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Anther culture for the production of haploid and doubled haploids in Jatropha curcas L. and its hybrids

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Abstract

Jatropha curcas L. has recently received considerable attention for the production of sustainable and affordable biofuels. Large scale cultivation of Jatropha requires homozygous parental genotypes to develop sustainable high yielding hybrids. Doubled haploid culture has been shown to offer the shortest method of producing homozygous plants and to permit greater selection efficiency since all the genes are fixed in a homozygous stage. In the present study, an efficient in vitro method for plant regeneration via anther culture of Jatropha was developed. Efforts were taken to induce callus from anthers of immature buds of seven elite genotypes and hybrids of Jatropha. Highest percent callus (77%) induction was achieved from anthers cultured on MS medium supplemented with BA (1.0 mg/l) and Picloram (1.0 mg/l), but callus obtained on media with BA (1.0 mg/l) and 2,4-D (1.0 mg/l), Kinetin (0.5 mg/l) and 2,4-D (2.0 mg/l), and BA (0.5 mg/l) and NAA (2.0 mg/l) showed regeneration of plants on medium containing BA (2.0 mg/l), Kinetin (0.5 mg/l) and NAA (0.5 mg/l). Rooting was achieved in 90% of the elongated shoots on half-strength MS medium with IBA (2.0 mg/l). All in vitro anther-derived plants were transferred to a greenhouse with 100% success in primary hardening and around 85% in secondary hardening. Plants established in the field are growing well as normal plants. Molecular and flow cytometry analyses of embryogenic callus revealed 5.7% and 3% samples are haploid and doubled haploid, respectively.

Key message

Anther culture protocol is developed for generation of haploids and doubled haploids in Jatropha curcas and its hybrids.

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Abbreviations

MS:

Murashige and Skoog

BA:

6-Benzyl amino purine

Kn:

Kinetin

NAA:

Alpha naphthalene acetic acid

2,4-D:

2,4-Dichlorophenoxyacetic acid

IBA:

Indole-3-butyric acid

DH:

Doubled haploid

SSR:

Simple sequence repeats

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Acknowledgement

The authors gratefully acknowledge the encouragement and constant support of the leadership Dr. Makarand Phadke and Dr. Ajit Sapre at the Reliance Industries Limited to carry out the research work. The authors are thankful to Dr. Saakshi Jalali for reviewing the manuscript.

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Authors and Affiliations

  1. Agronomy Biotechnology, Reliance Technology Group, Reliance Industries Ltd, Reliance Corporate Park, Thane Belapur Road, Ghansoli, Navi Mumbai, 400 701, India

    Neeta S. Madan, Savarimuthu Arockiasamy, J. V. Narasimham, Manoj Patil, Vijay Yepuri & Purbasha Sarkar

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  1. Neeta S. Madan
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  2. Savarimuthu Arockiasamy
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  3. J. V. Narasimham
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  4. Manoj Patil
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Contributions

AS and JVN conceived idea, designed the experiments and reviewed and edited manuscript. NM, MP and AS conducted Anther culture experiments; NM wrote and edited the manuscript. AS Supervised the entire study. VY conducted molecular marker analysis of samples and edited the manuscript. PS did optical microscopy of flower buds.

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Correspondence to Savarimuthu Arockiasamy.

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Communicated by Maria Antonietta Germanà.

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Madan, N.S., Arockiasamy, S., Narasimham, J.V. et al. Anther culture for the production of haploid and doubled haploids in Jatropha curcas L. and its hybrids. Plant Cell Tiss Organ Cult 138, 181–192 (2019). https://doi.org/10.1007/s11240-019-01616-4

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