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Expression of Fusarium pseudograminearum FpNPS9 in wheat plant and its function in pathogenicity

Abstract

Fusarium pseudograminearum-induced crown rot causes significant reduction to wheat production worldwide. To date, efforts to develop effective resistance to this disease have been hampered by the quantitative nature of resistance trait and a lack of understanding of the molecular pathogenesis. Non-ribosomal peptides have important roles in development, pathogenicity, and toxins in many plant pathogens, while less is known in F. pseudograminearum. In this work, we studied the expression and function of a nonribosomal peptide gene FpNPS9 in F. pseudograminearum. We determined the expression of FpNPS9 which was significantly up regulated during the infection of wheat. A deletion mutant Δfpnps9 produced in this study displayed a normal growth and conidiation phenotype, however, hyphae polar growth was obviously affected. Deoxynivalenol production in this mutant was significantly reduced and the infection of wheat coleoptiles and wheat spikelet was attenuated. The Δfpnps9 showed serious defects on the extension of infectious hyphae in plant and inhibition of roots elongation compared with the wild type. The complementation assay using a FpNPS9-GFP fusion construct fully restored the defects of the mutant. GFP signal was detected in the germinating conidia and infectious hyphae in coleoptiles of the infected plants. Interestingly, the signal was not observed when it was grown on culture medium, suggesting that the expression of FpNPS9 was regulated by an unknown host factor. This observation was supported by the result of qRT-PCR. In summary, we provided new knowledge on FpNPS9 expression in F. pseudograminearum and its function in F. pseudograminearum pathogenicity in wheat.

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Acknowledgements

We give thanks to Dr. Daniel Ebbole (Texas A&M University, College Station, TX, USA) for a critical reading of the manuscript. This work was supported by National Key R&D Program of China (Grant number 2017YFD0301104) and National Special Fund for Agro-scientific Research in the Public Interest (Grant number 201503112).

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Correspondence to Honglian Li.

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Fig. S1 Tween 20 restored the pathogenicity of the mutant Δfpnps9.a and b showed two repeated experiments results (TIFF 16577 kb)

Fig. S2 Assessment of complementation assay. The expression profiles of several genes in complemented strain during development and infectious stage using cDNA as templates. GFP: 650 bp using primer pair Pfl2-gfp-F/Pfl2-gfp-R in GFP fragment; Neo: 583 bp using primer pair Neo-F/Neo-R in Neo; NPS9 + GFP: 1154 bp using primer pair pKNTG-com-F/pKNTG-com-R spanning FpNPS9 and GFP gene, respectively. FpNPS9: 284 bp using primer pair NPS9-cDNA-F1/NPS9- cDNA-R1 in FpNPS9. Reference genes: 350 bp and 420 bp using primer pairs Fp-tubulin-F/R and Fp-actin-F/R in Fp, respectively (TIFF 6046 kb)

Fig. S3 The graphic view for FpNPS9 and FpNPS5 loci on chromosome 3 in F. pseudograminearum. The arrangement is the same with the orthologs in F. graminearum. The green shadow indicated the FpNPS9 and the red box showed the FpNPS5 (TIFF 13364 kb)

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Kang, R., Li, G., Zhang, M. et al. Expression of Fusarium pseudograminearum FpNPS9 in wheat plant and its function in pathogenicity. Curr Genet 66, 229–243 (2020). https://doi.org/10.1007/s00294-019-01017-2

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Keyword

  • Fusarium crown rot
  • Fusarium pseudograminearum
  • Nonribosomal peptide synthetase
  • Pathogenicity
  • Deoxynivalenol (DON)
  • FpNPS9