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The putative histone-like transcription factor FgHltf1 is required for vegetative growth, sexual reproduction, and virulence in Fusarium graminearum

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Abstract

The target of rapamycin (TOR) signaling pathway plays critical roles in regulating vegetative development and virulence in Fusarium graminearum. Previously, we have demonstrated that the putative type 2A phosphatase FgPpg1, a downstream component of the pathway, is important for hyphal growth, sporulation, DON biosynthesis and virulence. Here, we report the identification of FgHLTF1 putatively encoding a histone-like transcription factor by the transcriptome analysis of an ΔFgppg1 mutant. The FgHLTF1 expression was significantly down-regulated by the deletion of FgPPG1 or treatment with rapamycin. Analysis of an F. graminearum strain expressing green fluorescent protein (GFP) revealed that FgHltf1-GFP fusion protein mainly localized to the nucleus. Targeted gene deletion mutants of FgHLTF1 exhibited a significant reduction in vegetative growth, sexual reproduction and virulence. Moreover, the growth of the ΔFghltf1 mutants was restricted by hyperosmotic stresses. Unlike the wild-type strain, the mutants showed anomalous subcellular translocation of FgHog1-GFP under hyperosmotic conditions, suggesting that FgHLTF1 is associated with the high osmolarity glycerol response pathway. Taken together, we conclude that FgHLTF1 is transcriptionally regulated by the TOR signaling pathway and plays important roles in regulating vegetative growth, sexual reproduction, virulence and hyperosmotic stresses in F. graminearum.

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Acknowledgements

This work was supported by National Key Basic Research and Development Program (2013CB127802) to ZYW.

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Authors and Affiliations

  1. State Key Laboratory of Rice Biology and Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects, Institute of Biotechnology, Zhejiang University, Hangzhou, 310029, China

    Wuyun Lv, Jinjin Wu, Zhe Xu, Han Dai, Zhonghua Ma & Zhengyi Wang

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  1. Wuyun Lv
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  2. Jinjin Wu
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294_2019_953_MOESM1_ESM.psd

Figure S1. Targeted gene replacement of FgHLTF1. (A) Construction of the DNA fragment for the FgHLTF1 deletion. KO1-F/KO1-R and KO2-F/KO2-R were primers used for the verification of FgHLTF1 gene deletion. (B-E) Agarose gel electrophoresis of PCR analysis. B, PCR results with KO1-F/KO1-R primers indicated the presence of the FgHLTF1 gene or not. C, PCR results with KO2-F/KO2-R primers indicated the size difference of DNA fragment. D, Southern blot analysis. Genomic DNAs of the wild-type PH-1, ΔFghltf1 mutants and the complemented strain cΔFghltf1 were digested with Xmn I and BamH I and probed with a ~1.0 kb fragment (PSD 3842 KB)

294_2019_953_MOESM2_ESM.psd

Figure S2. FgHLTF1 is involved in the cell wall degrading enzyme production. (A) The wild-type strain PH-1, the ΔFghltf1 mutants and the complemented strain cΔFghltf1 were grown on CMC and CM plates. (B) The percentage of mycelial radial growth inhibition was quantified. Linear bars in each column denote standard errors of three experiments. An asterisk indicates a significant difference in the inhibition of mycelial growth (P < 0.05) (PSD 10251 KB)

294_2019_953_MOESM3_ESM.psd

Figure S3. The localization of FgHltf1-GFP and FgHog1-RFP in the wild-type strain PH-1. The hyphae were incubated in liquid CM medium (A) with or without 0.7 M NaCl treatment for 2 h (B). Nuclei were stained with DAPI. Scale bars = 5 µm (PSD 2294 KB)

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Lv, W., Wu, J., Xu, Z. et al. The putative histone-like transcription factor FgHltf1 is required for vegetative growth, sexual reproduction, and virulence in Fusarium graminearum. Curr Genet 65, 981–994 (2019). https://doi.org/10.1007/s00294-019-00953-3

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