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Degradation and half-life of DNA present in biomass from a genetically-modified organism during land application

  • Fermentation, Cell Culture and Bioengineering - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

White biotechnology has made a positive impact on the chemical industry by providing safer, more efficient chemical manufacturing processes that have reduced the use of toxic chemicals, harsh reaction conditions, and expensive metal catalysts, which has improved alignment with the principles of Green Chemistry. The genetically-modified (GM) biocatalysts that are utilized in these processes are typically separated from high-value products and then recycled, or eliminated. Elimination routes include disposal in sanitary landfills, incineration, use as a fuel, animal feed, or reuse as an agricultural soil amendment or other value-added products. Elimination routes that have the potential to impact the food chain or environment have been more heavily scrutinized for the fate and persistence of biological products. In this study, we developed and optimized a method for monitoring the degradation of strain-specific DNA markers from a genetically-modified organism (GMO) used for the commercial production of 1,3-propanediol. Laboratory and field tests showed that a marker for heterologous DNA in the GM organism was no longer detectable by end-point polymerase chain reaction (PCR) after 14 days. The half-life of heterologous DNA was increased by 17% (from 42.4 to 49.7 h) after sterilization of the soil from a field plot, which indicated that abiotic factors were important in degradation of DNA under field conditions. There was no evidence for horizontal transfer of DNA target sequences from the GMO to viable organisms present in the soil.

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References

  1. Allentoft ME, Collins M, Harker D, Haile J, Oskam CL, Hale ML, Campos PF, Samaniego JA, Gilbert TP, Willerslev E, Zhang G, Scofield RP, Holdaway RN, Bunce M (2013) The half-life of DNA in bone: measuring decay kinetics in 158 dated fossils. Proc R Soc B 279:4724–4733

    Article  Google Scholar 

  2. Banuelos GS, Hanson BD (2010) Use of selenium-enriched mustard and canola seed meals as potential bioherbicides and green fertilizer in strawberry production. Hortic Sci 45:1567–1572

    Google Scholar 

  3. Bending GD, Lincoln SD (2000) Inhibition of soil nitrifying bacteria communities and their activities by glucosinolate hydrolysis products. Soil Biol Biochem 32:1361–1369

    Article  Google Scholar 

  4. Brankatschk R, Bodenhausen N, Zeyer J, Bürgmann H (2013) Simple absolute quantification method correcting for quantitative pcr efficiency variations for microbial community samples. Appl Environ Microbiol 78(13):4481–4489

    Google Scholar 

  5. Cervin, MM, Soucaille P, Valle F. (2008) Process for the biological production of 1,3-propanediol with high yield. US Patent 7,371,558. Filed 6 Oct 2003, and issued 13 May 2008

  6. Darzynkiewicz Z, Juan G, Li X, Gorczyca W, Murakami T, Traganos F (1997) Cytometry in cell necrobiology: analysis of apoptosis and accidental cell death (necrosis). Cytometry 27:1–20

    Article  CAS  PubMed  Google Scholar 

  7. Dizdaroglu M, Jaruga P (2013) Mechanisms of free radical-induced DNA damage to DNA. Free Radic Res 46(4):382–419

    Article  Google Scholar 

  8. Dubeau L, Chandler LA, Gralow JR, Nichols PW, Jones PA (1986) Southern blot analysis of DNA extracted from formalin-fixed pathology specimens. Cancer Res 46:2964–2969

    CAS  PubMed  Google Scholar 

  9. Eglinton G, Logan GA, Ambler RP, Boon JJ, Perizonius WRK (1991) Molecular preservation. Phil Trans R Soc Lond B 333:315–328

    Article  CAS  Google Scholar 

  10. Garibyan L, Avashia N (2013) Polymerase chain reaction. J Investig Dermatol Mar 133(3):e6. doi:10.1038/jid

    CAS  Google Scholar 

  11. Greaves MP, Wilson MJ (1970) The degradation of nucleic acids and montmorillonite-nucleic-acid complexes by soil microorganisms. Soil Biol Biochem 2:257–268

    Article  CAS  Google Scholar 

  12. Heid CA, Stevens J, Livak KJ, Williams PM (1996) Real time quantitative PCR. Genome Res 6:986–994

    Article  CAS  PubMed  Google Scholar 

  13. Huesing JE, Andres D, Braverman MP, Burns A, Felsot AS, Harrigan GG, Hellmich RL, Reynolds A, Shelton AM, van Rijssen WJ, Morris EJ, Eloff JN (2016) Global adoption of genetically modified (GM) crops: challenges for the public sector. J Agric Food Chem 64(2):394–402. doi:10.1021/acs.jafc.5b05116

    Article  CAS  PubMed  Google Scholar 

  14. Joshi MU, Pittman HK, Haisch CE, Verbanac KM (2008) Real-time PCR to determine transgene copy number and to quantitate the biolocalization of adoptively transferred cells from EGFP-transgenic mice. Biotechniques 45(3):247–258

    Article  CAS  PubMed  Google Scholar 

  15. Kerr BJ, Shurson GC (2013) Strategies to improve fiber utilization in swine. J Anim Sci Biotechnol 4:11. doi:10.1186/2049-1891-4-11

  16. Lindahl T (1993) Instability and decay of the primary structure of DNA. Nature 362:709–715

    Article  CAS  PubMed  Google Scholar 

  17. Lindahl T, Andersson A (1972) Rate of chain breakage at apurinic sites in double-stranded deoxyribonucleic acid. Biochemistry 11:3618–3623

    Article  CAS  PubMed  Google Scholar 

  18. Lindahl T, Nyberg B (1972) Rate of depurination of native deoxyribonucleic acid. Biochemistry 11:3610–3618

    Article  CAS  PubMed  Google Scholar 

  19. Lorenz MG, Wackernagel W (1987) Adsorption of DNA to sand and variable degradation rates of adsorbed DNA. Appl Environ Microbiol 53:2948–2952

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Lee C, Kim J, Shin SG, Hwang S (2006) Absolute and relative QPCR quantification of plasmid copy number in Escherichia coli. J Biotechnol 133(3):273–280

    Article  Google Scholar 

  21. Moore A (2011) Fertilizer potential of biofuel byproducts. In: Marco Aurelio Dos Santos Bernardes (ed) Biofuel production-recent developments and prospects. InTech, Rijeka

  22. Nelson KA, Motavelli PP, Smoot RL (2009) Utility of dried distillers grain as a fertilizer source for corn. J Agric Sci 1:3–13

    Google Scholar 

  23. Ogram A, Sayler GS, Gustin D, Lewis RJ (1988) DNA adsorption to soils and sediments. Environ Sci Technol 22:982–984

    Article  CAS  PubMed  Google Scholar 

  24. Romanowski G, Lorenz MG, Sayler G, Wackernagel W (1992) Persistence of free plasmid DNA in soil monitored by various methods, including a transformation assay. Appl Environ Microbiol 58(9):3013–3019

    Google Scholar 

  25. Shurson G, Spiehs M, Whitney M (2004) The use of maize distillers dried grains with solubles in pig diets. Pig News Inf 25(2):75N–83N

    Google Scholar 

  26. Suzuki M, Amano N, Kakinuma J, Tateno M (1997) Use of a 3D structure data base for understanding sequence-dependent conformational aspects of DNA. J Mol Biol 274(3):421–435

    Article  CAS  PubMed  Google Scholar 

  27. Wang AS, Hu P, Hollister EB (2012) Impact of indian mustard (Brassica juncea) and flax (Linum usitatissimum) seed meal applications on soil carbon, nitrogen, and microbial dynamics. Appl Environ Soil Sci 2012:1–14. doi:10.1155/2012/251609

  28. Westendorf ML, Wohlt JE (2002) Brewing by-products: their use as animal feeds. Vet Clin North Am Food Anim Pract 18(2):233–252

    Article  PubMed  Google Scholar 

  29. Whelan JA, Russell NB, Whelan MA (2003) A method for the absolute quantification of cDNA using real-time PCR. J Immunol Methods 278(1–2):261–269

    Article  CAS  PubMed  Google Scholar 

  30. Whitney MH, Shurson GC (2004) Growth performance of nursery pigs fed diets containing increasing levels of corn distillers dried grains with solubles originating from a modern Midwestern ethanol plant. J Anim Sci 82:132–138

    Article  Google Scholar 

  31. Xu L, Chen H, Hu X, Zhang R, Zhang Z, Luo ZW (2006) Average gene length is highly conserved in prokaryotes and eukaryotes and diverges only between the two kingdoms. Mol Biol Evol 23(6):1107–1108

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors would like to thank the Tennessee Department of Agriculture and the University of Tennessee for assistance in conducting these research field trials.

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Correspondence to James A. Zahn.

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Halter, M.C., Zahn, J.A. Degradation and half-life of DNA present in biomass from a genetically-modified organism during land application. J Ind Microbiol Biotechnol 44, 213–220 (2017). https://doi.org/10.1007/s10295-016-1876-x

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  • DOI: https://doi.org/10.1007/s10295-016-1876-x

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