Abstract
Glyphosate is a non-selective organophosphate herbicide that is widely used in agriculture, but its effects on soil microbial communities are highly variable and often contradictory, especially for high dose applications. We applied glyphosate at two rates: the recommended rate of 50 mg active ingredient kg−1 soil and 10-fold this rate to simulate multiple glyphosate applications during a growing season. After 6 months, we investigated the effects on the composition of soil microbial community, the catabolic activity and the genetic diversity of the bacterial community using phospholipid fatty acids (PLFAs), community level catabolic profiles (CLCPs), and 16S rRNA denaturing gradient gel electrophoresis (DGGE). Microbial biomass carbon (Cmic) was reduced by 45%, and the numbers of the cultivable bacteria and fungi were decreased by 84 and 63%, respectively, under the higher glyphosate application rate. According to the PLFA analysis, the fungal biomass was reduced by 29% under both application rates. However, the CLCPs showed that the catabolic activity of the gram-negative (G−) bacterial community was significantly increased under the high glyphosate application rate. Furthermore, the DGGE analysis indicated that the bacterial community in the soil that had received the high glyphosate application rate was dominated by G− bacteria. Real-time PCR results suggested that copies of the glyphosate tolerance gene (EPSPS) increased significantly in the treatment with the high glyphosate application rate. Our results indicated that fungi were impaired through glyphosate while G− bacteria played an important role in the tolerance of microbiota to glyphosate applications.
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References
Andréa DMM, Bonanholuchini T, Carlosbazarin L, Papini S, Matallo S, Barifousesavoy M, Tedeschi VL (2003) Influence of repeated applications of glyphosate on its persistence and soil bioactivity. Pesqui Agropecu Bras 38:1329–1335
Ayoubi PJ, Harker AR (1998) Whole-cell kinetics of trichloroethylene degradation by phenol hydroxylase in a Ralstonia eutropha JMP134 derivative. Appl Environ Microbiol 64:4353–4356
Bai SH, Ogbourne SM (2016) Glyphosate: environmental contamination, toxicity and potential risks to human health via food contamination. Environ Sci Pollut Res 23:18988–19001
Berg J, Tom-Petersen A, Nybroe O (2005) Copper amendment of agricultural soil selects for bacterial antibiotic resistance in the field. Lett Appl Microbiol 40:146–151
Bergström L, Börjesson E, Stenström J (2011) Laboratory and lysimeter studies of glyphosate and aminomethylphosphonic acid in a sand and a clay soil. J Environ Qual 40:98–108
Bonnet JL, Bonnemoy F, Dusser M, Bohatier J (2007) Assessment of the potential toxicity of herbicides and their degradation products to nontarget cells using two microorganisms, the bacteria Vibrio fischeri and the ciliate Tetrahymena pyriformis. Environ Toxicol 22:78–91
Busse MD, Ratcliff AW, Shestak CJ, Powers RF (2001) Glyphosate toxicity and the effects of long-term vegetation control on soil microbial communities. Soil Biol Biochem 33:1777–1789
Candela L, Álvarez-Benedí J, De Melo MC, Rao P (2007) Laboratory studies on glyphosate transport in soils of the Maresme area near Barcelona, Spain: transport model parameter estimation. Geoderma 140:8–16
Chang EH, Chiu CY (2015) Changes in soil microbial community structure and activity in a cedar plantation invaded by moso bamboo. Appl Soil Ecol 91:1–7
Chen JH, Liu XY, Zheng JW, Zhang B, Lu HF, Chi ZZ, Pan GX, Li LQ, Zheng JF, Zhang XH (2013) Biochar soil amendment increased bacterial but decreased fungal gene abundance with shifts in community structure in a slightly acid rice paddy from Southwest China. Appl Soil Ecol 71:33–44
Cobos-Vasconcelos DDL, Santoyo-Tepole F, Juárez-Ramírez C, Ruiz-Ordaz N, Galíndez-Mayer CJJ (2006) Cometabolic degradation of chlorophenols by a strain of Burkholderia in fed-batch culture. Enzyme Microb Technol 40:57–60
Dahlin S, Witter E (1998) Can the low microbial biomass C-to-organic C ratio in an acid and a metal contaminated soil be explained by differences in the substrate utilization efficiency and maintenance requirements? Soil Biol Biochem 30:633–641
Duke SO (2017) The history and current status of glyphosate. Pest Manag Sci. https://doi.org/10.1002/ps.4652
Duke SO, Powles SB (2008) Glyphosate: a once-in-a-century herbicide. Pest Manag Sci 64:319–325
Duke SO, Lydon J, Koskinen WC, Moorman TB, Chaney RL, Hammerschmidt R (2012) Glyphosate effects on plant mineral nutrition, crop rhizosphere microbiota, and plant disease in glyphosate-resistant crops. J Agr Food Chem 60:10375–10397
Edvantoro BB, Naidu R, Megharaj M, Singleton I (2003) Changes in microbial properties associated with long-term arsenic and DDT contaminated soils at disused cattle dip sites. Ecotox Environ Safe 55:344–351
Frostegård A, Bååth E (1996) The use of phospholipid fatty acid analysis to estimate bacterial and fungal biomass in soil. Biol Fert Soils 22:59–65
Frostegård Å, Bååth E, Tunlio A (1993) Shifts in the structure of soil microbial communities in limed forests as revealed by phospholipid fatty acid analysis. Soil Biol Biochem 25:723–730
Garland JL, Mills AL (1991) Classification and characterization of heterotrophic microbial communities on the basis of patterns of community-level sole-carbon-source utilization. Appl Environ Microbiol 57:2351
Haney R, Senseman S, Krutz L, Hons F (2002) Soil carbon and nitrogen mineralization as affected by atrazine and glyphosate. Biol Fert Soils 35:35–40
Hosseini Bai S, Xu Z, Blumfield TJ, Wild CH, Chen C (2014) Soil carbon and nitrogen dynamics in the first year following herbicide and scalping in a revegetation trial in south-east Queensland, Australia. Environ Sci Pollut Res 21:5167–5176
Iker M, Josem B, Isabel A, Lur E, Carlos G (2009) Effects of glyphosate on rhizosphere soil microbial communities under two different plant compositions by cultivation-dependent and -independent methodologies. Soil Biol Biochem 41:505–513
Imfeld G, Vuilleumier S (2012) Measuring the effects of pesticides on bacterial communities in soil: a critical review. Eur J Soil Biol 49:22–30
Jacob GS, Garbow JR, Hallas LE, Kimack NM, Kishore GM, Schaefer J (1988) Metabolism of glyphosate in Pseudomonas sp. strain LBr. Appl Environ Microbiol 54:2953–2958
Jacobsen CS, Hjelmsø MH (2014) Agricultural soils, pesticides and microbial diversity. Curr Opin Biotech 27:15–20
Lancaster SH, Hollister EB, Senseman SA, Gentry TJ (2010) Effects of repeated glyphosate applications on soil microbial community composition and the mineralization of glyphosate. Pest Manag Sci 66:59–64
Lapygina EV, Lysak LV, Zviagintsev DG (2002) Tolerance of soil bacterial complexes to salt shock. Microbiology 71:143–147
Lee SD (2007) Devosia subaequoris sp. nov., isolated from beach sediment. Int J Syst Evol Microbiol 57:2212–2215
Li YF, Zhang JJ, Chang SX, Jiang PK, Zhou GM, Fu SL, Yan ER, Wu JS, Lin L (2013) Long-term management effects on soil organic carbon pools and chemical composition in Moso bamboo (Phyllostachys pubescens) forests in subtropical China. Forest Ecol Manag 303:121–130
Li YC, Li YF, Chang SX, Xu QF, Guo ZY, Gao Q, Qin ZY, Yang YF, Chen JH, Liang X (2017a) Bamboo invasion of broadleaf forests altered soil fungal community closely linked to changes in soil organic C chemical composition and mineral N production. Plant Soil 418:507–521
Li YC, Li YF, Chang SX, Liang X, Qin H, Chen JH, Xu QF (2017b) Linking soil fungal community structure and function to soil organic carbon chemical composition in intensively managed subtropical bamboo forests. Soil Biol Biochem 107:19–31
Li YF, Hu SD, Chen JH, Müller K, Li YC, Fu WJ, Lin ZW, Wang HL (2018) Effects of biochar application in forest ecosystems on soil properties and greenhouse gas emissions: a review. J Soils Sediments 18:546–563
Liang C, Fujinuma R, Balser TC (2008) Comparing PLFA and amino sugars for microbial analysis in an Upper Michigan old growth forest. Soil Biol Biochem 40:2063–2065
Margesin R, Jud M, Tscherko D, Schinner F (2009) Microbial communities and activities in alpine and subalpine soils. FEMS Microbiol Ecol 67:208–218
Mcauliffe KS, Hallas LE, Kulpa CF (1990) Glyphosate degradation by Agrobacterium radiobacter isolated from activated sludge. J Ind Microbiol Biotechnol 6:219–221
Newman MM, Hoilett N, Lorenz N, Dick RP, Liles MR, Ramsier C, Kloepper JW (2016a) Glyphosate effects on soil rhizosphere-associated bacterial communities. Sci Total Environ 543:155–160
Newman MM, Lorenz N, Hoilett N, Lee NR, Dick RP, Liles MR, Ramsier C, Kloepper JW (2016b) Changes in rhizosphere bacterial gene expression following glyphosate treatment. Sci Total Environ 553:32–41
Nguyen DB, Rose MT, Rose TJ, Morris SG, van Zwieten L (2016) Impact of glyphosate on soil microbial biomass and respiration: a meta-analysis. Soil Biol Biochem 92:50–57
Pizarro H, Vera M, Vinocur A, Pérez G, Ferraro M, Helman RM, dos Santos Afonso M (2016) Glyphosate input modifies microbial community structure in clear and turbid freshwater systems. Environ Sci Pollut Res 23:5143–5153
Rick WY, Thomas SM (2001) Microbial nitrogen cycles: physiology, genomics and applications. Curr Opin Microbiol 4:307–312
Shehata AA, Schrödl W, Aldin AA, Hafez HM, Krüger M (2013) The effect of glyphosate on potential pathogens and beneficial members of poultry microbiota in vitro. Curr Microbiol 66:350–358
Stratton GW, Stewart KE (1992) Glyphosate effects on microbial biomass in a coniferous forest soil. Environ Toxicol 7:223–236
Sun YC, Chen YC, Tian ZX, Li FM, Wang XY, Zhang J, Xiao ZL, Lin M, Gilmartin N, Dowling DN (2005) Novel AroA with high tolerance to glyphosate, encoded by a gene of Pseudomonas putida 4G-1 isolated from an extremely polluted environment in China. Appl Environ Microbiol 71:4771–4776
Tian YS, Xiong AS, Xu J, Zhao W, Gao F, Fu XY, Xu H, Zheng JL, Peng RH, Yao QH (2010) Isolation from Ochrobactrum anthropi of a novel class II 5-enopyruvylshikimate-3-phosphate synthase with high tolerance to glyphosate. Appl Environ Microbiol 76:6001–6005
Toledohernández C, Bonesgonzález A, Ortizvázquez OE, Sabat AM, Bayman P (2007) Fungi in the sea fan Gorgonia ventalina: diversity and sampling strategies. Coral Reefs 26:725–730
Tong Z, Bischoff M, Nies L, Applegate B, Turco RF (2007) Impact of fullerene (C60) on a soil microbial community. Environ Sci Technol 41:2985–2991
Vance ED, Brooks PC, Jenkinson DS (1987) An extraction method for measuring soil microbial biomass. Soil Biol Biochem 19:703–707
Vazquez P, Holguin G, Puente ME, Lopezcortez A, Bashan Y (2000) Phosphate-solubilizing microorganisms associated with the rhizosphere of mangroves in a semiarid coastal lagoon. Biol Fert Soils 30:460–468
Wang MC, Gong M, Zang HB, Hua XM, Yao J, Pang YJ, Yang YH (2006) Effect of methamidophos and urea application on microbial communities in soils as determined by microbial biomass and community level physiological profiles. J Environ Sci Health B 41:399–413
Wang MC, Liu YH, Wang Q, Gong M, Hua XM, Pang YJ, Hu S, Yang YH (2008) Impacts of methamidophos on the biochemical, catabolic, and genetic characteristics of soil microbial communities. Soil Biol Biochem 40:778–788
Weaver MA, Krutz LJ, Zablotowicz RM, Reddy KN (2007) Effects of glyphosate on soil microbial communities and its Mississippi soil. Pest Manag Sci 63:388–393
White DC, Findlay RH (1988) Biochemical markers for measurement of predation effects on the biomass, community structure, nutritional status, and metabolic activity of microbial biofilms. Hydrobiologia 159:119–132
Wu VC, Fung DY, Kang DH, Thompson LK (2001) Evaluation of thin agar layer method for recovery of acid-injured foodborne pathogens. J Food Prot 64:1067–1071
Zhang CP, Hu XQ, Luo JY, Wu ZY, Wang L, Li B, Wang YL, Sun GC (2015) Degradation dynamics of glyphosate in different types of citrus orchard soils in China. Molecules 20:1161–1175
Zhou JZ, Bruns MA, Tiedje JM (1996) DNA recovery from soils of diverse composition. Appl Environ Microbiol 62:316–322
Zhou M, Xu HL, Wei XL, Ye ZQ, Wei LP, Gong WM, Wang YQ, Zhu Z (2006) Identification of a glyphosate-resistant mutant of rice 5-enolpyruvylshikimate 3-phosphate synthase using a directed evolution strategy. Plant Physiol 140:184–195
Funding
This work was financially supported by the Program for Changjiang Scholars and Innovative Research Team in University (IRT_14R27), the National Natural Science Foundation of China (NSFC) (31372140, 30971871, 40371071), and the Fundamental Research Funds for the Central Universities (No. 020814380002).
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Liu, Y., Li, Y., Hua, X. et al. Glyphosate application increased catabolic activity of gram-negative bacteria but impaired soil fungal community. Environ Sci Pollut Res 25, 14762–14772 (2018). https://doi.org/10.1007/s11356-018-1676-0
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DOI: https://doi.org/10.1007/s11356-018-1676-0
Keywords
- Soil microbial community
- PLFA
- Biolog
- DGGE
- Glyphosate