scholarly journals Natural Genetic Variation of Xanthomonas campestris pv. campestris Pathogenicity on Arabidopsis Revealed by Association and Reverse Genetics

mBio ◽  
2013 ◽  
Vol 4 (3) ◽  
Author(s):  
Endrick Guy ◽  
Anne Genissel ◽  
Ahmed Hajri ◽  
Matthieu Chabannes ◽  
Perrine David ◽  
...  
Keyword(s):  
Xanthomonas Campestris ◽  
Genetic Basis ◽  
Reverse Genetics ◽  
Causal Agent ◽  
Aflp Markers ◽  
Avirulence Gene ◽  
Black Rot ◽  
Virulence Determinants ◽  
Content Type ◽  
Type Iii

ABSTRACTThe pathogenic bacteriumXanthomonas campestrispv. campestris, the causal agent of black rot of Brassicaceae, manipulates the physiology and the innate immunity of its hosts. Association genetic and reverse-genetic analyses of a world panel of 45X. campestrispv. campestris strains were used to gain understanding of the genetic basis of the bacterium’s pathogenicity toArabidopsis thaliana. We found that the compositions of the minimal predicted type III secretome varied extensively, with 18 to 28 proteins per strain. There were clear differences in aggressiveness of thoseX. campestrispv. campestris strains on twoArabidopsisnatural accessions. We identified 3 effector genes (xopAC,xopJ5, andxopAL2) and 67 amplified fragment length polymorphism (AFLP) markers that were associated with variations in disease symptoms. The nature and distribution of the AFLP markers remain to be determined, but we observed a low linkage disequilibrium level between predicted effectors and other significant markers, suggesting that additional genetic factors make a meaningful contribution to pathogenicity. Mutagenesis of type III effectors inX. campestrispv. campestris confirmed thatxopACfunctions as both a virulence and an avirulence gene inArabidopsisand thatxopAMfunctions as a second avirulence gene on plants of the Col-0 ecotype. However, we did not detect the effect of any other effector in theX. campestrispv. campestris 8004 strain, likely due to other genetic background effects. These results highlight the complex genetic basis of pathogenicity at the pathovar level and encourage us to challenge the agronomical relevance of some virulence determinants identified solely in model strains.IMPORTANCEThe identification and understanding of the genetic determinants of bacterial virulence are essential to be able to design efficient protection strategies for infected plants. The recent availability of genomic resources for a limited number of pathogen isolates and host genotypes has strongly biased our research toward genotype-specific approaches. Indeed, these do not consider the natural variation in both pathogens and hosts, so their applied relevance should be challenged. In our study, we exploited the genetic diversity ofXanthomonas campestrispv. campestris, the causal agent of black rot on Brassicaceae (e.g., cabbage), to mine for pathogenicity determinants. This work evidenced the contribution of known and unknown loci to pathogenicity relevant at the pathovar level and identified these virulence determinants as prime targets for breeding resistance toX. campestrispv. campestris in Brassicaceae.

scholarly journals Chemical Targeting and Manipulation of Type III Secretion in the Phytopathogen Xanthomonas campestris for Control of Disease

2019 ◽  
Vol 86 (3) ◽  
Author(s):  
Le Zhou ◽  
Cheng Wang ◽  
Guo-Hua Wang ◽  
Zai-Wa Wei ◽  
Qiu-Xia Fu ◽  
...  
Keyword(s):  
Disease Control ◽  
Small Molecules ◽  
Type Iii Secretion ◽  
Xanthomonas Campestris ◽  
Black Rot ◽  
Content Type ◽  
Type Iii ◽  
Rot Disease ◽  
Antivirulence Agents ◽  
Further Development

ABSTRACT Xanthomonas campestris pv. campestris is the causative agent of black rot disease in crucifer plants. This Gram-negative bacterium utilizes the type III secretion system (T3SS), encoded by the hrp gene cluster, to aid in its resistance to host defenses and the ability to cause disease. The T3SS injects a set of proteins known as effectors into host cells that come into contact with the bacterium. The T3SS is essential for the virulence and hypersensitive response (HR) of X. campestris pv. campestris, making it a potential target for disease control strategies. Using a unique and straightforward high-throughput screening method, we examined a large collection of diverse small molecules for their potential to modulate the T3SS without affecting the growth of X. campestris pv. campestris. Screening of 13,129 different compounds identified 10 small molecules that had a significant inhibitory influence on T3SS. Moreover, reverse transcription-quantitative PCR (qRT-PCR) assays demonstrated that all 10 compounds repress the expression of the hrp genes. Interestingly, the effect of these small molecules on hrp genes may be through the HpaS and ColS sensor kinase proteins that are key to the regulation of the T3SS in planta. Five of the compounds were also capable of inhibiting X. campestris pv. campestris virulence in a Chinese radish leaf-clipping assay. Furthermore, seven of the small molecules significantly weakened the HR in nonhost pepper plants challenged with X. campestris pv. campestris. Taken together, these small molecules may provide potential tool compounds for the further development of antivirulence agents that could be used in disease control of the plant pathogen X. campestris pv. campestris. IMPORTANCE The bacterium Xanthomonas campestris pv. campestris is known to cause black rot disease in many socioeconomically important vegetable crops worldwide. The management and control of black rot disease have been tackled with chemical and host resistance methods with variable success. This has motivated the development of alternative methods for preventing this disease. Here, we identify a set of novel small molecules capable of inhibiting X. campestris pv. campestris virulence, which may represent leading compounds for the further development of antivirulence agents that could be used in the control of black rot disease.

scholarly journals The Predicted Lytic Transglycosylase HpaH from Xanthomonas campestris pv. vesicatoria Associates with the Type III Secretion System and Promotes Effector Protein Translocation

2016 ◽  
Vol 85 (2) ◽  
Author(s):  
Jens Hausner ◽  
Nadine Hartmann ◽  
Michael Jordan ◽  
Daniela Büttner
Keyword(s):  
Type Iii Secretion ◽  
Protein Function ◽  
Xanthomonas Campestris ◽  
Effector Proteins ◽  
Transport Pathway ◽  
Content Type ◽  
Bacterial Membranes ◽  
Type Iii ◽  
Lytic Transglycosylase

ABSTRACT The pathogenicity of the Gram-negative plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria depends on a type III secretion (T3S) system, which spans both bacterial membranes and translocates effector proteins into plant cells. The assembly of the T3S system presumably involves the predicted lytic transglycosylase (LT) HpaH, which is encoded adjacent to the T3S gene cluster. Bacterial LTs degrade peptidoglycan and often promote the formation of membrane-spanning macromolecular protein complexes. In the present study, we show that HpaH localizes to the bacterial periplasm and binds to peptidoglycan as well as to components of the T3S system, including the predicted periplasmic inner rod proteins HrpB1 and HrpB2 as well as the pilus protein HrpE. In vivo translocation assays revealed that HpaH promotes the translocation of various effector proteins and of early substrates of the T3S system, suggesting a general contribution of HpaH to type III-dependent protein export. Mutant studies and the analysis of reporter fusions showed that the N-terminal region of HpaH contributes to protein function and is proteolytically cleaved. The N-terminally truncated HpaH cleavage product is secreted into the extracellular milieu by a yet-unknown transport pathway, which is independent of the T3S system.

scholarly journals Functional Characterization of the Type III Secretion Substrate Specificity Switch Protein HpaC from Xanthomonas campestris pv. vesicatoria

2011 ◽  
Vol 79 (8) ◽  
pp. 2998-3011 ◽  
Author(s):  
Steve Schulz ◽  
Daniela Büttner
Keyword(s):  
Amino Acids ◽  
Substrate Specificity ◽  
Type Iii Secretion ◽  
Protein Function ◽  
Xanthomonas Campestris ◽  
Functional Characterization ◽  
Effector Proteins ◽  
Content Type ◽  
Type Iii ◽  
In The Wild

ABSTRACTPathogenicity ofXanthomonas campestrispv.vesicatoriadepends on a type III secretion (T3S) system which translocates effector proteins into eukaryotic cells and is associated with an extracellular pilus and a translocon in the host plasma membrane. T3S substrate specificity is controlled by the cytoplasmic switch protein HpaC, which interacts with the C-terminal domain of the inner membrane protein HrcU (HrcUC). HpaC promotes the secretion of translocon and effector proteins but prevents the efficient secretion of the early T3S substrate HrpB2, which is required for pilus assembly. In this study, complementation assays with serial 10-amino-acid HpaC deletion derivatives revealed that the T3S substrate specificity switch depends on N- and C-terminal regions of HpaC, whereas amino acids 42 to 101 appear to be dispensable for the contribution of HpaC to the secretion of late substrates. However, deletions in the central region of HpaC affect the secretion of HrpB2, suggesting that the mechanisms underlying HpaC-dependent control of early and late substrates can be uncoupled. The results of interaction and expression studies with HpaC deletion derivatives showed that amino acids 112 to 212 of HpaC provide the binding site for HrcUCand severely reduce T3S when expressed ectopically in the wild-type strain. We identified a conserved phenylalanine residue at position 175 of HpaC that is required for both protein function and the binding of HpaC to HrcUC. Taking these findings together, we concluded that the interaction between HpaC and HrcUCis essential but not sufficient for T3S substrate specificity switching.

scholarly journals Complete Genome Sequences of Two Strains of Xanthomonas campestris pv. campestris Isolated in Japan

2020 ◽  
Vol 9 (2) ◽  
Author(s):  
Kasumi Takeuchi ◽  
Ichiro Mitsuhara
Keyword(s):  
Chinese Cabbage ◽  
Complete Genome ◽  
Xanthomonas Campestris ◽  
Black Rot ◽  
Genome Sequences ◽  
Content Type ◽  
Xanthomonas Campestris Pv Campestris

Here, we report the complete genome sequences of two strains of Xanthomonas campestris pv. campestris (MAFF106712 and MAFF302021), which cause black rot in crucifer crops, isolated from Chinese cabbage and cauliflower, respectively, in Japan. The MAFF106712 chromosome was 5,002,720 bp, with a G+C content of 65.2%, and harbored one plasmid of 78,747 bp. The MAFF302021 chromosome was 5,048,651 bp, with a G+C content of 65.1%.

scholarly journals Visualizing the Translocation and Localization of Bacterial Type III Effector Proteins by Using a Genetically Encoded Reporter System

2016 ◽  
Vol 82 (9) ◽  
pp. 2700-2708 ◽  
Author(s):  
Jayde A. Gawthorne ◽  
Laurent Audry ◽  
Claire McQuitty ◽  
Paul Dean ◽  
John M. Christie ◽  
...  
Keyword(s):  
Plant Pathogens ◽  
Effector Proteins ◽  
Host Cells ◽  
Dependent Manner ◽  
Virulence Determinants ◽  
Entry Site ◽  
Host Cell Membrane ◽  
Content Type ◽  
Type Iii ◽  
Bacterial Type

ABSTRACTBacterial type III secretion system (T3SS) effector proteins are critical determinants of infection for many animal and plant pathogens. However, monitoring of the translocation and delivery of these important virulence determinants has proved to be technically challenging. Here, we used a genetically engineered LOV (light-oxygen-voltage) sensing domain derivative to monitor the expression, translocation, and localization of bacterial T3SS effectors. We found theEscherichia coliO157:H7 bacterial effector fusion Tir-LOV was functional following its translocation and localized to the host cell membrane in discrete foci, demonstrating that LOV-based reporters can be used to visualize the effector translocation with minimal manipulation and interference. Further evidence for the versatility of the reporter was demonstrated by fusing LOV to the C terminus of theShigella flexnerieffector IpaB. IpaB-LOV localized preferentially at bacterial poles before translocation. We observed the rapid translocation of IpaB-LOV in a T3SS-dependent manner into host cells, where it localized at the bacterial entry site within membrane ruffles.

Development of race-specific molecular marker for Xanthomonas campestris pv. campestris race 3, the causal agent of black rot of crucifers

2018 ◽  
Vol 98 (5) ◽  
pp. 1119-1125 ◽  
Author(s):  
Khandker Shazia Afrin ◽  
Md Abdur Rahim ◽  
Mehede Hassan Rubel ◽  
Sathishkumar Natarajan ◽  
Jae-Young Song ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Xanthomonas Campestris ◽  
Bacterial Species ◽  
Causal Agent ◽  
Black Rot ◽  
Specific Marker ◽  
Dna Fragments ◽  
Specific Primers ◽  
Scar Primer ◽  
Xanthomonas Campestris Pv Campestris

Race-specific molecular markers were established to distinguish Xanthomonas campestris pv. campestris (Xcc) race 3, the causal agent of black rot disease of crucifers. The available genome sequences of Xcc races were aligned and identified three DNA fragments specific to Xcc race 3. The identified race-specific DNA fragments namely XccR3-49, XccR3-52, and XccR3-55 were used for designing the race-specific primers to detect and identify Xcc race 3. The specificity of race-specific primers was tested against the genomic DNA extracted from Xcc (races 1–7), Xcc strains, Xc pathovars, and other bacterial species. XccR3-49, a specific sequence characterized amplified region (SCAR) primer set, gave a single band with 867 bp length for Xcc race 3 only. The remaining two markers XccR3-52 and XccR3-55 showed polymorphic amplification with amplicon sizes of 1889 and 2109 bp for Xcc race 3, respectively. Additionally, the SCAR primer set detected Xcc race 3 rapidly and efficiently in artificially infected cabbage leaves with bio-PCR. This result showed that the newly developed race-specific markers can successfully and efficiently detect and identify Xcc race 3 from Xanthomonas campestris pv. campestris races, Xanthomonas species/pathovars, as well as other plant pathogenic bacteria (Pseudomonas syringae pv. maculicola and Erwinia carotovora subsp. carotovora). Up to now, this is the first report describing the race-specific marker for the detection of Xcc race 3.

scholarly journals Mutations That Impact the Enteropathogenic Escherichia coli Cpx Envelope Stress Response Attenuate Virulence in Galleria mellonella

2012 ◽  
Vol 80 (9) ◽  
pp. 3077-3085 ◽  
Author(s):  
S. Leuko ◽  
T. L. Raivio
Keyword(s):  
Escherichia Coli ◽  
Stress Response ◽  
Type Iii Secretion ◽  
Galleria Mellonella ◽  
Infection Model ◽  
Virulence Determinants ◽  
Content Type ◽  
Type Iii ◽  
Envelope Stress

ABSTRACTIn this paper, we show that the larvae of the greater wax moth,Galleria mellonella, can be used as a model to study enteropathogenicEscherichia coli(EPEC) virulence.G. mellonellalarvae are killed after infection with EPEC type strain E2348/69 but not by an attenuated derivative that expresses diminished levels of the major virulence determinants or by a mutant specifically defective in type III secretion (T3S). Infecting EPEC inhabit the larval hemocoel only briefly and then become localized to melanized capsules, where they remain extracellular. Previously, it was shown that mutations affecting the Cpx envelope stress response lead to diminished expression of the bundle-forming pilus (BFP) and the type III secretion system (T3SS). We demonstrate that mutations that activate the Cpx pathway have a dramatic effect on the ability of the bacterium to establish a lethal infection, and this is correlated with an inability to growin vivo. Infection with allE. colistrains led to increased expression of the antimicrobial peptides (AMPs) gloverin and cecropin, although strain- and AMP-specific differences were observed, suggesting that theG. mellonellahost perceives attenuated strains and Cpx mutants in unique manners. Overall, this study shows thatG. mellonellais an economical, alternative infection model for the preliminary study of EPEC host-pathogen interactions, and that induction of the Cpx envelope stress response leads to defects in virulence.

Pathovar specific molecular detection of Xanthomonas campestris pv. campestris, the causal agent of black rot disease in cabbage

2019 ◽  
Vol 41 (3) ◽  
pp. 318-328 ◽  
Author(s):  
Mehede H. Rubel ◽  
Sathishkumar Natarajan ◽  
Mohammad R. Hossain ◽  
Ujjal K. Nath ◽  
Khandker S. Afrin ◽  
...  
Keyword(s):  
Xanthomonas Campestris ◽  
Molecular Detection ◽  
Causal Agent ◽  
Black Rot ◽  
Rot Disease ◽  
Xanthomonas Campestris Pv Campestris
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