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Involvement of two glycoside hydrolase family 19 members in colony morphotype and virulence in Flavobacterium columnare

  • Aquaculture and Fisheries
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Abstract

Flavobacterium columnare is the pathogenic agent of columnaris disease in aquaculture. Using a recently developed gene deletion strategy, two genes that encode the Glyco_hydro_19 domain (GH19 domain) containing proteins, ghd-1 and ghd-2, were deleted separately and together from the F. columnare G4 wild type strain. Surprisingly, the single-, Δghd-1 and Δghd-2, and double-gene mutants, Δghd-1 Δghd -2, all had rhizoid and non-rhizoid colony morphotypes, which we named Δghd-1, Δghd-2, Δghd-1 Δghd-2, and NΔghd-1, NΔghd-2, and NΔghd-1 Δghd-2. However, chitin utilization was not detected in either these mutants or in the wild type. Instead, skimmed milk degradation was observed for the mutants and the wild type; the non-rhizoid strain NΔghd-2 exhibited higher degradation activity as revealed by the larger transparent circle on the skimmed milk plate. Using zebrafish as the model organism, we found that non-rhizoid mutants had higher LD50 values and were less virulent because zebrafish infected with these survived longer. Transcriptome analysis between the non-rhizoid and rhizoid colony morphotypes of each mutant, i.e., NΔ ghd -1 versus (vs) Δghd-1, NΔghd-2 vs Δghd-2, and NΔghd-1 Δghd-2 vs Δghd-1 Δghd-2, revealed a large number of differentially expressed genes, among which 39 genes were common in three of the pairs compared. Although most of these genes encode hypothetical proteins, a few molecules such as phage tail protein, rhs element Vgr protein, thiol-activated cytolysin, and TonB-dependent outer membrane receptor precursor, expression of which was down-regulated in non-rhizoid mutants but up-regulated in rhizoid mutants, may play a role F. columnare virulence.

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Authors and Affiliations

  1. State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China

    Xiaolin Zhang  (张晓林), Nan Li  (李楠), Ting Qin  (秦婷) & Pin Nie  (聂品)

  2. University of Chinese Academy of Sciences, Beijing, 100049, China

    Xiaolin Zhang  (张晓林)

  3. College of Fisheries, Jimei University, Xiamen, 361021, China

    Bei Huang  (黄贝)

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  1. Xiaolin Zhang  (张晓林)
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  2. Nan Li  (李楠)
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  3. Ting Qin  (秦婷)
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  4. Bei Huang  (黄贝)
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  5. Pin Nie  (聂品)
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Correspondence to Pin Nie  (聂品).

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Supported by the Chinese Academy of Sciences (No. XDA08010207), the National Key Technology R&D Program of China (No. 2012BAD25B02), and the State Key Laboratory of Freshwater Ecology and Biotechnology (No. 2016FBZ04)

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Zhang, X., Li, N., Qin, T. et al. Involvement of two glycoside hydrolase family 19 members in colony morphotype and virulence in Flavobacterium columnare. Chin. J. Ocean. Limnol. 35, 1511–1523 (2017). https://doi.org/10.1007/s00343-017-6160-z

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