Hostname: page-component-cd9895bd7-fscjk Total loading time: 0 Render date: 2024-12-27T19:51:11.588Z Has data issue: false hasContentIssue false
  • English
  • Français

Bacterial and fungal communities in the rhizosphere of field-grown genetically modified pine trees (Pinus radiata D.)

Published online by Cambridge University Press:  11 November 2010

Jana Lottmann ,
Maureen O’Callaghan ,
David Baird  and
Christian Walter
Jana Lottmann*
Affiliation:
AgResearch Limited, Biocontrol and Biosecurity Section, Private Bag 4749, Christchurch 8140, New Zealand
Maureen O’Callaghan
Affiliation:
AgResearch Limited, Biocontrol and Biosecurity Section, Private Bag 4749, Christchurch 8140, New Zealand
David Baird
Affiliation:
VSN (NZ) Limited, 40 McMahon Drive, Christchurch, New Zealand
Christian Walter
Affiliation:
Scion – Next Generation Biomaterials, Te Papa Tipu Innovation Park, Private Bag 3020, Rotorua, New Zealand
*
* Corresponding author: [email protected]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

This study assessed the impact of Pinus radiata (D. Don) genetically modified (GM) by biolistic insertion of the LEAFY and nptII genes on rhizosphere microbial communities of field grown trees. Rhizosphere soil was sampled quarterly for two consecutive years. A culture-independent approach was used to characterise the microbial communities based on PCR and denaturing gradient gel electrophoresis (DGGE) of 16S/18S rDNA gene fragments, and internal transcribed spacer (ITS) fragments amplified from total rhizosphere DNA. Trees from two independent transformation events were sampled, together with non-modified control trees of the same parental genotype. DGGE profiles of rhizosphere general Bacteria did not differ between GM and control trees with one exception (summer 2006 sample). For Alphaproteo- and Actinobacteria, significant differences between treatments were detected in one out of eight samplings. Small seasonal shifts could be detected in all bacterial communities. General fungal and ectomycorrhizal communities did not differ significantly between GM and control trees with the exception of summer 2006, when ectomycorrhizal communities associated with GM trees from one transformation event differed from those associated with control trees. Small seasonal shifts of general fungal and ectomycorrhizal communities were seen over the two-year sampling period. More detailed analysis of microbial communities at one sampling date (using amplified rDNA restriction analysis (ARDRA) and 16S/18S rDNA sequencing) revealed significant differences in four ARDRA groups between one GM treatment and the control (bacteria), and significant differences in one ARDRA group between the two GM treatments (fungi). When data from all sampling dates are considered together, the low incidence of statistical differences in the microbial communities associated with the genetically modified and control trees suggests that there was no significant impact of this genetic modification on rhizosphere microbial communities.

Keywords

Bacterial and fungal communitiesectomycorrhizal fungiDGGEARDRAdiversitygenetic engineering (GE)transgenic treesbiosafety
Type
Research Article
Information
Environmental Biosafety Research , Volume 9 , Issue 1 , January 2010 , pp. 25 - 40
Copyright
© ISBR, EDP Sciences, 2010

References

Andreote, FD, Mendes, R, Dini-Andreote, F, Rossetto, PB, Labate, CA, Pizzirani-Kleiner, AA, van Elsas, JD, Azevedo, JL, Araújo, WL (2008) Transgenic tobacco revealing altered bacterial diversity in the rhizosphere during early plant development. Antonie Van Leeuwenhoek 93: 415–424 Google ScholarPubMed
Andreote, FD, Carneiro, RT, Salles, JF, Marcon, J, Labate, CA, Azevedo, JL, Araújo, WL (2009) Culture-independent assessment of Rhizobiales-related Alphaproteobacteria and the diversity of Methylobacterium in the rhizosphere and rhizoplane of transgenic eucalyptus. Microbial Ecol . 57: 82–93 CrossRefGoogle ScholarPubMed
Batista, R, Saibo, N, Lourenco, T, Oliveira, MM (2008) Microarray analyses reveal that plant mutagenesis may induce more transcriptome changes than transgene insertion. PNAS 105: 3640–3645 Google Scholar
Bishop-Hurley, S, Zabkiewicz, RJ, Grace, LJ, Gardner, RC, Wagner, A, Walter, C (2001) Conifer genetic engineering: transgenic Pinus radiata (D. Don) and Picea abies (Karst) plants are resistant to the herbicide Buster. Plant Cell Reports 20: 235–243 Google Scholar
Chow, ML, Radomski, CC, McDermott, JM, Davies, J, Axelrood, PE (2002) Molecular characterization of bacterial diversity in Lodgepole pine (Pinus contorta) rhizosphere soils from British Columbia forest soils differing in disturbance and geographic source. FEMS Microb. Ecol. 42: 347–357 Google ScholarPubMed
Chu-Chou, M, Grace, LJ (1988) Mycorrhizal fungi of Radiata pine in different forests of the North and South Islands in New Zealand. Soil Biol. Biochem . 20: 883–886 CrossRefGoogle Scholar
Costa, R, Götz, M, Mrotzek, N, Lottmann, J, Berg, G, Smalla, K (2005) Effects of site and plant species on rhizosphere community structure as revealed by molecular analysis of microbial guilds. FEMS Microbiol. Ecol. 56: 236–249 CrossRefGoogle ScholarPubMed
Costa, R, Gomes, NCM, Krögerrecklenfort, E, Opelt, K, Berg, G, Smalla, K (2007) Pseudomonas community structure and antagonistic potential in the rhizosphere: insights gained by combining phylogenetic and functional gene-based analyses. Environ. Microbiol. 9: 2260–2273 CrossRefGoogle Scholar
Dunfield, KE, Germida, JJ, (2001) Diversity of bacterial communities in the rhizosphere and root interior of field-grown genetically modified Brassica napus . FEMS Microb. Ecol. 82: 1–9 CrossRefGoogle Scholar
Dunfield, KE, Germida, JJ (2004) Impact of genetically modified crops on soil and plant associated microbial communities. Environ. Quality 33: 806–815 Google ScholarPubMed
Filion, M, Hamelin, RC, Bernier, L, St-Arnaud, M (2004) Molecular profiling of rhizosphere microbial communities associated with healthy and diseased black spruce (Picea mariana) seedlings grown in a nursery. Appl. Environ. Microbiol. 70: 3541–3551 CrossRefGoogle Scholar
Gardes, M, Bruns, TD (1993) ITS primers with enhanced specificity for basidiomycetes – application to the identification of mycorrhizae and rusts. Mol. Ecol. 2: 113–118 Google Scholar
Glasbey, CA, Vali, L, Gustafsson, JS (2005) A statistical model for unwarping of 1-D electrophoresis gels. Electrophoresis 26: 4237–4242 Google ScholarPubMed
Gomes, NCM, Heuer, H, Schönfeld, J, Costa, R, Hagler-Mendonca, 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: 167–180 Google Scholar
Gower JC, (1985) Measures of similarity, dissimilarity and distance. In Kotz S, Johnson NL, Read CB, eds, Encyclopaedia of Statistical Sciences 5, Wiley, New York
Grace, LJ, Charity, JA, Gresham, B, Kay, N, Walter, C (2005) Insect resistant transgenic Pinus radiata . Plant Cell Reports 24: 103–111 Google Scholar
Graff, A, Conrad, R (2005) Impact of flooding on soil bacterial communities associated with poplar (Populus sp.) trees. FEMS Microbiol. Ecol. 53: 401–415 CrossRefGoogle Scholar
Grayston, SJ, Griffith, GS, Mawdsley, JL, Campbell, CD, Bardgett, RD (2001) Accounting for variability in soil microbial communities of temperate upland grassland ecosystems. Soil Biol. Biochem. 33: 533–551 CrossRefGoogle Scholar
Gyamfi, S, Pfeifer, U, Stierschneider, M, Sessitsch, A (2002) Effects of transgenic glufosinate-tolerant oilseed rape (Brassica napus) and the associated herbicide application on eubacterial and Pseudomonas communities in the rhizosphere. FEMS Microb. Ecol. 41: 181–190 Google Scholar
Hampp, R, Ecke, M, Schaeffer, C, Wallenda, T, Wingler, A, Kottke, I, Sundberg, B (1996) Axenic mycorrhization of wild type and transgenic hybrid aspen expressing T-DNA indole acetic acid-biosynthetic genes. Trees - Structure and function 11: 59–64 Google Scholar
Henderson, AR, Walter, C (2006) Genetic engineering in Conifer Plantation Forestry. Silvae Genetica 55: 253–262 Google Scholar
Heuer, H, Krsek, M, Baker, P, Smalla, K, Wellington, EM (1997) Analysis of actinomycete communities by specific amplification of genes encoding 16S rRNA and gel-electrophoretic separation in denaturing gradients. Appl. Environ. Microbiol. 63: 3233–3241 Google ScholarPubMed
Heuer H, Wieland G, Schönfeld J, Schönwälder A, Gomes NCM, Smalla K (2001) Bacterial community profiling using DGGE or TGGE analysis. In Rouchelle P, ed, Environmental molecular microbiology: protocols and applications, Horizon Scientific Press, Wymondham, pp 177–190
Heuer, H, Kroppenstedt, RM, 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: 1325–1335 CrossRefGoogle Scholar
Hotelling H (1947) Multivariate quality control. In Eisenhart C, Hastay MW, Wallis WA, eds, Techniques of Statistical Analysis, McGraw-Hill, New York
Izumi, H, Cairney, JW, Killham, K, Moore, E, Alexander, IJ, Anderson, IC (2008) Bacteria associated with ectomycorrhizas of slash pine (Pinus elliottii) in south-eastern Queensland, Australia. FEMS Microbiol. Letters 282: 196–204 Google Scholar
Kaldorf, M, Fladung, M, Muhs, HJ, Buscot, F (2002) Mycorrhizal colonization of transgenic aspen in a field trial. Planta 214: 653–660 Google Scholar
Kataoka, R, Taniguchi, T, Ooshima, H, Futai, K (2008) Comparison of the bacterial communities established on the mycorrhizae formed on Pinus thunbergii root tips by eight species of fungi. Plant Soil 304: 267–275 Google Scholar
Lamarche, J, Hamelin, RC (2007) No evidence of an impact on the rhizosphere diazotroph community by the expression of Bacillus thuringiensis Cry1Ab toxin by Bt white spruce. Appl. Environ. Microbiol. 73: 6577–6583 CrossRefGoogle ScholarPubMed
Landeweert, R, Leeflang, P, Smit, E, Kuyper, T (2005) Diversity of an ectomycorrhizal fungal community studied by a root tip and total soil DNA approach. Mycorrhiza 15: 1–6 Google ScholarPubMed
Lane DJ (1991) 16S/23S rRNA sequencing. In Stackebrandt E, Goodfellow M, eds, Nucleic Acid Techniques in Bacterial Systematics, Hoboken, Wiley, New York, pp 115–175
LeBlanc, PM, Hamelin, RC, Filion, M (2007) Alteration of soil rhizosphere communities following genetic transformation of white spruce. Appl. Environ. Microbiol. 73: 4128–4134 Google ScholarPubMed
Lottmann, J, Heuer, H, Smalla, K, Berg, G (1999) Influence of transgenic T4-lysozyme producing potato plants on potentially beneficial plant-associated bacteria. FEMS Microbiol. Ecol. 29: 365–377 CrossRefGoogle Scholar
Lottmann, J, Heuer, H, De Vries, J, Mahn, A, Düring, K, Wackernagel, W, Smalla, K, Berg, G (2000) Establishment of introduced antagonistic bacteria in the rhizosphere of transgenic potatoes and their effect on the bacterial community. FEMS Microbiol. Ecol. 33: 41–49 CrossRefGoogle ScholarPubMed
Lukow, T, Dunfield, PF, Liesack, W (2000) Use of T-RFLP technique to assess spatial and temporal changes in the bacterial community structure within an agricultural soil planted with transgenic and non-transgenic potato plants. FEMS Microbiol. Ecol. 32: 241–247 CrossRefGoogle Scholar
Muyzer, G, Smalla, K (1998) Application of denaturing gradient gel electrophoresis (DGGE) and temperature gradient gel electrophoresis (TGGE) in microbial ecology. Antonie Van Leeuwenhoek 73: 127–141 Google Scholar
Muyzer, G, de Waal, EC, Uitterlinden, AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl. Environ. Microbiol. 59: 695–700 Google ScholarPubMed
Nübel, U, Engelen, B, Felske, 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: 5636–5643 Google Scholar
O’Callaghan, M, Gerard, EM, Bell, NL, Waipara, NW, Aalders, LT, Baird, DB, Conner, AJ (2008) Microbial and nematode communities associated with potatoes genetically modified to express the antimicrobial peptide magainin and unmodified potato cultivars. Soil Biol. Biochem. 40: 1446–1459 Google Scholar
Oliver, KL, Hamelin, RC, Hintz, WE (2008) Effects of transgenic hybrid aspen overexpressing polyphenol oxidase on rhizosphere diversity. Appl. Envion. Microbiol. 74: 5340–5348 CrossRefGoogle Scholar
Opelt, K, Chobot, V, Hadacek, F, Schönmann, S, Eberl, L, Berg, G (2007) Investigations of the structure and function of bacterial communities associated with Sphagnum mosses. Environ. Microbiol. 9: 2795–2809 CrossRefGoogle Scholar
Parke, JL, Gurian-Sherman, D (2001) Diversity of the Burkholderia cepacia complex and implications for risk assessment of biological control strains. Annu. Rev. Phytopathol. 39: 225–258 CrossRefGoogle Scholar
Payne RW, Harding SA, Murray DA, Soutar DM, Baird DB, Welham SJ, Kane AF, Gilmour AR, Thompson R, Webster R, Tunnicliffe, Wilson G (2007) The Guide to GenStat Release 10, Part 2: Statistics. VSN International, Hemel Hempstead
Plaingam, N, Somrithipol, S, Jones, EBG (2003) Infundibulomyces: a new genus of coelomycetes from Thailand. Can. J. Bot. 81: 732–737 Google Scholar
Priha, O, Grayston, SJ, Hiukka, R, Pennanen, T, Smolander, A (2001) Microbial community structure and characteristics of the organic matter in soils under Pinus sylvestris, Picea abies and Betula pendula at two forest sites. Biol. Fertil. Soils 33: 17–24 Google Scholar
Seppänen, SK, Pasonen, HL, Vauramo, S, Vahala, J, Toikka, M, Kilpeläinen, I, Setälä, H, Teeri, TH, Timonen, S, Pappinen, A (2007) Decomposition of the leaf litter and mycorrhiza forming ability of silver birch with a genetically modified lignin biosynthesis pathway. Appl. Soil Ecol. 36: 100–106 Google Scholar
Shannon CE, Weaver W (1949) The mathematical theory of communication. University of Illinois Press, Urbana, pp 3–24
Smalla, K, Wieland, G, Buchner, A, Zock, A, Parzy, J, Kaiser, S, Roskot, N, Heuer, H, Berg, G (2001) Bulk and rhizosphere soil bacterial communities studied by denaturing gradient gel electrophoresis: plant-dependent enrichment and seasonal shifts revealed. Appl. Environ. Microbiol. 67: 4742–4751 Google ScholarPubMed
Smith SE, Read DJ (1997) Mycorrhizal Symbiosis. Second edition, Academic Press, London
Timonen, S, Hurek, T (2006) Characterisation of bacterial populations associating with Pinus sylvestris – Suillus bovinus mycorrhizospheres. Can. J. Microbiol. 52: 769–778 Google Scholar
Vainio, EJ, Hantula, J (2000) Direct analysis of wood-inhabiting fungi using denaturing gradient gel electrophoresis of amplified ribosomal DNA. Mycol. Res. 104: 927–936 Google Scholar
Valenzuela, S, Balocchi, C, Rodriguez, J (2006) Transgenic trees and forest biosafety. Electronic J. Biotechnol. 9: 335–339 Google Scholar
Van Frankenhuyzen, K, Beardmore, T (2004) Current status and environmental impact of transgenic forest trees. Can. J. Forest Res. 34: 1163–1180 Google Scholar
Wagner A, Ralph J, Akiyama T, Flint H, Phillips L, Torr K, Nanayakkara B, Te Kiri L (2007) Exploring lignification in conifers by silencing hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyltransferase in Pinus radiata. PNAS 104: 11856–11861
Walter, C (2004) Genetic engineering in conifer forestry: Technical and social considerations. In Vitro Cell Dev. Biol. Plant 40: 434–441 Google Scholar
Walter C, Grace LJ (2000) Genetic engineering of conifers for plantation forestry: Pinus radiata transformation. In Jain SM, Minocha SC, eds, Molecular biology of woody plants, Kluwer, Dordrecht, pp 79–104
Walter, C, Grace, LJ, Wagner, A, White, DWR, Walden, AR, Donaldson, SS, Hinton, H, Gardner, RC, Smith, DR (1998) Stable transformation and regeneration of transgenic plants of Pinus radiata D. Don. Plant Cell Reports 17: 460–468 Google Scholar
Weisburg, WG, Barns, SM, Pelletier, DA, Lane, DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J. Bacteriol. 173: 697–703 Google ScholarPubMed
White TJ, Bruns TD, Lee S, Taylor J (1990) Analysis of phylogenetic relationships by amplification and direct sequencing of ribosomal RNA genes. In Innis MA, Gelfand DH, Sninsky JJ, White TJ, eds, PCR Protocols: a Guide to Methods and Applications, Academic Press, New York, pp 315–322