Articles
IN VITRO SHOOT REGENERATION FROM LEAF TISSUE OF APPLE (CULTIVAR “ORINE”): HIGH SHOOT PROLIFERATION USING CARRY OVER EFFECT OF TDZ
Article number
520_30
Pages
291 – 300
Language
Abstract
To develop an efficient in vitro system to regenerate shoots from the apple cultivar “Orine” (Malus x domestica Borkh.), several experiments were conducted using leaf tissue as explant. “Orine” is one of the leading apple cultivars in Japan.
Leaf segments were obtained from in vitro raised apple plants.
The orientation and maturity of the tissue on the regenerative capacity were investigated.
The explants were tested for shoot initiation on basic Murashige and Skoog (MS) medium supplemented with different combinations and concentrations of growth regulators.
Initiation of compact embryoid-like structures as well as slight callus formation was visible on the cut edges of the explant in presence of 20 μM TDZ (thidiazuron) and 0.1 μM NAA (naphthaleneacetic acid) in dark.
Among the other cytokinins tested TDZ has proven again its high cytokinin activity for in vitro manipulation of woody plant species.
Proliferation of the embryoid-like structures started when the cultures were transferred to light.
Inclusion of 1.4 μM gibberellic acid (GA3) in the medium promoted shoot elongation.
Proliferating shoot masses were subcultured and maintained on MS medium without growth regulators.
In fact, it was interesting to observe enhanced shoot proliferation in this medium, probably due to “carry over” effect of TDZ. Plants regenerated were transferred to ½ MS (sucrose 2%) with IBA (indolebutyric acid) at 1.25 μM for root development.
The rooted plants were acclimatized and placed in the greenhouse for further analysis of morphology.
Leaf segments were obtained from in vitro raised apple plants.
The orientation and maturity of the tissue on the regenerative capacity were investigated.
The explants were tested for shoot initiation on basic Murashige and Skoog (MS) medium supplemented with different combinations and concentrations of growth regulators.
Initiation of compact embryoid-like structures as well as slight callus formation was visible on the cut edges of the explant in presence of 20 μM TDZ (thidiazuron) and 0.1 μM NAA (naphthaleneacetic acid) in dark.
Among the other cytokinins tested TDZ has proven again its high cytokinin activity for in vitro manipulation of woody plant species.
Proliferation of the embryoid-like structures started when the cultures were transferred to light.
Inclusion of 1.4 μM gibberellic acid (GA3) in the medium promoted shoot elongation.
Proliferating shoot masses were subcultured and maintained on MS medium without growth regulators.
In fact, it was interesting to observe enhanced shoot proliferation in this medium, probably due to “carry over” effect of TDZ. Plants regenerated were transferred to ½ MS (sucrose 2%) with IBA (indolebutyric acid) at 1.25 μM for root development.
The rooted plants were acclimatized and placed in the greenhouse for further analysis of morphology.
Authors
N. Gamage, T. Nakanishi
Keywords
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