Protoplasts were isolated from hypocotyls of 7-d-old seedlings of three genotypes of Brassica carinata after enzymatic digestion in cellulase R-10 (0.5%) and pectolyase Y-23 (0.025%). The protoplasts were stabilized with 0.4 M mannitol used as osmoticum, and were cultured in darkness in Kao's liquid
Efficient plant regeneration from hypocotyl protoplasts of broccoli (Brassica oleraceaL. ssp.italicaPlenck)
β Scribed by H. M. Kao; W. A. Keller; S. Gleddie; G. G. Brown
- Publisher
- Springer
- Year
- 1990
- Tongue
- English
- Weight
- 825 KB
- Volume
- 9
- Category
- Article
- ISSN
- 0721-7714
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β¦ Synopsis
We have assessed the capacity of cultured protoplasts from two tissue sources of several commercially-grown broccoli cultivars to regenerate plants. A procedure that employs hypocotyl protoplasts and a culture medium with a high NAA:2,4-D auxin ratio was developed. The procedure permits highly efficient formation of colonies that regenerate shoots at frequencies of 8-17% with two of the four cultivars tested. The time required for the development of plants from protoplasts was 8-11 weeks. No mtDNA rearrangements were observed among any of 17 analysed regenerants. Double-stranded RNAs were detected in mitochondrial DNA (mtDNA) preparations of some, but not all, regenerants of one of the cultivars.
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