1. PHYTOREMEDIATION OF EXPLOSIVES
USING TRANSGENIC PLANTS
By Greg O’Keefe

2. CONTENTS
 Introduction
 Materials and Methods
 Results
 Discussion
 Future Trends
 Conclusion

3. INTRODUCTION
 Explosive contamination in environment growing
concern
 Threatening to humans, animals, ecosystems
 Due to improper disposal and lack of suitable
remediation methods
 Incineration, composting, soil slurry bioremediation are
inadequate
 Ammunition production and warehouse facilities,
waste disposal and destruction facilities, firing
ranges, impact areas
 Approx. 50 million acres contaminated in US alone

4. INTRODUCTION
 Phytoremediation using transgenic plants new and
novel method
 Bacterial genes transformed into plants
 Capable of degrading multiple types of explosives
 In situ remediation, renewable resources,
environmentally friendly materials

5. INTRODUCTION
(1)

6. INTRODUCTION
 TNT and RDX two most popular explosives
 TNT
 Mutagen
 Dermatitis, vomiting, liver damage
 RDX
 Convulsions in mammals
 Vertigo, unconsciousness, vomiting
 Crosses blood brain barrier

7. MATERIALS AND METHODS
 Four Major Steps:
1. Construction of Vector
2. Transformation
3. Growth of Transgenic Plants
4. Analysis of Explosives

8. CONSTRUCTION OF VECTOR
oConstruction of vector followed by PCR
oAmplification of target gene

9. TRANSFORMATION
 Introducing vector into plant
 Standard agrobacterium-mediated transformation
 Floral dip method for Arabidopsis thaliana

10. GROWTH OF TRANSGENIC PLANTS
 Grown in explosive contaminated media or soil
 Necessary for control media/soil with no explosives
present
 Grown along wild type plants
 Samples of media/soil taken at intervals
 Must analyze explosive content over time

11. ANALYSIS OF EXPLOSIVES

12. RESULTS
 Plant growth and toxicity:
(3)

13. RESULTS
(4)

14. RESULTS
 Transformation of RDX
(5)

15. RESULTS
 Transformation of TNT
(6)

16. DISCUSSION
 Transgenic plants have significantly greater
resistance to toxic effects of explosives
 Transgenic plants efficiently remove TNT and RDX
from soil/media/water
 Problem with size of plants and their roots
 More research on explosive metabolites
 Regulatory framework

17. FUTURE TRENDS
 Introducing multiple transgenes into single species
 More research on trees
 Phytoremediation in conjunction with
rhizoremediation and nanotechnology
 Introduce plants in greenhouse and in the field

18. CONCLUSION
 Phytoremediation using transgenic plants for
degradation of explosives
 In situ, low cost, renewable resources, lack of specific
equipment, no introduction of new chemicals to
environment
 Increased resistance to explosives, efficient uptake and
transformation from soil/media/water
 Development is slow to due regulatory framework
 Plants need to be used in greenhouse setting and in the
field
 To increase efficiency:
 Introduce multiple transgenes, integrate with
rhizoremediation and nanotechnology

19. REFERENCES
1. http://seth-smith.org.uk/helena/cv/hssthesis.pdf (Seth-Smith HMB (2002) Microbial degradation of
RDX. PhD Dissertation, University of Cambridge)
2. Pitchel, J. (2012, March 19). Distribution and Fate of Military Explosives and Propellants in Soil: A
Review. Hindawi. Retrieved October 21, 2014, from
http://www.hindawi.com/journals/aess/2012/617236/
3. Hannink, N., Rosser, S., French, C., Basran, A., Murray, J., Nicklin, S., et al. (2001, October 12).
Phytodetoxification of TNT by transgenic plants expressing a bacterial nitroreductase.. National Center
for Biotechnology Information. Retrieved September 29, 2014, from
http://www.ncbi.nlm.nih.gov/pubmed/11731787
4. Dillewijn, P. v., Couselo, J., Corredoria, E., Delgado, A., Willitch, R., Bellester, A., et al. (2007, August
27). Bioremediation of 2,4,6-Trinitrotoluene by Bacterial Nitroreductase Expressing Transgenic Aspen. -
Environmental Science & Technology (ACS Publications). Retrieved September 29, 2014, from
http://pubs.acs.org/doi/abs/10.1021/es801231w
5. Jackson, R., Rylott, E., Fournier, D., Hawari, J., & Bruce, N. (2007, October 16). Exploring the
biochemical properties and remediation applications of the unusual explosive-degrading P450 system
XplA/B. National Center for Biotechnology Information. Retrieved September 29, 2014, from
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2040458/
 Hannink, N. K., Subramanian, M., Rosser, S. J., Basran, A., Murray, J. A. H., Shanks, J. V., & Bruce, N.
C. (2007). Enhanced transformation of TNT by tobacco plants expressing a bacterial nitroreductase.
International Journal of Phytoremediation, 9(4-6), 385-401. Retrieved from
http://search.proquest.com/docview/200169777?accountid=39340