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Identification of Pathogenicity-Related Genes in Biofilm-Defective Acidovorax citrulli by Transposon Tn5 Mutagenesis

文献类型： 外文期刊

作者： Luo, Jinyan 1 ; Qiu, Wen 2 ; Chen, Lei 1 ; Anjum, Syed Ishtiaq 2 ; Yu, Menghao 2 ; Shan, Changlin 2 ; Ilyas, Mehmoona; 1 ;

作者机构： 1.Shanghai Extens & Serv Ctr Agr Technol, Dept Plant Quarantine, Shanghai 201103, Peoples R China

2.Zhejiang Univ, Inst Biotechnol, State Key Lab Rice Biol, Hangzhou 310058, Zhejiang, Peoples R China

3.Kohat Univ Sci & Technol, Dept Zool, Kohat 26000, Pakistan

4.Zhoushan Entry Exit Inspect & Quarantine Bur, Dept Plant Quarantine, Hangzhou 310012, Zhejiang, Peoples R China

5.Univ Sargodha, Dept Biotechnol, Sargodha 40100, Pakistan

6.Zhejiang Acad Agr Sci, Minist Agr, Key Lab Detect Pesticide Residues, State Key Lab Breeding B

关键词： bacterial fruit blotch;Tn5;Acidovorax citrulli;pathogenesis;biofilm

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）

ISSN： 1422-0067

年卷期： 2015 年 16 卷 12 期

页码：

收录情况： SCI

摘要： Biofilm formation is important for virulence of a large number of plant pathogenic bacteria. Indeed, some virulence genes have been found to be involved in the formation of biofilm in bacterial fruit blotch pathogen Acidovorax citrulli. However, some virulent strains of A. citrulli were unable to format biofilm, indicating the complexity between biofilm formation and virulence. In this study, virulence-related genes were identified in the biofilm-defective strain A1 of A. citrulli by using Tn5 insertion, pathogenicity test, and high-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR). Results from this study indicated that 22 out of the obtained 301 mutants significantly decreased the virulence of strain A1 compared to the wild-type. Furthermore, sequence analysis indicated that the obtained 22 mutants were due to the insertion of Tn5 into eight genes, including Aave 4244 (cation diffusion facilitator family transporter), Aave 4286 (hypothetical protein), Aave 4189 (alpha/beta hydrolase fold), Aave 1911 (IMP dehydrogenase/GMP reductase domain), Aave 4383 (bacterial export proteins, family 1), Aave 4256 (Hsp70 protein), Aave 0003 (histidine kinase, DNA gyrase B, and HSP90-like ATPase), and Aave 2428 (pyridoxal-phosphate dependent enzyme). Furthermore, the growth of mutant Aave 2428 was unaffected and even increased by the change in incubation temperature, NaCl concentration and the pH of the LB broth, indicating that this gene may be directly involved in the bacterial virulence. Overall, the determination of the eight pathogenicity-related genes in strain A1 will be helpful to elucidate the pathogenesis of biofilm-defective A. citrulli.

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