Transformation and regeneration of Brassica rapa using Agrobacterium tumefaciens

Transformation and regeneration of Brassica rapa using Agrobacterium tumefaciens

Transformation and regeneration procedures for obtaining transgenic Brassica rapa ssp. oleifera plants are described. Regeneration frequencies were increasedby using silver nitrate and by adjusting the duration of exposure to 2,4-D. For transformation, Agrobacterium tumefaciens strain EHA101 contain...

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Bibliographic Details
Published in:Plant cell reports Vol. 11; no. 10; pp. 499 - 505
Main Authors: RADKE, S. E, TURNER, J. C, FACCIOTI, D
Format: Journal Article
Language:English
Published: Berlin Springer 01-09-1992
Subjects:
Biological and medical sciences
Biotechnology
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Methods. Procedures. Technologies
Transgenic animals and transgenic plants
Transgenic plants
Vegetals
Brassica rapa
Enzyme
β-D-Glucuronidase
Agrobacterium tumefaciens
Tissue culture
Kanamycin kinase
Genetic transfer
Regeneration
Plant
Hypocotyl
Cruciferae
Dicotyledones
Angiospermae
Bacteria
Spermatophyta
Transgenic plant
Rhizobiaceae
Silver Nitrates
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Summary:Transformation and regeneration procedures for obtaining transgenic Brassica rapa ssp. oleifera plants are described. Regeneration frequencies were increasedby using silver nitrate and by adjusting the duration of exposure to 2,4-D. For transformation, Agrobacterium tumefaciens strain EHA101 containing a binary plasmid with the neomycin phosphotransferase gene (NPT II) and the b-glucuronidase gene (GUS) was cocultivated with hypocotyl explants from the oilseed B. rapa cvs. Tobin and Emma. Transformed plants were obtained within three months of cocultivation. Transformation frequencies for the cultivars Tobin and Emma were 1-9%. Evidence for transformation was shown by NPT II dot blot assay, the GUS fluorometric assay, Southern analysis, and segregation of the kanamycin-resistance trait in the progeny. The transformation and regeneration procedure described here has been used routinely to transform two cultivars of B. rapa and 18 cultivars of B. napus.
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ISSN:0721-7714
1432-203X
DOI:10.1007/bf00236265