The present study aims to identify the diverse endophytic fungi residing in the roots of Dendrobium moniliforme and their role in plant growth and development. Nine endophytic fungi were isolated from the root sections and characterized by molecular technique. Quantification of the indole acetic acid (IAA) compound by these endophytes was done. Further, Chemical profiling of R11 and R13 fungi was done by Gas Chromatography-Mass Spectroscopy (GC-MS). Asymbiotic seed derived protocorms of Rhynchostylis retusa was used for the plant growth assay to investigate the growth promoting activities of the fungal elicitor prepared from the isolated fungi from D. moniliforme. Among the isolated fungi, the relative dominant fungus was Fusarium sp. The R13 and R6 fungi were identified only at the genus level which concludes the fungi are of new species or strain. The indole acetic acid production was relatively higher in R10. Bioactive compound diversity was observed in the organic extract of R11 and R6. The presence of phenolic compound and essential oil suggest their contribution for the antimicrobial and antioxidant properties to their host plant, D. moniliforme. The plant growth assay result concluded, the fungal elicitor prepared from R10, Colletotrichum alatae was the best among all other for the plant growth activities.
The Vanda pumila is a monopodial orchid with beautiful flowers that are native to Thailand but now found across South Asia. The immature seeds of Vanda pumila were used for in vitro culture and then the protocorms developed were used as explants for seedling development and mass propagation. Protocorms were cultured on 1/2 MS (Murashige and Skoog, 1962) medium fortified separately with Kinetin (Kn), 6-Benzyl amino purine (BAP) and Gibberellic Acid (GA3) each in different concentrations as (0.5 mg/L, 1.0 mg/L and 2.0 mg/L) well as each on each concentrations of each medium supplemented with 5% and 10% coconut water (CW) respectively. The greatest number of shoots (9.50 ± 0.29 shoots per culture) was developed on 1/2 MS medium fortified with 1.0 mg/L Kn plus 10% CW and the longest shoots (0.78 ± 0.07 cm per culture) developed on 1/2 MS medium fortified with 2.0 mg/L BAP plus 10% CW. The shoots derived from protocorms were then developed on 1/2 MS medium fortified with three different rooting hormones viz. Indole-3-acetic acid (IAA), Indole-3-butyric acid (IBA) and α-Naphthalene acetic acid (NAA), each in four concentrations (0.5 mg/L, 1.0 mg/L, 1.5 mg/L and 2.0 mg/L) as well as 1.0 mg/L of each hormone supplemented with 10% CW. The 1/2 MS medium fortified with 0.5 mg/L IAA was found to be the most effective condition for the development of maximum number of root (5 ± 0.0 roots per culture) and root length (0.93 ± 0.07 cm). Hence, the present study could be useful for standardizing the protocol for mass propagation of the endangered orchid V. pumila.
The immature seeds of Dendrobiumchryseum, asympodial epiphytic orchid with yellow flowers, were cultured in vitro, and the resultant protocorms were used as explants for seedling development. Protocorms were cultured on½ M.S. medium fortified with Kinetin (Kn), 6-Benzylaminopurine (BAP), and Gibberellic Acid (GA3) in three concentrations (0.5mg/l, 1.0mg/land 2.0 mg/l) both alone and supplemented with 5% and 10% coconut water (C.W.). The highest number of shootsofD. chryseum developed on ½ - M.S. medium fortified with 2.0mg/lofKn and10% C.W. and the longest shootsdeveloped on ½ M.S. media fortified with 1.0mg/lGA3, and 10% C.W. The shoot derived from protocorms were placed in ½ M.S. medium fortified with three different rooting hormones, Indole -3- acetic acid (IAA), Indole -3-butyric acid (IBA) and α-Naphthalene acetic acid (NAA) in different concentrations alone as well as with each 1.0mg/l hormone combined with 10% C.W. The most effective of these media was ½ M.S. medium fortified with 1.5 mg/l IAA for rooting as well as for the production of longest roots. The present study could be useful for standardizing the protocol for mass propagation of the endangered orchid Dendrobiumchryseum.
From the medicinal orchid Dendrobium chryseum Rolfe, which is used in traditional and folk Chinese medicine, the protocorms were raised in Murashige and Skoog (MS) media in three strengths, full strength (FMS), half strength (1/2 MS), and quarter strength (1/4 MS), with or without the phytohormones 6-benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA) and coconut water (CW). The comparative cytotoxic activities of the wild and in vitro-raised protocorms were evaluated in human cervical carcinoma (HeLa) and human glioblastoma (U251) cell lines by MTT assay. In in vivo and in vitro, the methanol extracts of D. chryseum showed significant cytotoxic activities. Significant growth inhibition (%) and potent IC50 values were demonstrated in HeLa cell lines (49.79% (210.5 μg/mL) for in vitro-raised Dendrobium chryseum (DCT) versus 46.97% (226.5 μg/mL) for wild Dendrobium chryseum (DCW)). Similarly, activities against U251 cell lines exhibited also significant inhibition (28.76% (612.54 μg/mL) for DCW and 17.15% (1059.92 μg/mL) for DCT). The cytotoxic activities of both, wild and tissue-cultured samples, were superior in HeLa cells. In U251 cells, the wild sample was more active than the tissue-cultured one with a moderate cytotoxic effect. Hence, protocorm culture may therefore be a promising future tool for producing pharmacologically bioactive compounds in medicinal orchids. Such sustainable technology approach will minimize the pressure on the natural population of threatened but commercially important medicinal orchids.
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