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A strong root-specific expression system for stable transgene expression in bread wheat

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Abstract

Key message

A strong, stable and root-specific expression system was developed from a rice root-specific GLYCINE - RICH PROTEIN 7 promoter for use as an enabling technology for genetic manipulation of wheat root traits.

Abstract

Root systems play an important role in wheat productivity. Genetic manipulation of wheat root traits often requires a root-specific or root-predominant expression system as an essential enabling technology. In this study, we investigated promoters from rice root-specific or root-predominant expressed genes for development of a root expression system in bread wheat. Transient expression analysis using a GREEN FLUORESCENT PROTEIN (GFP) reporter gene driven by rice promoters identified six promoters that were strongly expressed in wheat roots. Extensive organ specificity analysis of three rice promoters in transgenic wheat revealed that the promoter of rice GLYCINE-RICH PROTEIN 7 (OsGRP7) gene conferred a root-specific expression pattern in wheat. Strong GFP fluorescence in the seminal and branch roots of wheat expressing GFP reporter driven by the OsGRP7 promoter was detected in epidermal, cortical and endodermal cells in mature parts of the root. The GFP reporter driven by the promoter of rice METALLOTHIONEIN-LIKE PROTEIN 1 (OsMTL1) gene was mainly expressed in the roots with essentially no expression in the leaf, stem or seed. However, it was also expressed in floral organs including glume, lemma, palea and awn. In contrast, strong expression of rice RCg2 promoter-driven GFP was found in many tissues. The GFP expression driven by these three rice promoters was stable in transgenic wheat plants through three generations (T1–T3) examined. These data suggest that the OsGRP7 promoter can provide a strong, stable and root-specific expression system for use as an enabling technology for genetic manipulation of wheat root traits.

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Abbreviations

GFP:

Green fluorescent protein

GRP:

Glycine-rich protein

MTL:

Metallothionein-like

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Acknowledgments

We are grateful to Smitha Louis and Dhara Bhatt for production of transgenic wheat plants and Terry Grant for maintenance of the controlled environment facility for growing wheat plants. We thank Dr. Emmanuel Delhaize for his critical reading of this manuscript.

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Authors and Affiliations

  1. CSIRO Agriculture, 306 Carmody Rd., St Lucia, QLD, 4067, Australia

    Gang-Ping Xue, Anne L. Rae, Janneke Drenth & C. Lynne McIntyre

  2. CSIRO Agriculture, Clunies Ross Street, Canberra, ACT, 2601, Australia

    Rosemary G. White & Terese Richardson

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  1. Gang-Ping Xue
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  2. Anne L. Rae
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  3. Rosemary G. White
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  4. Janneke Drenth
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  6. C. Lynne McIntyre
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Correspondence to Gang-Ping Xue.

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Communicated by P. Lakshmanan.

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Xue, GP., Rae, A.L., White, R.G. et al. A strong root-specific expression system for stable transgene expression in bread wheat. Plant Cell Rep 35, 469–481 (2016). https://doi.org/10.1007/s00299-015-1897-3

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  • DOI: https://doi.org/10.1007/s00299-015-1897-3

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