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Comparison of in vitro regeneration pathways in Punica granatum L.

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Abstract

When cotyledonary explants, excised from in vitro germinated seedlings, of pomegranate (Punica granatum L.) were incubated on solid Murashige and Skoog (1962) medium supplemented with 21 μM naptheleneacetic acid (NAA) and 9 μM 6-benzyladenine (BA), 80% of explants developed callus. A high frequency of shoot organogensis was obtained when explants were incubated on MS medium supplemented with 8 μM BA, 6 μM NAA, and 6 μM giberrellic acid (GA3). However, adding 24 μM silver nitrate (AgNO3) to this medium markedly enhanced shoot regeneration frequency (63%) and mean number of shoots per explant (11.26) and length of shoots (2.22 cm). Highest frequency of in vitro rooting, mean number of roots/shoot (4.32), and mean root length (2.71 cm) were obtained when regenerated shoots were transferred to half-strength MS medium supplemented with 0.02% activated charcoal. Well-rooted plantlets were acclimatized, and then transferred to soil medium. Moreover, when zygotic embryos of P. granatum, excised from seeds collected at 16 weeks following full bloom, were incubated on MS medium containing 30 g l−1 sucrose, 15% coconut water, 21 μM NAA, and 9 μM BA, they developed the highest frequency of embryogenic callus, clumps with globular embryos, and mean number of both globular and heart-shaped embryos per callus clump. Subjecting zygotic embryo explants to six-week dark incubation period was essential for embryogenic callus induction, and these were subsequently transferred to 16 h photoperiod for further growth and development of somatic embryos. Germination of somatic embryos was observed when these were transferred to MS medium was supplemented with 60 g l−1 sucrose.

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Authors and Affiliations

  1. Department of Biotechnology, Dr. Y.S. Parmar University of Horticulture and Forestry, Solan, HP, 173230, India

    Kamlesh Kanwar, Jomy Joseph & Raj Deepika

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  1. Kamlesh Kanwar
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  2. Jomy Joseph
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  3. Raj Deepika
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Correspondence to Kamlesh Kanwar.

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Kanwar, K., Joseph, J. & Deepika, R. Comparison of in vitro regeneration pathways in Punica granatum L.. Plant Cell Tiss Organ Cult 100, 199–207 (2010). https://doi.org/10.1007/s11240-009-9637-4

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