Gu Yi, Lu Huiqi, Shao Ying, Fu Dandan, Wu Jianmei, Hu Jiangang, Tu Jian, Song Xiangjun, Qi Kezong
Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, Anhui Agricultural University, Hefei 230036, Anhui, China.
Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, Anhui Agricultural University, Hefei 230036, Anhui, China.
Res Vet Sci. 2022 Dec 31;153:144-152. doi: 10.1016/j.rvsc.2022.10.016. Epub 2022 Oct 28.
Avian pathogenic Escherichia coli (APEC) causes persistent infection of poultry and multi-system diseases, which seriously endanger the development of the poultry industry. Biofilm allows bacteria to adapt to the natural environment and plays an important role in resistance to the external environment and the pathogenicity of APEC, but the mechanism of its formation and regulatory network have not been clarified. In this study, we used a Tn5 transposon random mutation library constructed with APEC and identified ydiF, a gene that has not previously been recognized in E. coli biofilm formation. To confirm that the ydiF gene really can regulate the formation of APEC biofilm, the ydiF gene deletion strain was constructed using APEC81. Protein association networks prediction results show that ydiF is mainly associated with genes related to the metabolism of sugars and fatty acids. Deletion of the ydiF gene significantly reduces the formation of APEC biofilm and scanning electron microscopy indicated that the degree of adhesion between the bacteria was also reduced. The deletion of the ydiF gene also significantly reduced the motility of APEC81 and through transmission electron microscopy APEC81 was observed to have significantly fewer flagella. However, the colony morphology of APEC81 on Congo red and Coomassie brilliant blue media was unaffected. The results of fluorescence quantification showed that the deletion of the ydiF gene caused a down-regulation in the transcription of genes related to the second messenger, sugar metabolism, and quorum sensing. These results indicate that ydiF plays an important role in biofilm formation and the movement of APEC. In addition, it may be possible to regulate the formation of APEC biofilms by different methods such as by regulating the second messenger and metabolic system.
禽致病性大肠杆菌(APEC)可导致家禽持续感染和多系统疾病,严重危及家禽业的发展。生物膜使细菌能够适应自然环境,在抵抗外部环境和APEC的致病性方面发挥重要作用,但其形成机制和调控网络尚未阐明。在本研究中,我们使用用APEC构建的Tn5转座子随机突变文库,鉴定出ydiF,一个以前在大肠杆菌生物膜形成中未被识别的基因。为了证实ydiF基因确实可以调节APEC生物膜的形成,使用APEC81构建了ydiF基因缺失菌株。蛋白质关联网络预测结果表明,ydiF主要与糖和脂肪酸代谢相关的基因有关。ydiF基因的缺失显著降低了APEC生物膜的形成,扫描电子显微镜显示细菌之间的粘附程度也降低了。ydiF基因的缺失还显著降低了APEC81的运动性,通过透射电子显微镜观察到APEC81的鞭毛明显减少。然而,APEC81在刚果红和考马斯亮蓝培养基上的菌落形态未受影响。荧光定量结果表明,ydiF基因的缺失导致与第二信使、糖代谢和群体感应相关基因的转录下调。这些结果表明,ydiF在APEC的生物膜形成和运动中起重要作用。此外,可能可以通过不同方法调节APEC生物膜的形成,例如通过调节第二信使和代谢系统。