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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