Effect of Different Media and Mannitol Concentrations on Growth and Development of Vandopsis lissochiloides (Gaudich.) Pfitz. under Slow Growth Conditions

In vitro conservation of orchid germplasm provides an effective technique for ex situ conservation of orchid diversity. In this study, an efficient protocol for in vitro conservation of Vandopsis lissochiloides (Gaudich.) Pfitz. plantlet under slow growth conditions was investigated. Plantlets were cultured on different strength of Vacin and Went medium (½VW and ¼VW) supplemented with different concentrations of mannitol (0, 2, 4, 6 and 8%), sucrose (0 and 3%) and 50 g/L potato extract, 150 mL/L coconut water. The cultures were incubated at 25±2 °C and maintained under 20 µmol/m2s light intensity for 24 weeks without subculture. At the end of preservation period, the plantlets were subcultured to fresh medium for growth recovery. The results found that the highest leaf number per plantlet could be observed on ¼VW medium without adding sucrose and mannitol while the highest root number per plantlet was found on ½VW added with 3% sucrose without adding mannitol after 24 weeks of in vitro storage. The results showed that the maximum number of leaves (5.8 leaves) and roots (5.0 roots) of preserved plantlets were produced on ¼VW medium without adding sucrose and mannitol. Therefore, ¼VW medium without adding sucrose and mannitol was the best minimum growth conditions for medium-term storage of V. lissochiloides plantlets.

Influence of Organic Supplements on Shoot Multiplication Efficiency of Phaius tankervilleae var. alba

The influence of organic supplements on growth and multiplication efficiency of Phaius tankervilleae var. alba seedlings was investigated. 12 week-old seedlings were cultured on half-strength semi-solid Murashige and Skoog (MS) medium supplemented with 30 g/L sucrose, 8 g/L agar and various concentrations of coconut water (0, 50, 100, 150 and 200 mL/L) combined with potato extract (0, 25 and 50 g/L) and the pH was adjusted to 5.8 prior to autoclaving. The cultures were then kept under constant photoperiod (16 h light: 8 h dark) at 25 ± 2 °C for 12 weeks. The highest number of shoots (3.0 shoots/explant) was obtained when cultured on the medium added with 50 ml/L coconut water and 50 g/L potato extract whereas the highest number of leaves (5.9 leaves/explant) and roots (6.1 roots/explant) could receive on the medium supplemented with 150 ml/L coconut water and 50 g/L potato extract. with 150 ml/L coconut water and 50 g/L potato extract. Additionally, plantlets of P. tankervilleae var. alba were transferred to grow into seven different substrates i.e. soil, sand, coconut husk chip, soil-sand mix (1: 1), soil-coconut husk chip mix (1: 1), sand-coconut husk chip mix (1: 1) and soil-sand-coconut husk chip mix (1: 1: 1) for four weeks. The results found that acclimatized plants showed 100% of survivals when sand, coconut husk chip and sand-coconut husk chip mix are used as substrates. The number of leaves induced by sand-coconut husk chip mix was significantly higher than that planted in other substrates (P > 0.05). Meanwhile, no significant difference in new shoot formation among these substrates was observed (P < 0.05). This precursory developing protocol was likely to be applied for more large scale of plant production as well as conservation of germplasm of this orchid species.

An Effect of Organic Supplements on Stimulating Growth of Vanda and Mokara Seedlings in Tissue Culture

This study aimed to investigate effect of different organic supplements on growth of Vanda and Mokara seedlings. Vanda and Mokara seedlings approximately 0.2 and 0.3 cm. in height were sub-cultured onto VW supplemented with 150 ml/L coconut water, 100 g/L potato extract, 100 g/L ‘Gros Michel’ banana (AAA group) and 100 g/L ‘Namwa’ banana (ABB group). The explants were sub-cultured onto the same medium every month for 3 months. The best medium increased stem height to 0.52 and 0.44 Cm. in Vanda and Mokara respectively was supplemented with coconut water. The maximum fresh weight of Vanda (0.59 g) was found on medium supplemented with ‘Gros Michel’ banana while Mokara cultured on medium supplemented with Potato extract had the maximum fresh weight (0.27 g) and number of roots (5.20 roots/shoot) statistically different (p≤ 0.05) to other treatments. However, Vanda cultured on medium supplemented with ‘Namwa’ banana had the maximum number of roots (3.80 roots/shoot). Our results suggested that growth of different orchid genera was responded diversely to different organic supplements.