Genetic fidelity assessment of encapsulated in vitro tissues of Bacopa monnieri after 6 months of storage by using ISSR and RAPD markers

Authors: JOE VIRGIN LARGIA MUTHIAH, Karutha Pandian Shunmugiah, RAMESH MANIKANDAN

Abstract: The present study has investigated the efficiency of regrowth of Bacopa monnieri (L.) Pennell after 6 months of storage along with its clonal fidelity checking. Shoot tips and nodal segments of in vitro grown plantlets were encapsulated and stored at different temperatures and the highest regeneration was achieved by storing shoot tips at 4 °C for a period of 6 months. The response of shoot tips was found to be much better when compared to nodal segments in the case of forming healthy shoots and roots. The genetic stability of regrown plantlets was analysed by using RAPD and ISSR markers. Twenty ISSR primers produced 130 clear and reproducible amplicons, of which 125 bands were monomorphic. A total of 115 bands were produced by 25 RAPD markers with 94% monomorphism. This corresponds to a very low genetic variation pertaining to the clonal nature of the regrown plantlets with that of the mother plant. By means of HPLC biochemical analysis, bacoside A content in the regrown plants was also found to be comparable with that of the mother plant. Therefore, the protocol derived can be efficiently utilised for storing the encapsulated shoot tips of B. monnieri for about 6 months.

Keywords: Encapsulation, storage, regrowth, genetic fidelity, ISSR, RAPD

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