Citri Pv Research Articles

Development and comparative validation of genomic-driven PCR-based assays to detect Xanthomonas citri pv. citri in citrus plants

BackgroundAsiatic Citrus Canker, caused by Xanthomonas citri pv. citri, severely impacts citrus production worldwide and hampers international trade. Considerable regulatory procedures have been implemented to prevent the introduction and establishment of X. citri pv. citri into areas where it is not present. The effectiveness of this surveillance largely relies on the availability of specific and sensitive detection protocols. Although several PCR- or real-time PCR-based methods are available, most of them showed analytical specificity issues. Therefore, we developed new conventional and real-time quantitative PCR assays, which target a region identified by comparative genomic analyses, and compared them to existing protocols.ResultsOur assays target the X. citri pv. citri XAC1051 gene that encodes for a putative transmembrane protein. The real-time PCR assay includes an internal plant control (5.8S rDNA) for validating the assay in the absence of target amplification. A receiver-operating characteristic approach was used in order to determine a reliable cycle cut-off for providing accurate qualitative results. Repeatability, reproducibility and transferability between real-time devices were demonstrated for this duplex qPCR assay (XAC1051-2qPCR). When challenged with an extensive collection of target and non-target strains, both assays displayed a high analytical sensitivity and specificity performance: LOD95% = 754 CFU ml− 1 (15 cells per reaction), 100% inclusivity, 97.2% exclusivity for XAC1051-2qPCR; LOD95% = 5234 CFU ml− 1 (105 cells per reaction), 100% exclusivity and inclusivity for the conventional PCR. Both assays can detect the target from naturally infected citrus fruit. Interestingly, XAC1051-2qPCR detected X. citri pv. citri from herbarium citrus samples. The new PCR-based assays displayed enhanced analytical sensitivity and specificity when compared with previously published PCR and real-time qPCR assays.ConclusionsWe developed new valuable detection assays useful for routine diagnostics and surveillance of X. citri pv. citri in citrus material. Their reliability was evidenced through numerous trials on a wide range of bacterial strains and plant samples. Successful detection of the pathogen was achieved from both artificially and naturally infected plants, as well as from citrus herbarium samples, suggesting that these assays will have positive impact both for future applied and academic research on this bacterium.

Transgenic Citrange troyer rootstocks overexpressing antimicrobial potato Snakin-1 show reduced citrus canker disease symptoms

Citrus canker is a major disease caused by Xanthomonas citri pv. citri. Snakin-1 is an antimicrobial peptide, which was previously shown to be effective against different bacterial and fungal diseases in potato, wheat and lettuce when expressed in transgenic plants. We generated transgenic Citrange Troyer citrus rootstocks constitutively expressing this peptide and 5 different transgenic lines were challenged against virulent X. citri isolates. Challenge assays conducted in vitro using detached leaves and in planta by infiltration revealed a significant reduction of the number and size of canker lesions in some of the transgenic lines.

Synthesis and biological evaluation of stilbene-based peptoid mimics against the phytopathogenic bacterium Xanthomonas citri pv. citri.

The emergence of drug-resistant phytopathogenic bacteria and the need for new types of biological disease-control agents have accelerated efforts toward searching for alternative candidates with a low propensity for resistance development. In this study, a new series of stilbene-based peptoid mimics were synthesized, and their biological activities were evaluated against citrus pathogenic bacteria in vitro and in vivo. Antibacterial bioassay results showed that the dicationic peptoid mimics 9a and 9b displayed excellent bioactivity against Xanthomonas citri pv. citri, with the minimum inhibitory concentration values of 25 μM, which were superior to those of commercial copper biocides Delite (200 μM) and Kasumin Bordeaux (100 μM). In vivo bioassay further confirmed their control efficacy against plant bacterial diseases. In addition, the antibacterial mechanism of action elucidated their membrane-disruption effects resulting in the leakage of the bacterial membranes, which was similar to that of antimicrobial peptides. Moreover, the inhibition effect on biofilm formation of peptoid mimics has also been demonstrated. Stilbene-based peptoid mimics synthesized in this study showed promising antibacterial activity with a potent membrane-disruptive mechanism. The results suggested that stilbene-based peptoid mimics have the potential as a candidate new type of bactericide for citrus disease protection.

Bacterial Black Spot (BBS) of Mango in Florida

Bacterial black spot, caused by the bacterium Xanthomonas citri pv. mangiferaeindicae, is a relatively new (~2015) bacterial disease in Florida that has the potential to limit mango production of some cultivars. This new 6-page fact sheet provides the current knowledge and status of the disease potential on various cultivars. Written by Jonathan Crane and Romina Gazis, and published by the UF/IFAS Horticultural Sciences Department.https://edis.ifas.ufl.edu/hs1369

Common bean resistance to Xanthomonas is associated with upregulation of the salicylic acid pathway and downregulation of photosynthesis

BackgroundCommon bacterial blight (CBB) caused by Xanthomonas phaseoli pv. phaseoli and Xanthomonas citri pv. fuscans is one of the major threats to common bean crops (Phaseolus vulgaris L.). Resistance to CBB is particularly complex as 26 quantitative resistance loci to CBB have been described so far. To date, transcriptomic studies after CBB infection have been very scarce and the molecular mechanisms underlying susceptibility or resistance are largely unknown.ResultsWe sequenced and annotated the genomes of two common bean genotypes being either resistant (BAT93) or susceptible (JaloEEP558) to CBB. Reciprocal BLASTp analysis led to a list of 20,787 homologs between these genotypes and the common bean reference genome (G19833), which provides a solid dataset for further comparative analyses. RNA-Seq after inoculation with X. phaseoli pv. phaseoli showed that the susceptible genotype initiated a more intense and diverse biological response than the resistant genotype. Resistance was linked to upregulation of the salicylic acid pathway and downregulation of photosynthesis and sugar metabolism, while susceptibility was linked to downregulation of resistance genes and upregulation of the ethylene pathway and of genes involved in cell wall modification.ConclusionsThis study helps better understanding the mechanisms occurring during the early colonization phase of common bean by Xanthomonas and unveils new actors potentially important for resistance and susceptibility to CBB. We discuss the potential link between the pathways induced during bean colonization and genes induced by transcription activator-like effectors (TALEs), as illustrated in other Xanthomonas pathovars.

Sources, Spectrum, Genetics, and Inheritance of Phaseolus vulgaris Resistance Against Xanthomonas citri pv. fuscans.

Common bean (Phaseolus vulgaris) is one of the most consumed agricultural products in the world. Its production is affected by common bacterial blight (CBB) caused by Xanthomonas citri pv. fuscans and X. phaseoli pv. phaseoli. In this work, we investigated the spectrum, genetics, and inheritance of common bean resistance to X. citri pv. fuscans. Inoculation of nine selected cultivars with an X. citri pv. fuscans strain showed that BRS Radiante and IAPAR 16 were resistant. These two cultivars were also resistant to six X. phaseoli pv. phaseoli strains of different geographic origins, demonstrating their broad-spectrum resistances. BRS Radiante sustained smaller X. citri pv. fuscans populations than two susceptible cultivars. Stomatal densities of IAPAR 16 and BRS Radiante were significantly higher than or not different from susceptible cultivars. BRS Radiante showed the lowest general combining ability values and the combination BRS Radiante × Carioca MG the lowest specific combining ability (SCA) values, revealing the capacity of BRS Radiante to increase resistance to X. citri pv. fuscans. Positive and negative parental SCA values indicated dominant and recessive genes involved in X. citri pv. fuscans resistance. Resistance of the BRS Radiante × Carioca MG cross segregated in a 9:7 ratio in the F2 population, indicating that it is governed by two complementary dominant genes. Maximum likelihood analysis showed that the resistance of BRS Radiante to X. citri pv. fuscans is conferred by a gene of major effect with contribution of additional polygenes. This study contributes with important knowledge on the resistance against CBB in Brazilian common bean cultivars as well as with molecular tools for confirmation of common bean hybrids.

Bacterial blight resistance in cotton: genetic basis and molecular mapping

Bacterial blight (BB, caused by Xanthomonas citri pv. malvacearum, Xcm) is a worldwide disease of cotton (Gossypium spp.). The disease has been effectively controlled through the use of BB resistant cultivars and planting of acid-delinted seed. However, a resurgence of BB has been noted in the US in recent years due to the spread of Xcm race 18 and growing of susceptible transgenic cultivars, which calls for a renewed effort to develop new BB resistant cultivars. However, there has been a paucity of information in genetics and breeding for BB resistance since the 1990s due to the lack of research efforts. This review was prepared to fill this void with an objective to provide detailed results from past qualitative and quantitative genetic studies on BB resistance, including genetic designs and specific germplasm used for conducting research. More than 20 major resistance B genes (B1 to B8, B9K, B9L, B10K, B10L, B11, B12, BIn, Bn, Bs, and more than 4 unnamed genes), with at least two polygene complexes (BSm and BDm), have been identified. One B gene may be resistant to a single or multiple Xcm races, and pyramiding of several B genes can enhance resistance to a single or multiple Xcm races. Allelic relationships among some of the genes are currently unknown. Quantitative genetics has been employed to estimate heritability, gene effects, additive and dominance variances, and effective number of genes for BB resistance. The studies suggest that the additive effect and additive variance play a predominant role in BB resistance, while the dominant effect and variance play a reduced role in resistance. Heritability estimates are moderate to high depending on environmental errors, and 1–2 effective numbers of genes have been estimated, consistent with Mendelian genetic studies. Studies in molecular mapping of several BB resistance genes (B2, B3, b6, and B12) have been conducted with the focus on B12 as it is resistant to races 1 through 19. Portable DNA markers have been developed and used in marker-assisted selection for BB resistance. Finally, areas where there is a lack of information and controversies are identified and assessed. This review provides an updated comprehensive account of the genetic basis for BB resistance in cotton.

Discovery of novel bis-sulfoxide derivatives bearing acylhydrazone and benzothiazole moieties as potential antibacterial agents

On the basis of the active substructure combination principle, 24 novel synthesis of novel bis-sulfoxide derivatives bearing acylhydrazone and benzothiazole moieties as potential antibacterial agents were designed and synthesized. The bioactivity assay results showed that many compounds had significant in vitro inhibitory effects against Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas citri pv. citri (Xac). Notably, compound 4b had the best in vitro antibacterial activity against Xoo at an half-maximal effective concentration value of 11.4 μg/mL, which was superior to those of thiodiazole copper (TDC) and bismerthiazol (BMT). Compared with TDC and BMT, compound 4b was more effective in vivo controlling rice bacterial leaf blight with curative and protection activities of 42.5% and 40.3%, respectively. In addition, compound 4b can influence biofilm formation, inhibit extracellular polysaccharide production, and ultimately reduce the pathogenicity of Xoo. All the results indicated that bis-sulfoxide derivatives bearing acylhydrazone and benzothiazole moieties can be used for the development of small-molecule pesticides with high antibacterial activity.

Antibacterial Activities of Novel Dithiocarbamate-Containing 4H-Chromen-4-one Derivatives.

To aid the development of novel antibacterial agents that possess a innovative mechanism of action, we built a series of novel dithiocarbamate-containing 4H-chromen-4-one derivatives. We evaluated the activities of the derivatives against three plant pathogens Xanthomonas oryzae pv oryzae (X. oryzae pv o.), Ralstonia solanacearum (R. solanacearum), and Xanthomonas axonopodis pv citri (X. axonopodis pv c.). The results of the antibacterial bioassay showed that most of the target compounds displayed good inhibitory effects against X. oryzae pv o. and X. axonopodis pv c. Remarkably, compound E6 showed the best in vitro antibacterial activity against X. axonopodis pv c., with an EC50 value of 0.11 μg/mL, which was better than those of thiodiazole copper (59.97 μg/mL) and bismerthiazol (48.93 μg/mL). Compound E14 exhibited the best in vitro antibacterial activity against X. oryzae pv o., with an EC50 value of 1.58 μg/mL, which was better than those of thiodiazole copper (83.04 μg/mL) and bismerthiazol (56.05 μg/mL). Scanning electron microscopy analysis demonstrated that compounds E6 and E14 caused the rupture or deformation of the cell membranes for X. axonopodis pv c. and X. oryzae pv o., respectively. In vivo antibacterial activity test and the defensive enzymes activity test results indicated that the compound E14 could reduce X. oryzae pv o. more effectively than thiodiazole-copper or bismerthiazol.

Identification of a virulence tal gene in the cotton pathogen, Xanthomonas citri pv. malvacearum strain Xss-V2\u201318

BackgroundBacterial blight of cotton (BBC), which is caused by the bacterium Xanthomonas citri pv. malvacearum (Xcm), is a destructive disease in cotton. Transcription activator-like effectors (TALEs), encoded by tal-genes, play critical roles in the pathogenesis of xanthomonads. Characterized strains of cotton pathogenic Xcm harbor 8–12 different tal genes and only one of them is functionally decoded. Further identification of novel tal genes in Xcm strains with virulence contributions are prerequisite to decipher the Xcm-cotton interactions.ResultsIn this study, we identified six tal genes in Xss-V2–18, a highly-virulent strain of Xcm from China, and assessed their role in BBC. RFLP-based Southern hybridization assays indicated that Xss-V2–18 harbors the six tal genes on a plasmid. The plasmid-encoded tal genes were isolated by cloning BamHI fragments and screening clones by colony hybridization. The tal genes were sequenced by inserting a Tn5 transposon in the DNA encoding the central repeat region (CRR) of each tal gene. Xcm TALome evolutionary relationship based on TALEs CRR revealed relatedness of Xss-V2–18 to MSCT1 and MS14003 from the United States. However, Tal2 of Xss-V2–18 differs at two repeat variable diresidues (RVDs) from Tal6 and Tal26 in MSCT1 and MS14003, respectively, inferred functional dissimilarity. The suicide vector pKMS1 was then used to construct tal deletion mutants in Xcm Xss-V2–18. The mutants were evaluated for pathogenicity in cotton based on symptomology and growth in planta. Four mutants showed attenuated virulence and all contained mutations in tal2. One tal2 mutant designated M2 was further investigated in complementation assays. When tal2 was introduced into Xcm M2 and expressed in trans, the mutant was complemented for both symptoms and growth in planta, thus indicating that tal2 functions as a virulence factor in Xcm Xss-V2–18.ConclusionsOverall, the results demonstrated that Tal2 is a major pathogenicity factor in Xcm strain Xss-V2–18 that contributes significantly in BBC. This study provides a foundation for future efforts aimed at identifying susceptibility genes in cotton that are targeted by Tal2.

Diversity and copper resistance of Xanthomonas affecting citrus

Since citrus canker was first described in the early 1900’s, four major types and three species of pathogenic xanthomonads associated to citrus were identified based on the characteristic symptoms and host range. Type A, caused by X. citri subsp. citri, develops typical raised cankered lesions and is pathogenic on most commercial citrus species and cultivars. Types B and C are caused by X. citri pv. aurantifolii which elicits canker lesions. Besides the typical type A, the more aggressive and most widely spread type, other subtypes have been described, such as type A*, type Aw, type A-Manatee, type A-Miami, type A-minus pthA, and type-A etrog. The type B strains are found only in Argentina and are more pathogenic on lemon and less aggressive on sweet orange, tangerines and grapefruit. The type C strains are found only in Brazil and are pathogenic only on Key lime. X. alfalfae subsp. citrumelonis, formerly known as type E, causes citrus bacterial spot and is more aggressive on trifoliate citrus and its hybrids. Lesions caused by citrus bacterial spot are flat or sunken and not raised. Several subtypes of X. citri subsp. citri have also been identified. Besides the differences in pathogenicity, xanthomonads may also vary in their sensitivity to copper. Copper resistance has been reported for strains of X. citri subsp. citri and X. alfalfae subsp. citrumelonis and is conferred by the gene clusters copLAB or copABCD. Resistance to copper has serious implications for the use of copper-based bactericides, which are the most important chemicals for the control of diseases caused by Xanthomonas on citrus.

Combined effect of salt stress and Xanthomonas axonopodis pv citri on citrus (Citrus aurantifolia)

Xanthomonas axonopodis pv citri (Xac) and salt stress are two crucial hindrances to citrus production. The effect of continuous salt application and Xac infection in citrus has been investigated. Citrus plants were exposed to salt stress by irrigating with 50 mM, 100 mM, 150 mM, and 200 mM NaCl solution on weekly basis and challenged by Xac. Salt stress affected the defense response of Citrus plants to Xac and therefore lesion diameter and disease severity were gradually increased at higher salt concentration. Meanwhile, accumulation of Na+ and Cl− in the leaves were also increased with the increase of salt concentration. Besides, physiological performance (PP) of plants was estimated based on the parameters such as net assimilation rate, chlorophyll content, stomatal conductance, transpiration rate and intercellular CO2 concentration. The PP of sole Xac treated plants was gradually increased and maintained up to end of the experiment, whereas plants treated with Xac+50 mM and Xac+100 mM NaCl showed the highest PP up to 30 days after inoculation and then decreased. However, the PP of Xac+150 mM and Xac+200 mM NaCl treated plants gradually decreased till the end of experiment. Similarly, the PP of 200 mM NaCl treated plants declined continuously. Interestingly, the PP in 50 mM and 100 mM NaCl treated plants was higher initially and then decreased at 30 DAI to 40 DAI. This study revealed that citrus canker disease development was enhanced by salt stress. In addition, the physiological performance of the plants was enhanced by Xac and Xac + moderate salt stress but then demolished under severe salt stress.

Antibiotic activities of propanolamine containing 1,4-benzoxazin-3-ones against phytopathogenic bacteria.

Various 1,4-benzoxazin-3-one derivatives containing propanolamine groups have been shown to exhibit good antibacterial activity against Pseudomonas syringae pv actinidiae (Psa), X. axonopodis pv citri (Xac) and Xanthomonas oryzae pv oryzae (Xoo). 1,4-benzoxazin-3-one 4n showed the best inhibitory effects against Psa, Xac and Xoo, exhibiting in vitro EC50 values of 4.95, 4.71 and 8.50 μg mL−1, respectively. These potencies were superior to the corresponding EC50 values of the commercial antibiotics bismerthiazol (BT, 89.10, and 116.90 μg mL−1) and thiodiazole copper (TC, 127.30, 82.73 and 87.50 μg mL−1). Treatment on the bacterial leaf blight of rice revealed that compound 4n displayed better curative (51%) and protective (48%) activities for reducing rice BLB than either BT (41%, 39%) or TC (43%, 41%). Scanning electron microscopy (SEM) imaging of Xoo that had been treated with 1,4-benzoxazin-3-one 4n (50–100 μg mL−1) revealed that the bacterial cells had experienced extensive cell wall damage, which is the likely cause of its antimicrobial activity and bacterial death.

CRISPR elements provide a new framework for the genealogy of the citrus canker pathogen Xanthomonas citri pv. citri

BackgroundXanthomonads are an important clade of Gram-negative bacteria infecting a plethora of economically important host plants, including citrus. Knowledge about the pathogen’s diversity and population structure are prerequisite for epidemiological surveillance and efficient disease management. Rapidly evolving genetic loci, such as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR), are of special interest to develop new molecular typing tools.ResultsWe analyzed CRISPR loci of 56 Xanthomonas citri pv. citri strains of world-wide origin, a regulated pathogen causing Asiatic citrus canker in several regions of the world. With one exception, 23 unique sequences built up the repertoire of spacers, suggesting that this set of strains originated from a common ancestor that already harbored these 23 spacers. One isolate originating from Pakistan contained a string of 14 additional, probably more recently acquired spacers indicating that this genetic lineage has or had until recently the capacity to acquire new spacers. Comparison of CRISPR arrays with previously obtained molecular typing data, such as amplified fragment length polymorphisms (AFLP), variable-number of tandem-repeats (VNTR) and genome-wide single-nucleotide polymorphisms (SNP), demonstrated that these methods reveal similar evolutionary trajectories. Notably, genome analyses allowed to generate a model for CRISPR array evolution in X. citri pv. citri, which provides a new framework for the genealogy of the citrus canker pathogen.ConclusionsCRISPR-based typing will further improve the accuracy of the genetic identification of X. citri pv. citri outbreak strains in molecular epidemiology analyses, especially when used concomitantly with another genotyping method.

Rapid Detection of Xanthomonas citri pv. fuscans and Xanthomonas phaseoli pv. phaseoli in Common Bean by Loop-Mediated Isothermal Amplification.

A single loop-mediated isothermal amplification (LAMP) assay was developed for specific detection of both pathogens that cause bacterial blight in common bean, Xanthomonas phaseoli pv. phaseoli (Xpp) and Xanthomonas citri pv. fuscans (Xcf). The objective was to provide a simple, easy-to-use, specific, and sensitive method to investigate the presence of one or both pathogens in plant material and seeds for routine diagnosis. The detection limits for both pathogens were 10 CFU/ml for cell suspensions and 1 fg of DNA, whereas in conventional PCR, the primers detected up to 105 CFU/ml and 1 ng of DNA. Specificity was confirmed by testing DNA from bean leaves, other Xanthomonas species, common fungal and bacterial bean pathogens, and bacteria from the leaf microbiota. The method was tested with bean leaves inoculated with Xpp, and the pathogen could be detected from 4 h up to 15 days postinoculation, even before disease symptoms were visible. When the method was applied to bacterium detection (Xpp or Xcf) in seed lots from infected plants, the bacterium detection rate was 100% (24 of 24). The pathogens were detected in seeds incubated for just 1 h in saline solution (0.85%), reducing the time needed for bacterium detection. The LAMP assay could be useful as a tool in bean bacterial blight management. Rapid and sensitive detection of bacteria in bean seed lots would reduce the risks of planting highly contaminated seeds in environments favorable to blight multiplication.

Design, Synthesis, Antibacterial Evaluation, and Induced Apoptotic Behaviors of Epimeric and Chiral 18β-Glycyrrhetinic Acid Ester Derivatives with an Isopropanolamine Bridge against Phytopathogens.

Because only a handful of agrochemicals can manage bacterial infections, the discovery and development of innovative, inexpensive, and high-efficiency antibacterial agents targeting these infections are challenging. Herein, a series of novel epimeric and chiral 18β-glycyrrhetinic acid (GA) ester derivatives with various tertiary amine pendants were designed, synthesized, and screened for pharmacological activity. Results showed that some of the title compounds were conferred with significantly enhanced antibacterial activity toward phytopathogens Xanthomonas oryzae pv oryzae (A2, B1-B3, and C1, EC50 values within 3.81-4.82 μg/mL) and Xanthomonas axonopodis pv citri (B1, EC50 = 3.18 μg/mL; B2, EC50 = 2.76 μg/mL). These activities are superior to those of GA (EC50 > 400 μg/mL), thiodiazole copper, and bismerthiazol. Pharmacophore studies revealed that the synergistic combination of GA skeleton and tertiary amine scaffolds contributed to the biological actions. In vivo experiments displayed their promising applications in controlling bacterial infections. Antibacterial mechanism studies revealed that the title compounds could trigger apoptosis in the tested pathogens, evident by bacteria morphological changes observed in scanning electron microscopy images. This outcome should motivate the development of various apoptosis inducers against plant bacterial diseases by a novel mode of action compared to that of existing agricultural chemicals.

RNA-seq analysis provides insights into cold stress responses of Xanthomonas citri pv. citri

BackgroundXanthomonas citri pv. citri (Xcc) is a citrus canker causing Gram-negative bacteria. Currently, little is known about the biological and molecular responses of Xcc to low temperatures.ResultsResults depicted that low temperature significantly reduced growth and increased biofilm formation and unsaturated fatty acid (UFA) ratio in Xcc. At low temperature Xcc formed branching structured motility. Global transcriptome analysis revealed that low temperature modulates multiple signaling networks and essential cellular processes such as carbon, nitrogen and fatty acid metabolism in Xcc. Differential expression of genes associated with type IV pilus system and pathogenesis are important cellular adaptive responses of Xcc to cold stress.ConclusionsStudy provides clear insights into biological characteristics and genome-wide transcriptional analysis based molecular mechanism of Xcc in response to low temperature.

Identification of Resistance Sources Among Traditional Cotton Genotypes from Burkina Faso Against Xanthomonas Citri Pv. Malvacearum

Bacterial blight of Cotton caused by Xanthomonas citri. pv malvacearum, is a severe disease occurring in all cotton-growing areas. In several countries, control has been achieved through the use of resistant cotton genotypes associated with agricultural methods limiting disease spread. The main objective of the present study was to identify resistance sources against races 18 and 20 of Xanthomonas citri pv. malvacearum in artificial inoculation conditions within a collection of local cotton cultivars. Our results show that two conventional and commercialized varieties, FK64 and FK37, and traditional accessions belonging to the species hirsutism, barbadense and arborerum harbor resistance against X. citri pv. malvacearum race 18 from leaves, stem and boll inoculation. However, with race 18 strain, some cultivars conserve bacteria into stem and leave that remain asymptomatic 35 days after inoculation. No source of resistance was detected against race 20 of X. citri pv. malvacearum. For better management of cotton blight, the knowledge of the phenotypic diversity of pathogen strains, their spatial and temporal distribution is a prerequisite to guide the use of resistance gene in breeding program and their deployment in cotton growing areas in Burkina Faso.

Analyses of Seven New Genomes of Xanthomonas citri pv. aurantifolii Strains, Causative Agents of Citrus Canker B and C, Show a Reduced Repertoire of Pathogenicity-Related Genes.

Xanthomonas citri pv. aurantifolii pathotype B (XauB) and pathotype C (XauC) are the causative agents respectively of citrus canker B and C, diseases of citrus plants related to the better-known citrus canker A, caused by Xanthomonas citri pv. citri. The study of the genomes of strains of these related bacterial species has the potential to bring new understanding to the molecular basis of citrus canker as well as their evolutionary history. Up to now only one genome sequence of XauB and only one genome sequence of XauC have been available, both in draft status. Here we present two new genome sequences of XauB (both complete) and five new genome sequences of XauC (two complete). A phylogenomic analysis of these seven genome sequences along with 24 other related Xanthomonas genomes showed that there are two distinct and well-supported major clades, the XauB and XauC clade and the Xanthomonas citri pv. citri clade. An analysis of 62 Type III Secretion System effector genes showed that there are 42 effectors with variable presence/absence or pseudogene status among the 31 genomes analyzed. A comparative analysis of secretion-system and surface-structure genes showed that the XauB and XauC genomes lack several key genes in pathogenicity-related subsystems. These subsystems, the Types I and IV Secretion Systems, and the Type IV pilus, therefore emerge as important ones in helping explain the aggressiveness of the A type of citrus canker and the apparent dominance in the field of the corresponding strain over the B and C strains.

Isolation and characterisation of carbapenem-resistant Xanthomonas citri pv. mangiferaeindicae-like strain gir from the faecal material of giraffes.

The purpose of this study was to determine if giraffes (Giraffa camelopardalis) living in captivity at the Jacksonville Zoo and Gardens, Jacksonville, FL were colonised with carbapenem-resistant bacteria and, if found, to identify underlying genetic mechanisms contributing to a carbapenem-resistant phenotype. Faecal samples from seven giraffes were examined for carbapenem-resistant bacteria. Only one isolate (a Xanthomondaceae) was found to be carbapenem-resistant by antibiotic susceptibility testing. This isolate was selected for additional characterization, including whole genome sequencing (WGS). Based on average nucleotide identity, the bacterium was identified as Xanthomonas citri pv. mangiferaeindicae-like strain gir. Phenotypic carbapenemase tests and PCR for the most common carbapenemase genes produced negative results, suggesting that carbapenem resistance was mediated by another mechanism. Resistance gene profile analysis of WGS results confirmed these results. Among identified resistance genes, a chromosomal class A beta-lactamase with 71% identity to the penP beta-lactamase gene from Xanthomonas citri ssp. citri was identified, which could contribute to a carbapenem-resistant phenotype.