Transgenic Research

, Volume 22, Issue 5, pp 877–892 | Cite as

Impact of the ahas transgene and of herbicides associated with the soybean crop on soil microbial communities

  • Rosinei Aparecida Souza
  • Letícia Carlos Babujia
  • Adriana Pereira Silva
  • Maria de Fátima Guimarães
  • Carlos Arrabal Arias
  • Mariangela Hungria
Original Paper


Although Brazil has recently reached the position as the second largest producer of genetically modified soybean [Glycine max (L.) Merr.], there are few reports on the effects of transgenic crops and the associated use of specific herbicides on soil microbial communities, both under the edaphoclimatic conditions in Brazil, and in other producer regions in the southern hemisphere. The aim of this study was to evaluate the effects of transgenic soybean containing the ahas gene conferring resistance to herbicides of the imidazolinone group, and of the herbicides associated with transgenic soybeans on the soil microbial community. Twenty field experiments were carried out during three growing seasons (summer of 2006/2007, short-season of 2007 and summer of 2007/2008), in nine municipalities located in six Brazilian states and in the Federal District. The experiments were conducted using a completely randomized block design with four replicates and three treatments: (1) conventional (non-transgenic) soybean cultivar Conquista with conventional herbicides (bentazone + acifluorfen-sodium and other herbicides, depending on the level of infestation in each region); (2) near-isogenic transgenic Cultivance (CV127) containing the ahas gene, with conventional herbicides; (3) transgenic Cultivance with specific herbicide of the imidazolinone group (imazapyr). As the objective of the study was to verify impacts of the transgene and herbicides on the soil microbial community of the whole area and not only a punctual rhizospheric effects, samples were taken at the 0–10 cm layer prior to cropping and at R2 soybean growth stage, between plant rows. Quantitative (microbial biomass C and N, MB-C and MB-N) and qualitative (DGGE of the 16S rDNA region) parameters of soil microbial community were evaluated. No qualitative or quantitative differences were found that could be attributed to the transgene ahas. A comparison of Cultivance soybean with conventional and imidazolinone-group herbicides applications also failed to reveal differences that could be attributed to the specific use of imazapyr, even after three consecutive croppings at the same site. Finally, no differences were detected between conventional (Conquista and conventional herbicides) and transgenic soybean managements (Cultivance and imazapyr). However, marked differences were observed in MB-C and MB-N between the different sites and times of year and, for the 16S rDNA-DGGE profiles, between different sites. In conclusion, microbial community evaluations were found to be sensitive and viable for monitoring different technologies and agricultural management methods, but no differences could be attributed to the ahas transgene for three consecutive cropping seasons.


Soil microbial biomass DGGE Soil microbial diversity Glycine max Imidazolinones Environmental monitoring Transgenic soybean 

Supplementary material

11248_2013_9691_MOESM1_ESM.docx (236 kb)
Supplementary material 1 (DOCX 236 kb)


  1. Andrade DS, Hamakawa PJ (1994) Estimativa do número de células viáveis de rizóbio no solo e em inoculantes por infecção em plantas. In: Hungria M, Araujo RS (eds) Manual de métodos empregados em estudos de microbiologia agrícola. EMBRAPA-SPI, Brasília, Brazil, pp 63–94. Available at
  2. Aragão FJL, Sarokin L, Vianna GR, Rech EL (2000) Selection of transgenic meristematic cells utilizing a herbicidal molecule results in the recovery of fertile transgenic soybean [Glycine max (L.) Merril] plants at a high frequency. Theor Appl Genet 101:1–6CrossRefGoogle Scholar
  3. Araújo ASF, Monteiro RTR, Abarkeli RB (2003) Effect of glyphosate on the microbial activity of two Brazilian soils. Chemosphere 52:799–804PubMedCrossRefGoogle Scholar
  4. Ávila LA (2007) Efeitos do algodão Bt (Bollgard Evento 531) na comunidade bacteriana da rizosfera. MSc. thesis, Universidade de São Paulo, São Paulo, BrazilGoogle Scholar
  5. Balota EL, Colozzi-Filho A, Andrade DS, Hungria M (1998) Biomassa microbiana e sua atividade em solos sob diferentes sistemas de preparo e sucessão de culturas. R Bras Ci Solo 22:641–649Google Scholar
  6. Balota EL, Colozzi-Filho A, Andrade DS, Dick RP (2003) Microbial biomass in soils under different tillage and crop rotation systems. Biol Fertil Soils 38:15–20CrossRefGoogle Scholar
  7. Balota EL, Colozzi-Filho A, Andrade DS, Dick RP (2004) Long-term tillage and crop rotation effects on microbial biomass and C and N mineralization in a Brazilian Oxisol. Soil Till Res 77:137–145CrossRefGoogle Scholar
  8. Bartlett RJ, Ross DN (1998) Colorimetric determination of oxidizable carbon in acid soil solutions. Soil Sci Soc Am J 52:1191–1192CrossRefGoogle Scholar
  9. Bending GD, Turner MK, Rayns F, Marx MC, Wood M (2004) Microbial and biochemical soil quality indicators and their potential for differentiating areas under contrasting agricultural management regimes. Soil Biol Biochem 36:1785–1792CrossRefGoogle Scholar
  10. Bohm GMB, Rombaldi CV (2010) Transformação genética e aplicação de glifosato na microbiota do solo, fixação biológica de nitrogênio, qualidade e segurança de grãos de soja geneticamente modificada. Ci Rural 40:213–221CrossRefGoogle Scholar
  11. Bremner JM (1965) Total nitrogen. In: Black CA (ed) Methods of soil analysis. American Society of Agronomy, Madison, pp 1149–1178Google Scholar
  12. Brighenti AM, Adegas FS, Bortoluzi ES, Almeida LA, Voll E (2002) Tolerância de genótipos de soja aos herbicidas trifluralin e imazaquin. Planta Daninha 20:63–69CrossRefGoogle Scholar
  13. Brookes PC, Kragt JF, Powlson DS, Jenkinson DS (1985a) Chloroform fumigation and the release of soil nitrogen. Soil Biol Biochem 17:831–835CrossRefGoogle Scholar
  14. Brookes PC, Kragt JF, Powlson DS, Jenkinson DS (1985b) Chloroform fumigation and the release of soil nitrogen: a rapid direct extraction method to measure microbial biomass nitrogen in soil. Soil Biol Biochem 17:837–847CrossRefGoogle Scholar
  15. Bruinsma M, Kowalchuk GA, van Veen JA (2003) Effects of genetically modified plants on microbial communities and processes in soil. Biol Fertil Soils 37:329–337Google Scholar
  16. Busse MD, Ratcliff GA, Shestak CJ, Powers RF (2001) Glyphosate toxicity and the effects of long-term vegetation control and soil on soil microbial communities. Soil Biol Biochem 33:1777–1789CrossRefGoogle Scholar
  17. CTNBIO (Comissão Técnica Nacional de Biossegurança) (1998) Comunicado n.º54, de 29 de setembro de 1998. Accessed 6 Feb 2012
  18. Devare MH, Jones CM, Thies JE (2004) Effect of Cry3Bb transgenic corn and tefluthrin on the soil microbial community: biomass, activity and diversity. J Environ Qual 33:837–843PubMedCrossRefGoogle Scholar
  19. Donegan KK, Palm CJ, Fieland VJ, Porteus LA, Ganio LM, Schaller DL, Bucao LQ, Seidler RJ (1995) Changes in levels, species and DNA fingerprints of soil microorganisms associated with cotton expressing the Bacillus thuringiengis var. kurstaki endotoxin. Appl Soil Ecol 2:111–124CrossRefGoogle Scholar
  20. Dunfield KE, Germida JJ (2001) Diversity of bacterial communities in the rhizosphere and root interior of field-grown genetically modified Brassica napus. FEMS Microbiol Ecol 38:1–9CrossRefGoogle Scholar
  21. Dunfield KE, Germida JJ (2003) Seasonal changes in the rhizosphere microbial communities associated with field-grown genetically modified canola (Brassica napus). Appl Environ Microbiol 69:7310–7318PubMedCrossRefGoogle Scholar
  22. Dunfield KE, Germida JJ (2004) Impact of genetically modified crops on soil- and plant-associated microbial comunities. J Environ Qual 38:806–815CrossRefGoogle Scholar
  23. Fang M, Kremer RJ, Motavalli PP, Davis G (2005) Bacterial diversity in rhizosphere of nontransgenic and transgenic corn. Appl Environ Microbiol 71:4132–4136PubMedCrossRefGoogle Scholar
  24. Fehr WR, Caviness CE (1977) Stages of soybean development. Agriculture and Home Economics Experiment Station, Iowa State University of Science and Technology, Ames, USA. (Special Report 80)Google Scholar
  25. Feigl F, Anger V (1972) Spot-test in inorganic analysis, 6th edn. Elsevier, AmsterdamGoogle Scholar
  26. Flint JL, Witt WW (1997) Microbial degradation of imazaquin and imazethapyr. Weed Sci 45:586–591Google Scholar
  27. Flores S, Saxena D, Stotzky G (2005) Transgenic Bt plants decompose less in soil than non-Bt plants. Soil Biol Biochem 37:1073–1082CrossRefGoogle Scholar
  28. Franchini JC, Crispino CC, Souza RA, Torres E, Hungria M (2007) Microbiological parameters as indicators of soil quality under various soil management and crop rotation systems in southern Brazil. Soil Till Res 92:18–29CrossRefGoogle Scholar
  29. Gomes NCM, Heuer H, Schonfeld J, Costa R, Mendonça-Hagler L, Smalla K (2001) Bacterial diversity of the rhizosphere of maize (Zea mays) grown in tropical soil studied by temperature gradient gel electrophoresis. Plant Soil 232:167–180CrossRefGoogle Scholar
  30. Haney RL, Senseman SA, Hons FM (2002) Bioremediation and biodegradation: effect of roundup ultra on microbial activity and biomass from selected soils. J Environ Qual 31:730–735PubMedCrossRefGoogle Scholar
  31. Heuer H, Kroppensted TRM, Lottmann J, Berg G, Smalla K (2002) Effects of T4 lysozyme release from transgenic potato roots on bacterial rhizosphere communities are negligible relative to natural factors. Appl Environ Microbiol 68:1325–1335PubMedCrossRefGoogle Scholar
  32. Hungria M, Campo RJ, Mendes IC (2003) Benefits of inoculation of common bean (Phaseolus vulgaris) crop with efficient and competitive Rhizobium tropici strains. Biol Fertil Soils 39:88–93CrossRefGoogle Scholar
  33. Hungria M, Franchini JC, Brandão-Junior O, Kaschuk G, Souza RA (2009) Soil microbial activity and crop sustainability in a long-term experiment with three soil-tillage and two crop-rotation systems. Appl Soil Ecol 42:288–296CrossRefGoogle Scholar
  34. Jaccard P (1912) The distribution of flora in the alpine zone. New Phytol 11:37–50CrossRefGoogle Scholar
  35. James C (2010) Global status of commercialized Biotech/GM crops: 2010. ISAAA, Ithaca, p 16Google Scholar
  36. Jenkinson DS, Powlson DS (1976) The effects of biocide treatment on metabolism in soil. V A method for measuring soil biomass. Soil Biol Biochem 8:209–213CrossRefGoogle Scholar
  37. Kaschuk G, Alberton O, Hungria M (2010) Three decades of soil microbial biomass studies in Brazilian ecosystems: lessons learned about soil quality and indications for improving sustainability. Soil Biol Biochem 42:1–13CrossRefGoogle Scholar
  38. Kaschuk G, Alberton O, Hungria M (2011) Quantifying effects of different agricultural land uses on soil microbial biomass and activity in Brazilian biomes: inferences to improve soil quality. Plant Soil 338:467–481CrossRefGoogle Scholar
  39. Kennedy AC (1999) Bacterial diversity in agroecosystems. Agric Ecosyst Environ 74:65–76CrossRefGoogle Scholar
  40. Kinney CA, Mandernack KW, Mosier AR (2005) Laboratory investigations into the effects of the pesticides mancozeb, chlorothalonil, and prosulfuron on nitrous oxide and nitric oxide production in fertilized soil. Soil Biol Biochem 7:837–850CrossRefGoogle Scholar
  41. Knupp AM, Martins CM, Faria JC, Rumjanek NG, Xavier GR (2009) Comunidade bacteriana como indicadora do efeito de feijoeiro geneticamente modificado sobre organismos não alvo. Pesq Agropec Bras 44:1692–1699CrossRefGoogle Scholar
  42. Koranda M, Schnecker J, Kaiser C, Fuchslueger L, Kitzler B, Stange CF, Sessitsch A, Zechmeister-Boltenstern S, Richter A (2011) Microbial processes and community composition in the rhizosphere of European beech—the influence of plant C exudates. Soil Biol Biochem 43:551–558PubMedCrossRefGoogle Scholar
  43. Kraemer AF, Marchesan E, Avila LA, Machado SLO, Grohs M (2009) Destino ambiental dos herbicidas do grupo das imidazolinonas—revisão. Planta Daninha 27:629–639CrossRefGoogle Scholar
  44. Kremer RJ, Monteiro RTR, Tornisielo VL (2005) Glyphosate affects soybean root exudation and rhizosphere microorganisms. Int J Environ Anal Chem 85:1165–1174CrossRefGoogle Scholar
  45. Li Q, Allen HL, Wollum AG II (2004) Microbial biomass and bacterial functional diversity in forest soils: effects of organic matter removal, compaction, and vegetation control. Soil Biol Biochem 36:571–579CrossRefGoogle Scholar
  46. Liphadzi KB, Al-Khatib K, Bensch CN, Stahlman PW, Dille JA, Todd T, Rice CW, Horak MJ, Head G (2005) Soil microbial and nematode communities as affected by glyphosate and tillage practices in a glyphosate-resistant cropping system. Weed Sci 53:536–545CrossRefGoogle Scholar
  47. Lottmann J, O’Callaghan M, Baird D, Walter C (2010) Bacterial fungal communities in the rhizosphere of field grown genetically modified pine trees (Pinus radiate D). Environ Biosafety Res 9:25–40PubMedCrossRefGoogle Scholar
  48. Mendes IC, Souza LV, Resck DVS, Gomes AC (2003) Propriedades biológicas em agregados de um LE sob plantio convencional e direto no cerrado. R Bras Ci Solo 27:435–443Google Scholar
  49. Milling A, Smalla K, Maidl FX, Schloter M, Munch JC (2004) Effects of transgenic potatoes with an altered starch composition on the diversity of soil and rhizosphere bacteria and fungi. Plant Soil 266:23–39CrossRefGoogle Scholar
  50. Nūbel U, Engelen B, Felsk A, Snaidr J, Wieshuber A, Amann RI, Ludwig W, Backhaus H (1996) Sequence heterogeneities of genes encoding 16S rRNAs in Paenibacillus polymyxa detected by temperature gradient gel electrophoresis. J Bacteriol 178:5636–5643PubMedGoogle Scholar
  51. OECD (Organisation for Economic Co-operation and Development) (2010) Safety assessment of transgenic organisms. OECD Publishing, OECD. (OECD Consensus Documents, 4)Google Scholar
  52. Pereira JL, Picanço MC, Silva AA, Santos EA, Tomé HVV, Olarte JB (2008) Effects of glyphosate and endosulfan on soil microorganisms in soybean crop. Planta Daninha 26:825–830CrossRefGoogle Scholar
  53. Petter FA, Cargnelutti Filho A, Barroso ALL, Pacheco LP (2007) Manejo de herbicidas na cultura da soja Roundup Ready. Planta Daninha 25:557–566CrossRefGoogle Scholar
  54. Rech EL, Vianna GR, Aragão FJ (2008) High-efficiency transformation by biolistics of soybean, common bean and cotton transgenic plantas. Nature Prot 3:410–418CrossRefGoogle Scholar
  55. Santos JB, Jakelaitis A, Silva AA, Vivian R, Costa MD, Silva AF (2005) Atividade microbiana do solo após aplicação de herbicidas em sistemas de plantio direto e convencional. Planta Daninha 23:683–691CrossRefGoogle Scholar
  56. SAS (SAS Institute Inc) (1999) Proprietary of software, version 6, 4th edn. SAS Institute, CaryGoogle Scholar
  57. Shaner DL, Singh BK (1993) Phytotoxicity of acetohydroxyacid synthase inhibitors is not due to accumulation of 2-ketobutyrate and/or 2-aminobutyrate. Plant Physiol 103:1221–1226PubMedGoogle Scholar
  58. Shen RF, Cai H, Gong WH (2006) Transgenic Bt cotton has no apparent effect on enzymatic activities or functional diversity of microbial communities in rhizosphere soil. Plant Soil 285:149–159CrossRefGoogle Scholar
  59. Silva AP, Franchini JC, Babujia LC, Souza RA, Hungria M (2010) Microbial biomass under different soil and crop managements in short- to long-term experiments performed In Brazil. Field Crops Res 119:20–26Google Scholar
  60. Sneath PHA, Sokal RR (1973) Numerical taxonomy. Freeman, San Francisco, p 573Google Scholar
  61. Souza RA, Hungria M, Franchini JC, Chueire LMO, Barcellos FG, Campo RJ (2008a) Avaliação qualitativa e quantitativa da microbiota do solo e da fixação biológica do nitrogênio pela soja. Pesq Agropec Bras 43:71–82Google Scholar
  62. Souza RA, Hungria M, Franchini JC, Maciel CD, Campo RJ, Zaia DAM (2008b) Conjunto mínimo de parâmetros para avaliação da microbiota do solo e da fixação biológica do nitrogênio pela soja. Pesq Agropec Bras 43:83–91CrossRefGoogle Scholar
  63. Tan S (2006) Herbicidal inhibitors of amino acid biosynthesis and herbicide-tolerant crops. Amino Acids 30:195–204PubMedCrossRefGoogle Scholar
  64. Topp E, Vallaeys T, Soulas G (1997) Pesticides: microbial degradation and effects on microorganisms. In: van Elsas JD, Trevors JT, Wellington EMH (eds) Modern soil microbiology. Marcel Dekker, New York, pp 547–575Google Scholar
  65. Tuffi Santos LD, Ferreira FA, Barros NF, Siqueira CH, Santos IC, Machado AFL (2005) Exsudação radicular do glyphosate por Brachiaria decumbens e seus efeitos em plantas de eucalipto e na respiração microbiana do solo. Planta Daninha 23:143–152CrossRefGoogle Scholar
  66. Vance ED, Brookes PC, Jenkinson DS (1987) An extraction method for measuring soil microbial biomass C. Soil Biol Biochem 19:703–707CrossRefGoogle Scholar
  67. Wallis PD, Haynes RJ, Hunter CH, Morris CD (2010) Effect of land use and management on soil bacterial biodiversity as measured by PCR-DGGE. Appl Soil Ecol 46:147–150CrossRefGoogle Scholar
  68. Wang Q, Zhou D, Cang L (2009) Microbial and enzyme properties of apple orchard soil as affected by long-term application of copper fungicide. Soil Biol Biochem 41:1504–1509CrossRefGoogle Scholar
  69. Wardle DA (1992) A comparative assessment of factors which influence microbial biomass carbon and nitrogen levels in soil. Biol Rev 67:321–358CrossRefGoogle Scholar
  70. Wardle DA (1995) Impacts of disturbance on detritus food webs in agro-ecosystems of contrasting tillage and weed management practices. In: Begon M, Fitter AH (eds) Advances in ecological research. Academic Press, London, pp 105–185Google Scholar
  71. Wei XD, Zou HL, Chu LM, Liao CM, Lan CY (2006) Field released transgenic papaya affects microbial communities and enzyme activities in soil. Plant Soil 285:347–358CrossRefGoogle Scholar
  72. Wei L, Hao HL, Weixiang W, Qi KW, Ying XC, Janice ET (2008) Transgenic Bt rice does not affect enzyme activities and microbial composition in the rhizosphere during crop development. Soil Biol Biochem 40:475–486CrossRefGoogle Scholar
  73. Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703PubMedGoogle Scholar
  74. Zilli JE, Smiderle OJ, Neves MCP, Rumjanek NG (2007) População microbiana em solo cultivado com soja e tratado com diferentes herbicidas em área de cerrado no estado de Roraima. Acta Amaz 37:201–212CrossRefGoogle Scholar
  75. Zilli JE, Botelho GR, Neves MCP, Rumjanek NG (2008) Efeito de glyphosate e imazaquin na comunidade bacteriana do rizoplano de soja (Glycine max (L.) Merrill) e em características microbiológicas do solo. R Bras Ci Solo 32:633–642CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Rosinei Aparecida Souza
    • 1
    • 2
  • Letícia Carlos Babujia
    • 1
    • 3
  • Adriana Pereira Silva
    • 1
    • 2
  • Maria de Fátima Guimarães
    • 1
    • 2
  • Carlos Arrabal Arias
    • 1
  • Mariangela Hungria
    • 1
  1. 1.Embrapa SojaLondrinaBrazil
  2. 2.Department of AgronomyUniversidade Estadual de LondrinaLondrinaBrazil
  3. 3.Department of ChemistryUniversidade Estadual de MaringáMaringáBrazil

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