Paudel Mukti Ram, Sharma Sujata, Joshi Pusp Raj, Pant Basant, Wagner Sven H, Gurung Pritam, Pant Krishna Kumar, Pant Bijaya
Central Department of Botany, Tribhuvan University, Kirtipur, Kathmandu, Bagmati, Nepal.
Annapurna Research Center, Annapurna Neurological Institute and Allied Sciences, Maitighar, Kathmandu, Bagmati, Nepal.
BMC Complement Med Ther. 2025 Feb 18;25(1):61. doi: 10.1186/s12906-025-04810-4.
Dendrobium amoenum is known for its aesthetic and medicinal values but it is threatened due to loss of wild resources. Plant tissue culture promotes wild resource protection and paves the way for secondary metabolite production. In this study, protocorms developed via in-vitro seed cultivation were used for bioactive secondary metabolite production. The objectives of this study were to evaluate total phenolic and flavonoid contents, to identify the bioactive secondary metabolites, to explore the antioxidants and cytotoxic properties of in-vitro-derived protocorms extracts of D. amoenum.
Seeds of D. amoenum were cultivated on 10% coconut water, 0.25 and 0.5 mg/L BAP supplemented full-strength and half-strength MS medium to produce protocorms for the isolation of bioactive components. A distinct yellow fraction (DAYF), light-green fraction (DALGF), green fraction (DAGF), and dark-green fraction (DADGF) were obtained from methanol extract on a methanol-based Sephadex LH-20 column. The total phenol and flavonoid contents along with the antioxidant and cytotoxic properties of the fractions were evaluated. The compounds in active DAYF were identified using a GC-MS.
On a full-strength solid MS medium supplemented with 10% coconut water, approximately 95% of the seeds grew into protocorms, while 88.33% did so on a full-strength liquid MS medium. The DAYF had a total phenol content of 206.38 μg of GAE and a total flavonoid content of 101.88 μg of QE. Owing to these high contents, the DAYF inhibited 50% of the DPPH free radicals at a concentration of 63.73 μg/ml. Similarly, it also reduced the growth of HeLa cells by 50% at 67.03 μg/ml and U2OS cells by 50% at 207.40 μg/ml, while it was nontoxic to normal human epithelium cells. Bioactive phenolic compounds 2-methoxy-4-vinylphenol (1), 3,4-dimethoxy-phenol (2), 2-methoxy-4-(1-propenyl)-phenol (3), 2,6-dimethoxy-4-(2-propenyl)-phenol (4), 3-methoxy-1,2-benzenediol (5) were identified in the DAYF.
Protocorms of D. amoenum could serve as sources of bioactive secondary metabolites highlighting their potential in alternative medicine.
滇南石斛以其美学价值和药用价值而闻名，但由于野生资源的丧失而受到威胁。植物组织培养有助于保护野生资源，并为次生代谢产物的生产铺平道路。在本研究中，通过体外种子培养发育的原球茎被用于生物活性次生代谢产物的生产。本研究的目的是评估总酚和黄酮含量，鉴定生物活性次生代谢产物，探索滇南石斛体外衍生原球茎提取物的抗氧化和细胞毒性特性。
将滇南石斛种子接种在添加10%椰子水、0.25和0.5mg/L BAP的全强度和半强度MS培养基上，培养以产生用于分离生物活性成分的原球茎。从基于甲醇的Sephadex LH-20柱上的甲醇提取物中获得了一个明显的黄色馏分（DAYF）、浅绿色馏分（DALGF）、绿色馏分（DAGF）和深绿色馏分（DADGF）。评估了各馏分的总酚和黄酮含量以及抗氧化和细胞毒性特性。使用GC-MS鉴定活性DAYF中的化合物。
在添加10%椰子水的全强度固体MS培养基上，约95%的种子长成原球茎，而在全强度液体MS培养基上这一比例为88.33%。DAYF的总酚含量为206.38μg GAE，总黄酮含量为101.88μg QE。由于这些高含量，DAYF在浓度为63.73μg/ml时抑制了50%的DPPH自由基。同样，它在67.03μg/ml时使HeLa细胞生长减少50%，在207.40μg/ml时使U2OS细胞生长减少50%，而对正常人上皮细胞无毒。在DAYF中鉴定出生物活性酚类化合物2-甲氧基-4-乙烯基苯酚（1）、3,4-二甲氧基苯酚（2）、2-甲氧基-4-（1-丙烯基）苯酚（3）、2,6-二甲氧基-4-（2-丙烯基）苯酚（4）、3-甲氧基-1,2-苯二酚（5）。
滇南石斛原球茎可作为生物活性次生代谢产物的来源，突出了它们在替代医学中的潜力。
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