Hybridization and in vitro culture of an orchid hybrid Ascocenda ‘Kangla’

The present study focuses on hybridization program involving two species belonging to two different vandaceous genera, viz., Ascocentrum ampullaceum (Roxb.) Schltr. var. auranticum, a narrow endemic orchid of Manipur and Vanda coerulea Griff., an endangered orchid of Appendix I of CITES (Convention on International Trade in Endangered Species of Wild Fauna and Flora), to synthesize the primary hybrid genus with intermediate and improved characters in the F₁ generation. Observations on the crossability in the present bigeneric cross between V. coerulea and A. auranticum had been achieved with 60% success when V. coerulea was taken as female parent. Murashige and Skoog (MS) basal medium at half-strength was effective for the development of the hybrid seedlings of V. coerulea × A. auranticum followed by Vacin and Went (VW) and Knudson C (KC) media. The best response of seedling growth was observed on MS medium at half-strength supplemented with 2.3 μM kinetin + 0.5 μM α-naphthalene acetic acid with maximum shoot height (2.7 cm), leaf number (4.6) and root number (4.1) after 150 days of inoculation. The survival percentage and growth performance of the seedlings were found to be higher (80% survival) in potting substrate consisting of brick:charcoal in the ratio 2:1 mulched with moss (Sphagnum sp.) than in potting substrate consisting of brick:charcoal:tree fern in the ratio 2:1:1. The first flowering was observed in the hybrid seedlings of V. coerulea × A. auranticum after 2 years of transfer to the ex vitro environment. Morphologically, the flowers differed from that of the parents clearly showing the success of the hybridization experiment. Registration of the hybrid has been made with the Royal Horticultural Society with the nomenclature Ascocenda ‘Kangla’ (No. T128725).     

Rapid regeneration of plants of Dendrobium lituiflorum Lindl. (Orchidaceae) by using banana extract

Effects of banana extract (BE) and 6-benzylaminopurine (BAP) were evaluated on asymbiotic seed germination and an early differentiation of protocorms and plant regeneration of Dendrobium lituiflorum Lindl. High percentage germination was achieved by culturing seeds on modified Knudson C medium supplemented with 10% (v/v) BE. Rapid regeneration was observed within 60 days of culture on 10% (v/v) BE supplemented KC medium where maximum percentage propagules showed development of leaves and root formation. Propagules on BAP supplemented KC medium showed no further development beyond one leaf stage. In another experiment, culture of shoots on 12.5% (v/v) BE supplemented KC medium led to multiplication, shoot elongation as well as vigorous rooting. Shoots cultured on 10 μM BAP supplemented MS medium showed maximum multiplication but these were stunted. Plants with well expanded deep green leaves and elongated roots from BE media were first hardened in vitro followed by ex vitro hardening on cocopeat:perlite (9:1) in the greenhouse conditions and exhibited 90% survival. The study emphasizes the role of BE as a natural additive at different stages of development from seed germination to plant regeneration.     

Micropropagation of orchids: A review on the potential of different explants

Orchids are among the most diverse of the flowering plant families, with over 800 described genera and 25,000 species. Orchids are prized for their beautiful long lasting flowers exhibiting an incredible range of diversity in size, shape and colour. Today growing orchids is more than just a hobby, it is an international business covering around 8% of the world floriculture trade and has the potential to alter the economic landscape of a country. Large-scale multiplication of exquisite and rare hybrids using tissue culture techniques has helped orchids occupy a position as one of the top ten cut flowers. As orchids are outbreeders, their propagation using seeds leads to the production of heterozygous plants. Hence, protocols providing regeneration from various vegetative parts of the plants are needed. Though orchid micropropagation has shown spectacular development in the recent years, the wide spread use of micropropagation is believed to be still limited due to problems like exudation of phenolics from explants, transplantation to field, somaclonal variation etc. We endeavour to include the major investigations on explant-based orchid tissue culture starting from the pioneering works of Rotor [Rotor, G., 1949. A method of vegetative propagation of Phalaenopsis species and hybrids. Am. Orchid Soc. Bull. 18, 738–739] followed by Morel [Morel, G., 1960. Producing virus-free cymbidiums. Am. Orchid Soc. Bull. 29, 495–497] and Wimber [Wimber, D.E., 1963. Clonal multiplication of cymbidiums through tissue culture of the shoot meristem. Am. Orchid Soc. Bull. 32, 105–107] to date.