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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 138, Issue 1, pp 181–192 | Cite as

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

  • Neeta S. Madan
  • Savarimuthu ArockiasamyEmail author
  • J. V. Narasimham
  • Manoj Patil
  • Vijay Yepuri
  • Purbasha Sarkar
Original Article

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.

Keywords

Jatropha Anther culture Callus Regeneration Haploid In vitro flowering Ploidy 

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

Notes

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.

Author contribution

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.

Compliance with ethical standards

Conflict of interest

The authors declare that they do not have any conflict of interest.

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© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Agronomy Biotechnology, Reliance Technology GroupReliance Industries LtdNavi MumbaiIndia

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