Elsevier

Industrial Crops and Products

Volume 39, September 2012, Pages 62-68
Industrial Crops and Products

Thidiazuron (TDZ) induced plant regeneration from cotyledonary petiole explants of elite genotypes of Jatropha curcas: A candidate biodiesel plant

https://doi.org/10.1016/j.indcrop.2012.02.011Get rights and content

Abstract

Jatropha curcas, the energy plant has attained great attention in recent years because of its biodiesel production potential and medicinal value. This makes it imperative to search for techniques for its rapid propagation. In the present investigation an efficient and reproducible method for plant regeneration through direct shoot bud induction from cotyledonary petiole explants of elite genotypes (CSMCRI-JC-1, CSMCRI-JC-2 and CSMCRI-JC-3) of J. curcas was developed. The best shoot buds induction (51.19%) and number of shoot buds (9.75) per explant was observed when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 μM TDZ after 6 weeks. The induced shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM 6-benzyl aminopurine (BAP) and 5.5 μM α-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 μM BAP and 8.5 μM IAA was found to be best combination for shoot elongation and 3.01–3.61 cm elongation was achieved after 6 weeks. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. Orientation (horizontal or vertical) and source (in vitro or in vivo) of explants also significantly influenced plant regeneration. The elongated shoots could be rooted on half-strength MS medium supplemented with 0.25 mg/L activated charcoal and different concentrations and combinations of IBA, IAA and NAA. Half strength of MS medium with 5 μM IBA, 5.7 μM IAA and 11 μM NAA was found to be best for rooting. The rooted plants could be established in soil with more than 90% survival rate.

Highlights

Jatropha curcas, the energy plant has attained great attention in recent years because of its biodiesel production potential and medicinal value. ► An efficient method for plant regeneration through direct organogenesis from cotyledonary petiole explants of elite genotypes of J. curcas was developed. ► TDZ is most potent cytokinin for high-frequency plant regeneration.

Introduction

Tree-borne oilseeds are the best and potential alternative to mitigate the current and future energy crisis and also to transform the vast stretches of wasteland into green oil fields. The potential sources identified so far include Jatropha curcas, Pongamia pinnata, Madhuca latifolia, Azadirachta indica, Calophyllum inophyllum and Simarouba glauca. Among these J. curcas emerges as the most promising tree-borne oilseed on the basis of adaptability to wide range of edapho-climatic conditions, high oil content coupled with the suitability of Jatropha oil as a source of biodiesel. J. curcas, a multipurpose large shrub belongs to the family Euphorbiaceae, is native to South America and widely distributed in South and Central America, Africa and Asia. Biodiesel prepared from J. curcas has been sucessfully tested in both mobile and stationary engines without modification in any of the engine parts. Now there is a surge of interest in J. curcas as a biodiesel “miracle tree” to help alleviate the energy crisis and generate income in rural areas of developing countries (Francis et al., 2005).

Commercial plantations of this important crop are raised through seedlings and stem cuttings. The seed viability and rate of germination are low, and quality seed screening is another laborious task thus, propagation through seed may not provide quality planting material for sustainable use. It was also observed that huge quantities of seed is being used for raising of planting material, if any alternative propagation technique is developed the seed can be diverted for biodiesel preparation. Propagation can be also carried out by stem cutting but the limitation in generation of large scale planting material is (a) availability of sufficient quantity of material and (b) propagation is seasonal. Thus, conventional propagation through seeds is not reliable and vegetative cuttings are inadequate to meet the demand of large scale quality planting material. Therefore, J. curcas improvement programme by modern methods of agrobiotechnology are of interest worldwide. This has increased the importance of developing tissue culture methods to facilitate large scale production of quality planting material and for the improvement of the species using genetic engineering technique. Attempts have been made to regenerate J. curcas (Sujatha and Mukta, 1996, Wei et al., 2004, Sujatha et al., 2005, Rajore and Batra, 2007, Jha et al., 2007, Kumar, 2009, Kumar and Reddy, 2010, Kumar et al., 2010a, Kumar et al., 2010b, Kumar et al., 2010c, Kumar et al., 2011a, Kumar et al., 2011b) either through callus mediated regeneration or direct shoot morphogenesis. In all the above reports, the frequency of regeneration has been low and unsatisfactory. It is also reported that regeneration in J. curcas is highly genotype dependent (da Camara Machado et al., 1997, Kumar et al., 2008, Kumar and Reddy, 2010, Kumar et al., 2010a, Kumar et al., 2011b). Keeping the economical importance of J. curcas in mind and critical analysis of earlier reports, the objective of this study was to develop an in vitro regeneration method without formation of intervening callus in J. curcas for mass propagation and genetic improvement. This study also compared the regeneration efficiency of in vitro and in vivo cotyledonary petiole explants.

Section snippets

Plant material and source of explant

The seeds of elite genotypes (CSMCRI-JC-1, CSMCRI-JC-2 and CSMCRI-JC-3) of J. curcas were obtained from Central Salt & Marine Chemicals Research Institute (CSMCRI) experimental plantation on wastelands at Chorvadla, India (21°75″N, 72°14″E) for the present study. Seedcoats were removed and then surface sterilized with 0.1% mercuric chloride (HgCl2) for 15 min and rinsed five times in sterile distilled water. The sterilized decoated seeds were germinated on growth regulators free MS liquid medium

Effect of TDZ on shoot bud induction

The concentration of TDZ in the medium significantly influenced the response of shoot bud induction irrespective of genotype studied. The percentage of induction of shoot buds and the number of induced shoot buds per explant were directly proportional to the concentration of TDZ. Of the different concentrations of TDZ tested, the highest percentage of shoot bud induction (66.97%) and highest number of induced shoot buds (13.76) per explant were observed in the presence of 9.08 μM TDZ, among the

Discussion

An efficient and reproducible method for the plant regeneration of elite genotypes of J. curcas plants has been developed. The method employed direct regeneration of shoots from the in vitro and in vivo cotyledonary petiole explants, without the formation of an intervening callus using a MS containing TDZ. TThe concentration of TDZ in the medium, orientation, source of explants and genotype significantly influenced the regeneration response. Huetteman and Preece (1993) reported that TDZ is a

Acknowledgements

The authors gratefully acknowledge the Council of Scientific and Industrial Research, New Delhi for financial assistance. Statistical data analysis support from Mr. Keyur Shah and Mr. Amritpal Singh, Assistant professor, Anand Agricultural University, Anand is thankfully acknowledged.

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