Factors influencing regeneration from protoplasts of Asparagus densiflorus cv. Sprengeri |
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Authors: | M Benmoussa S Mukhopadhyay Y Desjardins |
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Institution: | (1) Centre de recherche en horticulture, Département de Phytologie, FSAA, Pavillon Envirotron, Université Laval, Québec, G1K 7P4, Canada Fax no. +1-418-656-7856 e-mail: yves.desjardins@plg.ulaval.ca, CA |
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Abstract: | This article describes conditions to optimize the yield of viable protoplasts from callus tissue of Asparagus densiflorus cv. Sprengeri and their subsequent regeneration into plantlets. Callus tissue was initiated by culturing spear sections (5–7
mm) on Murashige and Skoog (MS) medium supplemented with 0.8% (wt/vol) Bacto agar, 3% (wt/vol) sucrose, 0.5 mg/l each of nicotinic
acid, pyridoxine-HCl, and thiamine-HCl, 1 mg/l p-chlorophenoxyaceticacid (pCPA) and 1 mg/l 6-benzylaminopurine (BAP). The maximum protoplast yield was obtained in a mixture
of 1% (wt/vol) Cellulysin, 0.8% (wt/vol) Rhozyme HP 150 and 0.3% (wt/vol) Macerase, dissolved in cell protoplast wash salt
solution with 7 mm CaCl2
.2H2O, 3 mm MES, 0.6 m glucose, and 0.1 m mannitol. First divisions were observed after 3–4 days of initial culture. The plating efficiency was highest (7.8%) in half-strength
MS semisolid medium containing 1 g/l glutamine, 0.6 m glucose, 0.1 m mannitol, 0.5 mg/l folic acid, 0.05 mg/l biotin, 2 mg/l ascorbic acid, 1 mg/l α-naphthaleneacetic acid, 0.5 mg/l zeatin, and 0.1% (wt/vol) Gelrite. Protoplast-derived microcolonies and microcalli were
cultured on the same medium on which the primary callus culture was initiated. After 10–12 weeks, calli were transferred to
shoot regeneration medium containing MS salts, 1 mg/l BAP, 0.5 mg/l pCPA and 0.2% Gelrite. Shoots (3–4 cm) were then transferred
to MS rooting medium with 2 mg/l indole-3-butyric acid, and 0.2% Gelrite. Plantlets were obtained within 4–5 weeks.
Received: 9 August 1995 / Revision received: 27 June 1997 / Accepted: 17 July 1997 |
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Keywords: | Tissue culture Fusarium Somatic hybridization |
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