Alam M Firoz, Sinha Pinaki, Hakim M Lokman
Department of Botany, Rajshahi University, Rajshahi, Bangladesh.
Methods Mol Biol. 2010;589:21-8. doi: 10.1007/978-1-60327-114-1_3.
For high frequency clonal propagation of Vanda teres, nodal segments are cultured on VW medium supplemented with 2% sucrose, 2 mg/L Kinetin, 0.5 mg/L NAA, 2 g/L peptone, 1 g/L activated charcoal and 2.2 g/L gelrite. The cultures are incubated at 24 +/- 2 degrees C under fluorescent light 50 micromol/m(2)/s for a 16 h photoperiod per day. The PLBs (protocorm like bodies) are induced within 12 weeks of culture and are subcultured to proliferate on the fresh nutrient culture medium for 8 weeks. The clumps of the PLBs are dissected and cultured on VW medium containing 2% sucrose, 15% coconut water (CW), 2 g/L peptone, 150 mg/L L-glutamine and 1 g/L activated charcoal. The PLB sections elongate to form shoots, and new PLBs are induced from the base within 8 weeks of culture. For plantlets formation, the shoots are cultured on VW medium amended with 2% sucrose, 15% CW, 2 g/L peptone, 1 g/L activated charcoal, 50 g/L banana pulp and 1 mg/L Indole-3-butyric acid (IBA). The regenerated plants are acclimatized and cultivated in the nursery, where they bloom within 3 years.
对于万代兰的高频克隆繁殖，将茎节段接种于添加了2%蔗糖、2 mg/L激动素、0.5 mg/L萘乙酸、2 g/L蛋白胨、1 g/L活性炭和2.2 g/L吉兰糖胶的VW培养基上。培养物在24±2℃、光照强度为50 μmol/m²/s的荧光灯下培养，每天光照16小时。原球茎状小体（PLBs）在培养12周内诱导形成，并转接至新鲜的营养培养基上增殖8周。将PLBs团块切割后接种于含有2%蔗糖、15%椰汁（CW）、2 g/L蛋白胨、150 mg/L L-谷氨酰胺和1 g/L活性炭的VW培养基上。PLB切片伸长形成芽，培养8周内从基部诱导产生新的PLBs。为了形成小植株，将芽接种于添加了以下成分的VW培养基上：2%蔗糖、15% CW、2 g/L蛋白胨、1 g/L活性炭、50 g/L香蕉果肉和1 mg/L吲哚-3-丁酸（IBA）。再生植株驯化后在苗圃中栽培，3年内开花。
