Abstract
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.
Highlights
Orchidaceae is a highly developed and extensively distributed monocotyledonous family consisting of terrestrial, saprophytic, and epiphytic species [1, 2]
They were inoculated on different strengths of Murashige and Skoog (MS) media (FMS, 1/2MS, and 1/4MS), FMS fortified with coconut water (CW) (5% and 10%) and MS medium supplemented with 0.5 mg/L BAP alone and in combination with 0.5 mg/L naphthaleneacetic acid (NAA), 1 mg/L BAP + 0.5 mg/L NAA, 1.5 mg/L BAP + 0.5 mg/L NAA, and 2 mg/L BAP + 0.5 mg/L NAA (Table 1)
E earliest response for in vitro seed germination was observed on 1/2 MS medium alone and on FMS medium supplemented with 5% and 10% CW which took three weeks of culture for germination (Table 2). is medium condition was found to be effective for the earlier development of protocorms which took four weeks of culture incubation (Table 1)
Summary
Orchidaceae is a highly developed and extensively distributed monocotyledonous family consisting of terrestrial, saprophytic, and epiphytic species [1, 2]. Dendrobium is the second largest genus of the family Orchidaceae in Nepal. Dendrobium chryseum is an epiphytic herb found throughout the western and eastern Himalayas at elevations from 1000 to 2150 m. It is popularly used in traditional and folk Chinese medicine for its antipyretic, eye benefiting, and immunomodulatory effects. While the chemical compound isolated from the stem of D. chryseum exhibited antioxidant activity [18], polysaccharides from the D. aurantiacum var denneanum (D. chryseum non Rolfe.) possess tumor-inhibitory and blood glucose-reducing effects in vivo [18].
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