As. Pac. J. Mol. Biol. & Biotech., Jan 2014 Vol. 1, 152-163
In vitro morphogenesis and RAPD analysis of Justicia tranquebariensis L.f.-an important medicinal plant
S. Raji1, M. Ayyanar2, P. Ponmanickam1 and T. Rajagopal1,3
1Post Graduate Department of Biotechnology, Ayya Nadar Janaki Ammal College (Autonomous), Sivakasi-26 124, Tamil Nadu, India.
2Post Graduate and Research Department of Botany, Pachaiyappa's College, Chennai-600 030, Tamil Nadu, India.
3Department of Zoology, Thiagarajar College (Autonomous), Madurai- 625 009, Tamil Nadu, India.
* Author for correspondence: T. Rajagopal
Post Graduate Department of Biotechnology, Ayya Nadar Janaki Ammal College (Autonomous), Sivakasi-626 124, Tamil Nadu, India.
Also: Department of Zoology, Thiagarajar College (Autonomous), Madurai- 625 009, Tamil Nadu, India.
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Abstract.
Justicia tranquebariensis L.f. is well known medicinal plants used in the treatment of respiratory problems. In the present study, a simple and reproducible protocol for rapid in vitro multiplication from nodal explants has been developed and randomly amplified polymorphic DNA (RAPD) markers were used for the first time for the detection of genetic polymorphism in this medicinal herb from in vitro grown plants with mother plants. Nodal explants of J. tranquebariensis showed better multiple shoot initiation. Among the different concentrations of 6-benzylaminopurine (BA), kinetin (KN), indole-3-acetic acid (IAA) and naphthalene acetic acid (NAA) tested, the maximum percentage of response (80%) was recorded on MS medium supplemented with KN (2.5 mg/l). Rooting was best achieved (80%) on half strength MS medium amended with IAA (1.0 mg/l). The plantlets regenerated in vitro with well developed shoots and roots were successfully transferred and established in plastic cups and pots containing soil mixture simultaneously with a 70% survival rate. Calli were induced from nodal and leaf explants with different concentrations of 2,4-dichlorophenoxy acetic acid (2,4-D), BA, NAA and optimum callus biomass (60%) was observed from leaf explants cultured on MS medium supplemented with 2, 4-D (1.0 mg/l). Ten arbitrary decamer primers have been used to amplify genomic DNA from in vitro raised field material and mother plant. Further, the genetic fidelity of micropropagated plants and callus was assessed by RAPD analysis and it showed that the similarity among the in vitro regenerated shoots and variation with the callus regenerated shoots. The results suggested that the multiple shoot induction and regeneration were regulated by appropriate cytokinin/auxin ratios rather than their relative concentrations. High multiplication frequency and molecular stability ensures the efficacy of the protocol developed for production and conservation of this important medicinal herb.