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A rapid and efficient Agrobacterium-mediated transient gene expression system for strawberry leaves and the study of disease resistance proteins

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Abstract

Transient gene expression is a simple and universal method and has been broadly applied in gene functional analyses in several plant species. However, few studies have reported the utilization and transient expression of strawberry leaves. Here, an Agrobacterium-mediated transient gene expression system was established in strawberry leaves. The strawberry leaf transient gene expression system was tested for protein expression, western blot assays, and protein subcellular localization. Furthermore, the Arabidopsis thaliana broad-spectrum disease resistance protein RPW8.2 was transiently expressed in strawberry leaves. Both the application of exogenous salicylic acid (SA) and the following infection by the strawberry powdery mildew pathogen, Podosphaera aphanis, resulted in increased expression of the RPW8.2 fusion protein. Reverse transcription quantitative PCR analysis showed that the transient expression of AtRPW8.2 in strawberry leaves could induce defense-related gene transcription levels. In conclusion, we demonstrated the potential of a rapid and efficient Agrobacterium-mediated transient gene expression system in strawberry leaves and its use for the study of disease resistant protein activity in strawberry leaves.

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Acknowledgements

We thank SX kindly provided the vectors of RPW8.2. The authors would also like to thank three anonymous reviewers for comments on the manuscript. The present study was financially supported by grants from the National Natural Science Foundation of China (Grant No. 31201657), the major program of Yangling Agricultural Hi-tech Industry Demo Zone synergy innovation for cooperation in production, study, and research (2016CXY-11), and the project for Agriculture Sci-Tech demonstration and extension of Yangling Agricultural Hi-tech Industry Demo Zone (TS-2016-18).

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Authors and Affiliations

  1. State Key Laboratory of Crop Stress Biology for Arid Areas, College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Meng-Yuan Cui, Wei Wei, Kuan Gao, Yin-Ge Xie, Ye Guo & Jia-Yue Feng

  2. Key Laboratory of Protected Horticulture Engineering in Northwest China, Ministry of Agriculture, Yangling, 712100, Shaanxi, China

    Meng-Yuan Cui, Wei Wei, Kuan Gao, Yin-Ge Xie & Jia-Yue Feng

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  1. Meng-Yuan Cui
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  2. Wei Wei
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  3. Kuan Gao
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  4. Yin-Ge Xie
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  5. Ye Guo
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  6. Jia-Yue Feng
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Contributions

JYF conceived the study. MYC performed all the experiments with the assistance of WW, KG, YGX, and YG. MYC, WW, and JYF analyzed the data. MYC and JYF contributed to writing the manuscript. All authors read and approved the final manuscript. The authors are grateful to Dr. Ke Duan of the Shanghai Academy of Agricultural Sciences for generously providing wild type Fragaria vesca plants.

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Correspondence to Jia-Yue Feng.

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Communicated by Maria Margarida Oliveira.

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Cui, MY., Wei, W., Gao, K. et al. A rapid and efficient Agrobacterium-mediated transient gene expression system for strawberry leaves and the study of disease resistance proteins. Plant Cell Tiss Organ Cult 131, 233–246 (2017). https://doi.org/10.1007/s11240-017-1279-3

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  • DOI: https://doi.org/10.1007/s11240-017-1279-3

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