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Fusaric acid is a virulence factor of Fusarium oxysporum f. sp. cubense on banana plantlets

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Abstract

Fusaric acid (FA) is highly phytotoxic and causes severe Fusarium oxysporum-induced vascular wilt. However, the molecular mechanisms of FA production in Fusarium oxysporum f. sp. cubense (FOC) are largely unknown. In this study, FoFUB4, a F. verticillioides FUB4 homolog was cloned from banana pathogenic fungus FOC and its deletion mutants were generated using homologous recombination. Our null mutation results showed that FoFUB4 did not affect fungal development and conidiation, but was essential for virulence on banana plantlets compared with wild type (WT) strain. Biochemical analysis showed that production of FA was not detected in mutant ΔFoFUB4 and FA production by WT was significantly reduced in Czapek Dox medium containing 6 compared with 120 mM NaNO3. Moreover, transcriptional analysis indicated that the expression levels of FoFUB4 were reduced in WT in Czapek Dox medium containing 6 compared with 120 mM NaNO3. Taken together, these results provide direct evidence for the vital roles of FA in virulence, and that nitrogen source regulated FA biosynthesis in FOC.

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Fig. 1: Diagram of gene replacement and complementation vector strategy.
Fig. 2: Identification of mutants ΔFoFUB4 and complemented transformants ΔFoFUB4-c with PCR.
Fig. 3: Determination of FA production.
Fig. 4: Phytotoxic effect of FA on banana plantlets.
Fig. 5: Virulence assay of WT, mutant ΔFoFUB4, complemented strain ΔFoFUB4-c and CK.
Fig. 6: Expression levels of FoFUB4 in WT in Czapek Dox medium containing 6 or 120 mM NaNO3.
Fig. 7: Production of FA in FOC influenced by nitrogen source.

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Acknowledgements

This study was supported by funding from the Natural Science Foundation of Hainan Province (No. 20163103), Basic Scientific Research Expenses for the Central Scientific Research Institutes (No. 2016hzs1J016; 2017hzs1J007) and National Natural Science Foundation of China (No. 31371900; 31471738; 31571957).

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Authors and Affiliations

  1. Key Laboratory of the Ministry of Agriculture for Integrated Pest Management on Tropical Crops, Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, 571101, China

    Zhaojian Ding, Laying Yang, Guofen Wang, Lijia Guo, Lei Liu, Jun Wang & Junsheng Huang

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  1. Zhaojian Ding
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  2. Laying Yang
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  3. Guofen Wang
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  4. Lijia Guo
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  5. Lei Liu
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  6. Jun Wang
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  7. Junsheng Huang
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Correspondence to Junsheng Huang.

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Section Editor: Marciel J. Stadnik

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ESM 1

(DOC 48 kb)

Fig. S1

Schematic representation of functional domains and intron positions of FoFUB4 in FOC. (TIF 112 kb)

High resolution image (GIF 1 kb)

Fig. S2

Colony morphology of WT, mutant ΔFoFUB4 and complemented strain ΔFoFUB4-c. Indicated strains were incubated at 28 °C for 4 days on PDA plates. (TIFF 956 kb)

High resolution image (GIF 72 kb)

Fig. S3

FoFUB4 does not affect biomass and conidiation of FOC. (A) Dry weight of biomass. Indicated strains were cultured in YPD at 28 °C and 160 rpm for 3 days. (B) Microconidial productivity. Indicated strains grown on PDA plates at 28 °C for 7 days. Microconidia were quantified under a microscope with the aid of a hemocytometer. Values represent means ± SD from three replicates. (TIF 89 kb)

High resolution image (GIF 4 kb)

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Ding, Z., Yang, L., Wang, G. et al. Fusaric acid is a virulence factor of Fusarium oxysporum f. sp. cubense on banana plantlets. Trop. plant pathol. 43, 297–305 (2018). https://doi.org/10.1007/s40858-018-0230-4

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  • DOI: https://doi.org/10.1007/s40858-018-0230-4

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