Assessment of Genetic Fidelity of Micropropagated Bacopa monnieri Plantlets Using ISSR Marker Assay

Krishna Mohan C., Abdul Kareem V.K., Rajasekharan* P.E.

Abstract


Bacopa monnieri (Linn.) Pennel referred as Brahmi is a small creeping herb, growing in wet damp and marshy places throughout India. In the present study, single nodal explants were collected from field growing B. monnieri and inoculated on MS (Murashige and Skoog) media with or without plant growth regulators (PGRs). A large number of plants were regenerated from cultures grown on both MS basal medium and MS+PGRs. In vitro cultures developed were conserved in both standard culture condition (SCC) and reduced culture condition (RCC). Certain morphological changes observed in those cultures grown on MS with NAA (β-Naphthalene acetic acid) and IBA (Indole-3-butyric acid) from those grown on MS basal medium. Inter Simple Sequence Repeat (ISSR) marker assay was employed to validate the genetic fidelity of these regenerated plantlets upto ten passages. Fifteen ISSR primers generated a total of 57 amplicons and out of which only one polymorphic band was generated and remaining all were monomorphic bands. The dendrogram analysis (Unweight Pair-group Method with Arithmetic Averages-UPGMA) revealed that there was no genetic inconsistency exists among the sub-cultured plantlets on MS basal medium incubated at both SCC and RCC and hardened vitro plants from its mother plant. But the cultures grown on MS with NAA and IBA showed a very little variation (linkage distance of 1.8 units). The results confirmed the clonal fidelity of the tissue culture-raised B. monnieri plantlets and corroborated the fact that multiplication through nodal cuttings using MS basal medium is the safest mode for multiplication to get true-to-type plants.

Keywords


Bacopa monnieri; Genetic Fidelity; ISSR marker





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