An efficient and reproducible Agrobacterium-mediated transformation method for hexaploid wheat (Triticum aestivum L.)

Hayta, Şadiye; Smedley, Mark; Demir, Selcen U.; Blundell, Robert; Hinchliffe, Alison; Atkinson, Nicola J.; Harwood, Wendy

doi:10.1186/s13007-019-0503-z

“…Transformation by overexpression of transcription factors such as maize Baby Boom and Wuschel2 has also yielded improved transformation efficiencies in monocots (Lowe et al, 2016), although there are no formal reports yet in wheat. Recently, an open-access wheat transformation system with transformation efficiencies of up to 25% was published (Hayta et al, 2019), albeit for a single cultivar.…”

Section: Transgenic Approachesmentioning

confidence: 99%

“…For CRISPR/Cas9 and other non-transient transgenic approaches several varieties may be used, although relatively few wheat varieties have been shown to display high enough transformation efficiencies to be practical. This means that traditionally most transgenic studies in wheat have been limited to a few varieties, such as 'Fielder ', Cadenza, 'Bobwhite', 'Kenong 199' and Kronos (Li et al, 2012;Richardson et al, 2014;Hayta et al, 2019). This is now changing thanks to work by groups at NIAB (UK), CAAS (China) and CSIRO (Australia) who have successfully transformed 39 (Wallington, 2015), 15 (Wang et al, 2017) and six (Richardson et al, 2014) varieties, respectively.…”

Section: Strategies For Usementioning

confidence: 99%

A roadmap for gene functional characterisation in wheat

Adamski

¹

,

Borrill

²

,

Brinton

³

et al. 2019

Preprint

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To adapt to the challenges of climate change and the growing world population, it is vital to increase global crop production. Understanding the function of genes within staple crops will accelerate crop improvement by allowing targeted breeding approaches. Despite the importance of wheat, which provides 20 % of the calories consumed by humankind, a lack of genomic information and resources has hindered the functional characterisation of genes in this species. The recent release of a high-quality reference sequence for wheat underpins a suite of genetic and genomic resources that support basic research and breeding. These include accurate gene model annotations, gene expression atlases and gene networks that provide background information about putative gene function. In parallel, sequenced mutation populations, improved transformation protocols and structured natural populations provide rapid methods to study gene function directly. We highlight a case study exemplifying how to integrate these resources to study gene function in wheat and thereby accelerate improvement in this important crop. We hope that this review provides a helpful guide for plant scientists, especially those expanding into wheat research for the first time, to capitalise on the discoveries made in Arabidopsis and other plants. This will accelerate the improvement of wheat, a complex polyploid crop, of vital importance for food and nutrition security.

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“…This is because no other cultivar has been found to give a similar or higher 31 transformation efficiency (Bartlett et al, 2008;Harwood et al, 2009;Harwood, 2012). Wheat, like 32 barley, is also transformable using Agrobacterium but despite successful attempts to increase 33 transformation efficiency (Hayta et al, 2019), the process is still cultivar dependant: cv Fielder being 34 one of the most responsive cultivars. Efficiencies as high as 86.7% can be obtained with barley cv 35…”

Section: Introduction 28mentioning

confidence: 99%

TRA1: a locus responsible for controllingAgrobacterium-mediated transformability in barley

Orman-Ligeza

¹

,

Harwood

²

,

Hedley

³

et al. 2019

Preprint

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AbstractIn barley (Hordeum vulgare L.), Agrobacterium-mediated transformation efficiency is highly dependent on genotype with very few cultivars being amenable to transformation. Golden Promise is the cultivar most widely used for barley transformation and developing embryos are the most common donor tissue. We tested whether barley mutants with abnormally large embryos were more or less amenable to transformation and discovered that mutant M1460 had a transformation efficiencies similar to that of Golden Promise. The large-embryo phenotype of M1460 is due to mutation at the LYS3 locus. There are three other barley lines with independent mutations at the same LYS3 locus, and one of these, Risø1508 has an identical missense mutation to that in M1460. However, none of the lys3 mutants except M1460 were transformable showing that the locus responsible for transformation efficiency, TRA1, was not LYS3 but another locus unique to M1460. To identify TRA1, we generated a mapping population by crossing M1460 to the cultivar Optic, which is recalcitrant to transformation. After four rounds of backcrossing to Optic, plants were genotyped and their progeny were tested for transformability. Some of the progeny lines were transformable at high efficiencies similar to those seen for the parent M1460 and some were not transformable, like Optic. A region on chromosome 2H inherited from M1460 is present in transformable lines only. We propose that one of the 225 genes in this region is TRA1.

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“…Recently, an open-access wheat transformation system with transformation efficiencies of up to 25% was published (Hayta et al, 2019), albeit for a single cultivar.…”

Section: Transgenic Approachesmentioning

confidence: 99%

“…For CRISPR/Cas9 and other non-transient transgenic approaches several varieties may be used, although relatively few wheat varieties have been shown to display high enough transformation efficiencies to be practical. This means that traditionally most transgenic studies in wheat have been limited to a few varieties, such as 'Fielder', Cadenza, 'Bobwhite', 'Kenong 199' and Kronos (Li et al, 2012;Richardson et al, 2014;Hayta et al, 2019). This is now changing thanks to work by groups at NIAB (UK), CAAS (China) and CSIRO (Australia) who have successfully transformed 39 (Wallington, 2015), 15 and six (Richardson et al, 2014) varieties, respectively.…”

Section: Strategies For Usementioning

confidence: 99%

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Supplemental Information 1: Table 2: Natural Variation Resources Available in Wheat

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To adapt to the challenges of climate change and the growing world population, it is vital to increase global crop production. Understanding the function of genes within staple crops will accelerate crop improvement by allowing targeted breeding approaches. Despite the importance of wheat, which provides 20 % of the calories consumed by humankind, a lack of genomic information and resources has hindered the functional characterisation of genes in this species. The recent release of a high-quality reference sequence for wheat underpins a suite of genetic and genomic resources that support basic research and breeding. These include accurate gene model annotations, gene expression atlases and gene networks that provide background information about putative gene function. In parallel, sequenced mutation populations, improved transformation protocols and structured natural populations provide rapid methods to study gene function directly. We highlight a case study exemplifying how to integrate these resources to study gene function in wheat and thereby accelerate improvement in this important crop. We hope that this review provides a helpful guide for plant scientists, especially those expanding into wheat research for the first time, to capitalise on the discoveries made in Arabidopsis and other plants. This will accelerate the improvement of wheat, a complex polyploid crop, of vital importance for food and nutrition security. any plant species, identify the correct wheat ortholog(s) using Ensembl Plants (https://plants.ensembl.org) and determine gene expression using expression browsers and gene networks. Suggestions for functional characterisation are provided including induced variation such as mutants, transgenics or Virus-Induced Gene Silencing (VIGs). In addition, publicly available populations incorporating natural variation are available. Finally steps for growing, genotyping and crossing plants are outlined. Links to detailed tutorials and further information are provided and can be found on www.wheat-training.com. 1

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An efficient and reproducible Agrobacterium-mediated transformation method for hexaploid wheat (Triticum aestivum L.)

Cited by 150 publications

References 49 publications

A roadmap for gene functional characterisation in wheat

A roadmap for gene functional characterisation in wheat

TRA1: a locus responsible for controllingAgrobacterium-mediated transformability in barley

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