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Pathogenesis-related proteins induced by agroinoculation-associated cell wall weakening can be obviated by spray-on inoculation or mannitol ex vivo culture

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Abstract

Agroinoculation transient expression systems are commonly accompanied with elevated pathogenesis-related (PR) protein production and leaf necrosis. We identified the major PR proteins in Nicotiana benthamiana in response to agroinoculation and determined that their occurrence was mainly due to agrobacterium infection and the method of inoculation, rather than due to viral vectors overexpressing foreign proteins. A spray-on inoculation method was optimized in this research and used to obviate the leaf necrosis and PR proteins induced by agrobacterium. Subsequently, this method also increased the yield and purity of the protein-of-interest. A further investigation of PR protein induction by a necrosis-inducing protein, Jun a 1, suggested that the plant pathogenic response was related to biochemical integrity of plant cell wall, which was also confirmed by an osmo-stabilizing mannitol ex vivo culture experiment. These findings provide insight into the response of plants to agroinoculation and suggested a connection between cell wall weakening and PR protein elicitation.

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Author notes

  1. Yiyang Zhou

    Present address: Department of Biochemistry and Molecular Biology, University of Texas, Medical Branch, Galveston, TX, 77550, USA

Authors and Affiliations

  1. Institute of Biomedical Studies, Baylor University, Waco, TX, 76798, USA

    Yiyang Zhou & Christopher M. Kearney

  2. Department of Biology, Baylor University, Waco, TX, 76798, USA

    Andrew M. Cox & Christopher M. Kearney

Authors
  1. Yiyang Zhou
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  2. Andrew M. Cox
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  3. Christopher M. Kearney
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Contributions

Y.Z. and C.K. designed the research. A.C. assisted with mass spectrometry data analysis. A.C. also composed Supplemental Tables 1–2, Supplemental Figures 1–4. Y.Z. wrote the manuscript with advice and revision from C.K. and with contribution and discussion from all of the co-authors.

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Correspondence to Yiyang Zhou.

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Zhou, Y., Cox, A.M. & Kearney, C.M. Pathogenesis-related proteins induced by agroinoculation-associated cell wall weakening can be obviated by spray-on inoculation or mannitol ex vivo culture. Plant Biotechnol Rep 11, 161–169 (2017). https://doi.org/10.1007/s11816-017-0439-6

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  • DOI: https://doi.org/10.1007/s11816-017-0439-6

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