Establishment of an efficient plant regeneration culture protocol and achievement of successful genetic transformation in Jatropha curcas L.

Abstract

An efficient and reproducible protocol is described for shoot-bud regeneration and Agrobacterium tumefaciens-mediated genetic transformation of J. curcas. Treating the explants with high concentrations (5-120 mg/L) of TDZ for short durations (5-80 min) before inoculation culture increased significantly the regeneration frequency and improved the quality of the regenerated buds. The highest shoot-buds induction rate (87.35%) was achieved when petiole explants were treated with 20 mg/L TDZ solution for 20 min and inoculated on hormone-free MS medium for 30 days. Regenerated shoots of 0.5 cm or a little longer were isolated and grafted to seedling stocks of the same species, and then the grafted plantlets were planted on half-strength MS medium containing 0.1 mg/L IBA and 2 mg/L sodium nitroprusside (SNP). This grafting strategy was found to be very effective, to obtain that healthy grafted plantlets ready for acclimatization within 20 days. By the above mentioned protocol and with general Agrobacterium - mediated genetic transformation methods only 65 days were needed to obtain intact transgenic plants.

Effect of different factors on plant regeneration from leaf explants of Jatropha curcas L.

“…Due to the high regeneration efficiency of leaf explants, it might be introduced into the studies on genetic transformation via Agrobacterium ‐mediated or particle bombardment. Since both transformation methods are used to obtain transgenic plants, it is necessary to induce adventitious bud regeneration through efficient plant tissue culture method as prerequisite for acquiring transgenic regenerated plantlets (Franco et al., 2016; Joshi et al., 2011; Kajikawa et al., 2012; Kumar et al., 2010; Liu et al., 2017; Qu et al., 2012). Therefore, the efficient tissue culture method proposed in this paper would be applied to study the improvement of varieties and the function of key genes in J. curcas .…”

Biotechnological Research Progress in Jatropha, a Biodiesel-Yielding Plant

“…Kumar and Reddy (2012) reported that TDZ supplemented medium was optimal for regeneration of jatropha from petiole explants ( Figure 1 ), in which MS medium supplemented with 2.27 µM TDZ exhibited a 51.19 percent response [ 16 ]. Similarly, leaf explants, when exposed to 20 mg/L TDZ for 20 min, induced 65 to 87 percentage response for shooting when cultured on basal MS media [ 17 , 18 ]. N-(2-chloro-4-pyridyl)-N-phenyl urea (CPPU) was employed for the induction of shoots in addition to the most regularly used plant growth regulators.…”

Effects of Adventitious Buds Regeneration From Three Kinds of Explants in Jatropha Curcas

“…In order to induce shoot-buds elongation, the regenerated adventitious buds were transferred with the parent body (explants) to MS medium supplemented with 0.2 mg/L kinetin (KT), 0.25 mg/L indole-3-acetic acid (IAA), 0.5 mg/L 6-benzyladenine (6-BA) and with different concentrations of gibberellic acid (GA3) (0, 0.1, 0.2, 0.4, 0.6 and 0.8 mg/L) for 15 days [9,11].…”