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.
Adebowale, K. O., Adedire, C. O. (2006) Chemical composition and insecticidal properties of the underutilized Jatropha curcas seed oil. Afr. J. Biotechnol. 5, 901–906.
Attaya, A. S., Geelen, D., Belal, A. E. H. (2012) Progress in Jatropha curcas tissue culture. Am-Eurasian J. Sustain. Agric. 6, 6–13.
Deore, A. C., Johnson, T. S. (2008) High-frequency plant regeneration from leaf-disc cultures of Jatropha curcas L.: an important biodiesel plant. Plant Biotech Rep. 2, 10–15.
Hamilton, C. M., Frary, A., Lewis, C. (1996) Stable transfer of intact high molecular weight DNA into plant chromosomes. Proc. Natl. Acad. Sci. 93, 9975–9979.
Höfgen, R., Willmitzer, L. (1988) Storage of competent cells for Agrobacterium transformation. Nucleic Acids Res. 16, 9877.
Jaganath, B., Subramanyam, K., Mayavan, S. (2014) An efficient in planta transformation of Jatropha curcas (L.) and multiplication of transformed plants through in vivo grafting. Protoplasm. 251, 591–601.
Jefferson, R. A. (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5, 387–405.
Jefferson, R. A., Kavanagh, T. A., Bevan, M. W. (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. E. M. B. O. J. 6, 3901–3907.
Liberalino, A. A. A., Bambirra, E. A., Moraes-Santos, T. (1988) Jatropha curcas L. seeds: chemical analysis and toxicity. Arq. Biol. Technol. 31, 539–550.
Khemkladngoen, N., Cartagena, J. A., Fukui, K. (2011) Physical wounding-assisted Agrobacteriummediated transformation of juvenile cotyledons of a biodiesel-producing plant, Jatropha curcas L. Plant Biotechnol. Rep. 5, 235–243.
Kumar, N., Anand, K. G. V., Reddy, M. P. (2011) In vitro regeneration from petiole explants of nontoxic Jatropha curcas. Ind. Crops Prod. 33, 146–151.
Kumar, N., Reddy, M. P. (2012) Thidiazuron (TDZ) induced plant regeneration from cotyledonary petiole explants of elite genotypes of Jatropha curcas: a candidate biodiesel plant. Ind. Crops Prod. 39, 62–68.
Liberalino, A. A. A., Bambirra, E. A., Moraes-Santos, T. (1988) Jatropha curcas L. seeds: chemical analysis and toxicity. Arquivos de biologia e tecnologi. 31, 539–550.
Liu, Y., Lu, J., Zhu, H. (2016) Efficient culture protocol for plant regeneration from cotyledonary petiole explants of Jatropha curcas L. Biotechnol. Biotec. Eq. 30, 907–914.
Liu, Y., Tong, X., Hui, W. (2015) Efficient culture protocol for plant regeneration from petiole explants of physiologically mature trees of Jatropha curcas L. Biotechnol. Biotec. Eq. 29, 479–488.
Liu, Y., Yin, X. G., Zhu, H. B. (2016) An efficient protocol for inducing regeneration in physic Nut (Jatropha curcas L.). Bangladesh J. Botan. 45, 87–93.
Liu, Y., Yu, L., Fu, Y. L. (2012) Development of an in vitro grafting method for the enhancement of growth of isolated shoots and buds in soybean (Glycine max L.). Biomed. Eng. Biotechnol. 1, 1003–1006.
Liu, Y., Yu, L., Zhang, Q. (2013) High concentration short duration treatment of benzyladenine stimulates adventitious bud regeneration from hypocotyl explants in soybean. Adv. Mater. Res. 647, 331–337.
Liu, Z. L., Liu, Y., Yang, Y. S. (2016) An Agrobacterium - mediated genetic transformation method of Jatropha curcas. Patent filed China and PCT. Authorization No. ZL201410275476.6, 2016-05-25. (in Chinese).
Lodhi, M. A., Ye, G. N., Weeden, N. F. (1994) A simple and efficient method for DNA extraction from grapevine cultivars and Vitis species. Plant Mol. Bio. Rep. 12, 6–13.
Lu W. (2003) Induction of callus from Jatropha curcas and rapid propagation. Chin. J. Appl. Environ. Biol. 9, 127–130.
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 15, 473–497.
Ötvös, K., Pasternak, T. P., Miskolczi, P. (2005) Nitric oxide is required for, and promotes auxinmediated activation of, cell division and embryogenic cell formation but does not influence cell cycle progression in alfalfa cell cultures. The Plant J. 43, 849–860.
Petri, C., Wang, H., Alburquerque, N. (2008) Agrobacterium-mediated transformation of apricot (Prunus armeniaca L.) leaf explants. Plant Cell Rep. 27, 1317–1324.
Petřivalský, M., Vaníčková, P., Ryzí, M. (2012) The effects of reactive nitrogen and oxygen species on the regeneration and growth of cucumber cells from isolated protoplasts. Plant Cell Tiss. Org. Cult. 108, 237–249.
Wei, Q., Lu, W. D., Liao, Y. (2004) Plant regeneration from epicotyl explant of Jatropha curcas. J. Plant Physiol. Mol. Biol. 30, 475–478.
Zhou, L., Ni, E., Yang, J. (2013) Rice OsGL1-6 is involved in leaf cuticular wax accumulation and drought resistance. PloS One 8, e65139.
About this article
Cite this article
Liu, Y., Liu, G., Yang, Y. et al. Establishment of an Efficient Plant Regeneration Culture Protocol and Achievement of Successful Genetic Transformation in Jatropha curcas L.. BIOLOGIA FUTURA 68, 428–442 (2017). https://doi.org/10.1556/018.68.2017.4.8
- Jatropha curcas
- Petiole explants
- Plant regeneration
- Genetic transformation