Articles
IN VITRO CALLUS INDUCTION AND PLANTLET REGENERATION PROTOCOL DEVELOPED FOR THE ORYZALIN TREATMENT OF ZAMIOCULCAS ZAMIIFOLIA (LODD.) ENGL. (ARACEAE)
Article number
813_26
Pages
201 – 208
Language
English
Abstract
A novel tissue culture protocol was developed for the oryzalin treatment of Zamioculcas zamiifolia (Lodd.) Engl. (Araceae) (Z. zamiifolia) callus tissue.
Leaflet and petiole explants were harvested from juvenile-like stock plants and disinfested with 95% ethanol, 0.65, 0.33 and 0.13% sodium hypochlorite.
Leaflet explants were trimmed to 1 x 1cm square and petioles trimmed to 2.5cm length.
Explants were cultured onto callus inducing medium composed of half strength MS macro- and micronutrients, half strength MS vitamins, 100mg l-1 myo-inositol, 0.2mg l-1 BA, 4mg l-1 2,4-D, 20g l-1 sucrose, and 3g l-1 gellan gum.
Cultures were transferred to fresh medium every 2 weeks, and stored in the dark at 25-27°C. Callus was observed on the explants about 4.5 weeks after cultures were initiated, and once a sufficient amount of callus had been produced, cultures were transferred to shoot induction medium composed of half strength MS macro- and micronutrients, half strength MS vitamins, 100mg l-1 myo-inositol, 1mg l-1 BA, 40g l-1 sucrose, and 3g l-1 gellan gum.
Cultures on shoot induction medium were kept in the light, and adventitious bud development was observed after 11 weeks on the medium.
Once the adventitious buds had elongated and the leaf sheath was about 2.5cm, cultures were transferred to shoot elongation medium with no plant growth regulators for further development.
Rooted plantlets, about 5cm in height, where then transferred to community pots in the greenhouse under 70% shade.
All plantlets transferred to the greenhouse developed normally.
The protocol developed was used for the oryzalin treatment of Z. zamiifolia callus in an experiment aimed at producing a tetraploid Z. zamiifolia plant in vitro.
Leaflet and petiole explants were harvested from juvenile-like stock plants and disinfested with 95% ethanol, 0.65, 0.33 and 0.13% sodium hypochlorite.
Leaflet explants were trimmed to 1 x 1cm square and petioles trimmed to 2.5cm length.
Explants were cultured onto callus inducing medium composed of half strength MS macro- and micronutrients, half strength MS vitamins, 100mg l-1 myo-inositol, 0.2mg l-1 BA, 4mg l-1 2,4-D, 20g l-1 sucrose, and 3g l-1 gellan gum.
Cultures were transferred to fresh medium every 2 weeks, and stored in the dark at 25-27°C. Callus was observed on the explants about 4.5 weeks after cultures were initiated, and once a sufficient amount of callus had been produced, cultures were transferred to shoot induction medium composed of half strength MS macro- and micronutrients, half strength MS vitamins, 100mg l-1 myo-inositol, 1mg l-1 BA, 40g l-1 sucrose, and 3g l-1 gellan gum.
Cultures on shoot induction medium were kept in the light, and adventitious bud development was observed after 11 weeks on the medium.
Once the adventitious buds had elongated and the leaf sheath was about 2.5cm, cultures were transferred to shoot elongation medium with no plant growth regulators for further development.
Rooted plantlets, about 5cm in height, where then transferred to community pots in the greenhouse under 70% shade.
All plantlets transferred to the greenhouse developed normally.
The protocol developed was used for the oryzalin treatment of Z. zamiifolia callus in an experiment aimed at producing a tetraploid Z. zamiifolia plant in vitro.
Authors
S.D. Vanzie-Canton, K.W. Leonhardt
Keywords
Z. zamiifolia, polyploidy, micropropagation, flow cytometry, cultivar development
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