Abstract
Efficient in vitro propagation of medicinally important endangered plant C. borivilianum has been achieved through somatic embryogenesis. Solid embryogenic medium [Murashige and Skoog medium containing 1.79mM NH4NO3, 10.72mM KNO3, 1.13μM 2,4-dichlorophenoxyacetic acid, 7.38μM 2-isopentenyladenine and 0.76mM proline] supplemented with polyethylene glycol and sucrose (3% each), exhibited 1.88-fold increase in embryo maturation compared to embryogenic medium containing 3% sucrose. Liquid embryogenic medium supported better somatic embryo production and maturation. Highest total (79) and mature (cotyledonary stage) somatic embryos (38) as well as highest germination (57.5%) was observed at inoculum density of 0.4g/40ml of liquid medium. 5.86 pH level exhibited optimal growth, maturation and germination of somatic embryos. Random amplified polymorphic DNA (RAPD) analysis of C. borivilianum plants regenerated through somatic embryogenesis revealed that they were genetically similar to the mother plant. The protocol established in the present study can be used for rapid mass multiplication of C. borivilianum in bioreactor employing liquid medium.
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