Transformation of Xanthomonas axonopodis pv. citri by electroporation

Amaral, Alexandre Morais do; Toledo, Cristiane P.; Baptista, Juliana Cristina; Machado, Marcos Antônio

doi:10.1590/s0100-41582005000300013

Cited by 21 publications

(11 citation statements)

References 13 publications

(13 reference statements)

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“…The two constructs, pK-DfliC and pK-flgE : : V, were used to transform the wild-type strain by electroporation (do Amaral et al, 2005). Transformed bacteria were selected on PYM agar medium supplemented with kanamycin (Müller et al, 1993).…”

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confidence: 99%

The Xanthomonas axonopodis pv. citri flagellum is required for mature biofilm and canker development

et al. 2011

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The Xanthomonas axonopodis pv. citri flagellum is required for mature biofilm and canker development Xanthomonas axonopodis pv. citri (Xac) is the causative agent of citrus canker. This bacterium develops a characteristic biofilm on both biotic and abiotic surfaces. To evaluate the participation of the single flagellum of Xac in biofilm formation, mutants in the fliC (flagellin) and the flgE (hook) genes were generated. Swimming motility, assessed on 0.25 % agar plates, was markedly reduced in fliC and flgE mutants. However, the fliC and flgE mutants exhibited a flagellarindependent surface translocation on 0.5 % agar plates. Mutation of either the rpfF or the rpfC gene, which both encode proteins involved in cell-cell signalling mediated by diffusible signal factor (DSF), led to a reduction in both flagellar-dependent and flagellar-independent surface translocation, indicating a regulatory role for DSF in both types of motility. Confocal laser scanning microscopy of biofilms produced in static culture demonstrated that the flagellum is also involved in the formation of mushroom-shaped structures and water channels, and in the dispersion of biofilms. The presence of the flagellum was required for mature biofilm development on lemon leaf surfaces. The absence of flagellin produced a slight reduction in Xac pathogenicity and this reduction was more severe when the complete flagellum structure was absent.

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confidence: 99%

The Xanthomonas axonopodis pv. citri flagellum is required for mature biofilm and canker development

et al. 2011

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“…and X . citri was cultured in NBY solid medium and then in NBY liquid medium according to Amaral et al [ 33 ] for 48 hours at 28°C and 180 rpm. S .…”

Section: Methodsmentioning

confidence: 99%

Evaluation of cytotoxicity features of antimicrobial peptides with potential to control bacterial diseases of citrus

et al. 2018

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Antimicrobial peptides (AMPs) can be found in various organisms, and could be considered an alternative for pesticides used to control plant pathogens, including those affecting citrus. Brazil is the largest producer and exporter of frozen concentrated orange juice in the world. However, the citrus industry has been affected by several diseases such as citrus canker and huanglongbing (HLB), caused by the bacteria Xanthomonas citri subsp. citri (X.citri) and Candidatus Liberibacter asiaticus (CaLas), respectively. In order to control these pathogens, putative AMPs were prospected in databases containing citrus sequences. Furthermore, AMPs already reported in the literature were also used for in vitro and in vivo assays against X.citri. Since CaLas cannot be cultivated in vitro, surrogates as Sinorhizobium meliloti and Agrobacterium tumefaciens were used. This study reports the evaluation of six AMPs obtained from different sources, two of them from Citrus spp. (citrus-amp1 and citrus-amp2), three from amphibians (Hylin-a1, K0-W6-Hy-a1 and Ocellatin 4-analogue) and one from porcine (Tritrpticin). Peptides K0-W6-Hy-a1, Ocellatin 4-analogue, and citrus-amp1 showed bactericidal activity against X.citri and S. meliloti and bacteriostatic effect on A. tumefaciens. These results were confirmed for X.citri in planta. In addition cytotoxicity evaluations of these molecules were performed. The AMPs that showed the lowest hemolytic activities were Triptrpticin, citrus-amp1 and citrus-amp2. Citrus-amp1 and citrus-amp2 not presented toxicity in experiments using in vivo model, G. mellonella and U87 MG cells. To verify the interaction of these AMPs with bacteria and erythrocyte cell membranes, vesicles mimicking these cells were built. Citrus-amp1 and Tritrpticin exhibited higher affinity to bacterial membranes, while Ocellatin 4-analogue and Hylin-a1 showed higher affinity to erythrocyte membranes; exclude their use in citrus. This work demonstrates an essential alternative, trough AMPs obtained from Citrus spp., which can be feasibly used to control bacterial pathogens.

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“…Electrocompetent cells of Xcc were prepared as previously described with some modifications. Briefly, an overnight subculture of Xcc cells (MAFF 301080) was used to seed 500 mL of NB medium.…”

Section: Methodsmentioning

confidence: 99%

Dynamic integration and excision of filamentous phage XacF1 in Xanthomonas citri pv. citri, the causative agent of citrus canker disease

et al. 2017

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Inovirus XacF1 (7325 nucleotides) is integrated into the genome of Xanthomonas citri pv. citri (Xcc) strains at the host dif site (attB) by the host XerC/D recombination system. The XacF1 attP sequence is located within the coding region of ORF12, a possible phage regulator. After integration, this open reading frame (ORF) is split into two pieces on the host genome. We examined dynamic integration/excision of XacF1 in Xcc strain MAFF 301080 and found that the integration started at 4 h postinfection (p.i.) and peaked at 12 h p.i. Thereafter, the ratio of integrated to free forms remained constant, suggesting equilibrium of integration and excision of XacF1 in the host genome. However, the integrated state became very unstable following a 5′‐deletion of ORF12 in XacF1, suggesting that ORF12 plays a key role in the integration cycle of XacF1 in Xcc strains.

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Transformation of Xanthomonas axonopodis pv. citri by electroporation

Cited by 21 publications

References 13 publications

The Xanthomonas axonopodis pv. citri flagellum is required for mature biofilm and canker development

The Xanthomonas axonopodis pv. citri flagellum is required for mature biofilm and canker development

Evaluation of cytotoxicity features of antimicrobial peptides with potential to control bacterial diseases of citrus

Dynamic integration and excision of filamentous phage XacF1 in Xanthomonas citri pv. citri, the causative agent of citrus canker disease

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