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An efficient and reproducible Agrobacterium-mediated transformation method for hexaploid wheat (Triticum aestivum L.).
Hayta, Sadiye; Smedley, Mark A; Demir, Selcen U; Blundell, Robert; Hinchliffe, Alison; Atkinson, Nicola; Harwood, Wendy A.
Afiliación
  • Hayta S; Department of Crop Genetics, John Innes Centre, Norwich Research Park, Norwich, Norfolk NR4 7UH UK.
  • Smedley MA; Department of Crop Genetics, John Innes Centre, Norwich Research Park, Norwich, Norfolk NR4 7UH UK.
  • Demir SU; Department of Crop Genetics, John Innes Centre, Norwich Research Park, Norwich, Norfolk NR4 7UH UK.
  • Blundell R; Department of Crop Genetics, John Innes Centre, Norwich Research Park, Norwich, Norfolk NR4 7UH UK.
  • Hinchliffe A; Department of Crop Genetics, John Innes Centre, Norwich Research Park, Norwich, Norfolk NR4 7UH UK.
  • Atkinson N; Department of Crop Genetics, John Innes Centre, Norwich Research Park, Norwich, Norfolk NR4 7UH UK.
  • Harwood WA; Department of Crop Genetics, John Innes Centre, Norwich Research Park, Norwich, Norfolk NR4 7UH UK.
Plant Methods ; 15: 121, 2019.
Article en En | MEDLINE | ID: mdl-31673278
BACKGROUND: Despite wheat being a worldwide staple, it is still considered the most difficult to transform out of the main cereal crops. Therefore, for the wheat research community, a freely available and effective wheat transformation system is still greatly needed. RESULTS: We have developed and optimised a reproducible Agrobacterium-mediated transformation system for the spring wheat cv 'Fielder' that yields transformation efficiencies of up to 25%. We report on some of the important factors that influence transformation efficiencies. In particular, these include donor plant health, stage of the donor material, pre-treatment by centrifugation, vector type and selection cassette. Transgene copy number data for independent plants regenerated from the same original immature embryo suggests that multiple transgenic events arise from single immature embryos, therefore, actual efficiencies might be even higher than those reported. CONCLUSION: We reported here a high-throughput, highly efficient and repeatable transformation system for wheat and this system has been used successfully to introduce genes of interest, for RNAi, over-expression and for CRISPR-Cas9 based genome editing.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Plant Methods Año: 2019 Tipo del documento: Article Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Plant Methods Año: 2019 Tipo del documento: Article Pais de publicación: Reino Unido