Abstract
The antineoplastic herb, Catharanthus roseus is a classified high-value low-volume medicinal herb which is in global attention of scientific research for modulation of its monoterpenoid indole alkaloids (MIA) pathway through genetic engineering. These secondary metabolites are generally stored in specific types of structures/compartments due to their cytotoxic nature and designated roles in plant defense response. However, their presence can hinder the genetic engineering process used to develop transgenic plants through de novo morphogenesis and regeneration of plants from cultured cells/tissues and hence, it always remained a critical impediment in transgenic research in C. roseus. The pre-plasmolysis treatment of leaf explants can help to tackle the recalcitrant nature of leaf explant and can support the direct regeneration response by ex-osmosis that minimizes the concentration of alkaloids. Therefore, this study was performed to chase the effect of osmotic conditions on recalcitrant leaves of C. roseus engaged in vitro plant regeneration and hypothesis of alkaloids ex-osmosis is confirmed by HPLC analysis.
Highlights
Catharanthus roseus is an important medicinal plant due to its property to synthesize two anticancer phytomolecules monoterpenoid indole alkaloids (MIAs) vinblastine and vincristine [10, 11]
De novo morphogenesis and regeneration of plants from cultured cells/tissues has always remained a critical impediment in transgenic research in Catharanthus roseus
To further improve the shoot bud regeneration response obtained on direct regeneration medium, the effect of a pre-plasmolytic treatment on leaf explant before subjecting them to a direct regeneration experiment was evaluated in the present study
Summary
Catharanthus roseus is an important medicinal plant due to its property to synthesize two anticancer phytomolecules monoterpenoid indole alkaloids (MIAs) vinblastine and vincristine [10, 11]. Suspension and hairy root culture systems failed to enhance antineoplastic MIAs due to their limited cellular-organ specific complexity, whereas C. roseus due to its recalcitrant nature was not found a very acquiescent system for whole plant genetic engineering through plant regeneration-based transgenic research [11]. Treating the leaves with pre-plasmolysis can help to confront their recalcitrant nature but can support the direct regeneration response by ex-osmosis of stored alkaloids and toxic molecules from leaves. This applied osmotic pressure will curtail the concentration of toxic phytomolecules inside the growing leaves and/or can influence the morphogenesis and organogenesis by quick uptake of plant growth promoters from culture media through an osmotic pull [14]. Acetonitrile to ammonium acetate (100 mM, pH 7.3) (50:50) was used and detection was done at 254 nm
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