High frequency in vitro plantlet regeneration in Solanum trilobatum L., an important ethno-medicinal plant and confirmation of genetic fidelity of R1 plantlets by using ISSR and RAPD markers

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Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Website:www.vegetosindia.org
Pub Email: contact@vegetosindia.org
Doi: 10.1007/s42535-019-00069-6
First Page: 508
Last Page: 520
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Keywords: Solanum trilobatum L. , Thidiazuron, Organogenesis, Acclimatization, ISSR, Genetic fidelity


Abstract

An efficient and reproducible protocol was developed for in vitro clonal propagation of Solanum trilobatum L., an ethno-medicinally important plant. Leaf, stem and cotyledon explants were used for callus induction and shoot regeneration via indirect organogenesis. Initially, maximum amounts (mg) of green friable callus (312.86 ± 0.50, 285.3 ± 0.40 and 305.13 ± 0.62) was induced from leaf, stem and cotyledon explants, respectively, on Murashige and Skoog (MS) medium supplemented with 13.57 μM L−1 of 2,4-dichloro phenoxy acetic acid (2,4-D) and 2.21 μM L−1 of 6-benzylaminopurine (BAP), after 4 weeks of culture. Highest mean number of shoots (69.2 ± 0.73, 34.1 ± 0.62 and 57.1 ± 0.62) were differentiated de novo from leaf, stem and cotyledon calluses, respectively, when cultured on MS medium amended with 2.27 μM L−1 of Thidiazuron (TDZ) and 2.68 μM L−1 of Naphthaleneacetic acid (NAA), after 4 weeks of culture. Maximum rooting response (97%) with mean number of roots (31.7 ± 0.61 roots per shoot) was observed from shoots when cultured on half strength MS medium supplemented with 4.90 μM L−1 of indole-3-butyric acid (IBA), after 4 weeks of culture. In vitro raised plantlets (R1) of S. trilobatum were hardened in plastic pots, acclimatized in green house and successfully transferred to field conditions with 82% survivability. ISSR and RAPD based PCR banding profile of the acclimated R1 plantlets was confirmed as synonymous to the mother plant.


Solanum trilobatum L. , Thidiazuron, Organogenesis, Acclimatization, ISSR, Genetic fidelity


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Acknowledgements

The first author is grateful to the UGC New Delhi for award of BSR fellowship. The authors are grateful to Prof. Sadanandam Abbagani, CSIR-Emeritus Scientist for his valuable suggestions. The authors gratefully acknowledge the Head, Department of Botany and Department of Biotechnology for providing research facilities. The authors grateful to UGC New Delhi for funds provided under SAP-DRS-III to Botany and SAP-DRS-II to Biotechnology Departments. The authors also grateful to DST-New Delhi for funds provided under DST-FIST to Botany and Biotechnology Departments. The authors thank Dr. Md. Mustafa for authenticating the plant material.


Author Information

Pendli Sreenu
Department of Botany, Kakatiya University, Warangal, India

Rohela Gulab Khan
Central Sericultural Research and Training Institute, Central Silk Board, Ministry of Textiles, Govt. of India, Pampore, India