Two new benzylbenzoate glucosides from Curculigo orchioides.

An extract from in vitro cultures of Curculigo orchioides grown as bulbils in shake flasks, afforded two new glucosides of substituted benzylbenzoate - curculigoside C (3) and curculigoside D (4) - together with two known compounds - curculigoside A (1) and curculigoside B (2). Their structures were elucidated on the basis of spectral evidence, in particular by using 2D NMR methods. Their vasoactive properties were assessed in isolated rat aortic rings.

A method for large-scale multiplication of Curculigo orchioides through bulbil formation from leaf explant in shake flask culture.

An efficient method has been developed for large-scale multiplication of Curculigo orchioides (Hypoxidaceae), an endangered medicinal plant, through direct bulbil formation from leaf explants in shake flask cultures. Leaf-segments (7 x 10 mm) were cultured in B5 liquid medium containing KNO3 (200 mgNL-1), (NH4)2SO4 (50 mgNL-1), benzyl adenosine (2.2 microM), adenine (0.11 mM), indole butyric acid (1.0 microM) and polyvinyl pyrrolidone (250 mgL-1). About 95% of explants produced maximum number of bulbils (546/flask at 6 weeks growth) in the medium. Shake flask cultures yielded 2737 bulbils/L medium whereas static cultures yielded 624 bulbils/L medium. Germination of bulbils was maximum (90.62%) on agar-gelled B5 medium containing benzyl adenosine (2.2 microM) and gibberellic acid (3.5 microM). Plantlets developed in vitro were successfully transferred to soil with a high rate of survivability (90%) and were comparable to natural population in growth and vigour.

Changes in phospholipids, fatty acids, oxidative enzymes, phenolics and protein levels during growth of normal and habituated tissues of Cocculus pendulus.

Cultures of C. pendulus were maintained on hormone free and hormone supplemented (NAA 1.0 mg/l and kinetin 0.5 mg/l) Murashige and Skoog medium. During the growth period, hormone free cultures had higher phenolic content, polyphenol oxidase activity and less protein content, peroxidase and IAA oxidase activity. Activity of all the three oxidising enzymes and phenolic content were high at 16 days growth. Total lipid content was higher (2.7-folds at 15 days) in hormone free cultures. Phospholipid content of both cultures was not markedly dissimilar except PC and DGDG contents. Thus it is evidenced that both the tissues were similar metabolically.

Effect of anti-oxidants and adsorbents on tissue browning associated metabolism in Cocculus pendulus callus cultures.

Explants and callus of C. pendulus produced intense brown substances in the medium which caused necrosis. Various anti-oxidants (ascorbic acid, cysteine and dithiothreitol) and adsorbents (activated charcoal and polyvinyl pyrrolidone) were used in different concentrations to prevent browning of the tissues. These in MS medium affected differently the growth, colour and texture of the tissues. It was concluded that both peroxidase and phenolase were involved in the browning. Increased peroxidase activity and decreased phenolase activity were probably due to more peroxidative oxidation of phenols and unavailability of substrate for phenolase activity. This resulted in faster growth of tissues, which further reduced the phenolase activity.