Efficient plant regeneration from hypocotyl protoplasts of broccoli (Brassica oleracea L. ssp. italica Plenck) |
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Authors: | H. M. Kao W. A. Keller S. Gleddie G. G. Brown |
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Affiliation: | (1) Department of Biology, McGill University, H3A 1B1 Montreal, Quebec, Canada;(2) Plant Biotechnology Institute, National Research Council of Canada, S7N 0W9 Saskatoon, Saskatchewan, Canada;(3) Plant Research Centre, Agriculture Canada, K1A 0C6 Ottawa, Ontario, Canada |
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Abstract: | 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.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - BAP 6-benzylaminopurine - GA3 gibberellic acid - IAA 3-indoleacetic acid - NAA 1-naphthaleneacetic acid - ZR zeatin riboside - CF colony formation - mtDNA mitochondrial DNA |
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