Articles
Features of in vitro regenerated microshoots from various explants in three persimmon cultivars
Article number
1254_3
Pages
13 – 18
Language
English
Abstract
Persimmon is one of the most valuable subtropical fruit plants and different biotechnological methods have been used in order to improve vegetative propagation of new cultivars and valuable plant material conservation.
The objective of this study was to investigate in vitro morphogenic capacity of vegetative buds and leaf cuttings in persimmon cultivars Zolotistaya, Nikitskaya Bordovaya and Ukrainka bred in Nikita Botanical Gardens.
Shoot segments with buds were surface-sterilized with 70% ethanol (1 min), 0.3-0.4% Cl2 (15 min) and 1% Thimerosal (10 min), and contamination frequency decreased to 15-20%. The explants in the culture vessels were kept in a growth room at 25±1°C under 16-h photoperiod supplied by cool-white fluorescent lamps giving 37.5 μmol m‑2 s‑1. After 4-6 weeks of growing on MS medium supplemented with 13.30-22.20 µM BAP and 0.49-1.28 µM IBA, about 1-2 microshoots per explant were obtained for all the studied cultivars.
Multiplication rate up to 3-6 adventitious microshoot per explant were increased by subculture.
Addition of TDZ to culture medium allowed callus formation and microshoot differentiation in leaf cuttings from in vitro-grown persimmon microshoots.
Calli were formed on MS medium subculture with 3, 6, 9 and 12 µM TDZ. Adventitious buds and microshoots were regenerated after 4-6 weeks from small and protuberant structures formed on the green callus surface.
Histological analysis demonstrated that 6.0 μM TDZ stimulated the formation of numerous areas of active meristematic cells and apical meristem cells.
Leaf primordia in epidermal and subepidermal layers of callus in cultivar Ukrainka were formed.
By indirect regeneration from morphogenic callus, about 7-8 microshoots per explant were obtained for the cultivar Zolotistaya, 4-6 for Ukrainka and 2-3 for Nikitskaya Bordovaya. Subculture on MS medium supplemented with 1.0 μM TDZ was favorable for microshoot elongation.
The objective of this study was to investigate in vitro morphogenic capacity of vegetative buds and leaf cuttings in persimmon cultivars Zolotistaya, Nikitskaya Bordovaya and Ukrainka bred in Nikita Botanical Gardens.
Shoot segments with buds were surface-sterilized with 70% ethanol (1 min), 0.3-0.4% Cl2 (15 min) and 1% Thimerosal (10 min), and contamination frequency decreased to 15-20%. The explants in the culture vessels were kept in a growth room at 25±1°C under 16-h photoperiod supplied by cool-white fluorescent lamps giving 37.5 μmol m‑2 s‑1. After 4-6 weeks of growing on MS medium supplemented with 13.30-22.20 µM BAP and 0.49-1.28 µM IBA, about 1-2 microshoots per explant were obtained for all the studied cultivars.
Multiplication rate up to 3-6 adventitious microshoot per explant were increased by subculture.
Addition of TDZ to culture medium allowed callus formation and microshoot differentiation in leaf cuttings from in vitro-grown persimmon microshoots.
Calli were formed on MS medium subculture with 3, 6, 9 and 12 µM TDZ. Adventitious buds and microshoots were regenerated after 4-6 weeks from small and protuberant structures formed on the green callus surface.
Histological analysis demonstrated that 6.0 μM TDZ stimulated the formation of numerous areas of active meristematic cells and apical meristem cells.
Leaf primordia in epidermal and subepidermal layers of callus in cultivar Ukrainka were formed.
By indirect regeneration from morphogenic callus, about 7-8 microshoots per explant were obtained for the cultivar Zolotistaya, 4-6 for Ukrainka and 2-3 for Nikitskaya Bordovaya. Subculture on MS medium supplemented with 1.0 μM TDZ was favorable for microshoot elongation.
Authors
N.N. Ivanova, Т.N. Kuzmina, I.V. Mitrofanova
Keywords
Diospyros kaki, vegetative buds, leaf cutting, morphogenic capacity, callus, indirect regeneration
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