Optimization of In Vitro Regeneration from Leaf Explants of Apple Cultivars Golden Delicious and Melrose

• Published in: HortScience Vol. 47; no. 8; pp. 1117 - 1122
• Main Authors: MITIC, Nevena, STANISIC, Mariana, MILOJEVIC, Jelena, TUBIC, Ljiljana, COSIC, Tatjana, NIKOLIC, Radomirka, NINKOVIC, Slavica, MILETIC, Rade
• Format: Journal Article
• Language: English
• Published: Alexandria, VA American Society for Horticultural Science 01-08-2012
• Subjects: Agronomy. Soil science and plant productions; Biological and medical sciences; Biotechnology; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; Genetics and breeding of economic plants; In vitro propagation: entire plant regeneration from tissues and cell cultures; Methods. Procedures. Technologies; Plant cells and fungal cells; Biotechnology; Horticulture; Rosaceae; Explant; Climacteric; Malus domestica; Temperate zone; Optimization; shoot multiplication; Dicotyledones; Angiospermae; Fruit tree; Incubation; pre-incubation in the dark; Pome fruit; Plant leaf; Micropropagation; Organogenesis; Regeneration; Gelling agent; Apple; Fruit crop; Vegetative apparatus; Spermatophyta; Shoot; Cultivar
• ISSN: 0018-5345; 2327-9834
• DOI: 10.21273/hortsci.47.8.1117

An efficient in vitro shoot regeneration method from leaf explants of apple cultivars Golden Delicious and Melrose by optimization of regeneration medium, explant type and orientation, dark pre-treatment, and gelling agent is presented. Murashige and Skoog’s regeneration medium containing 22 μM thidiazuron (TDZ) and 1.5 μM indole-3-butyric acid (IBA) (M2 medium) was superior for regeneration as well as for subsequent shoot multiplication in both cultivars, providing regeneration frequency of 95% or higher in the best combination with other factors. Pre-incubation in the dark proved to be an essential factor for regeneration. The use of agar as a gelling agent provides satisfactory regeneration frequency compared with media gelled with Phytagel TM . Leaf explants of cv. Melrose with adaxial surface in contact with M2 medium and those of cv. Golden Delicious orientated contrary regenerated the highest mean number of shoots per explant. Under optimal conditions, a maximal index of shoot-forming capacity of 11.44 and 6.30 for ‘Melrose’ and ‘Golden Delicious’, respectively, was achieved. Regenerated shoots were successfully rooted and acclimated ex vitro.