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Jatropha curcas is a non-food, perennial biofuel crop
geographically limited to the tropical and
subtropical world. To confer resistance to cold
stress, we transformed Jatropha using
Agrobacterium-mediated and biolistic
transformation methods. We used reporter genes
(GUS and GFP) and CBF3 gene with a stress
inducible promoter (rd29A) in the transformation
studies. Successful transformation with GUS and
GFP were demonstrated using GUS staining and
microscopic observation of green fluorescence
respectively. GUS transformants were PCR positive
and molecular analysis is being carried out on
putative CBF3 transformants.
Leaf and embryo explants of J. curcas were
transformed with GUS and GFP
Background
Native to Mexico and Central America and has been
cultivated in tropical and subtropical regions.
Belongs to family Euphorbiaceae.
Advantages:
Naturally drought resistant
A non-food biofuel crop
Can grow on marginal lands
High content of oil (30-40% of seed weight)
Production cost is low ($ 34-48/ barrel)
The oil yields 2,268 liters of oil per hectare, which
can be translated to 597 gallons of oil for every 2.5
acres.
Jatropha oil was tested as a jet fuel blend for Air
New Zealand (2007) and Continental Airlines on
January 7, 2009.
Challenge
It is susceptible to cold stress and can’t be grown in
temperate regions.
Other Uses
•Seeds are considered anthelmintic in Brazil.
•Latex contains an alkaloid known as “Jatrophine”
(having anti-cancerous properties.)
•Bark yields a dark blue dye used in the Philippines
for coloring cloth, finishing nets and lines.
•Leaves are used for fumigating houses against bed
bugs in Ghana.
ASTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
SUMMARY
GENETIC TRANSFORMATION OF JATROPHA CURCAS
Aurellia Whitmore1, Benham Tabatabai
2, Dr. Puthiyaparambil Josekutty
2, Dr. Sairam Rudrabhatla
2
1Southern University at New Orleans, 6400 Press Dr., New Orleans, LA 70126.
2Penn State Harrisburg, 777 West
Harrisburg Pike, Middletown, PA 17057 http://harrisburg.psu.edu/reu/sustainable-bioenergy
Agrobacterium-mediated transformation
1) Decontaminate
deshelled seed
1) Collect in
vitro
germinated
leaves.
2) Excise embryo/cotelydon 2) Lightly poke
3)Immerse explants into cultures.
4) Transfer explants to co cultivation media.
5) Place in dark 25° C for 72 hours
Biolistic Transformation
1) Decontaminat
e endosperm
of seed.
1) Collect in
vitro
germinated
leaves.
3) Coat gold particles (0.6 µM) with plasmid DNA
4) Load explants to regeneration medium.
5) Fire gene gun
6) Incubate in dark (leaf explants), in light (embryo
explants)
A. Agrobacterium Cell
B. Agrobacterium DNA
C. Ti Plasmid
D. Plant Cell
E. Plant Mitochondria
F. Plant Chloroplast
G. Plant Nucleus
a. T-DNA
b. Vir genes
c. Replication origin
Li M. R., Li H. Q., Pan X. , Wu G. J. (2008). Establishment of an Agrobacterium-mediated cotyledon disc transformation method for Jatropha curcas. Plant Cell
Tissue Organ and Culture, 92 (2):173-181.
Openshaw K. (2000). A review of Jatropha curcas: an oil plant of unfulfilled promise. Biomass Bioenergy, 19: 1-15.
Pan J, Fu Q, Zheng-Fu X . (2010). Agrobacterium tumefaciens-mediated transformation of biofuels plant Jatropha curcas using kanamycin selection. African
Journal of Biotechnology, 9 (39): 6477-6481.
Parajuli, Ranjan (2009).“Jatropha Curcas and Its Potential Applications; A Compilation Paper on Plantation and Application of Jatropha Curcas”
http://www.environmental-expert.com/Files/0/articles/73213/Jatropha.pdf
Purkayastha J., Sugla T., Paul A., Solleti S.K., Mazumdar P., Basu A., Mohommad A., Ahmed Z., Sahoo L. (2010) Efficient in vitro plant regeneration from shoot
apices and gene transfer by particle bombardment in Jatropha curcas. Biologia Plantarium, 54 : 13-20.
• PCR results for GUS and GFP expressions were positive
• Molecular analysis of putative CBF3 transformants is
being carried out.
Gradient PCR (GFP
expression)
PCR (GUS
expression)
Biolistic transformation Agrobacterium- mediated
transformation
GUS plasmid construct Biodiesel Crops
GFP expression from Agrobacterium-mediated
transformation
GUS expression from
Biolistic transformation
GUS expression from
Agrobacterium-mediated
transformation
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1.http://www.nepadbiosafety.net/for-
regulators/resources/subjects/biotechnology/plant-
transformation-bombardment
http://gas2.org/2008/10/02/jatropha-from-
haitian-voodoo-to-biodiesel-holy-grail/
REFERENCES
Acknowledgements
Thank you to The National Science Foundation (NSF) for their
fellowship, Pennsylvania State- Harrisburg, Ben Tabatabai, Dr.
Puthiyaparambil Josekutty, Dr. Sairam Rudrabhatla NSF-REU staff and
colleagues
