Agrobacterium tumefaciencs-mediated transformation of wheat using a superbinary vector and a polyamine-supplemented regeneration medium

Khanna, Harjeet and Daggard, Grant (2003) Agrobacterium tumefaciencs-mediated transformation of wheat using a superbinary vector and a polyamine-supplemented regeneration medium. Plant Cell Reports, 21 (5). pp. 429-436. ISSN 0721-7714

Abstract

Immature embryo-derived calli of spring wheat (Triticum aestivum L.) cv Veery5 were transformed using Agrobacterium tumefaciens strain LBA4404 carrying either binary vector pHK22 or superbinary vector pHK21, the latter carrying an extra set of vir genes – vir B, -C and -G. In both cases, transient β-glucuronidase (GUS) expression ranging from 35–63% was observed 3 days after co-cultivation, but 587 calli infected with pHK22/LBA4404 failed to produce a single stably transformed plant, whereas 658 calli infected with pHK21/LBA4404 gave rise to 17 transformants carrying both the GUS and bar genes. Regeneration media supplemented with 0.1 M spermidine improved the recovery of transformants from pHK21/LBA4404-infected calli from 7% to 24.2%, resulting in an increase in the overall transformation frequency from 1.2% to 3.9%. The results suggest that two important factors that could lead to an improvement in transformation frequencies of cereals like wheat are (1) the use of superbinary vectors and (2) modification of the polyamine ratio in the regeneration medium. Stable expression and inheritance of the transgenes was confirmed by both genetic and molecular analyses. T1 progeny showed segregation of the transgenes in a typical Mendelian fashion in most of the plants. Of the transformed plants, 35% showed singlecopy insertion of the transgene as shown by both Southern analysis and the segregation ratios.


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Item Type: Article (Commonwealth Reporting Category C)
Refereed: Yes
Item Status: Live Archive
Additional Information: Author's version not held.
Depositing User: Assoc Prof Grant Daggard
Faculty / Department / School: Historic - Faculty of Sciences - Department of Biological and Physical Sciences
Date Deposited: 20 Apr 2010 07:41
Last Modified: 02 Jul 2013 23:48
Uncontrolled Keywords: Agrobacterium tumefaciens; wheat; transformation; superbinary vector; polyamines
Fields of Research (FOR2008): 10 Technology > 1001 Agricultural Biotechnology > 100105 Genetically Modified Field Crops and Pasture
07 Agricultural and Veterinary Sciences > 0703 Crop and Pasture Production > 070305 Crop and Pasture Improvement (Selection and Breeding)
Socio-Economic Objective (SEO2008): E Expanding Knowledge > 97 Expanding Knowledge > 970106 Expanding Knowledge in the Biological Sciences
B Economic Development > 82 Plant Production and Plant Primary Products > 8205 Winter Grains and Oilseeds > 820507 Wheat
Identification Number or DOI: doi: 10.1007/s00299-002-0529-x GENETIC TRANSFORMATION AND
URI: http://eprints.usq.edu.au/id/eprint/7515

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