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Overexpression of a thaumatin-like protein gene from Vitis amurensis improves downy mildew resistance in Vitis vinifera grapevine

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Abstract

Downy mildew is a highly destructive disease in grapevine production. A gene encoding pathogenesis-related (PR) thaumatin-like protein was isolated from the downy mildew-resistant grapevine “Zuoshan-1,” a clonal selection from wild Vitis amurensis Rupr. The predicted thaumatin-like protein (VaTLP) has 225 amino acids and it is acidic, with a calculated isoelectric point of 4.8. The full length of the VaTLP gene was transformed into somatic embryogenic calli of V. vinifera ‘Thompson Seedless’ via Agrobacterium tumefaciens. Real-time RT-PCR confirmed that the VaTLP gene was expressed at a high level in the transgenic grapevines. Improved resistance of the transgenic lines against downy mildew was evaluated using leaf disks and whole plants inoculated with Plasmopara viticola, the pathogen causing grapevine downy mildew disease. Bioassay of the pathogen showed that both hyphae growth and asexual reproduction were inhibited significantly among the transgenic plants. Histological analysis also confirmed this disease resistance by demonstrating the inhibition and malformation of hyphae development in leaf tissue of the transgenic plants. These results indicated that the accumulation of VaTLP could enhance resistance to P. viticola in transgenic ‘Thompson Seedless’ grapevines.

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Acknowledgements

This research was supported by China Agriculture Research System (grant no. CARS-30-yz-2) and China Agricultural University Scientific Fund (grant no. 2012RC019). We are grateful to the friends in UC Davis who helped make this research possible: Qingfeng Huang for assistance in vector construction, Abhaya Dandekar for providing Agrobacterium tumefaciens and Kim J. Carney for supplying the embryogenic culture. We also express our gratitude to Wentao Xu in China Agricultural University for their microscopy facility.

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Authors and Affiliations

  1. The Viticulture and Enology Program, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China

    Rongrong He, Jiao Wu, Yali Zhang, Xinlong Li, Shaoli Liu, Chaoxia Wang & Jiang Lu

  2. Department of Viticulture and Enology, University of California, Davis, Davis, CA, 95616, USA

    Rongrong He, Cecilia B. Agüero & M. Andrew Walker

  3. Center for Viticulture and Enology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200024, China

    Jiang Lu

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  1. Rongrong He
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  2. Jiao Wu
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Correspondence to M. Andrew Walker or Jiang Lu.

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Rongrong He and Jiao Wu are contributed equally to this work.

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He, R., Wu, J., Zhang, Y. et al. Overexpression of a thaumatin-like protein gene from Vitis amurensis improves downy mildew resistance in Vitis vinifera grapevine. Protoplasma 254, 1579–1589 (2017). https://doi.org/10.1007/s00709-016-1047-y

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