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:

E-ISSN: 2229-4473.
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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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Author Information

Department of Botany, Kakatiya University, Warangal, India