In this study, bulbs of Narcissus tazetta ‘Chinensis’ were used as explants for micropropagation. Several disinfection protocols, callus medium and regeneration medium were tested in addition to the secondary metabolites content analysis. Four disinfection protocols were used. The most successful was: 1) disinfection of bulbs in 70% ethanol and 2% HgCl2, 10 minutes for each, outside cabinet, or 2) 70% ethyl alcohol (4 min), 20% clorox (5 min), 0.1% mercuric chloride (1 min) and 9% hydrogen peroxide (1 min) inside cabinet, washing with sterilized distilled water after every previous step. This technique is considered as a novel effective procedure. Indirect micropropagation on MS medium supplemented with 2 mg·L-1 2,4-D succeeded in narcissus callus induction which regenerated into shoots on MS medium supplemented with 1 mg.L-1 IBA + 1mg.L-1 TIBA + 1 gm.L-1 of activated charcoal. The same medium with activated charcoal sowed high yield in a direct micropropagation protocol. Average shoot and root per explant showed significant increase when using indirect micropropagation. Secondary metabolites from direct and indirect regeneration, using phytochemical screening detected increasing alkaloids content in callus only, while regenerated plantlets were the same as mother plants, as well as there were no significant changes in mother and regenerated plants in terms of total phenol and flavonoids content.
Hamdy M. Abdel-Rahman
Genetic Engineering and Biotechnology Division, Department of Genetics and Cytology, National Research Centre, 33 El Buhouth ST., 12622, Dokki, Giza, Egypt.
K. N. Rashed
Department of Pharmacognosy, National Research Centre, 33 El-Buhouth St.-Dokki, P.O. 12622, Giza, Egypt.
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