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Micropropagation of Paphiopedilum x Dalatense
Abstract
Paphiopedilum x dalatense is a beautiful orchid species with large flowers in variable colors and leaves covered with stripes and beautiful unseen mosaic spots. Recently, many people exploit this species, causing it becomes very rare. In this study, we studied the effects of various organic matter: potato, banana and tryptone, yeast powder, peptone on the growth and development of P. dalatense shoots as well as the effects of NAA and humic acid on in vitro rooting of this orchid were investigated. The research results showed that MS medium supplemented with 100 g/L banana in combination with 100 g/L potato (5,4 shoots/sample, 18,8 mm/shoot, 4,5 leaves/shoot, and shoots survival rate of 100%) or MS medium supplemented with 1 g/L peptone (4,19 shoots/sample, 15 mm/shoot, 4 leaves/bud, and 92% of shoots survival rate) were the best response for the shoot formation and development. In addition, the half strength MS culture medium supplemented with 1 mg/L NAA (5,2 leaves/sample, 4,6 roots/buds, 3,56 cm/root, and 100% rate for rooting) was the suitable medium for the in vitro rooting of P. dalatense. Being cultured on half strength MS medium supplemented with 2 mg/L humic acid, the rooting rate reached 100% with the greatest root number and the longest root (5 roots/shoots, 5,5 cm/root). The obtained results on the in vitro propagation on this orchid helps contribute to the conservation and increases the genotic pool of this precious wild orchid species, as well as the rapid multiplication of healthy plantlets serving the commercialization of precious orchid species.
... In particular, humic acid has antibacterial activity by inhibiting the growth of bacteria and fungi, thereby reducing mycotoxins (Islam et al., 2005). To date, there have been several studies on the investigation of the effect of humic acid on in vitro propagation of plants, for example, effects of auxins and humic acids on in vitro rooting of strawberry (Fragaria x ananassa Duch.) (Rzepka-Plevnes et al., 2011), the influence of humic acid (HA) on histological development, antioxidant enzyme changes and endogenous hormone levels during adventitious root formation in evergreen azalea microshoots (Elmongy et al., 2018) but only one for the orchid species Paphiopedilum x dalatense (Vinh et al., 2021). ...
... This may be a new direction for further studies to regenerate shoots from PLB in orchids of this genus. The results of the high regeneration rate and the number of shoots in this study are consistent with the results of the study on Paphiopedilum x dalatense (Vinh et al., 2021). In a study of R. imschootiana shoot multiplication derived from protocorm, MS medium supplemented with 1.5 mg/L KIN and 15% CW was observed to be the most suitable, specifically, shoot multiplication rate reached 15.33 shoots/explant after 12 weeks of culture (Dan et al., 2018). ...
... For R. imschootiana, when using humic acid, the quality of seedlings is good with dark green leaves, and strong root, and this will be a new direction in the propagation process of the genus Renanthera in the next research rounds. Vinh et al. (2021) in the Paphiopedilum x dalatense propagation study indicated that the growth and development of roots were maximum at 2.0 mg/L humic acid, which is consistent with our results. In addition, Mohamed et al. (2017) studied the propagation of evergreen Azalea and obtained the optimal rooting medium results when adding 1.0 mg/L of humic acid. ...
Renanthera imschootiana Rolfe is an extremely rare and endangered tropical epiphytic orchid. Studies on in vitro culture of R. imschootiana were conducted in order to conserve and increase the genetic pool of this wild orchid species. In this study, the effect of plant growth regulators (BA and NAA), ripe bananas in combination with potato, humic acid, and silver nanoparticles (AgNPs) were investigated to find a suitable condition for in vitro plant regeneration from R. imschootiana protocorm-like body (PLB). The results showed that after 45 days of culture, MS medium supplemented with 100 g ripe banana/liter and 100 g potato/liter was suitable for regenerating shoots from the protocorm-like body (PLB) (17.56 shoots/PLBs and 100.00% of shoot-regenerating PLBs); MS medium supplemented with 2.0 mg/L BA, 0.3 mg/L NAA and 4 ppm AgNPs was the most suitable for the growth of shoots after 60 days of culture (10.00 leaves/shoot and shoot length of 4.22 cm). The in vitro shoots were transferred to half-strength MS supplemented with humic acid and AgNPs to investigate the root formation of R. imschootiana. After 60 days of culture, the best rooting was obtained at 2.0 mg/L humic acid and 6 ppm AgNPs (8.33 roots/shoot; root length of 4.00 cm and 100.00% root formation). The findings suggest that the in vitro micropropagation from R. imschootiana protocorm-like body provides a useful alternative tool for the conservation of this endangered species.
... Increasing the NAA concentration to 1.5 mg/L inhibited root formation, reducing the number of roots. High concentrations of auxin induce cell division in plant tissues to form callus, thereby inhibiting root formation (Vinh et al., 2021). Similar results for root regeneration in 1/2 MS medium supplemented with 1.0 mg/L NAA for P. verticillatum shoots in vitro were reported by Bisht et al. (2012). ...
Polygonatum punctatum Royle ex Kunth is a rare medicinal plant species in Vietnam, with rhizomes used as a tonic for kidney yang deficiency, for treating joint pain, and for pain relief. Overexploitation of natural medicinal plants has driven the need for propagation techniques to meet increasing demand, compensate for habitat degradation, and conserve rare species. In this study, tissue culture techniques were applied to investigate the effects of different growth regulators on the in vitro regeneration of P. punctatum. The results showed that sterilizing young leaf and petiole samples with 0.1% HgCl2 for 6 minutes achieved optimal effectiveness. The MS medium supplemented with 1.5 mg/L BA and 0.5 mg/L 2,4-D was identified as the most suitable for inducing callus formation from young leaves and petioles. The MS medium supplemented with 2.0 mg/L BA and 0.2 mg/L NAA yielded the best results for shoot regeneration, while the 1/2 MS medium supplemented with 1.0 mg/L NAA was appropriate for root formation. The in vitro propagation results of P. punctatum contribute to the conservation and sustainable development of this valuable medicinal plant and aim towards the rapid multiplication of healthy plantlets for commercialization.
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