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Overexpression of the chitinase gene CmCH1 from Coniothyrium minitans renders enhanced resistance to Sclerotinia sclerotiorum in soybean

Abstract

Pathogenic fungi represent one of the major biotic stresses for soybean production across the world. Sclerotinia sclerotiorum, the causal agent of Sclerotinia stem rot, is a devastating fungal pathogen that is responsible for significant yield losses in soybean. In this study, the chitinase gene CmCH1, from the mycoparasitic fungus Coniothyrium minitans, which infects a range of ascomycetous sclerotia, including S. sclerotiorum and S. minor, was introduced into soybean. Transgenic plants expressing CmCH1 showed higher resistance to S. sclerotiorum infection, with significantly reduced lesion sizes in both detached stem and leaf assays, compared to the non-transformed control. Increased hydrogen peroxide content and activities of defense-responsive enzymes, such as peroxidase, superoxide dismutase, phenylalanine ammonia lyase, and polyphenoloxidase were also observed at the infection sites in the transgenic plants inoculated with S. sclerotiorum. Consistent with the role of chitinases in inducing downstream defense responses by the release of elicitors, several defense-related genes, such as GmNPR2, GmSGT-1, GmRAR1, GmPR1, GmPR3, GmPR12, GmPAL, GmAOS, GmPPO, were also significantly upregulated in the CmCH1-expressing soybean after inoculation. Collectively, our results demonstrate that overexpression of CmCH1 led to increased accumulation of H2O2 and up-regulation of defense-related genes and enzymes, and thus enhanced resistance to S. sclerotiorum infection while showing no detrimental effects on growth and development of soybean plants.

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Acknowledgements

This work was supported by grants from the Jilin Provincial Agricultural Science & Technology Innovation Project (CXGC2017JQ013) and China National Novel Transgenic Organisms Breeding Project (2016ZX08004-004). We thank Prof. Daohong Jiang of Huazhong Agricultural University for providing the CmCHI1 gene. We also thank Editage (www.editage.cn) for English language editing.

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YD and QL designed the experiments. XY and JY conducted the experiments and drafted the manuscript. GX, DG, and QZ conducted the Agrobacterium-mediated transformation experiments. WX and HL conducted the inoculation assay. LN and YZ participated in the qRT-PCR analyses. All authors participated in the manuscript revision.

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Correspondence to Qiyun Li or Yingshan Dong.

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Xiangdong Yang and Jing Yang have contributed equally to this work.

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M, DNA marker (2k); Ctl+, positive control; Nt, non-transformed plants Williams 82; Bk; Blank control; 1–7, T0 transgenic plants (PPTX 2921 kb)

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Yang, X., Yang, J., Li, H. et al. Overexpression of the chitinase gene CmCH1 from Coniothyrium minitans renders enhanced resistance to Sclerotinia sclerotiorum in soybean. Transgenic Res 29, 187–198 (2020). https://doi.org/10.1007/s11248-020-00190-2

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Keywords

  • Soybean
  • Chitinase
  • Sclerotinia sclerotiorum
  • Resistance
  • Coniothyrium minitans