to estimate bacterial and fungal biomass in soil. Biol Fert Soils, 22(1): 59-65.
[25] German D P, Weintraub M N, Grandy A S, et al. 2011. Corrigendum to “Optimization of hydrolytic and oxidative enzyme methods for ecosystem studies”. Soil Biol Biochem, 43(7): 1387-1397.
[26] Högberg M N, Högberg P, Myrold D D. 2007. Is microbial community composition in boreal forest soils determined by pH, C-to-N ratio, the trees, or all three? Oecologia, 150(4): 590-601.
[27] Högberg P, Fan H B, Quist M, et al. 2006. Tree growth and soil acidification in response to 30 years of experimental nitrogen loading on boreal forest. Global Change Biol, 12(3): 489-499.
[28] Jia Y L, Yu G R, He N P, et al. 2014. Spatial and decadal variations in inorganic nitrogen wet deposition in China induced by human activity. Sci Rep, 4(1): 3763-3763.
[29] Lamarque J F, Kiehl J T, Brasseur G P, et al. 2005. Assessing future nitrogen deposition and carbon cycle feedback using a multimodel approach: Analysis of nitrogen deposiont. J Geophys Res-atmos, 110(D19): 2657-2677.
[30] Liu K H, Fang Y T, Yu F M, et al. 2010. Soil acidification in response to acid deposition in three subtropical forests of subtropical China. Pedosphere, 20(3): 399-408.
[31] Liu Y R, Li X, Shen Q R, et al. 2013. Enzyme activity in water-stable soil aggregates as affected by long-term application of organic manure and chemical fertiliser. Pedosphere, 23(1): 111-119.
[32] Lu R K. 1999. Soil Agro-chemistrical Analysis. Beijing: China Agricultural Sciences Press, 107-147.
[33] Lu X K, Mo J M, Gilliam F S, et al. 2010. Effects of experimental nitrogen additions on plant diversity in an old-growth tropical forest. Global Change Biol, 16(10): 2688-2700.
[34] Lucas R W, Casper B B, Jackson J K, et al. 2007. Soil microbial communities and extracellular enzyme activity in the New Jersey Pinelands. Soil Biol Biochem, 39(10): 2508-2519.
[35] Marx M C, Kandeler E, Wood M, et al. 2005. Exploring the enzymatic landscape: distribution and kinetics of hydrolytic enzymes in soil particle-size fractions. Soil Biol Biochem, 37(1): 35-48.
[36] Mikha M M, Rice C W. 2004. Tillage and manure effects on soil and aggregate-associated carbon and nitrogen. Soil Sci Soc Am J, 68(3): 809-816.
[37] Mo J M, Brown S, Xue J H, et al. 2006. Response of litter decomposition to simulated N deposition in disturbed, rehabilitated and mature forests in subtropical China. Plant Soil, 282(1): 135-151.
[38] Muruganandam S, Israel D W, Robarge W P. 2009. Activities of nitrogen-mineralization enzymes associated with soil aggregate size fractions of three tillage systems. Soil Sci Soc Am J, 73(3): 751-759.
[39] Nie M, Pendall E, Bell C, et al. 2014. Soil aggregate size distribution mediates microbial climate change feedbacks. Soil Biol Biochem, 68: 357-365.
[40] Nilsson L O, Bååth E, Falkengren-Grerup U, et al. 2007. Growth of ectomycorrhizal mycelia and composition of soil microbial communities in oak forest soils along a nitrogen deposition gradient. Oecologia, 153(2): 375-384.
[41] Pregitzer K S, Burton A J, Zak D R, et al. 2010. Simulated chronic nitrogen deposition increases carbon storage in Northern Temperate forests. Glob Change Biol, 14(1): 142-153.
[42] Reuss J O, Johnson D W. 1986. Acid deposition and the acidification of soil and waters. Springer Verlag, NewYork.
[43] Rousk J, Brookes P C, Bååth E. 2009. Contrasting soil pH effects on fungal and bacterial growth suggest functional redundancy in carbon mineralization. Appl Environ Micbob, 75(6): 1589-1596.
[44] Saiya-Cork K R, Sinsabaugh R L, Zak D R. 2002. The effects of long term nitrogen deposition on extracellular enzyme activity in an Acer saccharum forest soil. Soil Biol Biochem, 34(9): 1309-1315.
[45] Schloter M, Dilly O, Munch J. 2003. Indicators for evaluating soil quality. Agr Ecosyst Environ, 98(1-3): 255-262.
[46] Schutter M E, Dick R P. 2002. Microbial community profiles and activities among aggregates of winter fallow and cover-cropped soil. Soil Sci Soc Am J, 66(1): 142-153.
[47] Sinsabaugh R L, Gallo M E, Lauber C, et al. 2005. Extracellular enzyme activities and soil organic matter dynamics for northern hardwood forests receiving simulated nitrogen deposition. Biogeochemistry, 75(2): 201-215.
[48] Six J, Conant R T, Paul E A, et al. 2002. Stabilization mechanisms of soil organic matter: implications for C-saturation of soils. Plant Soil, 241(2): 155-176.
[49] Six J, Elliott E, Paustian K. 2000. Soil structure and soil organic matter II. A normalized stability index and the effect of mineralogy. Soil Sci Soc Am J, 64(3): 1042-1049.
[50] Sowerby A, Emmett B, Beier C, et al. 2005. Microbial community changes in heathland soil communities along a geographical gradient: interaction with climate change manipulations. Soil Biol Biochem, 37(10): 1805-1813.
[51] Thoms C, Gattinger A, Jacob M, et al. 2010. Direct and indirect effects of tree diversity drive soil microbial diversity in temperate deciduous forest. Soil Biol Biochem, 42(9): 1558-1565.
[52] Turner B L, Hopkins D W, Haygarth P M, et al. 2002. β-Glucosidase activity in pasture soils. Appl Soil Ecol, 20(2): 157-162.
[53] Waldrop M P, Zak D R. 2006. Response of oxidative enzyme activities to nitrogen deposition affects soil concentrations of dissolved organic carbon. Ecosystems, 9(6): 921-933.
[54] Waldrop M P, Zak D R, Sinsabaugh R L, et al. 2004. Nitrogen deposition modifies soil carbon storage through changes in microbial enzymatic activity. Ecol App, 14(4): 1172-1177.
[55] Wardle D.A. 1992. A comparative assessment of factors which influence microbial biomass carbon and nitrogen levels in soil. Biological Biol Rev, 67(3): 321-358.
[56] Wen X F, Wang H M, Wang J L, et al. 2010. Ecosystem carbon exchanges of a subtropical evergreen coniferous plantation subjected to seasonal drought, 2003-2007. Biogeosciences, 7(1): 357-369.
[57] Xu Z W, Yu G R, Zhang X Y, et al. 2015. The variations in soil microbila communities, enzyme activities and their relationships with soil organic matter decomposition along the northern slope of Changbai Mountain. Appl Soil Ecol, 86(86): 19-29.
[58] Xu Z W, Zhang X Y, Xie J, et al. 2014. Total Nitrogen Concentrations in Surface Water of Typical Agro- and Forest Ecosystems in China, 2004-2009. PLoS ONE, 9(3): 1-9.
[59] Yu H Y, Ding W X, Luo J F, et al. 2012. Long-term effect of compost and inorganic fertilizer on activities of carbon-cycle enzymes in aggregates of an intensively cultivated sandy loam. Soil Use Manage, 28(3): 347-360.
[60] Zhang J B, Cai Z C, Zhu T B, et al. 2013. Mechanisms for the retention of inorganic N in acidic forest soils of southern China. Sci Rep, 3(6145): 2342.
[61] Zhang Q C, Shamsi I H, Xu D T, et al. 2012. Chemical fertilizer and organic manure inputs in soil exhibit a vice versa pattern of microbial community structure. Appl Soil Ecol, 57(1): 1-8.
[62] Zhao X, Xing G X. 2009. Variation in the relationship between nitrification and acidification of subtropical soils as affected by the addition of urea or ammonium sulfate. Soil Biol Biochem, 41(12): 2584-2587.