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3 Biotech

, 9:408 | Cite as

Biotransformation of 2,4-dinitrotoluene by the beneficial association of engineered Pseudomonas putida with Arabidopsis thaliana

  • Özlem AkkayaEmail author
  • Ebru Arslan
Original Article

Abstract

2,4-dinitrotoluene (2,4-DNT) is a priority environmental xenobiotic pollutant which has toxic, mutagenic, and carcinogenic properties. Thus, its biodegradation by applying recent approaches such as taking advantage of plant-bacteria interactions is crucial. In this work, the genes from Burkholderia sp. R34, necessary for 2,4-DNT degradation, were integrated into wild-type Pseudomonas putida (P. putida) KT2440 genome, and this strain, named KT.DNT, was inoculated to soil in in vitro conditions. To estimate the disappearance of 2,4-DNT in contaminated soil, samples were taken from different time intervals, extracted and analyzed using high-performance liquid chromatography (HPLC). Biotransformation of 2,4-DNT increased gradually and the degradation in soil after 14-days of treatment with the bacterium was found to be the 97.1%, indicating that the engineered strain could be a remarkable candidate for in situ bioremediation of 2,4-DNT-contaminated sites. In addition, in vitro interaction of this bacterium with a model plant, Arabidopsis thaliana (A. thaliana), enhanced lateral root and root hair formation together with dry root weight. Moreover, the initial 2,4-DNT concentration was decreased to 68% within 2 h with the plant-associated KT.DNT in liquid culture. Hence, the usage of this bacterium with plants could also be a promising application for the 2,4-DNT biotransformation.

Keywords

Pseudomonas putida 2,4-DNT Degradation Soil Arabidopsis thaliana 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

References

  1. Akkaya Ö, Pérez-Pantoja DR, Calles B, Nikel PI, de Lorenzo V (2018) The metabolic redox regime of Pseudomonas putida tunes its evolvability towards novel xenobiotic substrates. MBio 9:e01512–e01518PubMedPubMedCentralGoogle Scholar
  2. Bao Y, Lies DP, Fu H, Roberts GP (1991) An improved Tn7-based system for the single-copy insertion of cloned genes into chromosomes of Gram-negative bacteria. Gene 109:167–168PubMedGoogle Scholar
  3. Bondy-Denomy J, Davidson AR (2014) When a virus is not a parasite: the beneficial effects of prophages on bacterial fitness. J Microbiol 52:235–242PubMedGoogle Scholar
  4. Choi KH, Schweizer HP (2006) Mini-Tn7 insertion in bacteria with single attTn7 sites: example Pseudomonas aeruginosa. Nat Protoc 1:153–161PubMedGoogle Scholar
  5. Coenye T, Vandamme P (2003) Diversity and significance of Burkholderia species occupying diverse ecological niches. Environ Microbiol 5:719–729PubMedGoogle Scholar
  6. de las Heras A, Chavarría v, de Lorenzo V (2011) Association of dnt genes of Burkholderia sp. DNT with the substrate-blind regulator DntR draws the evolutionary itinerary of 2,4-dinitrotoluene biodegradation. Mol Microbiol 82(2):287–299Google Scholar
  7. Duque E, Marque SS, Ramos JL (1993) Mineralization of p-methyl-C-benzoate in soils by Pseudomonas putida (pWW0). Microb Releases 2(3):175–177PubMedGoogle Scholar
  8. Dutta SK, Hollowell GP, Hashem FM, Kuykendall DL (2003) Enhanced bioremediation of soil containing 2,4-dinitrotoluene by a genetically modified Sinorhizobium meliloti. Soil Biol Biochem 35(5):667–675Google Scholar
  9. Espinosa-Urgel M, Kolter R, Ramos JL (2002) Root colonization by Pseudomonas putida: love at first sight. Microbiology 148(2):341–343PubMedGoogle Scholar
  10. Fernández M, Niqui-Arroyo JL, Conde S, Ramos JL, Duque E (2012) Enhanced tolerance to naphthalene and enhanced rhizoremediation performance for Pseudomonas putida KT2440 via the NAH7 catabolic plasmid. Appl Environ Microbiol 78(15):5104–5110PubMedPubMedCentralGoogle Scholar
  11. Fernández M, Conde S, Duque E, Ramos JL (2013) In vivo gene expression of Pseudomonas putida KT2440 in the rhizosphere of different plants. Microb Biotechnol 6:307–313PubMedPubMedCentralGoogle Scholar
  12. Gao Y, Liu C, Ding Y, Sun C, Zhang R, Xian M, Zhao G (2014) Development of genetically stable Escherichia coli strains for poly(3-hydroxypropionate) production. PLoS One 9:e97845PubMedPubMedCentralGoogle Scholar
  13. Gong T, Liu R, Che Y, Xu X, Zhao F, Yu H, Song C, Liu Y, Yang C (2016) Engineering Pseudomonas putida KT2440 for simultaneous degradation of carbofuran and chlorpyrifos. Microb Biotechnol 9:792–800PubMedPubMedCentralGoogle Scholar
  14. Jahn M, Vorpahl C, Türkowsky D, Lindmeyer M, Bühler B, Harms H, Müller S (2014) Accurate determination of plasmid copy number of flow-sorted cells using droplet digital PCR. Anal Chem 86:5969–5976PubMedGoogle Scholar
  15. Jenkins TF, Walsh ME (1991) Field screening method for 2, 4-dinitrotoluene in soil. Cold Regions Research and Engineering Lab, HanoverGoogle Scholar
  16. Jiménez JI, Miñambres B, García JL, Díaz E (2002) Genomic analysis of the aromatic catabolic pathways from Pseudomonas putida KT2440. Environ Microbiol 4:824–841PubMedGoogle Scholar
  17. Kiiskila JD, Das P, Sarkar D, Datta R (2015) Phytoremediation of explosive-contaminated soils. Curr Pollut Rep 1:23–34Google Scholar
  18. Koch B, Jensen LE, Nybroe O (2001) A panel of Tn7-based vectors for insertion of the gfp marker gene or for delivery of cloned DNA into Gram-negative bacteria at a neutral chromosomal site. J Microbiol Meth 45:187–195Google Scholar
  19. Koma D, Yamanaka H, Moriyoshi K, Ohmoto T, Sakai K (2012) A convenient method for multiple insertions of desired genes into target loci on the Escherichia coli chromosome. Appl Microbiol Biotechnol 93:815–829PubMedGoogle Scholar
  20. Küce P, Coral G, Kantar C (2015) Biodegradation of 2,4-dinitrotoluene (DNT) by Arthrobacter sp. K1 isolated from a crude oil contaminated soil. Ann Microbiol 65(1):467–476Google Scholar
  21. Kundu D, Hazra C, Chaudhari A (2015) Isolation, screening and assessment of microbial isolates for biodegradation of 2, 4-and 2, 6-dinitrotoluene. Int J Curr Microbiol App Sci 4(1):564–574Google Scholar
  22. Lin J-M, Stark B, Webster D (2003) Effects of vitreoscilla hemoglobin on the 2,4-dinitrotoluene (2,4-DNT) dioxygenase activity of Burkholderia and on 2,4-DNT degradation in two-phase bioreactors. J Ind Microbiol Biotechnol 30(6):362–368PubMedGoogle Scholar
  23. Martinez-Garcia E, Jatsenko T, Kivisaar M, de Lorenzo V (2015) Freeing Pseudomonas putida KT2440 of its proviral load strengthens endurance to environmental stresses. Environ Microbiol 17:76–90PubMedGoogle Scholar
  24. Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor, New YorkGoogle Scholar
  25. Molina L, Ramos C, Duque E, Ronchel MC, Garcı́a JM, Wyke L, Ramos JL (2000) Survival of Pseudomonas putida KT2440 in soil and in the rhizosphere of plants under greenhouse and environmental conditions. Soil Biol Biochem 32(3):315–321Google Scholar
  26. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15:473–497Google Scholar
  27. Nanda AM, Thormann K, Frunzke J (2014) Impact of spontaneous prophage induction on the fitness of bacterial populations and host-microbe interactions. J Bacteriol 197:410–419PubMedGoogle Scholar
  28. Nelson KE, Weinel C, Paulsen IT, Dodson RJ, Hilbert H, Santos VAPM et al (2002) Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440. Environ Microbiol 4:799–808PubMedGoogle Scholar
  29. Nikel PI, Martínez-García E, de Lorenzo V (2014) Biotechnological domestication of Pseudomonads using synthetic biology. Nat Rev Microbiol 12(5):368PubMedGoogle Scholar
  30. Nikel PI, Chavarría M, Danchin A, de Lorenzo V (2016) From dirt to industrial applications: pseudomonas putida as a synthetic biology chassis for hosting harsh biochemical reactions. Curr Opin Chem Biol 34:20–29PubMedGoogle Scholar
  31. Nishino SF, Paoli GC, Spain JC (2000) Aerobic degradation of dinitrotoluenes and pathway for bacterial degradation of 2,6-dinitrotoluene. Appl Environ Microbiol 66(5):2139–2147PubMedPubMedCentralGoogle Scholar
  32. Pérez-Pantoja D, Nikel PI, Chavarría M, de Lorenzo V (2013) Endogenous stress caused by faulty oxidation reactions fosters evolution of 2,4-dinitrotoluene-degrading bacteria. PLoS Genet 9:e1003764PubMedPubMedCentralGoogle Scholar
  33. Pizarro-Tobías P, Udaondo Z, Roca A, Ramos JL (2015) Events in root colonization by Pseudomonas putida. Pseudomonas. Springer, Dordrecht, pp 251–286Google Scholar
  34. Planchamp C (2013) Direct and indirect effects of the rhizobacteria Pseudomonas putida KT2440 on maize plants. Dissertation, Université de NeuchâtelGoogle Scholar
  35. Planchamp C, Glauser G, Mauch-Mani B (2015) Root inoculation with Pseudomonas putida KT2440 induces transcriptional and metabolic changes and systemic resistance in maize plants. Front Plant Sci 13(5):719Google Scholar
  36. Podlipná R, Pospíšilová B, Vaněk T (2015) Biodegradation of 2, 4-dinitrotoluene by different plant species. Ecotoxicol Environ Saf 112:54–59PubMedGoogle Scholar
  37. Remans T, Thijs S, Truyens S, Weyens N, Schellingen K, Keunen E, Gielen H, Cuypers A, Vangronsveld J (2012) Understanding the development of roots exposed to contaminants and the potential of plant-associated bacteria for optimization of growth. Ann Bot 110(2):239–252PubMedPubMedCentralGoogle Scholar
  38. Rickert DE, Butterworth BE, Popp JA, Krahn DF (1984) Dinitrotoluene: acute toxicity, oncogenicity, genotoxicity, and metabolism. CRC Crit Rev Toxicol 13:217–234Google Scholar
  39. Rocheleau S, Kuperman RG, Martel M, Paquet L, Bardai G, Wong S, Sarrazin M, Dodard SG, Gong P, Hawari J, Checkai T, Sunahara GI (2006) Phytotoxicity of nitroaromatic energetic compounds freshly amended or weathered and aged in sandy loam soil. Chemosphere 62:545–558PubMedGoogle Scholar
  40. Rodgers JD, Bunce NJ (2001) Treatment methods for the remediation of nitroaromatic explosives. Water Res 35(9):2101–2111PubMedGoogle Scholar
  41. Rylott EL, Bruce NC (2009) Plants disarm soil: engineering plants for the phytoremediation of explosives. Trend Biotechnol 27:73–81Google Scholar
  42. Segura A, Ramos JL (2013) Plant–bacteria interactions in the removal of pollutants. Curr Opin Biotechnol 24(3):467–473PubMedGoogle Scholar
  43. Sims JL, Sims RC, Matthews JE (1990) Approach to bioremediation of contaminated soil. Hazard Waste Hazard Mater 7(2):117–149Google Scholar
  44. Spanggord RJ, Spain JC, Nishino SF, Mortelmans KE (1991) Biodegradation of 2,4- dinitrotoluene by a Pseudomonas sp. Appl Environ Microbiol 57:3200–3205PubMedPubMedCentralGoogle Scholar
  45. Stenuit BA, Agathos SN (2010) Microbial 2,4,6-trinitrotoluene degradation: could we learn from (bio)chemistry for bioremediation and vice versa? Appl Microbiol Biotechnol 88:1043–1064PubMedGoogle Scholar
  46. Suen WC, Spain JC (1993) Cloning and characterization of Pseudomonas sp. strain DNT genes for 2,4-dinitrotoluene degradation. J Bacteriol 175:1831–1837PubMedPubMedCentralGoogle Scholar
  47. Thijs S, Weyens N, Sillen W, Gkorezis P, Carleer R, Vangronsveld J (2014) Potential for plant growth promotion by a consortium of stress-tolerant 2,4-dinitrotoluene-degrading bacteria: isolation and characterization of a military soil. J Microbial Biotechnol 7:294–306Google Scholar
  48. Valvano MA, Keith KE, Cardona ST (2005) Survival and persistence of opportunistic Burkholderia species in host cells. Curr Opin Microbiol 8:99–105PubMedGoogle Scholar
  49. Walsh ME, Lambert DJ (2006) Extraction kinetics of energetic compounds from training range and army ammunition plant soils (No. erdc/crrel-tr-06-6). Engineering research and development center Cold Regions Research and Engineering Lab, HanoverGoogle Scholar
  50. Xu J, Jing N (2012) Effect of 2,4-dinitrotoluene exposure on enzyme activity, energy reserves and condition factors in common carp (Cyprinus carpio). J Hazard Mater 203–204:299–307PubMedGoogle Scholar

Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.Department of Molecular Biology and GeneticsGebze Technical UniversityKocaeliTurkey

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