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In vitro plant regeneration from cotyledonary explants of Cucumis melo L. var. cantalupensis and genetic stability evaluation using RAPD analysis

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Abstract

An efficient plant regeneration protocol useful in genetic studies for important breeding lines of cantaloupe melons was developed. Three cultivars (Charentais-T, Vedrantais and Isabelle) and three doubled-haploid melon lines (NAD, DH-L2 and DH-L6) were tested for their potential to regenerate under in vitro conditions. Cotyledon explants from 6-day-old seedlings were cultured on three Murashige and Skoog based medium supplemented with different combinations of 6-benzylaminopurine (BAP) and indoleacetic acid (IAA). After 5 weeks of culture all the genotypes were able to regenerate on all media, however the regeneration frequency varied widely among the cultivars and the medium combinations used. The Charentais-T cultivar developed the highest number of shoots/explant on media containing BAP 1.0-IAA 1.0 × 10−2 mg/L (4.1) and BAP 1.2-IAA 1.2 × 10−2 mg/L (4.4) while on medium supplemented with BAP 1.3-IAA 1.3 × 10−2 mg/L the Vedrantais genotype gave the best result (2.4). The work first highlighted a good morphogenetic response of the DH lines; among all the genotypes and combination used, NAD (65.6 %) and DH-L6 (85.4 and 50.0 %) had the highest yield of regenerated plants. The results confirmed that the regeneration competence was strongly affected by genotype. Plantlets with well-developed shoot and root systems were successfully acclimatized and exhibited normal morphology and growth characteristics. In order to confirm genetic stability, a PCR-based RAPD analysis was carried out using 20 decamer-primers. No polymorphic bands were observed among all the plants revealing the genetic homogeneity of the regenerants. Furthermore, the ploidy level of regenerated plants was confirmed by flow cytometry.

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Acknowledgments

The authors wish to thank Dr. Sara Sestili for her critical comments on the manuscript.

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  1. Consiglio per la Ricerca in Agricoltura e l’Analisi dell’Economia Agraria (CREA), Unità di Ricerca per l’Orticoltura, Via Salaria 1, 63077, Monsampolo Del Tronto, Italy

    Maria Silvia Sebastiani & Nadia Ficcadenti

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  1. Maria Silvia Sebastiani
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Correspondence to Maria Silvia Sebastiani.

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Sebastiani, M.S., Ficcadenti, N. In vitro plant regeneration from cotyledonary explants of Cucumis melo L. var. cantalupensis and genetic stability evaluation using RAPD analysis. Plant Cell Tiss Organ Cult 124, 69–79 (2016). https://doi.org/10.1007/s11240-015-0875-3

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