curcas In Vitro Propagation Camille González*1, Lauren Kong*2 Ben Tabatabai3, Kamal Chowdhury4,Matt Reitzel3, Shobha Potlakayala3, Alison Shuler 3,Sairam Rudrabhatla3 1University of Puerto Rico - Rio Piedras, Ponce de León Ave., San Juan, PR 00925 2Mills College, 5000 MacArthur Blvd., Oakland, CA 94613 3Penn State – Harrisburg, 777 W. Harrisburg Pike, Middletown, PA 17057 4Claflin University, 400 Magnolia St., Orangeburg, SC 29115

Callus Induction by Media Hypocotyl and Cotyledon Callus Varying in Carbon Source Induction Abstract 100 105

Materials and Methods 80 100 60 40 95 Sucrose 20 Maltose

Jatropha Embryo Percentage ofcalli 90 Maltose

Jatropha curcas, a member of the family, is 0 Percentage ofCalli Jatropha NBM and MC were surface sterilized and the JCI 1 JCI 2 JCI 3 JCI 5 85 a native to tropical regions. Its ability to grow in varied Media MC - H NBM - H MC - C NBM - C embryos were germinated on JEG5 growth media and incubated conditions and its high oil content make it one of the world’s A) Comparison between JCI media varying B) Comparison between hypocotyls and most promising non-food crops. The present study was under light. in carbon source for new growth . cotyledons with varying carbon and seed source. geared towards developing an efficient tissue culture protocol for Jatropha. Four different types of media, varying in After one to two weeks the cotyledons and hypocotyls of the concentration of growth regulators and carbon source, were resulting were excised and planted on Jatropha Callus Conclusion and Future Works used for callus induction. Preliminary results suggest that Induction 1-S/M media plates and maltose may be a preferable carbon source for callus induction incubated in a dark growth environment. More than 90% cotyledon and hypocotyl explants from from explants for certain growth regulators combination. Jatropha Callus Induction both seed sources produced callus. Hypocotyl explants In contrast, no considerable difference was observed for Leaf explants from a 3 year old Jatropha plant were collected, responded slightly better in sucrose medium. For leaf cotyledon and hypocotyl callus induction. By developing an surface sterilized and plated on explants, an interaction between growth regulator effective tissue culture and transformation protocol, cold Jatropha Callus Induction media concentration and carbon source was observed. The rate of tolerant Jatropha can be produced to grow in temperate alternating between 5 different calli formation observed in media JCI 5M (90% calli climate. concentration of growth regulators and/or different carbon formation), JCI 5S (70%), JCI 2M (72.5%), and JCI 2S (52%) was source. The plates were incubated in the dark to induce callus observed to be considerably higher than the other media. formation. These findings suggest that utilizing maltose instead of sucrose in combination with growth regulators present in JCI Introduction 2 and JCI 5 media might provide a more effective medium for Jatropha tissue culture. Linnaeus, native to tropical areas, is a Future works will involve regeneration of Jatropha plants perennial belonging to the Euphorbiaceae family (Gubitz, from callus, optimization of transformation protocol to 1999). Its status as a non-food crop, ability to grow quickly in Results introduce cold tolerance gene (CBF3), and regeneration of poor conditions, and its animal repelling toxicity make transgenic plants for growing in temperate climate. Jatropha a promising biofuel feedstock (Kumar & Sharma, 2008). Moreover, it bears within five years of planting, allowing it to produce large quantities of oil-yielding seeds References throughout its 50 year life span (Openshaw, 2000). In addition, seeds can be used in , insecticides, , medicine G.M. Gubitz a, M. Mittelbach b*, M. Trabi c (1999) Exploitation of the tropical oil seed plant and varnish (Sirisomboom et. al. 2007). Embryo Germination (2 weeks) Jatropha curcas L. Biosure Technology. 67, 73-82. Jones, N., Miller, J. H., (1992) Jatropha curcas: A multipurpose for problematic sites. Cold weather limits growth of Jatropha, restricting wide- Washington DC: The World Bank. scale cultivation on temperate land. Activation of cold Kumar, A., Sharma, S., An evaluation of multipurpose oil seed crop for industrial uses (Jatropha curcas): A review. Industrial Crop Production. (2008), doi:10.1016/j.indcrop. tolerance genes through genetic engineering in Jatropha would 2008.01.001 increase its survival under cold stress. This would expand its Openshaw K. (2000) A review of Jatropha curcas: an oil plant of unfulfilled promise. and . 19, 1-15. supply throughout the world, granting non-tropical regions access to this potential biofuel. For genetic engineering, Callus induction form cotelydon Callus induction form hypocotyl explant explant efficient tissue culture and transformation protocols are Acknowledgments essential. In this study Jatropha cotyledon, hypocotyl and leaf explants were cultured on callus induction media. These The National Science Foundation included variations in carbon source and concentration of The Pennsylvania State University – Harrisburg for providing the necessary resources and hormonal growth factors to determine optimal growth facilitiesject. The Central Pennsylvania Laboratory for and all its members for their assistance and conditions for Jatropha explants. Callus induction from leaf explant (8 weeks) collaboration on this pro * Contributed Equally