Journal of the International Aroid Society, Inc.
Volume 20, Number 1 (1997); Pages 29-36
Progress in protoplast Isolation and culture from axenic tissues of hybrid Anthurium
By Adelheid R. Kuehnle     (Buy)


Protoplasts capable of first divisions were isolated from Anthurium andraeanum Linden ex Andre hybrids using axenic etiolated shoots of the University of Hawaii Antburium cultivars 'Rudolph', UH1060, and UH1003. Enzymatic digestion by 1.5% (w/v) Cellulase R-lO, 0.5% Macerozyme R-10 and 0.5% Driselase in 0.5 M mannitol gave consistent yields between 1.6 X 104 to 2.2 X 104 protoplasts per gram fresh weight of etiolated shoots. In contrast, similar digestions using green leaves of micropropagated plantlets always resulted in mixtures of protoplasts and partially digested cells in ratios of about 1:1 to 1:3, respectively, with a mean yield of 3.8 X 105 protoplasts per gram fresh weight of leaves. Etiolated shoots exposed to a 14-day low light treatment gave consistently higher yields than those with no light exposure, with 3.8 X 104 and 1.8 X 104 protoplasts per gram fresh weight, respectively. Anthurium 'Rudolph' and UH1060 protoplasts were cultured in darkness at densities ranging from 2.5 X 104 to 7.0 X 104 protoplasts ml- ' in a basic tobacco protoplast culture medium with 1 mg' liter-l 6-benzylaminopurine, 1 mg·liter-' IX-naphthaleneacetic acid, and 0.1 mg·liter-' 2,4-dichlorophenoxyacetic acid either in liquid or embedded in 0.6% agarose. Antburium etiolated shoot protoplasts divided within 2 to 6 days and some sustained divisions to form microcolonies within 10 days under these conditions.


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