Analysis of the biofilm proteome of Xylella fastidiosa

Silva, Mariana S.; Souza, Alessandra Alves de; Takita, Marco Aurélio; Labate, Carlos Alberto; Machado, Marcos Antônio

doi:10.1186/1477-5956-9-58

Cited by 26 publications

(23 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Zinc is a structural and catalytic cofactor in a variety of proteins including Zn-metalloproteases important in digesting plant components for the nutrition of the bacterium [33], [34], [35], [36]. Based on observations that supplemental Zn inhibited biofilm formation in microtiter plates [17] we chose to supplement PD2 media with zinc at 0.25 mM and at 2.5 mM.…”

Section: Resultsmentioning

confidence: 99%

Xylella fastidiosa Differentially Accumulates Mineral Elements in Biofilm and Planktonic Cells

et al. 2013

View full text Add to dashboard Cite

Xylella fastidiosa is a bacterial plant pathogen that infects numerous plant hosts. Disease develops when the bacterium colonizes the xylem vessels and forms a biofilm. Inductively coupled plasma optical emission spectroscopy was used to examine the mineral element content of this pathogen in biofilm and planktonic states. Significant accumulations of copper (30-fold), manganese (6-fold), zinc (5-fold), calcium (2-fold) and potassium (2-fold) in the biofilm compared to planktonic cells were observed. Other mineral elements such as sodium, magnesium and iron did not significantly differ between biofilm and planktonic cells. The distribution of mineral elements in the planktonic cells loosely mirrors the media composition; however the unique mineral element distribution in biofilm suggests specific mechanisms of accumulation from the media. A cell-to-surface attachment assay shows that addition of 50 to 100 µM Cu to standard X. fastidiosa media increases biofilm, while higher concentrations (>200 µM) slow cell growth and prevent biofilm formation. Moreover cell-to-surface attachment was blocked by specific chelation of copper. Growth of X. fastidiosa in microfluidic chambers under flow conditions showed that addition of 50 µM Cu to the media accelerated attachment and aggregation, while 400 µM prevented this process. Supplementation of standard media with Mn showed increased biofilm formation and cell-to-cell attachment. In contrast, while the biofilm accumulated Zn, supplementation to the media with this element caused inhibited growth of planktonic cells and impaired biofilm formation. Collectively these data suggest roles for these minerals in attachment and biofilm formation and therefore the virulence of this pathogen.

show abstract

Section: Resultsmentioning

confidence: 99%

Xylella fastidiosa Differentially Accumulates Mineral Elements in Biofilm and Planktonic Cells

et al. 2013

View full text Add to dashboard Cite

show abstract

“…These include a the non-fimbrial adhesin, YapH [26], the FadL porin [27], citrate synthase [28], UDP-glucose dehydrogenase [19], the molecular chaperone DnaK [29-31], the elongation factor Ef-Tu [29,32], the polynucleotide phosphorylase [33] and a TonB-dependent receptor protein [19] (Table 2). These findings further validate our experimental results.…”

Section: Resultsmentioning

confidence: 99%

“…Ef-Tu was also over-expressed in the wild type strain of Lactobacillus crispatus as compared to an isogenic mutant that lost the aggregative phenotype and strengthening the claim for a role in adhesion [53]. Moreover, in the citrus pathogen Xylella fastidiosa , Ef-Tu was reported to be up-regulated in biofilms [32]. Recent work demonstrated that in X .…”

Section: Resultsmentioning

confidence: 99%

Insights into xanthomonas axonopodis pv. citri biofilm through proteomics

et al. 2013

View full text Add to dashboard Cite

BackgroundXanthomonas axonopodis pv. citri (X. a. pv. citri) causes citrus canker that can result in defoliation and premature fruit drop with significant production losses worldwide. Biofilm formation is an important process in bacterial pathogens and several lines of evidence suggest that in X. a. pv. citri this process is a requirement to achieve maximal virulence since it has a major role in host interactions. In this study, proteomics was used to gain further insights into the functions of biofilms.ResultsIn order to identify differentially expressed proteins, a comparative proteomic study using 2D difference gel electrophoresis was carried out on X. a. pv. citri mature biofilm and planktonic cells. The biofilm proteome showed major variations in the composition of outer membrane proteins and receptor or transport proteins. Among them, several porins and TonB-dependent receptor were differentially regulated in the biofilm compared to the planktonic cells, indicating that these proteins may serve in maintaining specific membrane-associated functions including signaling and cellular homeostasis. In biofilms, UDP-glucose dehydrogenase with a major role in exopolysaccharide production and the non-fimbrial adhesin YapH involved in adherence were over-expressed, while a polynucleotide phosphorylase that was demonstrated to negatively control biofilm formation in E. coli was down-regulated. In addition, several proteins involved in protein synthesis, folding and stabilization were up-regulated in biofilms. Interestingly, some proteins related to energy production, such as ATP-synthase were down-regulated in biofilms. Moreover, a number of enzymes of the tricarboxylic acid cycle were differentially expressed. In addition, X. a. pv. citri biofilms also showed down-regulation of several antioxidant enzymes. The respective gene expression patterns of several identified proteins in both X. a. pv. citri mature biofilm and planktonic cells were evaluated by quantitative real-time PCR and shown to consistently correlate with those deduced from the proteomic study.ConclusionsDifferentially expressed proteins are enriched in functional categories. Firstly, proteins that are down-regulated in X. a. pv. citri biofilms are enriched for the gene ontology (GO) terms ‘generation of precursor metabolites and energy’ and secondly, the biofilm proteome mainly changes in ‘outer membrane and receptor or transport’. We argue that the differentially expressed proteins have a critical role in maintaining a functional external structure as well as enabling appropriate flow of nutrients and signals specific to the biofilm lifestyle.

show abstract

“…A previous studies of the X. fastidiosa proteome revealed differences between the planktonic and biofilm stages in the expression of proteins related to metabolism, motility, attachment, and stress conditions (Silva et al, 2011) and showed the presence of 30 extracellular proteins related to survival and pathogenesis after growth in solid medium for 21 days (Smolka et al, 2003; Silva et al, 2011). In the present study, we identified 48 proteins from strain 9a5c and 44 proteins from strain J1a12, which resulted in the identification of 71 non-redundant proteins, of which 21 were identified in both strains ( Figures 3A,B ).…”

Section: Resultsmentioning

confidence: 95%

In vitro Determination of Extracellular Proteins from Xylella fastidiosa

et al. 2016

View full text Add to dashboard Cite

The phytopathogen Xylella fastidiosa causes economic losses in important agricultural crops. Xylem vessel occlusion caused by biofilm formation is the major mechanism underlying the pathogenicity of distinct strains of X. fastidiosa. Here, we provide a detailed in vitro characterization of the extracellular proteins of X. fastidiosa. Based on the results, we performed a comparison with a strain J1a12, which cannot induce citrus variegated chlorosis symptoms when inoculated into citrus plants. We then extend this approach to analyze the extracellular proteins of X. fastidiosa in media supplemented with calcium. We verified increases in extracellular proteins concomitant with the days of growth and, consequently, biofilm development (3–30 days). Outer membrane vesicles carrying toxins were identified beginning at 10 days of growth in the 9a5c strain. In addition, a decrease in extracellular proteins in media supplemented with calcium was observed in both strains. Using mass spectrometry, 71 different proteins were identified during 30 days of X. fastidiosa biofilm development, including proteases, quorum-sensing proteins, biofilm formation proteins, hypothetical proteins, phage-related proteins, chaperones, toxins, antitoxins, and extracellular vesicle membrane components.

show abstract

Analysis of the biofilm proteome of Xylella fastidiosa

Cited by 26 publications

References 34 publications

Xylella fastidiosa Differentially Accumulates Mineral Elements in Biofilm and Planktonic Cells

Xylella fastidiosa Differentially Accumulates Mineral Elements in Biofilm and Planktonic Cells

Insights into xanthomonas axonopodis pv. citri biofilm through proteomics

In vitro Determination of Extracellular Proteins from Xylella fastidiosa

Contact Info

Product

Resources

About