![]() ![]() aeruginosa PAO1 and Flg22-1, and Flg22-2 from P. (A) Sequence alignment of Flg22_Pa from P. Plant immunity induced by Flg22-1 and Flg22-2. All experiments were repeated three times with similar results. Different letters indicate statistically significant differences between different treatments (one-way ANOVA, Tukey’s test P < 0.05). (D) Quantitative analysis of primary root length, number of lateral roots, and lateral root density. kilonensis F113 and its fliC mutants on shoot and root growth in A. ![]() The fraction under each image indicates the number of times that the HR was inhibited compared to the number of test inoculations. After 6 h, an overlapping inoculation of 5 × 10 6 CFU/mL of the HR-inducing strain Pst DC3000 (lower circles) was made. benthamiana leaves with 1 × 10 8 CFU/mL of the test Pseudomonas strains (upper circles). (B) Challenge-inoculation HR assays for functional PTI were conducted by first infiltrating N. kilonensis F113, and the fliC mutants in N. Our findings on the divergence of duplicated flagellins provide a conceptual framework for better understanding the functional determinant flagellin and its peptide in multiple-flagellin plant-growth-promoting rhizobacteria.įlg22 Pseudomonas flagella flagellin motility plant immunity plant-microbe interactions.Ĭontribution of the duplicated flagellins to plant immunity and root growth. This work focuses on flagellin duplication in some plant-associated Pseudomonas. IMPORTANCE Flagellins have emerged as important microbial patterns. These findings suggest that Pseudomonas has evolved duplicate flagellins, with only FliC-1 contributing to motility and plant immune elicitation. Also, Flg22-2 epitopes in the other 11 Pseudomonas strains were presumed to have low binding affinity due to the same change in the 19th amino acid. A change in the 19th amino acid in Flg22-2 reduced its binding affinity to the FLS2/brassinosteroid insensitive 1-associated kinase 1 complex. Compared to the Flg22-2 epitope (relative to FliC-2), Flg22-1 (relative to FliC-1) induced stronger FLAGELLIN SENSING 2 (FLS2)-mediated microbe-associated molecular pattern-triggered immunity and significantly inhibited plant root growth. ![]() In silico prediction revealed that all fliC-2 genes in the 12 Pseudomonas strains have a poorly conserved promoter motif. The transcriptional expression of fliC-2 was significantly lower than that of fliC-1 in medium and in planta, most likely due to variations in promoter activity. Only the fliC-1 gene but not the fliC-2 gene in strain F113 is responsible for flagellar biogenesis, motility, and plant immune elicitation. We demonstrate that the two flagellins (FliC-1 and FliC-2) in 12 Pseudomonas strains, including F113, are evolutionarily distinct. Here, the model plant-growth-promoting bacterium Pseudomonas kilonensis F113 was used to decipher the divergent functions of duplicated flagellins. Although the functions of flagellins have been extensively studied in bacteria with a single flagellin-encoding gene, the function of multiple flagellin-encoding genes in a single bacterial species is largely unknown. Flagellins are the main constituents of the flagellar filaments that provide bacterial motility, chemotactic ability, and host immune elicitation ability. ![]()
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