标题：Regeneration of Cytologically Stable Plants Through Dedifferentiation, Redifferentiation, and Artificial Seeds in Spathoglottis plicata Blume. (Orchidaceae)
摘要：Spathoglottis plicataBlume. is a horticulturally important vulnerable ground orchid with beautiful flowers blooming round the year. High-frequency protocorm-like body (PLB) formation was established via callus culture from vegetative tissues ofin vitrogerminated seedlings ofS. plicata. Media containing MS salts and Gamborg's B5 vitamins supplemented with 1.0 mg⋅L−12,4-dichlorophenoxyacetic acid (2,4-D), 3.0 mg⋅L−1α-naphthaleneacetic acid (NAA), 1.0 mg⋅L−1kinetin (KIN), and 10% (v/v) ‘Aloe veragel’ (AvG) were effective in fragile calli induction. A maximum of (22.3 ± 0.52) PLBs were induced from about 250 mg callus within 45–55 days in the presence of 2.0 mg⋅L−1NAA and 3.0 mg⋅L−16-benzylaminopurine (BAP). Briefly, 3.0% sodium alginate was found to be most suitable for the formation of an appropriate shape and good germination rates (86.7%) of artificial seeds. Out of three different temperatures (4, 15, and 24 °C), the best result was achieved at 4 °C with 66.7% germinability even after 90 days of storage. Plantlets were acclimatized with 86.6% survival rate and 76.3% of these plants produced flowers within 12–15 months of field transfer. Chromosomal studies revealed cytological stability of all regenerants containing 2n = 40 chromosomes as in the parental plants. The present protocol can be applied reliably for the purposes of large-scale commercial propagation and short-term conservation of this orchid.
