A Simple Procedure for Mesophyll Protoplast Culture and Plant Regeneration in Brassica oleracea L. and Brassica napus L

Narpal Deep Kaur, Miroslava Vyvadilová, Miroslav Klíma and Miroslav Bechyně
Additional contact information
Narpal Deep Kaur: Institute of Tropics and Subtropics, Czech University of Agriculture in Prague, Prague, Czech Republic
Miroslava Vyvadilová: Research Institute of Crop Production, Prague-Ruzyně, Czech Republic
Miroslav Klíma: Research Institute of Crop Production, Prague-Ruzyně, Czech Republic
Miroslav Bechyně: Institute of Tropics and Subtropics, Czech University of Agriculture in Prague, Prague, Czech Republic

Czech Journal of Genetics and Plant Breeding, 2006, vol. 42, issue 3, 103-110

Abstract: An improved protocol for Brassica protoplast culture and plant regeneration was developed. Isolated protoplasts from four-weeks-old in vitro shoot tip culture of Brassica oleracea var. botrytis cv. Siria F1 and Brassica napus doubled haploid of breeding line OP-1 were cultured at a density of 9.8-11.2 × 104 protoplasts/ml in darkness at 25°C in a modified medium containing 2% glucose, 0.25 mg/l 2,4-D, 1 mg/l BAP and 1 mg/l NAA. The first divisions of protoplasts were observed on the third day of culture in B. oleracea and on the fourth day in B. napus. The protoplast cultures were diluted with low osmotic medium on 7th and 11th day. The frequency of dividing cells was about 80% in B. oleracea and 50% in B. napus. After one month, the microcalli of approximately 0.5-1 mm in size were transferred into an induction medium with various combinations of growth regulators. Minimum duration of enzyme treatment time and extended dark period in the initial phase of culture increased the survival rate of protoplasts. Organogenesis started when the calli enlarged in size on an induction medium (1 mg/l NAA, 0.02 mg/l GA3, 1 mg/l 2iP) with 2% sucrose and 0.8% agar. Regeneration frequency of calli was found to be 69-75% in B. oleracea and 2-3% in B. napus. Well-developed shoots were transferred for rooting to a half-strength MS medium without growth regulators. More than 100 B. oleracea regenerants were transferred into soil, and they produced normal heads and set seeds. This very simple procedure is efficient and suitable mainly for B. oleracea var. botrytis and represents a background for fusion experiments.

Keywords: Brassica napus L.; Brassica oleracea L.; organogenesis; protoplast culture; regeneration (search for similar items in EconPapers)
Date: 2006
References: View references in EconPapers View complete reference list from CitEc
Citations:

Downloads: (external link)
http://cjgpb.agriculturejournals.cz/doi/10.17221/3649-CJGPB.html (text/html)
http://cjgpb.agriculturejournals.cz/doi/10.17221/3649-CJGPB.pdf (application/pdf)
free of charge

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:caa:jnlcjg:v:42:y:2006:i:3:id:3649-cjgpb

DOI: 10.17221/3649-CJGPB

Access Statistics for this article

Czech Journal of Genetics and Plant Breeding is currently edited by Ing. Markéta Knížková, (Executive Editor)

More articles in Czech Journal of Genetics and Plant Breeding from Czech Academy of Agricultural Sciences
Bibliographic data for series maintained by Ivo Andrle ().