Efficient culture protocol for plant regeneration from cotyledonary petiole explants ofJatropha curcasL.

Abstract

Yin (2016) Efficient culture protocol for plant regeneration from cotyledonary petiole explants of Jatrophacurcas L., Biotechnology & Biotechnological Equipment, 30:5, 907-914, DOI: 10.1080/13102818.2016 A high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from cotyledonary petiole explants of Jatropha curcas L. has been developed. The cotyledonary petiole explants of J. curcas cultured directly on medium supplemented with thidiazuron (TDZ) induce regeneration of poor quality shoot buds that have a low regeneration frequency. However, treating the explants with high concentrations (10À60 mg/L) of TDZ solution for certain time periods (5À80 min) significantly increased the regeneration frequency and improved the quality of the regenerated shoot buds. The best shoot buds induction (88.42%) and number of shoot buds (12.67) per explant were observed when in vitro explants were treated with 20 mg/L TDZ solution for 20 min before being transferred on hormone-free medium after 30 days. Regeneration was also influenced by the orientation (horizontal or vertical) of the explants on the medium, and by the origin of the cotyledonary petioles (in vitro or in vivo) used for the preparation of explants. We performed subsequent experiments for elongation and rooting of the regenerated shoot buds. Addition of L-arginine to the medium was conducive to the elongation of the shoot buds. A concentration of 7.5 mg/L L-arginine yielded the best results. The elongated shoots could initiate roots to become intact plantlets in half-strength Murashige and Skoog medium containing 0.1 mg/L indole-3-butyric acid. After acclimatization, these plantlets could be transplanted to the soil and the growth was normal. Therefore, application of the methods described here helped to increase plant regeneration efficiency.

High Concentration of Thidiazuron Stimulates Adventitious Bud Regeneration from Cotyledon Explants in Jatropha curcas

“…Our study suggested that cytokinin might be required for a short duration during induction of shoot-buds formation after high concentration of TDZ treatment.The success of experimental results given in this paper suggested that the induction of adventitious buds formation might not requires cytokinin during the whole culture period; when the process of cell division for the formation of adventitious buds was triggered, the presence of cytokinin was no longer necessary, and in the contrary, long presence as incorporated in the medium might have only negative effects. What's more, it has been well documented in many text books and academic journals that cytokinin has the effects of apical suppression and hinders the growth of roots [26,27].…”

“…Seeds coded M-19 of J. curcas were grown and collected from the farm in Haikou, Hainan province of China [27].…”

“…The percentage of induction of shoot buds and the number of shoot buds per explant were recorded after 30 days of culture. For shoot buds elongation, the regenerated shoot buds were transferred along with the mother tissues (explants) to MS medium supplemented with 0.4 mg/L gibberellic acid (GA3), 0.5 mg/L 6-benzylaminopurine (BA), 0.2 mg/L kinetin (KT) and 0.25 mg/L indole-3-acetic acid (IAA) (Sigma-Aldrich Co., St. Louis, MO, USA) [23,27]. The length of the elongated shoots was recorded after 15 days of culture.…”

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

“…Seeds of J. curcas coded M-19 [15] were surface-sterilized for 60 s with 75% (v/v) ethanol after removing the outer seed coat, immersed in 2% (v/v) sodium hypochlorite (NaClO) for 20 min and finally rinsed 5 times in sterile distilled water. The embryos were removed from the seed and incubated on hormone-free Murashige and Skoog salt (MS) medium [22].…”

Increase of Regeneration Efficiency in JATROPHA CURCAS by In Vitro Culture Methods

“…Elongated shoots (at least 10 mm) were cut from the maternal tissue and inoculated into fresh half-strength MS medium contained 0.1 mg/L indole-3-butyric acid (IBA) for 20 days of culture [9]. Plantlets were transplanted carefully to plastic bottles with soil and sterile sand at the ratio of 1:1 and placed lid with the transparent plastic film for 15 days of culture.…”