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Chinese Journal of Oceanology and Limnology

, Volume 35, Issue 6, pp 1511–1523 | Cite as

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

  • Xiaolin Zhang (张晓林)
  • Nan Li (李楠)
  • Ting Qin (秦婷)
  • Bei Huang (黄贝)
  • Pin Nie (聂品)
Aquaculture and Fisheries

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.

Keywords

Flavobacterium columnare GH19 domain gene deletion rhizoid colony non-rhizoid colony 

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Copyright information

© Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Xiaolin Zhang (张晓林)
    • 1
    • 2
  • Nan Li (李楠)
    • 1
  • Ting Qin (秦婷)
    • 1
  • Bei Huang (黄贝)
    • 3
  • Pin Nie (聂品)
    • 1
  1. 1.State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of HydrobiologyChinese Academy of SciencesWuhanChina
  2. 2.University of Chinese Academy of SciencesBeijingChina
  3. 3.College of FisheriesJimei UniversityXiamenChina

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