Shoot regeneration from cotyledonary leaf explants of Jatropha curcas: a biodiesel plant

Abstract

A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds were transferred to MS medium containing 10 lM kinetin (Kn), 4.5 lM BAP, and 5.5 lM a-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 lM BAP and 8.5 lM IAA was found to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing different concentrations and combinations of IBA, IAA, and NAA for 4 days, followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l -1 activated charcoal. Elongated shoot treated with 15 lM IBA, 5.7 lM IAA, and 11 lM NAA resulted in highest percent rooting. The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement of J. curcas through genetic modification.

Plant regeneration of non-toxic Jatropha curcas—impacts of plant growth regulators, source and type of explants

“…The in vitro and field-grown mature leaf explants, in vitro and glasshouse-grown seedlings cotyledonary leaf explants were cultured on MS medium supplemented with 3% sucrose and various concentrations of thidiazuron (TDZ) or BAP individually and in combination with 5 μM indole-3-butyric acid (IBA) to find the optimum culture medium for regeneration of shoot bud (Table 1) Kumar and Reddy 2010;Kumar et al 2010a). Leaf explants were inoculated on to the medium in 200×38 mm culture tubes (Borosil, India).…”

“…The regenerated shoot buds were transferred on to MS medium supplemented with 3% sucrose, 10 μM kinetin (Kn), 4.5 μM BAP and 5.5 μM α-naphthaleneacetic acid (NAA) for 4 weeks for shoot proliferation Kumar and Reddy 2010;Kumar et al 2010a). Shoots were individually separated and further tested for their elongation on MS medium supplemented with 3% sucrose and different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA and IBA (Table 3).…”

“…44 filter paper for 4 days (Fig. 1i) Kumar and Reddy 2010;Kumar et al 2010a). This auxins treated elongated shoots were trans-ferred to growth regulators free half strength MS solid medium supplemented with 0.25 mg/L activated charcoal.…”

“…To meet the large-scale demand and ensure easy supply of this elite material, there is a need to develop mass multiplication techniques. Attempts have been made to regenerate toxic variety of J. curcas (Sujatha and Mukta 1996;Rajore and Batra 2007;Jha et al 2007;Deore and Johnson 2008;Misra et al 2010;Kumar and Reddy 2010;Kumar et al 2010a). However, there is very few reports on non-toxic variety of J. curcas where regeneration was obtained from leaf with the presence of intermediary callus (Sujatha et al 2005) and petiole (Kumar et al 2010b(Kumar et al , 2011.…”

“…However, there is very few reports on non-toxic variety of J. curcas where regeneration was obtained from leaf with the presence of intermediary callus (Sujatha et al 2005) and petiole (Kumar et al 2010b(Kumar et al , 2011. It has also been reported that regeneration in J. curcas is highly genotype dependent (da Camara Machado et al 1997;Kumar 2008;Kumar and Reddy 2010;Kumar et al 2010a). The objective of this study was to develop a direct plant regeneration method from mature and cotyledonary leaf explants of non-toxic J. curcas for subsequent use in mass multiplication and genetic improvement.…”