Abstract
Ubiquitination plays important roles in disease resistance in plants. We report the identification and functional characterization of the RING-type ubiquitin ligase gene VpUR9 from Chinese wild Vitis pseudoreticulata accession Baihe-35-1. VpUR9, encodes 164 amino acids and possesses a RING conserved motif. It is homologously cloned from the cDNA library of the high powdery mildew (Erysiphe necator [Schw.] Burr) resistant V. pseudoreticulata accession Baihe-35-1 inoculated with E. necator. The gene is induced in response to powdery mildew and salicylic acid. VpUR9 fused with FLAG-tag controlled by 35S promoter was transformed into 15 regenerated V. vinifera L. cv. Red Globe lines via Agrobacterium tumefaciens-mediated transformation. Twelve of these lines were confirmed by Western blot of FLAG-tag. As a result, the powdery mildew-resistance of Red Globe transformed with VpUR9 was repressed. Furthermore, the expression of some disease-resistant related genes (NPR1, PR1, PR10 and PAL) of the transgenic Red Globe declined compared with wild type grapes when inoculated with powdery mildew or salicylic acid. When treated with jasmonic acid methyl ester, its PR1 gene expression decreased, while the expressions of NPR1, PR10 and PAL all increased, contrasting with the wild type grape.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (Grant No. 31171924). The authors thank Dr. Alexander (Sandy) Lang from RESCRIPT Co. New Zealand for useful comments and language editing which have greatly improved the manuscript.
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YW and JZ designed research. LD, XX, YY, CZ, YX and JZ performed experiments. LD, XX, YY, CZ, YX and JZ analyzed the results. LD and YY wrote the manuscript and YW revised the manuscript. All authors read approved the manuscript.
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Communicated by Sergey V Dolgov.
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Dai, L., Xie, X., Yang, Y. et al. VpUR9, a novel RING-type ubiquitin ligase gene from Vitis pseudoreticulata, is involved in powdery mildew response in transgenic V. vinifera plants. Plant Cell Tiss Organ Cult 131, 41–49 (2017). https://doi.org/10.1007/s11240-017-1260-1
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DOI: https://doi.org/10.1007/s11240-017-1260-1