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IBBR publication #2022

Somatic embryogenesis of muskmelon (Cucumis melo L.) and genetic stability assessment of regenerants using flow cytometry and ISSR markers

Raji MR, Lotfi M, Tohidfar M, Zahedi B, Carra A, Abbate L, Carimi F

Protoplasma 170 (3): 115. (2018)
doi: 10.1007/s00709-017-1194-9

A new protocol for in vitro regeneration through direct somatic embryogenesis for two muskmelon cultivars (Cucumis melo L., ’Mashhadi’ and ’Eivanaki’) is reported. Somatic embryos were obtained culturing 4 and 8-day-old cotyledons, seeds and hypocotyls on MS medium supplemented with three different hormonal combinations never tested so far for melon (NOA+TDZ, NOA+BAP and 2.4-D+4-CPPU). Results were compared with those obtained when explants were cultivated in the presence of 2.4-D+BAP, previously used on melon. Embryogenesis occurred more successfully in 4-day-old cotyledons and seeds than hypocotyls and 8-day-old cotyledons. The best result was achieved with NOA+BAP. Genotypes significantly affected embryogenesis. The number of embryos in ’Eivanaki’ was significantly higher than ’Mashhadi’. Embryo proliferation when explants were maintained in jars (9.3%) was found to be higher compared to petri dishes. For the first time, genetic stability of melon regenerated plants was evaluated using ISSR markers. Polymerase chain reaction (PCR) products demonstrated a total of 102 well-resolved bands, and regenerants were 93% similar compared to the mother plant. Somaclonal changes during embryogenesis were evaluated by flow cytometry, showing 91% of the same patterns in regenerated plants. The results suggest that the new hormone components are effective when applied for in vitro embryogenesis of muskmelon as they show a high frequency in regeneration and genetic homogeneity.

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