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The non-edible plant Jatropha curcas L. is one of the most promising feedstock for sustainable biodiesel production as it is not a source of edible vegetable oils, produces high amounts of oil (approx. 30-60% in dry seeds) and does not require high-cost maintenance. However, as with other undomesticated crops, the cultivation of J. curcas presents several drawbacks, such as low productivity and susceptibility to pests. Hence, varietal improvement by genetic engineering is essential if J. curcas is to become a viable alternative source of biodiesel. There is to date no well-established and efficient transformation system for J. curcas. In this study, we tested various physical wounding treatments, such as sonication and sandvortexing, with the aim of developing an efficient Agrobacterium-mediated transformation for J. curcas. The highest stable transformation rate (53%) was achieved when explants were subjected to 1 min of sonication followed by 9 min of shaking in Agrobacterium suspension. The transformation frequency achieved using this protocol is the highest yet reported for J. curcas.

Abstracte seed oil of jatropha (Jatropha curcas L.) is a source of biodiesel fuel. Although jatropha can grow in semiarid lands unsuitable for the food production, its oil productivity in such conditions is unsatisfactory at present. erefore, it is desirable to improve the oil productivity of jatropha even in semi-arid lands by enhancing its drought tolerance. Genetic engineering is promising to dramatically improve plant traits. Although we previously reported a transformation method, which involves wounding of tissue explants in order to increase the chance of Agrobacterium infection, for jatropha, it remains a challenge to enhance the shoot regeneration and root induction processes. Here, we report the generation of three kinds of transgenic jatropha plants in an attempt to improve their drought tolerance. e rst one overexpresses the PPAT gene, which encodes an enzyme that catalyzes the CoA biosynthetic pathway; the second overexpresses the NF-YB gene, which encodes a subunit of the NF-Y transcription factor; and the last overexpresses the GSMT and DMT genes, which encode enzymes that catalyze production of glycine betaine. We also report a modi ed protocol that improves the e ciency of shoot regeneration and root induction in transgenic jatropha plantlets.

The efficient transformation of plants with large DNA molecules containing a set of useful genes would provide vast possibilities for the genetic improvement of agricultural as well as nonagricultural plants. The development of the bioactive beads (BABs) transformation method has proven useful for introduction of large DNA molecules into plant cells. In this chapter, the BABs transformation method used for the transformation of a 100-kb BAC DNA construct containing wheat genes into rice will be presented. Furthermore, the improved production method for BABs will be described. With the conventional method for producing BABs, the bead size varies, and the larger beads tend to carry fewer DNA molecules than the smaller beads. Thus, in order to facilitate the preparation of BABs with more uniform sizes, a simple set-up -composed of a sine wave sound generator and microsyringe pump was fabricated. Using this bead-maker set-up, uniform and smaller beads could be produced which enhance the transformation efficiency.