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

Article

Khanna, Harjeet and Daggard, Grant. 2003. "Agrobacterium tumefaciens-mediated transformation of wheat using a superbinary vector and a polyamine-supplemented regeneration medium." Plant Cell Reports. 21 (5), pp. 429-436. https://doi.org/10.1007/s00299-002-0529-x
Article Title

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

ERA Journal ID2645
Article CategoryArticle
AuthorsKhanna, Harjeet (Author) and Daggard, Grant (Author)
Journal TitlePlant Cell Reports
Journal Citation21 (5), pp. 429-436
Number of Pages8
Year2003
PublisherSpringer
Place of PublicationGermany
ISSN0721-7714
1432-203X
Digital Object Identifier (DOI)https://doi.org/10.1007/s00299-002-0529-x
Web Address (URL)https://link.springer.com/article/10.1007/s00299-002-0529-x
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 single-copy insertion of the transgene as shown by both Southern analysis and the segregation ratios.

KeywordsAgrobacterium tumefaciens; wheat; transformation; superbinary vector; polyamines
Contains Sensitive ContentDoes not contain sensitive content
ANZSRC Field of Research 2020300105. Genetically modified field crops and pasture
300406. Crop and pasture improvement (incl. selection and breeding)
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Byline AffiliationsDepartment of Biological Physical Sciences
Faculty of Sciences
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