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Comparative mapping of powdery mildew resistance gene Pm21 and functional characterization of resistance-related genes in wheat

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The powdery mildew resistance gene Pm21 was physically and comparatively mapped by newly developed markers. Seven candidate genes were verified to be required for Pm21 -mediated resistance to wheat powdery mildew.

Abstract

Pm21, a gene derived from wheat wild relative Dasypyrum villosum, has been transferred into common wheat and widely utilized in wheat resistance breeding for powdery mildew. Previously, Pm21 has been located to the bin FL0.45–0.58 of 6VS by using deletion stocks. However, its fine mapping is still a hard work. In the present study, 30 gene-derived 6VS-specific markers were obtained based on the collinearity among genomes of Brachypodium distachyon, Oryza and Triticeae, and then physically and comparatively mapped in the bin FL0.45–0.58 and its nearby chromosome region. According to the maps, the bin FL0.45–0.58 carrying Pm21 was closely flanked by the markers 6VS-03 and 6VS-23, which further narrowed the orthologous regions to 1.06 Mb in Brachypodium and 1.38 Mb in rice, respectively. Among the conserved genes shared by Brachypodium and rice, four serine/threonine protein kinase genes (DvMPK1, DvMLPK, DvUPK and DvPSYR1), one protein phosphatase gene (DvPP2C) and two transcription factor genes (DvGATA and DvWHY) were confirmed to be required for Pm21-mediated resistance to wheat powdery mildew by barley stripe mosaic virus-induced gene silencing (BSMV-VIGS) and transcriptional pattern analyses. In summary, this study gives new insights into the genetic basis of the Pm21 locus and the disease resistance pathways mediated by Pm21.

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Acknowledgments

This research was supported by Grants from the National Natural Science Foundation of China (31471497, 31300138), the Natural Science Foundation of Jiangsu Province (BK20130503, BK20140500), the Foundation of Key Laboratory of Wheat Biology and Genetic Improvement on Low & Middle Yangtze River Valley Winter Wheat Region (2015YAAS-OP-001), and the Foundation of Jiangsu University (13JDG103). The authors are grateful to Professor PD Chen (the Cytogenetics Institute, Nanjing Agricultural University) for supporting this research.

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    1. School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, Jiangsu, China

      Huagang He, Yaoyong Ji & Feng Wang

    2. School of Environment, Jiangsu University, Zhenjiang, 212013, Jiangsu, China

      Shanying Zhu

    3. Key Laboratory of Wheat Biology and Genetic Improvement On Low and Middle Yangtze River Valley Wheat Region (Ministry of Agriculture), Yangzhou Academy of Agricultural Sciences, Yangzhou, 225007, Jiangsu, China

      Huagang He, Zhengning Jiang, Renhui Zhao & Tongde Bie

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    Huagang He and Shanying Zhu: These authors contributed equally to this work.

    Electronic supplementary material

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    Supplementary material 1. Primers for 6VS-specific markers. F: forward primer; R: reverse primer (PDF 96 kb)

    122_2016_2668_MOESM2_ESM.pdf

    Supplementary material 2. Primers for VIGS and quantitative real-time PCR. F: forward primer, R: reverse primer. Primers designed to amplify specific gene fragments for VIGS, and perform quantitative real-time PCR are designated by V and Q, respectively. Restriction sites are shown in italic (GAATTC for EcoRI and GTCGAC for SalI) (PDF 65 kb)

    122_2016_2668_MOESM3_ESM.pdf

    Supplementary material 3. Arabidopsis homologs of the candidate D. villosum proteins. The data were obtained by BLAST against The Arabidopsis Information Resource (TAIR) using the corresponding wheat proteins as query sequences. a, The protein sequence of wheat ERF, not found in the annotated wheat genome, was predicted by ORF Finder from Chinese spring cDNA AK333697. The coverage of matched sequence of wheat ERF against Arabipdopsis homolog AtERF was only 10 % (PDF 25 kb)

    122_2016_2668_MOESM4_ESM.pdf

    Supplementary material 4. Second structures of wheat proteins homologous to the candidate D. villosum proteins predicted by the SMART program. The bar represents the numbers of amino acid of wheat proteins (PDF 75 kb)

    122_2016_2668_MOESM5_ESM.pdf

    Supplementary material 5. Silencing efficiency analysis of the candidate genes by using qPCR, using BSMV:00-infected sample as the control. The transcriptional values of the tested genes in BSMV:00-infected materials were set to 1. The error bars indicate SD (PDF 27 kb)

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    He, H., Zhu, S., Jiang, Z. et al. Comparative mapping of powdery mildew resistance gene Pm21 and functional characterization of resistance-related genes in wheat. Theor Appl Genet 129, 819–829 (2016). https://doi.org/10.1007/s00122-016-2668-4

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