Plant regeneration from cotyledon protoplasts of Brassica carinata |
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Authors: | S K Jaiswal N Hammatt S S Bhojwani E C Cocking M R Davey |
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Institution: | (1) Plant Genetic Manipulation Group, Department of Botany, University of Nottingham, NG7 2RD Nottingham, UK;(2) Present address: Institute of Horticultural Research, East Malling, ME19 6BJ Maidstone, Kent, UK;(3) Department of Botany, University of Delhi, 110007 Delhi, India |
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Abstract: | Protoplasts isolated from cotyledons of Brassica carinata, underwent sustained division when cultured at 5.0 × 104 ml-1 in modified 8p medium (KM8P) with 1.0% (w/v) Seaplaque agarose. Cell colonies produced callus when agarose droplets, in which the protoplasts were embedded, were transferred to K8 medium with 0.6% (w/v) Sigma Type I or Type VII agarose at day 16, giving a plating efficiency of 1.6%. Seventy percent of the protoplast derived-tissues produced shoot buds after subculture to MS medium containing 3.0% (w/v) sucrose, 1.125 mgl-1 BAP, 0.035 mgl-1 GA and 0.6% (w/v) Type I agarose, resulting in shoot formation from 1.1% of the protoplasts originally plated. Protoplast-derived colonies transferred to hormone-free MS medium with 1.0% (w/v) sucrose and 0.6% (w/v) Type I agarose produced roots. The latter gave rise to shoots after excision from the parent callus and culture on MS medium with 3.0% sucrose, 0.225 mgl-1 BAP, and 0.6% (w/v) Type I agarose. Shoots regenerated directly from protoplast-derived calli, or indirectly from roots, developed prolific root systems when placed on hormone-free MS medium with 1.0% (w/v) sucrose and 0.6% (w/v) Type I agarose.Abbreviations BAP
6-benzylaminopurine
- CH
casein hydrolysate
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GA
gibberellic acid
- K
kinetin
- NAA
-naphthaleneacetic acid
- MES
2(N-morpholino)ethanesulphonic acid, 2,iP-6( , -dimethylallyamino) purine
- IAA
indole-3-acetic acid
- Z
zeatin
- ZR
zeatin riboside |
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Keywords: | Abyssinian mustard Brassica carinata cotyledon protoplasts plant regeneration |
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