Journal article
Evidence for stable transformation of wheat by floral dip in Agrobacterium tumefaciens
Plant cell reports, Vol.28(6), pp.903-913
06/2009
Handle:
https://hdl.handle.net/2376/104000
PMCID: PMC2688021
PMID: 19308413
Abstract
Hexaploid wheat, one of the world’s most important staple crops, remains a challenge for genetic transformation. We are developing a floral transformation protocol for wheat that does not require tissue culture. This paper presents three transformants in the hard red germplasm line Crocus that have been characterized thoroughly at the molecular level over three to six generations. Wheat spikes at the early boot stage, i.e. the early, mid or late uninucleate microspore stages, were immersed in an infiltration medium of strain C58C1 harboring pDs(Hyg)35S, or strain AGL1 harboring pBECKSred. pDs(Hyg)35S contains the
NPTII
and
hph
selectable markers, and transformants were detected using paromomycin spray at the whole plant level, NPTII ELISAs, or selection on medium with hygromycin. Strain AGL1, harboring pBECKSred, which contains the maize anthocyanin regulators,
Lc
and
C1
, and the
NPTII
gene, was also used to produce a Crocus transformant. T1 and T2 seeds with red embryos were selected; T1 and T2 plants were screened by sequential tests for paromomycin resistance and NPTII ELISAs. The transformants were low copy number and showed Mendelian segregation in the T2. Stable transmission of the transgenes over several generations has been demonstrated using Southern analysis. Gene expression in advanced progeny was shown using Reverse Transcriptase-PCR and ELISA assays for NPTII protein expression. This protocol has the potential to reduce the time and expense required for wheat transformation.
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Details
- Title
- Evidence for stable transformation of wheat by floral dip in Agrobacterium tumefaciens
- Creators
- Janice M Zale - Plant Sciences Department, The University of Tennessee, 2431 Joe Johnson Drive, Knoxville, TN 37996-4561 USAS Agarwal - Plant Sciences Department, The University of Tennessee, 2431 Joe Johnson Drive, Knoxville, TN 37996-4561 USAS Loar - Plant Sciences Department, The University of Tennessee, 2431 Joe Johnson Drive, Knoxville, TN 37996-4561 USAC. M Steber - USDA/ARS Wheat Genetics, Physiology and Disease Evaluation Unit, Washington State University, 209 Johnson Hall, Pullman, WA 99164-6420 USA
- Publication Details
- Plant cell reports, Vol.28(6), pp.903-913
- Academic Unit
- Crop and Soil Sciences, Department of
- Publisher
- Springer-Verlag; Berlin/Heidelberg
- Identifiers
- 99900546988201842
- Language
- English
- Resource Type
- Journal article