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Year : 2022  |  Volume : 18  |  Issue : 77  |  Page : 207-215

Genetic clonal fidelity assessment of rhizome-derived micropropagated Acorus calamus L. – A medicinally important plant by random amplified polymorphic DNA and inter-simple sequence repeat markers

1 Department of Biotechnology, Manipur University, Canchipur, Imphal- 795003, Manipur, India
2 Department of Life Sciences, Presidency University, Kolkata, West Bengal, India
3 Department of Botany, Deen Dayal Upadhyaya College, University of Delhi, New Delhi, India

Correspondence Address:
Potshangbam Nongdam
Department of Biotechnology, School of Life Science, Manipur University, Canchipur, Manipur
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/pm.pm_408_21

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Background: Acorus calamus – a critical medicinal plant, is overexploited, leading to population reduction. Establishing an efficient in vitro protocol is essential for the large-scale production of genetically identical plants. Objectives: Development of fast and reliable in vitro regeneration protocol for A. calamus and clonal fidelity assessment of the regenerants using molecular markers. Materials and Methods: Plants were regenerated on Murashige and Skoog medium with different concentrations of growth regulators in two phases – shooting and rooting. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were employed to evaluate the genetic stability of in vitro clones. Results: 6 Benzylaminopurine (BAP) at 1.6 and 2.4 mgL−1 was effective for shoot induction, while root induction was superior in indole-3-butyric acid-incorporated medium at 2.5 mgL. Thirteen RAPD and 16 ISSR primers produced 59 and 96 clear, unambiguous, and reproducible bands, respectively. Both the markers revealed a high monomorphism of 96.79% and 95.63% among the regenerants. Nei's genetic distance analysis disclosed a close genetic association (0.000–0.068) among the genotypes. Conclusion: ISSR was better than RAPD markers in clonal fidelity assessment of the regenerants. The in vitro protocol developed is reliable and suitable for the rapid propagation of true-to-type A. calamus plants.

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