Functional characterization and transcriptional analysis of icd2 gene encoding an isocitrate dehydrogenase of Xanthomonas campestris pv. campestris

Chiang, Ying-Chuan; Liao, Chao-Tsai; Du, Shin-Chiao; Hsiao, Yi‐Min

doi:10.1007/s00203-017-1370-5

Cited by 9 publications

(7 citation statements)

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“…As several factors having a role in bacterial attachment have been reported to involved in pathogenicity in X. campestris pv. campestris [10, 11, 13, 14], we aimed to determine whether the lolA mutation affected the virulence of X. campestris pv. campestris .…”

Section: Resultsmentioning

confidence: 99%

“…In X. campestris pv. campestris , several factors reported to influence biofilm formation and bacterial adhesion also have roles in pathogenicity, namely extracellular mannanase ManA [6], GGDEF domain protein XC_0249 [9], guanylate cyclase XC_0250 [9], isocitrate dehydrogenase encoded by icd2 [10], major outer membrane protein MopB [11], the RavA/RavR two-component system [12], tail-specific protease encoded by prc [13], and UTP-glucose-1-phosphate uridylyltransferase encoded by galU [14].…”

Section: Introductionmentioning

confidence: 99%

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Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris

2019

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BackgroundThe gram-negative Xanthomonas campestris pv. campestris is the pathogenic bacterium that causes black rot disease in crucifers. The virulence determinants of this bacterium include extracellular enzymes, exopolysaccharides, and biofilm formation. Here, one transposon mutant of X. campestris pv. campestris strain 17 that affects biofilm formation was isolated, and subsequent analyses led to the identification of the lolA gene, which encodes an outer membrane lipoprotein chaperone.ResultsThe lolA mutant exhibited significant reductions in bacterial attachment, extracellular enzyme production, virulence, and tolerance in the presence of myriad membrane-perturbing agents. These phenotypic changes of the mutant could be complemented to the wild-type level through the intact lolA gene. Proteomic analysis revealed that 109 proteins were differentially expressed after lolA mutation. These differentially expressed proteins were categorized in various functional groups and were mainly associated with the membrane component, were involved in transport, and contained receptor activity. Through reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis, deletion of lolA was determined to have caused significantly reduced expression of genes that encode the major extracellular enzymes, the biofilm-related proteins, and the virulence-related proteins. The RT-qPCR analysis also indicated that the expression of several genes that encode putative outer membrane lipoproteins and TonB-dependent receptors was reduced after lolA mutation.ConclusionsThis is the first report to define the lolA gene as a virulence factor and to contribute to the functional understanding of, and provide new information concerning, the role of lolA in Xanthomonas. Furthermore, the results of this study provide and extend new insights into the function of lolA in bacteria.Electronic supplementary materialThe online version of this article (10.1186/s12866-019-1387-9) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris

2019

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“…Furthermore, CRYL1 has been shown to be highly expressed in mammalian livers and kidneys, organs directly involved in lipid metabolism (Chen et al, 2003), and thus this gene in SEL BSF may fulfil a similar role, particularly during the feeding period of this species. Additionally, the gene isocitrate dehydrogenase 1 ( IDH1 , LOC119648135) encodes a rate‐limiting enzyme in the tricarboxylic acid cycle which is involved in cellular energy metabolism (Chiang et al, 2017), and has shown evidence that expression of this gene fulfils a protective function, supressing intracellular and mitochondrial reactive oxidative stress levels (Yang et al, 2017). Increased expression of both of these genes (among others) in SEL F3 larvae (Figure 10) in conjunction with significant positive correlations with larval mass (Table S6) illustrates the functional metabolic shift in this experimental line.…”

Section: Discussionmentioning

confidence: 99%

Gene expression differentials driven by mass rearing and artificial selection in black soldier fly colonies

Hull

Greenwood

Lloyd³

et al. 2022

Insect Molecular Biology

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The micro-evolutionary forces that shape genetic diversity during domestication have been assessed in many plant and animal systems. However, the impact of these processes on gene expression, and consequent functional adaptation to artificial environments, remains under-investigated. In this study, whole-transcriptome dynamics associated with the early stages of domestication of the black soldier fly (BSF), Hermetia illucens, were assessed. Differential gene expression (DGE) was evaluated in relation to (i) generational time within the cultured environment (F2 vs. F3), and (ii) two selection strategies [no artificial selective pressure (NS); and selection for greater larval mass (SEL)]. RNA-seq was conducted on 5th instar BSF larvae (n = 36), representing equal proportions of the NS (F2 = 9; F3 = 9) and SEL (F2 = 9; F3 = 9) groups. A multidimensional scaling plot revealed greater gene expression variability within the NS and F2 subgroups, while the SEL group clustered separately with lower levels of variation.Comparisons between generations revealed 898 differentially expressed genes (DEGs; FDR-corrected p < 0.05), while between selection strategies, 213 DEGs were observed (FDR-corrected p < 0.05). Enrichment analyses revealed that metabolic, developmental, and defence response processes were over-expressed in the comparison between F2 and F3 larvae, while metabolic processes were the main differentiating factor between NS and SEL lines. This illustrates the functional adaptations that occur in BSF colonies across generations due to mass rearing; as well as highlighting genic dynamics associated with artificial selection for production traits that might inform future selective breeding strategies.

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“…Since several factors playing a role in bacterial attachment are involved in pathogenicity in Xcc [12][13][14]17,18], we aimed to determine whether the wxcX gene is associated with the virulence of Xcc. The wxcX mutant revealed reduced virulence compared with the wild type (Fig.…”

Section: Wxcx Is Required For the Full Virulence Of Xccmentioning

confidence: 99%

“…The role of biofilm formation in the virulence of plant pathogenic bacteria has been reported for a range of bacterial infections in plants, such as Erwinia amylovora , the causal agent of fire blight, as well as Xylella fastidiosa , the causal agent of oak leaf scorch . In Xcc, several factors having a role in biofilm formation or bacterial adhesion are involved in virulence, including exopolysaccharide , extracellular mannanase ManA , GGDEF domain proteins XCC2731 and XC_0249 , guanylate cyclase XC_0250 , isocitrate dehydrogenase encoded by icd2 , major outer‐membrane protein MopB , RavA/RavR two‐component system , RNA‐binding protein RsmA , tail‐specific protease encoded by prc , and UTP‐glucose‐1‐phosphate uridylyltransferase encoded by galU .…”

Section: Introductionmentioning

confidence: 99%

WxcX is involved in bacterial attachment and virulence in Xanthomonas campestris pv. campestris

et al. 2018

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Xanthomonas campestris pv. campestris (Xcc) is the causative agent of black rot in crucifers. Here, one EZ-Tn5 transposon mutant of Xcc, altered in bacterial attachment, was isolated. Further analysis revealed that the transposon was inserted in the wxcX gene (encodes a hypothetical protein) of the transposon mutant. Sequence analysis revealed that WxcX is highly conserved in Xanthomonas, but none has been characterized. In this study, it was indicated that mutation of wxcX resulted in enhanced bacterial attachment, reduced virulence on the host cabbage, and increased sensitivity to sodium dodecyl sulfate. The affected phenotypes of the wxcX mutant could be complemented to wild-type levels by the intact wxcX gene. Site-directed mutagenesis revealed that E408 and E411 are critical amino acid residues for WxcX function in bacterial attachment. Taken together, our results demonstrate the roles of wxcX in attachment, virulence, and tolerance to sodium dodecyl sulfate in Xanthomonas for the first time.

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Functional characterization and transcriptional analysis of icd2 gene encoding an isocitrate dehydrogenase of Xanthomonas campestris pv. campestris

Cited by 9 publications

References 57 publications

Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris

Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris

Gene expression differentials driven by mass rearing and artificial selection in black soldier fly colonies

WxcX is involved in bacterial attachment and virulence in Xanthomonas campestris pv. campestris

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