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increased agrobacterium-mediated transformation and rooting efficiencies in canola (brassica napus l.) from hypocotyl segment explantsAn efficient protocol for the production of transgenic Brassica napus cv. Westar plants was developed by optimizing two important parameters: preconditioning time and co-cultivation time. Agrobacterium tumefaciens-mediated transformation was performed using hypocotyls as explant tissue. Two variants of a green fluorescent protein (GFP)-encoding gene--mGFP5-ER and eGFP--both under the constitutive expression of the cauliflower mosaic virus 35S promoter, were used for the experiments. Optimizing the preconditioning time to 72 h and co-cultivation time with Agrobacterium to 48 h provided the increase in the transformation efficiency from a baseline of 4% to 25%. With mGFP5-ER, the transformation rate was 17% and with eGFP it was 25%. Transgenic shoots were selected on 200 mg/l kanamycin. Rooting efficiency was 100% on half-strength Murashige and Skoog medium with 10 g/l sucrose and 0.5 mg/l indole butyric acid in the presence of kanamycin.
Document ID
20040087661
Document Type
Reprint (Version printed in journal)
Authors
Cardoza, V.
(2431 Center Drive Knoxville, TN 37996-4561, United States)
Stewart, C. N.
Date Acquired
August 21, 2013
Publication Date
February 1, 2003
Publication Information
Publication: Plant cell reports
Volume: 21
Issue: 6
ISSN: 0721-7714
Subject Category
Life Sciences (General)
Distribution Limits
Public
Copyright
Other
Keywords
NASA Discipline Life Sciences Technologies
Non-NASA Center