Abstract
Finger millet [Eleusine coracana (L.) Gaertn.] is an important cereal because of its mineral-nutrition value. With the increasing demand, there is a pressing need to conserve it through biotechnological approaches. High-frequency somatic embryogenesis from seed-derived callus of E. coracana was developed on Murashige–Skoog (MS) medium supplemented with a combination of auxins [Indole-3-acetic acid (IAA), 2,4-Dichlorophenoxy acetic acid (2,4-D)] and cytokinins [6-Benzylaminopurine (BAP), kinetin (KN)] in different concentrations, ranging from 0.1 to 5.0 mg L−1. Seeds cultured on this medium produced three different types of primary callus. Type I callus was very compact and dark brown, type II callus was light brownish and type III callus appeared whitish and light brown. All three types of calli had differential proliferation responses. Type II compact brown calli were obtained on the MS medium supplemented with 1.0 and 1.5 mg 2,4-Dichlorophenoxy acetic acid L−1 and 0.5 mg kinetin L−1. Friable yellowish embryogenic calli with a large number of somatic embryos were developed within 60 days after being transferred to auxins and cytokinin (1.0 and 1.5 mg 2,4-Dichlorophenoxy acetic acid L−1 and 0.5 mg Kinetin L−1) along with 200 mg casein hydrolysate L−1. Germination of somatic embryos on a half-strength MS medium supplemented with 0.1% Kinetin led to the development of healthy plantlets within 30 days. Genetic fingerprinting using random amplified polymorphic DNA (RAPD) revealed high levels of genetic fidelity. The study provides methods and hormonal concentrations required to develop somatic embryos in E. coracana for its genetic improvement and conservation.
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Abbreviations
- RAPD:
-
Random amplified polymorphic DNA
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- NAA:
-
1-Naphthalene acetic acid
- IAA:
-
Indole-3-acetic acid
- BAP:
-
6-Benzylaminopurine
- KN:
-
Kinetin
- CH:
-
Casein hydrolysate
- Glu:
-
Glutamine
- TDZ:
-
Thidiazuron
- EtBr:
-
Ethidium bromide
- NaoAc:
-
Sodium acetate
- EDTA:
-
Ethylenediaminetetraacetic acid
- CTAB:
-
Cetyl trimethyl ammonium bromide
- TAE:
-
Tris–acetate-EDTA
- FAO:
-
Food and Agriculture Organization
- SDS:
-
Sodium dodecyl sulfate
- MS media:
-
Murashige and Skoog media
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Acknowledgements
The authors thank Gandhi Krishi Vignan Kendra, Bangalore for providing finger millet seed samples of various genotypes. We also acknowledge Jawaharlal Nehru Memorial Fund, New Delhi and Department of Science & Technology, Promotion of University Research and Scientific Excellence (DST-PURSE), New Delhi, and Rashtriya Uchchattar Shiksha Abhiyan (RUSA) Ministry of Human Resource Development, Government of India for their financial support. We thank Dr. Ramu S Vemanna, Regional Centre for Biotechnology for the revision of the research paper.
Funding
This work was supported by Jawaharlal Nehru Memorial Fund, New Delhi (SU-1/124/2018-19/92) and Department of Science & Technology, Promotion of University Research and Scientific Excellence (DST-PURSE), New Delhi for their financial support.
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JV set up the experiment, carried out the entire experimental work, and analyzed the data. VR assisted the experiment. TS helped in preparing manuscript draft and interpretation of data. CS and GD (Research Guide) guided in setting up the experiment, data analysis, and manuscript preparation. All authors have read and approved the final manuscript.
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Venkatesan, J., Ramu, V., Sethuraman, T. et al. Assessing the genetic fidelity of somatic embryo-derived plantlets of finger millet by random amplified polymorphic DNA analysis. Biotechnol Lett 44, 1379–1387 (2022). https://doi.org/10.1007/s10529-022-03305-3
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DOI: https://doi.org/10.1007/s10529-022-03305-3