In vitro seed germination, proliferation, and ISSR marker-based clonal fidelity analysis of Shorea tumbuggaia Roxb.: an endangered and high trade medicinal tree of Eastern Ghats

Abstract

An in vitro plant tissue culture approach provides a feasible alternative for extensive mass manufacturing and conservation of elite and critically endangered medicinal plant species. The presented work describes a proficient in vitro seed germination and proliferation procedure for the conservation of an endangered high trade medicinal tree, Shorea tumbuggaia Roxb. Given the precarious position of this species, effective conservation is highly indispensable for its continued existence, primarily for ecological reasons and also for forestry purposes as this tree species is a valuable source of timber and oleoresin. In the present study, over 90% in vitro seed germination was achieved. Multiple shoot formation was established from seedlings with a maximum of 4.60 ± 0.30 multiple shoots per explant using MS (Murashige and Skoog) medium supplemented with a combination of 6.66 μM 6-benzylaminopurine and 0.454 μM thidiazuron. Rooting of multiple shoot cultures of 85% was achieved on half strength MS medium supplemented with 2.24 μM indole 3-butyric acid with 6.48 ± 0.58 roots per plantlet. Rooted plantlets showed more than 70% acclimatization in potting conditions and were established successfully in a greenhouse, with morphological similarities to mother plants. Clonal fidelity assessment of micropropagated plants with ten inter-simple sequence repeat primers reproduced a total of 24 monomorphic bands. The presented work represents the first report of in vitro proliferation, conservation, and clonal fidelity assessment of tissue culture generated S. tumbuggaia plantlets.