Abstract
To analyze the persistence of the 35S promoter, nos terminator, and hpt, we buried the leaves and rootstocks of transgenic watermelons (Citrullus lanatus (Thunb.) Matsum. & Nakai) in 10 cm of soil. Qualitative and quantitative PCR analyses showed that the amount of transgenes in leaf samples was greatly decreased, by 70%, after 1 month, and only 2.5% remained after 2 months. No transgenes were detected in the leaves after 3 months. For buried rootstock samples, transgenes also degraded quickly, but a very small amount was still detectable up to 3 months later. In our investigation of possible gene transfer from decomposing transgenic watermelon to soil bacteria, only the 35S promoter was detected. However, further examination using colony dot hybridization tests indicated that such a transfer did not occur.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Badosa E, Moreno C, Montesinos E (2004) Lack of detection of ampicillin resistance gene transfer from Bt176 transgenic corn to culturable bacteria under field conditions. FEMS Microbiol Ecol 48:169–178
Bertolla F, Simonet P (1999) Horizontal gene transfer in the environment: natural transformation as a putative process for gene transfers between transgenic plants and microorganisms. Res Microbiol 150:375–384
Blum SAE, Lorenz MG, Wackernagel W (1997) Mechanism of retarded DNA degradation and prokaryotic origin of DNases in nonsterile soils. Syst Appl Microbiol 20:513–521
Davies J (1994) Inactivation of antibiotics and the dissemination of resistance genes. Science 264:375–382
de Vries J, Wackernagel W (1998) Detection of nptII (kanamycin resistance) genes in genomes of transgenic plants by marker-rescue transformation. Mol Genet Genom 257:606–613
Eden PA, Schmidt TM, Blakemore RP, Pace NR (1991) Phylogenetic analysis of Aquaspirillum magnetotacticum using polymerase chain reaction-amplified 16S rRNA-specific DNA. Intl J Syst Bacteriol 41:324–325
Gallori E, Bazzicalupo M, Dal Canto L, Fani R, Nannipieri P, Vettori C, Stotzky G (1994) Transformation of Bacillus subtilis by DNA bound on clay in non-sterile soil. FEMS Microbiol Ecol 15:119–126
Gebhard F, Smalla K (1998) Transformation of Acinetobacter sp. strain BD413 by transgenic sugar beet DNA. Appl Environ Microb 64:1550–1554
Hay I, Morency M, Séguin A (2002) Assessing the persistence of DNA in decomposing leaves of genetically modified poplar trees. Can J For Res 32:977–982
Hoffmann T, Golz C, Schieder O (1994) Foreign DNA sequences are received by a wild-type strain of Aspergillus niger after co-culture with transgenic higher plants. Curr Genet 27:70–76
Kim C-G, Lee B, Kim DI, Park JE, Kim HJ, Park KW, Yi H, Jeong SC, Yoon WK, Harn CH, Kim HM (2008) Detection of gene flow from GM to non-GM watermelon in a field trial. J Plant Biol 51:74–77
Lee B, Kim C-G, Park J-Y, Park KW, Yi H, Harn CH, Kim HM (2007) Assessment of the persistence of DNA in decomposing leaves of CMVP0-CP transgenic chili pepper in the field conditions. Kor J Environ Agric 26:319–324
Lorenz MG, Wackernagel W (1987) Adsorption of DNA to sand and variable degradation rates of adsorbed DNA. Appl Environ Microb 53:2948–2952
Lorenz MG, Wackernagel W (1994) Bacterial gene transfer by natural genetic transformation in the environment. Microbiol Rev 58:563–602
Motavalli PP, Kremer RJ, Fang M, Means NE (2004) Impact of genetically modified crops and their management on soil microbially mediated plant nutrient transformations. J Environ Qual 33:816–824
Nielsen KM, Gebhard F, Smalla K, Bones AM, Van Elsas JD (1997a) Evaluation of possible horizontal gene transfer from transgenic plants to the soil bacterium Acinetobacter calcoaceticus BD413. Theor Appl Genet 95:815–821
Nielsen KM, van Weerelt MDM, Berg TN, Bones AM, Hagler AN, van Elsas JD (1997b) Natural transformation and availability of transforming DNA to Acinetobacter calcoaceticus in soil microcosms. Appl Environ Microb 63:1945–1952
Nielsen KM, Bones AM, Smalla K, van Elsas JD (1998) Horizontal gene transfer from transgenic plants to terrestrial bacteria—a rare event? FEMS Microbiol Rev 22:79–103
Nielsen KM, van Elsas JD, Smalla K (2000) Transformation of Acinetobacter sp. strain BD413 (pFG4nptII) with transgenic plant DNA in soil microcosms and effects of kanamycin on selection of transformants. Appl Environ Microb 66:1237–1242
Paget E, Simonet P (1994) On the track of natural transformation in soil. FEMS Microbiol Ecol 15:109–118
Park SM, Lee JS, Jegal S, Jeon BY, Jung M, Park YS, Han SL, Shin YS, Her NH, Lee JH, Lee MY, Ryu KH, Yang SG, Harn CH (2005) Transgenic watermelon rootstock resistant to CGMMV (cucumber green mottle mosaic virus) infection. Plant Cell Rep 24:350–356
Park SM, Kwon JH, Lim MY, Shin YS, Her NH, Lee JH, Ryu KH, Harn CH (2007) CGMMV tolerance test of CGMMV-CP transgenic watermelon rootstock and establishment of transgenic line. J Plant Biotechnol 34:11–17
Romanowski G, Lorenz MG, Wackernagel W (1991) Adsorption of plasmid DNA to mineral surfaces and protection against DNase I. Appl Environ Microb 57:1057–1061
Romanowski G, Lorenz MG, Wackernagel W (1993) Use of polymerase chain reaction and electroporation of Escherichia coli to monitor the persistence of extracellular plasmid DNA introduced into natural soils. Appl Environ Microb 59:3438–3446
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Smalla K, Borin S, Heuer H, Gebbard F, van Elsas JD, Nielsen KM (2000) Horizontal transfer of antibiotic resistance genes from transgenic plants to bacteria—are there new data to fuel the debate? Proceedings of the 6th International Symposium on the Biosafety of Genetically Modified Organisms, Saskatoon, SK, CA, pp 146–154
Stotzky G (2000) Persistence and biological activity in soil of insecticidal proteins from Bacillus thuringiensis and of bacterial DNA bound on clays and humic acids. J Environ Qual 29:691–705
Tam T-Y, Mayfield CI, Inniss WE (1983) Microbial decomposition of leaf material at 0°C. Microb Ecol 9:355–362
Widmer F, Seidler RJ, Watrud LS (1996) Sensitive detection of transgenic plant marker gene persistence in soil microcosm. Mol Ecol 5:603–613
Widmer F, Seidler RJ, Donegan KK, Reed GL (1997) Quantification of transgenic plant marker gene persistence in the field. Mol Ecol 6:1–7
Acknowledgements
This research was supported by grants from the KRIBB Research Initiative Program, the Crop Functional Genomics Center, and the National Academy of Agricultural Science and the Biogreen 21 Program of the RDA in Korea.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Lee, B., Park, JY., Park, K.W. et al. Evaluating the Persistence of DNA from Decomposing Transgenic Watermelon Tissues in the Field. J. Plant Biol. 53, 338–343 (2010). https://doi.org/10.1007/s12374-010-9121-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12374-010-9121-z