Arifullah Mohammed

In Vitro Production of Echioidinin, 7-O-Methywogonin from Callus Cultures of Andrographis lineata and Their Cytotoxicity on Cancer Cells

Andrographis lineata is an herbal medicinal plant used in traditional medicine as a substitute for Andrographis paniculata. Here, using mature leaf explants of A. lineata we demonstrate for the first time the callus induction established on MS medium containing 1.0 mg l–1 IAA. Dried callus was subjected to solvent extraction with acetone. Further the acetone residue was separated by silica gel column chromatography, crystallized and characterized on the basis of nuclear magnetic resonance (proton and c13) and liquid chromatographic mass spectroscopy. This analysis revealed the occurrence of two known flavones namely, 7-O-methylwogonin (MW) and Echioidinin (ED). Furthermore, these compounds were tested for their cytotoxicity against leukemic cell line, CEM. We identify that ED and MW induced cytotoxicity in a time- and concentration-dependent manner. Further increase in the LDH release upon treatment with ED and MW further confirmed our cytotoxicity results against leukemic cell line. Strikingly, MW was more potent than ED when compared by trypan blue and MTT assays. Our results recapitulate the utility of callus cultures for the production of plant specific bioactive secondary metabolites instead of using wild plants. Together, our in vitro studies provide new insights of A. lineata callus cultures serving as a source for cancer chemotherapeutic agents.

In vitro plant regeneration, flowering and fruiting from nodal explants of Andrographis lineata nees (Acanthaceae)

In the present study, in vitro propagation of tribal endemic medicinal plant, Andrographis lineata has been established using mature nodal explants. High frequency of regeneration (91.4%) was achieved on MS medium containing BA (3.0 mg L–1) along with IAA (0.2 mg L–1). Strikingly, irrespective of the season and collection period, we observed axillary flower induction and fruit formation from the above in vitro cultures supplemented with BA and NAA. Transition from the vegetative to the reproductive phase occurred within 2 months of culture, and importantly was influenced by factors such as sucrose and cytokinins. In vitro-regenerated flowers were morphologically identical to in vivo flowers. Furthermore, our scanning electron microscopic studies revealed that pollen external morphology of both in vitro and in vivo flowers were similar. The flowers self-fertilized and produced fruits in vitro. Elongated shoots rooted well on half-strength MS basal medium, and were successfully acclimatized to the garden conditions. Altogether, our established protocol can be utilized in plant breeding for the purpose of ex situ conservation, quick flowering, and fruit set.

In vitro callus induction and plantlet regeneration of Indian red wood, Soymida febrifuga A. Juss (Roxb.)

were screened for their callus induction and morphogenic potential by culturing on Murashige and Skoog (MS) medium supplemented with naphthalene acetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), indole3acetic acid (IAA) alone or in combination with benzyladenine (BA), kinetin (KN), 2-iP (2-isopentynyl adenine), coconut milk. Multiple shoots were regenerated from cotyledonary node, shoot tip explants derived callus by one stage process where caulogenesis occurred on the primary callus. However cotyledons, leaves and root segments gave rise to adventitious shoots from callus by two stage method of indirect organogenesis. High frequency of shoot differentiation was observed from cotyledon and leaf on MS medium fortified with BA (8.9 to 13.3