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LuxR-type regulators play pivotal roles in regulating numerous bacterial processes, including bacterial motility and virulence, thereby exerting a significant influence on bacterial behavior and pathogenicity. Xanthomonas oryzae pv. oryzicola, a rice pathogen, causes bacterial leaf streak. Our research has identified VmsR, which is a response regulator of the two-component system (TCS) that belongs to the LuxR family. These findings of the experiment reveal that VmsR plays a crucial role in regulating pathogenicity, motility, biofilm formation, and the production of extracellular polysaccharides (EPSs) in Xoc GX01. Notably, our study shows that the vmsR mutant exhibits a reduced swimming motility but an enhanced swarming motility. Furthermore, this mutant displays decreased virulence while significantly increasing EPS production and biofilm formation. We have uncovered that VmsR directly interacts with the promoter regions of fliC and fliS, promoting their expression. In contrast, VmsR specifically binds to the promoter of gumB, resulting in its downregulation. These findings indicate that the knockout of vmsR has profound effects on virulence, motility, biofilm formation, and EPS production in Xoc GX01, providing insights into the intricate regulatory network of Xoc. [ABSTRACT FROM AUTHOR]

This study was conducted on Xanthomonas axonopodis pv, which causes significant economic losses in the agricultural sector. Here, we study a common bacterial blight disease caused by the phaseoli (XaP) bacterial pathogen on Üstün42 and Akbulut bean genera. In this study, a total of 4000 images, healthy and diseased, were used for both bean breeds. These images were classified by AlexNet, VGG16, and VGG19 models. Later, reclassification was performed by applying pre-processing to the raw images. According to the results obtained, the accuracy rates of the pre-processed images classified by the VGG19, VGG16 and AlexNet models were determined as 0.9213, 0.9125 and 0.8950, respectively. The models were then hybridized with LSTM and BiLSTM for raw and pre-processed images and new models were created. When the performance of these hybrid models was evaluated, it was found that the models hybridized with LSTM were more successful than the simple models, while the models hybridized with BiLSTM gave better results than the models hybridized with LSTM. In particular, the VGG19+BiLSTM model attracted attention by achieving 94.25% classification accuracy with pre-processed images. This study emphasizes the effectiveness of image processing techniques in agriculture in the field of disease detection and is important as a new dataset in the literature for evaluating the performance of hybridized models. [ABSTRACT FROM AUTHOR]

Bacterial spot is a serious disease caused by several species of Xanthomonas affecting pepper and tomato production worldwide. Since the strategies employed for disease management have been inefficient and pose a threat for environmental and human health, the development of alternative methods is gaining relevance. The aim of this study is to isolate and characterize lytic phages against Xanthomonas pathogens. Here, we isolate two jumbo phages, named XaC1 and XbC2, from water obtained from agricultural irrigation channels by the enrichment technique using X. vesicatoria as a host. We determined that both phages were specific for inducing the lysis of X. vesicatoria strains, but not of other xanthomonads. The XaC1 and XbC2 phages showed a myovirus morphology and were classified as jumbo phages due to their genomes being larger than 200 kb. Phylogenetic and comparative analysis suggests that XaC1 and XbC2 represent both different and novel genera of phages, where XaC1 possesses a low similarity to other phage genomes reported before. Finally, XaC1 and XbC2 exhibited thermal stability up to 45 °C and pH stability from 5 to 9. All these results indicate that the isolated phages are promising candidates for the development of formulations against bacterial spot, although further characterization is required. [ABSTRACT FROM AUTHOR]

The use of resistant varieties is an important strategy for managing tomato bacterial spot. The objective of this study was to evaluate the reaction of tomato genotypes to Xanthomonas euvesicatoria pv. perforans. The experiment was conducted in a randomized block design with 10 genotypes and four repetitions. The genotypes consisted of hybrids (UFU-1, UFU-2, UFU-3), wild resistant accession Solanum pennellii, commercial susceptible cultivar Santa Clara and homozygous lines (UFU-5, UFU-6, UFU-11, UFU-12, UFU-15). The UFU B8 isolate of X. euvesicatoria pv. perforans was used. The bacterial suspension was prepared and adjusted in a spectrophotometer OD550 = 0.5 (1 × 109 CFU mL−1). Inoculation occurred 10 days after transplantation. Disease severity was assessed at 3, 6, 9, 12 and 15 days after inoculation, and the area under the disease progress curve (AUDPC) was calculated. There was a significant difference between the genotypes regarding the severity of the disease at 3, 6 and 12 days after inoculation. Lineages 5, 6 and 12, the hybrid UFU-1 and the wild accession S. pennellii showed the lowest severity of the disease, being promising for promoting genetic improvement programs aimed at resistance to the bacteria. [ABSTRACT FROM AUTHOR]

Xanthomonas oryzae pv. oryzicola (Xoc) is a notorious plant pathogen. Like most bacterial pathogens, Xoc has evolved a complex regulatory network to modulate the expression of various genes related to pathogenicity. Here, we have identified TfmR, a transcriptional regulator belonging to the TetR family, as a key player in the virulence mechanisms of this phytopathogenic bacterium. We have demonstrated genetically that tfmR is involved in the hypersensitive response (HR), pathogenicity, motility and extracellular polysaccharide production of this phytopathogenic bacterium. Our investigations extended to exploring TfmR's interaction with RpfG and HrpX, two prominent virulence regulators in Xanthomonas species. We found that TfmR directly binds to the promoter region of RpfG, thereby positively regulating its expression. Notably, constitutive expression of RpfG partly reinstates the pathogenicity compromised by TfmR-deletion mutants. Furthermore, our studies revealed that TfmR also exerts direct positive regulation on the expression of the T3SS regulator HrpX. Similar to RpfG, sustained expression of HrpX partially restores the pathogenicity of TfmR-deletion mutants. These findings underscore TfmR's multifaceted role as a central regulator governing key virulence pathways in Xoc. Importantly, our research sheds light on the intricate molecular mechanisms underlying the regulation of pathogenicity in this plant pathogen. [ABSTRACT FROM AUTHOR]

Extensive microbial interactions occur within insect hosts. However, the interactions between the Huanglongbing (HLB) pathogen and endosymbiotic bacteria within the Asian citrus psyllid (ACP, Diaphorina citri Kuwayama) in wild populations remain elusive. Thus, this study aimed to detect the infection rates of HLB in the ACP across five localities in China, with a widespread prevalence in Ruijin (RJ, 58%), Huidong (HD, 28%), and Lingui (LG, 15%) populations. Next, microbial communities of RJ and LG populations collected from citrus were analyzed via 16S rRNA amplicon sequencing. The results revealed a markedly higher microbial diversity in the RJ population compared to the LG population. Moreover, the PCoA analysis identified significant differences in microbial communities between the two populations. Considering that the inter-population differences of Bray–Curtis dissimilarity in the RJ population exceeded those between populations, separate analyses were performed. Our findings indicated an increased abundance of Enterobacteriaceae in individuals infected with HLB in both populations. Random forest analysis also identified Enterobacteriaceae as a crucial indicator of HLB infection. Furthermore, the phylogenetic analysis suggested a potential regulatory role of ASV4017 in Enterobacteriaceae for ACP, suggesting its possible attractant activity. This research contributes to expanding the understanding of microbial communities associated with HLB infection, holding significant implications for HLB prevention and treatment. [ABSTRACT FROM AUTHOR]

Walnut (Juglans regia L.) is a monoecious species and although it exhibits self-compatibility, it presents incomplete overlap of pollen shed and female receptivity. Thus, cross-pollination is prerequisite for optimal fruit production. Cross-pollination can occur naturally by wind, insects, artificially, or by hand. Pollen has been recognized as one possible pathway for Xanthomonas arboricola pv. juglandis infection, a pathogenic bacterium responsible for walnut blight disease. Other than the well-known cultural and chemical control practices, artificial pollination technologies with the use of drones could be a successful tool for walnut blight disease management in orchards. Drones may carry pollen and release it over crops or mimic the actions of bees and other pollinators. Although this new pollination technology could be regarded as a promising tool, pollen germination and knowledge of pollen as a potential pathway for the dissemination of bacterial diseases remain crucial information for the development and production of aerial pollinator robots for walnut trees. Thus, our purpose was to describe a pollination model with fundamental components, including the identification of the "core" pollen microbiota, the use of drones for artificial pollination as a successful tool for managing walnut blight disease, specifying an appropriate flower pollination algorithm, design of an autonomous precision pollination robot, and minimizing the average errors of flower pollination algorithm parameters through machine learning and meta-heuristic algorithms. [ABSTRACT FROM AUTHOR]

Plants must adapt to the complex effects of several stressors brought on by global warming, which may result in interaction and superposition effects between diverse stressors. Few reports are available on how drought stress affects Xanthomonas albilineans (Xa) infection in sugarcane (Saccharum spp. hybrids). Drought and leaf scald resistance were identified on 16 sugarcane cultivars using Xa inoculation and soil drought treatments, respectively. Subsequently, four cultivars contrasting to drought and leaf scald resistance were used to explore the mechanisms of drought affecting Xa–sugarcane interaction. Drought stress significantly increased the occurrence of leaf scald and Xa populations in susceptible cultivars but had no obvious effect on resistant cultivars. The ROS bursting and scavenging system was significantly activated in sugarcane in the process of Xa infection, particularly in the resistant cultivars. Compared with Xa infection alone, defense response via the ROS generating and scavenging system was obviously weakened in sugarcane (especially in susceptible cultivars) under Xa infection plus drought stress. Collectively, ROS might play a crucial role involving sugarcane defense against combined effects of Xa infection and drought stress. [ABSTRACT FROM AUTHOR]

The ability of bacteriophages to destroy bacteria has made them the subject of extensive research. Interest in bacteriophages has recently increased due to the spread of drug-resistant bacteria, although genomic research has not kept pace with the growth of genomic data. Genomic analysis and, especially, the taxonomic description of bacteriophages are often difficult due to the peculiarities of the evolution of bacteriophages, which often includes the horizontal transfer of genes and genomic modules. The latter is particularly pronounced for temperate bacteriophages, which are capable of integration into the bacterial chromosome. Xanthomonas phage PBR31 is a temperate bacteriophage, which has been neither described nor classified previously, that infects the plant pathogen Xanthomonas campestris pv. campestris. Genomic analysis, including phylogenetic studies, indicated the separation of phage PBR31 from known classified bacteriophages, as well as its distant relationship with other temperate bacteriophages, including the Lederbervirus group. Bioinformatic analysis of proteins revealed distinctive features of PBR31, including the presence of a protein similar to the small subunit of D-family DNA polymerase and advanced lysis machinery. Taxonomic analysis showed the possibility of assigning phage PBR31 to a new taxon, although the complete taxonomic description of Xanthomonas phage PBR31 and other related bacteriophages is complicated by the complex evolutionary history of the formation of its genome. The general biological features of the PBR31 phage were analysed for the first time. Due to its presumably temperate lifestyle, there is doubt as to whether the PBR31 phage is appropriate for phage control purposes. Bioinformatics analysis, however, revealed the presence of cell wall-degrading enzymes that can be utilised for the treatment of bacterial infections. [ABSTRACT FROM AUTHOR]

There exists a wide range of plant pathogens that are commonly referred to as seed-borne pathogens due to their dominant mode of spread. Treating seeds to eliminate such pathogens is therefore very important in contemporary seed production. In the present study, eight types of nanoparticles were evaluated for their effectiveness against Xanthomonas hortorum pv. carotae, a seed-borne pathogen that affects plants of the Apiaceae family. Initially, parameters considering the inhibitory and bactericidal activity of individual nanoparticles were evaluated under in vitro conditions. In this way, three nanoparticles based on copper, silver, and silver/selenium composite were identified as being the most effective. Subsequently, their ability to eliminate Xanthomonas hortorum pv. carotae from artificially infected carrot seeds was tested. This was achieved through the qPCR quantification of the pathogen in 14-day-old plantlets developed from seeds inoculated with Xhc. Based on the obtained results, copper-based nanoparticles were the most effective, resulting in an approximately 10-fold decrease in the occurrence of Xhc in plantlets compared to the untreated control. Taking into account the fact that X. hortorum pathovars also attack other important horticultural crops, the presented results may have a much wider scope than just carrot seeds. [ABSTRACT FROM AUTHOR]

Banana Xanthomonas wilt, caused by Xanthomonas vasicola pv. musacearum (Xvm), is a devastating disease that results in total yield loss of affected plants. Resistance to the disease is limited in Musa acuminata, but it has been identified so far in the zebrina subspecies. This study identified markers associated with tolerance to Xvm in Monyet, a tetraploid banana from the zebrina subspecies which was identified to be partially resistant to the bacterium. We used a triploid progeny of 135 F1 hybrids resulting from a cross between Monyet (Xvm partially resistant) and Kokopo (diploid and Xvm susceptible). The F1 hybrids were screened in pots for resistance to Xvm. The population was genotyped using the genotyping-by-sequencing platform of Diversity Array Technology (DArTSeq). The adjusted means of the phenotypic data were combined with the allele frequencies of the genotypic data in continuous mapping. We identified 25 SNPs associated with resistance to Xvm, and these were grouped into five quantitative traits loci (QTL) on chromosomes 2, 3, 6, and 7. For each marker, we identified the favorable allele and the additive effect of replacing the reference allele with the alternative allele. The comparison between weevil borer (Cosmopolites sordidus (Germar)) and Xvm QTL revealed one QTL shared between the two biotic stresses at the distal end of chromosome 6 but with a repulsion linkage. This linkage should be broken down by generating more recombinants in the region. We also identified 18 putative alleles in the vicinity of the SNPs associated with resistance to Xvm. Among the 18 putative genes, two particularly putative genes, namely, Ma06_g13550 and Ma06_g36840, are most likely linked to disease resistance. This study is a basis for marker-assisted selection to improve banana resistance to banana Xanthomonas wilt, especially in East and Central Africa where the disease is still devastating the crop. [ABSTRACT FROM AUTHOR]

The genus Xanthomonas primarily comprises phytopathogenic species. By carrying out deep phylo-taxonogenomics, we recently reported that the genera Xylella, Stenotrophomonas, and Pseudoxanthomonas are misclassified and belong to the genus Xanthomonas. Considering the importance of Xanthomonas/Xylella as plant pathogens and to further determine the taxonomic and phylogenetic breadth of this genus, we extended our earlier study by including all the reported genera and families in the order. This investigation revealed that at least four more genera belong to the genus Xanthomonas, with a notable case being Lysobacter, after which the family and order are named. Similarly, our investigation also allowed us to reveal the expanded taxonomic breadth of the related genus Rhodanobacter. This finding of a major related genus that lacks plant pathogenic species will allow for taxonomy-based comparative studies. The phylo-taxonogenomic revelations were further supported by complete 16S rRNA-based sequence boundaries proposed for genus delineation. Accordingly, we propose a taxonomic revision of these major and closely related genera along with their constituent families within the order Lysobacteraceae (Xanthomonadaceae). The identification of a major related genus lacking plant pathogenic species will be important in investigating the origin and success of pathogenic species/lineages in the genus Xanthomonas. [ABSTRACT FROM AUTHOR]

The present study was focused on the pathosystem pepper plants (Capsicum annuum L.)-phytopathogenic bacterium X. euvesicatoria (wild strain 269p)-bacteriophage BsXeu269p/3 and the possibility of bacteriophage-mediated biocontrol of the disease. Two new model systems were designed for the monitoring of the effect of the phage treatment on the infectious process in vivo. The spread of the bacteriophage and the pathogen was monitored by qPCR. A new pair of primers for phage detection via qPCR was designed, as well as probes for TaqMan qPCR. The epiphytic bacterial population and the potential bacteriolytic effect of BsXeu269p/3 in vivo was observed by SEM. An aerosol-mediated transmission model system demonstrated that treatment with BsXeu269p/3 reduced the amount of X. euvesicatoria on the leaf surface five-fold. The needle-pricking model system showed a significant reduction of the amount of the pathogen in infectious lesions treated with BsXeu269p/3 (av. 59.7%), compared to the untreated control. We found that the phage titer is 10-fold higher in the infection lesions but it was still discoverable even in the absence of the specific host in the leaves. This is the first report of in vivo assessment of the biocontrol potential of locally isolated phages against BS pathogen X. euvesicatoria in Bulgaria. [ABSTRACT FROM AUTHOR]

As agricultural practices become more sustainable, adopting more sustainable practices will become even more relevant. Searching for alternatives to chemical compounds has been the focus of numerous studies, and bacteriocins are tools with intrinsic biotechnological potential for controlling plant diseases. We continued to explore the biotechnological activity of the bacteriocin Gluconacin from Gluconacetobacter diazotrophicus, PAL5 strain, by investigating this protein's antagonism against important tomato phytopathogens and demonstrating its effectiveness in reducing bacterial spots caused by Xanthomonas perforans. In addition to this pathogen, the bacteriocin Gluconacin demonstrated bactericidal activity in vitro against Ralstonia solanacearum and Pseudomonas syringae pv. tomato, agents that cause bacterial wilt and bacterial spots, respectively. Bacterial spot control tests showed that Gluconacin reduced disease severity by more than 66%, highlighting the biotechnological value of this peptide in ecologically correct formulations. [ABSTRACT FROM AUTHOR]

Simple Summary: Cassava is one of the most important crops worldwide. However, bacterial blight of cassava, as its most important disease, has caused substantial economic losses to the cassava industry. The main method of controlling cassava bacterial blight has been through chemical control, but this has polluted the environment significantly. In this context, biological control using microorganisms not only has the capacity to inhibit the disease, but also is environmentally friendly, showing great potential. Therefore, there is an urgent need to study the endophytic biological control method of cassava bacterial blight. On this basis, in the current study, we conducted a series of in vivo and in vitro experiments to isolate, screen and identify a biocontrol Bacillus ME9 with an antagonistic effect against Xpm11, the pathogen that causes cassava bacterial blight. In this study, the results also demonstrated that B. ME9 has a strong colonization ability, and revealed that its genome consists of a variety of genes related to antibacterial lipopeptides, which may be directly related to its antibacterial ability. B. ME9 had a strong antagonistic affect against Xpm11, and showed a certain effect on a variety of pathogens, which is expected to be further developed in its application as a commercial bacteriological agent. In recent years, the bacterial blight of cassava has caused substantial economic losses to the Chinese cassava industry. Chemical control methods have become the primary approach to control this disease; however, their widespread usage and harmful residues have raised concerns about environmental pollution. In order to avoid this, it is urgent to seek a green ecological method to prevent and control it. Biological control through the utilization of microorganisms not only effectively inhibits the disease, but also gives consideration to environmental friendliness. Therefore, investigating an endophytic biological control method for cassava bacterial blight is of great importance. In this study, cassava leaf tissues were used as test specimens in order to isolate endophytic bacteria by using dilution and separation methods. Bacillus ME9, derived from cassava endophytic bacteria, exhibits good antagonism against a diverse range of pathogens, including Xpm11. Its genome consists of a series of genes encoding antibacterial lipopeptides, which may be directly related to its antibacterial capabilities. Furthermore, inoculation resulted in a substantial change in the diversity of the endophytic bacterial community, characterized by improved diversity, and displayed an obvious inhibition of pathogenic bacterial growth, demonstrating successful colonization within plants. The results laid a foundation and provided theoretical support for the development and utilization of cassava endophytic bacterial diversity and endogenous disease control strategies. [ABSTRACT FROM AUTHOR]

Gram-negative bacteria belonging to the Xanthomonas genus include plant pathogens representing a major challenge in the field of agriculture for a wide variety of economically important crops, such as tomato, pepper, and lettuce. Due to the massive usage of agrochemicals, Xanthomonas spp. are developing resistance to copper pesticides typically used to control microbial infections. An interesting alternative approach to control bacterial phytopathogens consists of using eco-friendly biocontrol agents, often beneficial microorganisms. Here, following the targeted, broad-spectrum screening of thousands of microorganisms isolated from different environmental locations, we isolated Bacillus velezensis strain 71 and Paenibacillus peoriae strain To99 displaying potent antagonistic activity against Xanthomonas spp. We found that oxydifficidin and polymyxin A secreted by B. velezensis 71 and P. peoriae To99, respectively, are mainly responsible for the anti-Xanthomonas activity. We further evaluated the performance of cell suspensions and cell-free supernatants of these isolates in controlling tomato bacterial spot disease in growth chamber and greenhouse conditions to validate the in vitro results. The overall results demonstrate the potential of treatments based on the secondary metabolites from both isolates and their cells as an alternative to copper-based chemicals to control leaf spot diseases caused by Xanthomonas spp. phytopathogens. [ABSTRACT FROM AUTHOR]

Simple Summary: Candidatus Liberibacter asiaticus (CLas) is a putative causal agent of Huanglongbing, a serious citrus pathogen. Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is the primary vector of CLas. Florida only has CLas, and the only vector of CLas in Florida is D. citri. We explored patterns in the proportion of psyllids carrying CLas by analyzing psyllids collected from seven field sites from 2008 through February 2012. There was an upsurge in prevalence in the fall and, in some groves, a second peak in the spring. We also found that females were more common in the first and last months of the year. D. citri has different abdominal colors that we grouped into blue/green and gray/brown. Gray/brown psyllids had a higher CLas prevalence than blue/green individuals. We suggest several explanations for these patterns, but the two most likely involve a hypothesized CLas gradient in the first few millimeters of new growth and differences in immune response related to sex and color. In application, the results confirm that psyllid management is always necessary because psyllids carry CLas year-round. The seven study sites had seven levels of management, from abandoned to intensively managed. In general, more aggressively managed groves had fewer psyllids with CLas. A well-managed organic grove was an exception, having low CLas prevalence but abundant psyllids. It is possible that the low CLas prevalence at this grove was due to management techniques or that this area was infected more recently. Candidatus Liberibacter asiaticus (CLas) is one of the putative causal agents of huanglongbing, which is a serious disease in citrus production. The pathogen is transmitted by Diaphorina citri Kuwayama (Hemiptera: Psyllidae). As an observational study, six groves in central Florida and one grove at the southern tip of Florida were sampled monthly from January 2008 through February 2012 (50 months). The collected psyllids were sorted by sex and abdominal color. Disease prevalence in adults peaked in November, with a minor peak in February. Gray/brown females had the highest prevalence, and blue/green individuals of either sex had the lowest prevalence. CLas prevalence in blue/green females was highly correlated with the prevalence in other sexes and colors. Thus, the underlying causes for seasonal fluctuations in prevalence operated in a similar fashion for all psyllids. The pattern was caused by larger nymphs displacing smaller ones from the optimal feeding sites and immunological robustness in different sex–color morphotypes. Alternative hypotheses were also considered. Improving our understanding of biological interactions and how to sample them will improve management decisions. We agree with other authors that psyllid management is critical year-round. [ABSTRACT FROM AUTHOR]

Bacterial blight, caused by Xanthomonas arboricola pv. juglandis, is a significant disease affecting walnut production worldwide. Outbreaks are most severe in spring, and closely tied to host phenology and weather conditions. Pathogen infections are mainly observed in catkins, female flowers, leaves, and fruits. In this study, the effect of wetness duration and temperature on walnut infections by X. arboricola pv. juglandis was determined through two independent experiments conducted under controlled environmental conditions. The combined effect of both climatic parameters on disease severity was quantified using a third-order polynomial equation. The model obtained by linear regression and backward elimination technique fitted well to the data (R2 = 0.94 and R2adj = 0.93). The predictive capacity of the forecasting model was evaluated on pathogen-inoculated walnut plants exposed to different wetness duration–temperature combinations under Mediterranean field conditions. Observed disease severity in all events aligned with predicted infection risk. Additionally, the relationship between leaf and fruit age and the disease severity was quantified and modelled. A prediction model, which has been referred to as the WalBlight-risk model, is proposed for evaluation as an advisory system for timing bactericide sprays to manage bacterial blight in Mediterranean walnut orchards. [ABSTRACT FROM AUTHOR]

Citrus canker, caused by the bacterium Xanthomonas citri (Xcc), is one of the most devastating diseases for the citrus industry. Xylose is a constituent of the cell wall of plants, and the ability of Xcc to use this carbohydrate may play a role in virulence. Xcc has two genes codifying for xylose isomerase (XI), a bifunctional enzyme that interconverts D-xylose into D-xylulose and D-glucose into D-fructose. The aim of this work was to investigate the functional role of the two putative XI ORFs, XAC1776 (xylA1) and XAC4225 (xylA2), in Xcc pathogenicity. XI-coding genes of Xcc were deleted, and the single mutants (XccΔxylA1 or XccΔxylA2) or the double mutant (XccΔxylA1ΔxylA2) remained viable. The deletion of one or both XI genes (xylA1 and/or xylA2) increased the aggressiveness of the mutants, causing disease symptoms. RT-qPCR analysis of wild strain and xylA deletion mutants grown in vivo and in vitro revealed that the highest expression level of hrpX and xylR was observed in vivo for the double mutant. The results indicate that XI depletion increases the expression of the hrp regulatory genes in Xcc. We concluded that the intracellular accumulation of xylose enhances Xcc virulence. [ABSTRACT FROM AUTHOR]

β-Carbonic anhydrase (βCA) is very important for plant growth and development, but its function in immunity has also been examined. In this study, we found that the expression level of Solanum lycopersicum βCA1 (SlβCA1) was significantly upregulated in plants treated with Xanthomonas euvesicatoria 85-10. The protein was localized in the nucleus, cell membrane and chloroplast. Using tomato plants silenced with SlβCA1, we demonstrated that SlβCA1 plays an active role in plant disease resistance. Moreover, we found that the elicitor PopW upregulated the expression of SlβCA1, while the microbe-associated molecular pattern response induced by PopW was inhibited in TRV-SlβCA1. The interaction between PopW and SlβCA1 was confirmed. Here, we found that SlβCA1 was positively regulated during PopW-induced resistance to Xanthomonas euvesicatoria 85-10. These data indicate the importance of SlβCA1 in plant basic immunity and its recognition by the Harpin protein PopW as a new target for elicitor recognition. [ABSTRACT FROM AUTHOR]

Biological control of plant diseases has gained attraction for controlling various bacterial diseases at a field trial stage. An isolated endophytic bacterium, Bacillus velezensis 25 (Bv-25), from Citrus species had strong antagonistic activity against Xanthomonas citri subsp. citri (Xcc), which causes citrus canker disease. When Bv-25 was incubated in Landy broth or yeast nutrient broth (YNB), the ethyl acetate extract of Landy broth exhibited higher levels of antagonistic activity against Xcc compared to that of YNB. Therefore, the antimicrobial compounds in the two ethyl acetate extracts were detected by high performance liquid chromatography–mass spectrometry. This comparison revealed an increase in production of several antimicrobial compounds, including difficidin, surfactin, fengycin, and Iturin-A or bacillomycin-D by incubation in Landy broth. RNA sequencing for the Bv-25 grown in Landy broth were performed, and the differential expressions were detected for the genes encoding the enzymes for the synthesis of antimicrobial compounds, such as bacilysin, plipastatin or fengycin, surfactin, and mycosubtilin. Combination of metabolomics analysis and RNA sequencing strongly suggests that several antagonistic compounds, especially bacilysin produced by B. velezensis, exhibit an antagonistic effect against Xcc. [ABSTRACT FROM AUTHOR]

Leaf scald caused by Xanthomonas albilineans (Xa) is a major bacterial disease in sugarcane that represents a threat to the global sugar industry. Little is known about the population structure and genetic evolution of this pathogen. In this study, 39 Xa strains were collected from 6 provinces in China. Of these strains, 15 and 24 were isolated from Saccharum spp. hybrid and S. officinarum plants, respectively. Based on multilocus sequence analysis (MLSA), with five housekeeping genes, these strains were clustered into two distinct phylogenetic groups (I and II). Group I included 26 strains from 2 host plants, Saccharum spp. hybrid and S. officinarum collected from 6 provinces, while Group II consisted of 13 strains from S. officinarum plants in the Zhejiang province. Among the 39 Xa strains, nucleotide sequence identities from 5 housekeeping genes were: ABC (99.6–100%), gyrB (99.3–100%), rpoD (98.4–100%), atpD (97.0–100%), and glnA (97.6–100%). These strains were clustered into six groups (A–F), based on the rep-PCR fingerprinting, using primers for ERIC2, BOX A1R, and (GTG)5. UPGMA and PCoA analyses revealed that group A had the most strains (24), followed by group C with 11 strains, while there was 1 strain each in groups B and D–F. Neutral tests showed that the Xa population in S. officinarum had a trend toward population expansion. Selection pressure analysis showed purification selection on five concatenated housekeeping genes from all tested strains. Significant genetic differentiation and infrequent gene flow were found between two Xa populations hosted in Saccharum spp. hybrids and S. officinarum. Altogether, these results provide evidence of obvious genetic divergence and population structures among Xa strains from China. [ABSTRACT FROM AUTHOR]

The application of Trichoderma is a form of biological control that has been effective in combating Xanthomonas oryzae pv. oryzae, the causative agent of the devastating disease known as bacterial blight of rice. In this present study, four strains of Trichoderma, viz., T. paraviridescens (BDISOF67), T. erinaceum (BDISOF91), T. asperellum (BDISOF08), and T. asperellum (BDISOF09), were collected from the rice rhizosphere and used to test their potentiality in reducing bacterial blight. The expression patterns of several core defense-related enzymes and genes related to SA and JA pathways were studied to explore the mechanism of induced resistance by those Trichoderma strains. The results primarily indicated that all Trichoderma were significantly efficient in reducing the lesion length of the leaf over rice check variety (IR24) through enhancing the expression of core defense-related enzymes, such as PAL, PPO, CAT, and POD activities by 4.27, 1.77, 3.53, and 1.57-fold, respectively, over control. Moreover, the results of qRT-PCR exhibited an upregulation of genes OsPR1, OsPR10, OsWRKY45, OsWRKY62, OsWRKY71, OsHI-LOX, and OsACS2 after 24 h of inoculation with all tested Trichoderma strains. However, in the case of RT-PCR, no major changes in OsPR1 and OsPR10 expression were observed in plants treated with different Trichoderma strains during different courses of time. Collectively, Trichoderma induced resistance in rice against X. oryzae pv. oryzae by triggering these core defense-related enzymes and genes associated with SA and JA pathways. [ABSTRACT FROM AUTHOR]

Isolation for antibacterial compounds from natural plants is a promising approach to develop new pesticides. In this study, two compounds were obtained from the Chinese endemic plant Piper austrosinense using bioassay-guided fractionation. Based on analyses of 1H-NMR, 13C-NMR, and mass spectral data, the isolated compounds were identified as 4-allylbenzene-1,2-diol and (S)-4-allyl-5-(1-(3,4-dihydroxyphenyl)allyl)benzene-1,2-diol. 4-Allylbenzene-1,2-diol was shown to have strong antibacterial activity against four plant pathogens, including Xanthomonas oryzae pathovar oryzae (Xoo), X. axonopodis pv. citri (Xac), X. oryzae pv. oryzicola (Xoc) and X. campestris pv. mangiferaeindicae (Xcm). Further bioassay results exhibited that 4-allylbenzene-1,2-diol had a broad antibacterial spectrum, including Xoo, Xac, Xoc, Xcm, X. fragariae (Xf), X. campestris pv. campestris (Xcc), Pectobacterium carotovorum subspecies brasiliense (Pcb) and P. carotovorum subsp. carotovorum (Pcc), with minimum inhibitory concentration (MIC) values ranging from 333.75 to 1335 μmol/L. The pot experiment showed that 4-allylbenzene-1,2-diol exerted an excellent protective effect against Xoo, with a controlled efficacy reaching 72.73% at 4 MIC, which was superior to the positive control kasugamycin (53.03%) at 4 MIC. Further results demonstrated that the 4-allylbenzene-1,2-diol damaged the integrity of the cell membrane and increased cell membrane permeability. In addition, 4-allylbenzene-1,2-diol also prevented the pathogenicity-related biofilm formation in Xoo, thus limiting the movement of Xoo and reducing the production of extracellular polysaccharides (EPS) in Xoo. These findings suggest the value of 4-allylbenzene-1,2-diol and P. austrosinense could be as promising resources for developing novel antibacterial agents. [ABSTRACT FROM AUTHOR]

Diaphorina citri Kuwayama (D. citri) is an insect vector of phloem-limited 'Candidatus Liberibacter asiatus' (CLas), the presumed pathogen of citrus Huanglongbing (HLB). Recently, our lab has preliminarily found it acquired and transmitted Citrus tristeza virus (CTV), which was previously suggested to be vectored by species of aphids. However, the influences of one of the pathogens on the acquisition and transmission efficiency of the other pathogen remain unknown. In this study, CLas and CTV acquisition and transmission by D. citri at different development stages under field and laboratory conditions were determined. CTV could be detected from the nymphs, adults, and honeydew of D. citri but not from the eggs and exuviates of them. CLas in plants might inhibit CTV acquisition by D. citri as lower CTV–positive rates and CTV titers were detected in D. citri collected from HLB-affected trees compared to those from CLas–free trees. D. citri were more likely to obtain CTV than CLas from host plants co-infected with the two pathogens. Intriguingly, CTV in D. citri facilitated the acquisition and transmission of CLas, but CLas carried by D. citri had no significant effect on the transmission of CTV by the same vector. Molecular detection and microscopy methods confirmed the enrichment of CTV in the midgut after a 72-h acquisition access period. Collectively, these results raise essential scientific questions for further research on the molecular mechanism of pathogen transmission by D. citri and provide new ideas for the comprehensive prevention and control of HLB and CTV. [ABSTRACT FROM AUTHOR]

Bacterial spot of tomato (BST) is a disease that severely afflicts tomato crops, especially in geographic areas such as the Southeastern U.S., where the environmental conditions favor rapid disease development. Farmers usually use chemical treatments such as copper–mancozeb mixtures and acibenzolar-S-methyl, among other methods, to manage BST. However, these chemical treatments generally fail to improve marketable yields, thus raising the question of whether the BST treatments are economical. We evaluated the efficacy and profitability of bactericide treatments consisting of copper-mancozeb, acibenzolar-S-methyl, and streptomycin, as well as three inoculation levels of Xanthomonas euvesicatoria pv. perforans, on the management of BST in Florida. Across three separate field trials, BST severity was inversely correlated with marketable tomato yields; however, bactericide treatments provided no statistical improvement in marketable yields. By accounting for yield and the BST treatment costs, our profitability analysis showed that the BST treatments did not pay off economically; the net returns of these treatments were statistically equivalent to the untreated controls. [ABSTRACT FROM AUTHOR]

Phytopathogenic bacteria represent a risk to global food production by impacting a variety of crops. The aim of this study was to characterize the contents of bioactive constituents in extracts from Ginkgo biloba L. leaves and fruits and test their activity against six phytopathogenic bacteria that affect horticultural crops. Gas chromatography–mass spectrometry (GC−MS) was used for the chemical profiling of the aqueous methanol extracts, and their bacteriostatic activity against Clavibacter michiganensis subsp. michiganensis, Pseudomonas cichorii, Pseudomonas syringae pv. pisi, Pseudomonas syringae pv. syringae, Pseudomonas syringae pv. tomato, and Xanthomonas vesicatoria (formerly Xanthomonas campestris pv. vesicatoria) was determined in vitro through the agar dilution method. The protective effect of the leaf extract was tested in vivo against the most relevant bacterial pathogens based on their economic/scientific importance, i.e., C. michiganensis subsp. michiganensis and P. syringae pv. pisi, in tomato (Solanum lycopersicum L.) and pea (Pisum sativum L.) plants, respectively, under greenhouse conditions. The GC−MS characterization of G. biloba extracts revealed the presence of dihydro-4-hydroxy-2(3H)-furanone, 2,4-dimethyl-3-hexanol, catechol, 3-O-methyl-D-fructose, 4,6-di-O-methyl-α-D-galactose, methyl 2-O-methyl-α-D-xylofuranoside, and 3-methyl mannoside. In vitro growth inhibition tests showed that, while the fruit extract had no activity, the leaf extract exhibited minimum inhibitory concentrations between 500 and 1000 μg mL−1, which may be attributed to the presence of 2,4-dimethyl-3-hexanol and catechol. In vivo tests of the leaf extract demonstrated full protection in tomato and pea plants at 1000 and 1500 μg∙mL−1, respectively. The results indicate that G. biloba leaves may be employed as a biorational source for integrated pest management in horticulture. [ABSTRACT FROM AUTHOR]

The Gram-negative bacterial genus Xanthomonas includes numerous infra-specific taxa known as pathovars, which are defined primarily on host range and disease symptoms. With the advent of molecular sequence data, many pathovars have been transferred from X. campestris into other Xanthomonas species to better harmonise taxonomy and phylogeny. We performed whole-genome shotgun sequencing on pathotype strains of the following X. campestris pathovars: blepharidis, carissae, clerodendri, convolvuli, coriandri, daturae, euphorbiae, fici, heliotropii, ionidii, lawsoniae, mirabilis, obscurae, paulliniae, pennamericanum, spermacoces, uppalii, vernoniae, viegasii and zingibericola. These genomes showed more than 98% average nucleotide identity with the type-strain of X. euvesicatoria and less than 88% with the type-strain of X. campestris. We propose the transfer of these pathovars into X. euvesicatoria and present an emended species description for X. euvesicatoria. [ABSTRACT FROM AUTHOR]

Rice is an important staple food for more than half of the world's population. Though the genetic potential of commonly cultivated varieties of rice is diminished due to various biotic and abiotic constraints, bacterial leaf blight (BLB) of rice caused by Xanthomonas oryzae pv. oryzae (Xoo) is considered one of its most destructive diseases in India. Based on morpho-cultural characteristics, bacterial pathogens isolated from the leaves of a rice plant showing typical BLB symptoms were identified as Xanthomonas oryzae pv. oryzae. Morphological studies revealed that the pathogen is Gram-negative, a short rod, with rounded ends, single or in pairs, light yellow, circular, whitish yellow to straw-colored, convex, yellow, slightly raised, motile with a single polar flagellum, capsulate and non-spore-forming. Biochemical tests, viz., the Gram reaction, KOH test and catalyst test, showed a positive reaction for all the isolates. Twenty isolates of Xoo were collected from the basmati-growing areas of the Jammu, Samba and Kathua districts in the Jammu sub-tropics during 2019, and their pathogenicity was confirmed on five susceptible rice cultivars, viz., Basmati-370, Pusa-1121, TN-1, SJR and Jaya, by the leaf-clipping method, and subsequently, Koch's postulate was established in each case. Seven Xoo pathotypes, viz., Pathotype 1, Pathotype 2, Pathotype 3, Pathotype 4, Pathotype 5, Pathotype 6 and Pathotype 7, were identified from the total sample of 20 isolates. Pathotype 2 was the most dominant (100%), followed by Pathotype 5 (44.44%), Pathotype 4 (40%), Pathotype 6 (40%), Pathotype 7 (33.33%), Pathotype 3 (22.22%) and Pathotype 1 (20%), in the Jammu sub-tropics. In Jammu district, Pathotype 5 was highly distributed (44.44%) followed by Pathotype 7 (33.33%) and Pathotype 3 (22.22%). Pathotype 4 and Pathotype 6 each showed a 40 percent distribution in Kathua district, followed by Pathotype 1 (20%). Only one pathotype, i.e., Pathotype 2, was recorded in Samba district with a 100 percent distribution. Five genes, viz., Xa13, Xa4, Xa13 and Xa5 + Xa13, showed complete resistance, whereas Xa4, Xa5, Xa7, Xa8, Xa21, Xa4 + Xa5 and Xa4 + Xa21 showed susceptible response against the test isolates. It was observed that most of the single BLB-resistant genes were moderately to highly susceptible to almost all the Xoo isolates, whereas combinations of BLB resistance genes possessed high resistance against all the Xoo isolates. The studies revealed that diverse pathogenic variations existed in the Xoo population in the basmati-growing region of Jammu and Kashmir. Based on the response exhibited by Xoo isolates on differential lines, seven pathotypes (Pathotype 1–7) were identified, and their virulence spectrum on rice differentials showed the occurrence of 5, 3, 10, 10, 20, 10 and 15 percent, respectively, in the Jammu sub-tropics. To develop durable and sustainable resistant cultivars, it is essential to identify predominate race(s) in a specific geographical area and continuously monitor the virulence pattern there. [ABSTRACT FROM AUTHOR]

This study was carried out to determine the life span of Xanthomonas euvesicatoria (Xeu) in seeds, soil and plant residues and the first inoculum source that cause the onset of the disease. The study was carried out in the province of Tokat. Rifampicin-resistant Xeu isolate was inoculated into soil, seeds and plant residues. As a result of the studies, it was determined that Xeu remained viable in inoculated seeds until the 220th day (7 months), and lived in the soil for 105 days in summer and 14 days in winter. In plant residues, it was determined that the the life span in summer period was 60 days at 0 cm depth, 30 days at 20 cm depth; 30 days at 0 cm depth and 15 days at 20 cm depth in that of winter period. According to the results of this study, it was determined that the Xeu did not survive for a long time in plant residues and soil in Tokat province and thus was not carried to the next year, on the other hand the seed was the most important source of inoculum. [ABSTRACT FROM AUTHOR]

Bacteriophages have greatly engaged the attention of scientists worldwide due to the continuously increasing resistance of phytopathogenic bacteria to commercially used chemical pesticides. However, the knowledge regarding phages is still very insufficient and must be continuously expanded. This paper presents the results of the isolation, characterization, and evaluation of the potential of 11 phage isolates as natural predators of a severe phytopathogenic bacterium—Xanthomonas euvesicatoria. Phages were isolated from the rhizosphere of tomato plants with symptoms of bacterial spot. The plaque morphology of all isolates was determined on a X. euvesicatoria lawn via a plaque assay. Three of the isolates were attributed to the family Myoviridae based on TEM micrographs. All phages showed good long-term viability when stored at 4 °C and −20 °C. Three of the phage isolates possessed high stability at very low pH values. Fifty-five-day persistence in a soil sample without the presence of the specific host and a lack of lytic activity on beneficial rhizosphere bacteria were found for the phage isolate BsXeu269p/3. The complete genome of the same isolate was sequenced and analyzed, and, for the first time in this paper, we report a circular representation of a linear but circularly permuted phage genome among known X. euvesicatoria phage genomes. [ABSTRACT FROM AUTHOR]

Simple Summary: Controlling Asian citrus psyllid (Diaphorina citri), the insect vector of Candidatus Liberibacter asiaticus (CLas), is an important part of Huanglongbing management. Specific viruses were recently discovered in a psyllid host where they may act as biological agents for minimizing psyllid populations or CLas transmission. We analyzed the psyllid gut virome from five regions in Florida and identified four D. citri-associated viruses at different compositions. The presence of these specific viruses in the gut indicated potential interactions between these viruses and CLas. Our results provided evidence that D. citri-associated viruses located in the host gut and their potential activities may be useful for pest management. Asian citrus psyllid (Diaphorina citri) transmits the bacterial pathogen Candidatus Liberibacter asiaticus (CLas), the putative causative agent of citrus Huanglongbing disease (HLB). Insect-specific viruses can act against insects as their natural enemies, and recently, several D. citri-associated viruses were discovered. The insect gut plays an important role as not only a pool for diverse microbes but also as a physical barrier to prevent the spread of pathogens such as CLas. However, there is little evidence of the presence of D. citri-associated viruses in the gut and of the interaction between them and CLas. Here, we dissected psyllid guts collected from five growing regions in Florida, and the gut virome was analyzed by high throughput sequencing. Four insect viruses, including D. citri-associated C virus (DcACV), D. citri densovirus (DcDV), D. citri reovirus (DcRV), and D. citri flavi-like virus (DcFLV), were identified, and their presence in the gut, including an additional D. citri cimodo-like virus (DcCLV), were confirmed with PCR-based assays. Microscopic analysis showed that DcFLV infection leads to morphological abnormalities in the nuclear structure in the infected psyllid gut cells. The complex and diverse composition of microbiota in the psyllid gut suggests a possible interaction and dynamics between CLas and the D. citri-associated viruses. Our study identified various D. citri-associated viruses that localized in the psyllid gut and provided more information that helps to evaluate the potential vectors for manipulating CLas in the psyllid gut. [ABSTRACT FROM AUTHOR]

Xanthomonas oryzae pv. oryzicola (Xoo) is a plant pathogenic bacterium that can cause rice bacterial blight disease, which results in a severe reduction in rice production. Antimicrobial-dependent microbial controlling is a useful way to control the spread and outbreak of plant pathogenic bacteria. However, the abuse and long-term use of antimicrobials also cause microbial antimicrobial resistance. As far as known, the mechanism of antimicrobial resistance in agricultural plant pathogenic bacteria still lacks prospecting. In this study, we explore the mechanism of Zhongshengmycin (ZSM)-resistance in Xoo by GC-MS-based metabolomic analysis. The results showed that the down-regulation of the TCA cycle was characteristic of antimicrobial resistance in Xoo, which was further demonstrated by the reduction of activity and gene expression levels of key enzymes in the TCA cycle. Furthermore, alanine was proven to reverse the ZSM resistance in Xoo by accelerating the TCA cycle in vivo. Our results are essential for understanding the mechanisms of ZSM resistance in Xoo and may provide new strategies for controlling this agricultural plant pathogen at the metabolic level. [ABSTRACT FROM AUTHOR]

Xanthomonas spp. are important plant pathogens that seriously endanger crop yields and food security. RpfF is a key enzyme that is involved in the synthesis of diffusible signal factor (DSF) signals and predominates in the signaling pathway regulating quorum sensing (QS) in Xanthomonas. Currently, novel RpfF enzyme-based quorum sensing agents have been proposed as a promising strategy for the development of new pesticides. However, few reports are available that comprehensively summarize the progress in this field. Therefore, we provide a comprehensive review of the recent advances in DSF-mediated QS and recently reported inhibitors that are proposed as bactericide candidates to target the RpfF enzyme and control plant bacterial diseases. [ABSTRACT FROM AUTHOR]

Pepper (Capsicum annuum L.) carrying the gds (corresponding to bs5) gene can prevent the development of bacterial leaf spot disease without HR. However, little is known regarding the development of the resistance mechanism encoded by gds, especially its influence on the bacterium. Here, the effect of gds was compared with pattern-triggered immunity (PTI), another form of asymptomatic resistance, to reveal the interactions and differences between these two defense mechanisms. The level of resistance was examined by its effect on the bacterial growth and in planta expression of the stress and pathogenicity genes of Xanthomonas euvesicatoria. PTI, which was activated with a Pseudomonas syringae hrcC mutant pretreatment, inhibited the growth of Xanthomonas euvesicatoria to a greater extent than gds, and the effect was additive when PTI was activated in gds plants. The stronger influence of PTI was further supported by the expression pattern of the dpsA bacterial stress gene, which reached its highest expression level in PTI-induced plants. PTI inhibited the hrp/hrc expression, but unexpectedly, in gds plant leaves, the hrp/hrc genes were generally expressed at a higher level than in the susceptible one. These results imply that different mechanisms underlie the gds and PTI to perform the symptomless defense reaction. [ABSTRACT FROM AUTHOR]

Xanthomonas citri pv. citri (Xcc) (X. citri subsp. citri) type A is the causal agent of citrus bacterial canker (CBC) on most Citrus spp. and close relatives. Two narrow-host-range strains of Xcc, Aw and A*, from Florida and Southwest Asia, respectively, infect only Mexican lime (Citrus aurantifolia) and alemow (C. macrophylla). In the initial stage of infection, these xanthomonads enter via stomata to reach the apoplast. Herein, we investigated the differences in chemotactic responses for wide and narrow-host-range strains of Xcc A, X. euvesicatoria pv. citrumelonis (X. alfalfae subsp. citrumelonis), the causal agent of citrus bacterial spot, and X. campestris pv. campestris, the crucifer black rot pathogen. These strains of Xanthomonas were compared for carbon source use, the chemotactic responses toward carbon compounds, chemotaxis sensor content, and responses to apoplastic fluids from Citrus spp. and Chinese cabbage (Brassica pekinensis). Different chemotactic responses occurred for carbon sources and apoplastic fluids, depending on the Xanthomonas strain and the host plant from which the apoplastic fluid was derived. Differential chemotactic responses to carbon sources and citrus apoplasts suggest that these Xanthomonas strains sense host-specific signals that facilitate their location and entry of stomatal openings or wounds. [ABSTRACT FROM AUTHOR]

In order to trace back the spread of bacterial spot pathogens within and among countries, Vancheva et al. [[4]] developed the first multilocus variable number of tandem repeat analyses (MLVA) scheme for pepper- and tomato-pathogenic strains of I X. euvesicatoria i and applied this new tool to analyse a representative set of 88 I X. euvesicatoria i pepper strains from Bulgaria and North Macedonia. To understand the possible risks of introducing I X. citri i pathogens, Licciardello et al. [[14]] studied the susceptibility to these pathogens not only of classical citrus crop plants, but also of a diversity of ornamental plants from the I Rutaceae i family. Present, emerging or re-emerging plant diseases due to infection by bacteria of the I Lysobacteraceae i (syn: I Xanthomonadaceae i ) family are continually challenging food security and cause significant losses to the economies of European countries each year. [Extracted from the article]

Pomegranate bacterial blight caused by Xanthomonas axonopodis pv. punicae (Xap) is a highly destructive disease. In the absence of host resistance to the disease, we aimed to evaluate the biocontrol potential of endophytic bacteria against Xap. Thus, in this study, we isolated endophytes from pomegranate plants, identified them on the basis of 16S rDNA sequencing, tested them against Xap, and estimated the endophyte-mediated host defense response. The population of isolated endophytes ranged from 3 × 106 to 8 × 107 CFU/g tissue. Furthermore, 26 isolates were evaluated for their biocontrol activity against Xap, and all the tested isolates significantly reduced the in vitro growth of Xap (15.65% ± 1.25% to 56.35% ± 2.66%) as compared to control. These isolates could reduce fuscan, an uncharacterized factor of Xap involved in its aggressiveness. Lower blight incidence (11.6%) and severity (6.1%) were recorded in plants sprayed with endophytes 8 days ahead of Xap spray (Set-III) as compared to control plants which were not exposed to endophytes (77.33 and 50%, respectively%) during in vivo evaluation. Moreover, significantly high phenolic and chlorophyll contents were estimated in endophyte-treated plants as compared to control. The promising isolates mostly belonged to the genera Bacillus, Burkholderia, and Lysinibacillus, and they were deposited to the National Agriculturally Important Microbial Culture Collection, India. [ABSTRACT FROM AUTHOR]

Xanthomonas bacteria are known phytopathogens difficult to control in the field, which cause great losses in many economically important crops. Genomic islands are fragments acquired by horizontal transference that are important for evolution and adaptation to diverse ecological niches. Virulence and pathogenicity islands (PAIs) enhance molecular mechanisms related to host adaptation. In this work, we have analyzed 81 genomes belonging to X. campestris, and a complex group of X. citri, X. axonopodis, and X. fuscans belonging to nine different pathovars and three subspecies, to analyze and compare their genomic contents. Xanthomonas pan-genome is open and has a massive accessory genome. Each genome showed between three and 15 exclusive PAIs, well conserved through strains of the same pathovar or subspecies. X. axonopodis pv. anacardii had higher general similarity to X. citri subsp. citri and X. fuscans subsp. aurantifolii, with which a few PAIs were shared. Genomic synteny was even for almost all strains, with few rearrangements found in X. axonopodis pv. anacardii. The prophage regions identified in the genomes were mostly questionable or incomplete, and PAI13 in X. campestris pv. campestris ATCC33913 matched a prophage region of 19 transposable elements. Finally, PAIs in Xanthomonas are pathovar-specific, requiring individual strategies of combat. [ABSTRACT FROM AUTHOR]

Bacterial spot of tomato continues to pose a significant problem to tomato production worldwide. In Florida, bacterial spot of tomato caused by Xanthomonas perforans is one of the most important diseases responsible for tomato yield loss. This disease is difficult to control, and new strategies are continually being investigated to combat the devastating effect of this disease. Recent efforts focusing on essential oils based on small molecules have spurred interests in the utilization of this class of chemicals for disease management. In this study, we evaluated the efficacy of eugenol for the management of bacterial spot of tomato caused by X. perforans. In the greenhouse experiments, eugenol applied as a foliar spray significantly (p < 0.5) reduced bacterial spot disease compared to the untreated control. In the field experiments, the area under the disease progress curve (AUDPC) was significantly (p < 0.5) lower in the plots treated with eugenol or eugenol combined with the surfactant Cohere than in the untreated control plots, and it was comparable to the copper-based treatments. To provide additional insights into the possible pathways of eugenol activities, we applied a liquid chromatography mass spectrometry (LC-MS)-based metabolomic study using a thermo Q-Exactive orbitrap mass spectrometer with Dionex ultra high-performance liquid chromatography (UHPLC) on X. perforans strain 91–118 treated with eugenol. Our results showed that eugenol affected metabolite production in multiple pathways critical to bacterial survival. For example, treatment of cells with eugenol resulted in the downregulation of the glutathione metabolism pathway and associated metabolites, except for 5-oxoproline, which accumulation is known to be toxic to living cells. While the peaks corresponding to the putatively identified sarmentosin showed the most significant impact and reduced in response to eugenol treatment, branched-chain amino acids, such as L-isoleucine, increased in production, suggesting that eugenol may not negatively affect the protein biosynthesis pathways. The results from our study demonstrated the efficacy of eugenol in the management of bacterial spot of tomato under greenhouse and field conditions and identified multiple pathways that are targeted. [ABSTRACT FROM AUTHOR]

Soybean is a pivotal staple crop worldwide, supplying the main food and feed plant proteins in some countries. In addition to interacting with mutualistic microbes, soybean also needs to protect itself against pathogens. However, to grow inside plant tissues, plant defense mechanisms ranging from passive barriers to induced defense reactions have to be overcome. Pathogenic but also symbiotic micro-organisms effectors can be delivered into the host cell by secretion systems and can interfere with the immunity system and disrupt cellular processes. This review summarizes the latest advances in our understanding of the interaction between secreted effectors and soybean feedback mechanism and uncovers the conserved and special signaling pathway induced by pathogenic soybean cyst nematode, Pseudomonas, Xanthomonas as well as by symbiotic rhizobium. [ABSTRACT FROM AUTHOR]

Rice bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae (Xoo) is responsible for a significant reduction in rice production. Due to the small impact on the environment, biogenic nanomaterials are regarded as a new type of antibacterial agent. In this research, three colloids of silver nanoparticles (AgNPs) were synthesized with different biological materials such as Arctium lappa fruit, Solanum melongena leaves, and Taraxacum mongolicum leaves, and called Al-AgNPs, Sm-AgNPs and Tm-AgNPs, respectively. The appearance of brown colloids and the UV-Visible spectroscopy analysis proved the successful synthesis of the three colloids of AgNPs. Moreover, FTIR and XRD analysis revealed the formation of AgNPs structure. The SEM and TEM analysis indicated that the average diameters of the three synthesized spherical AgNPs were 20.18 nm, 21.00 nm, and 40.08 nm, respectively. The three botanical AgNPs had the strongest bacteriostatic against Xoo strain C2 at 20 μg/mL with the inhibition zone of 16.5 mm, 14.5 mm, and 12.4 mm, while bacterial numbers in a liquid broth (measured by OD600) decreased by 72.10%, 68.19%, and 65.60%, respectively. Results showed that the three AgNPs could inhibit biofilm formation and swarming motility of Xoo. The ultrastructural observation showed that Al-AgNPs adhered to the surface of bacteria and broke the bacteria. Overall, the three synthetic AgNPs could be used to inhibit the pathogen Xoo of rice bacterial leaf blight. [ABSTRACT FROM AUTHOR]

Citrus canker is a quarantined disease caused by the bacterial plant pathogen Xanthomonas citri subsp. citri (Xcc), which causes persistent surface damage, leaf and fruit drop, and tree decline in citrus plants. The citrus cultivar Citron C-05 (Citrus medica L.) is a disease-resistant genotype identified after years of screening at the National Center for Citrus Improvement (Changsha), which displays allergic, necrotic, and disease-resistant responses to Xcc. In this study, the BAK1 gene was identified in this cultivar to be a disease resistance gene involved in plant-microbe interaction between citrus and Xcc. Functional investigations of this gene revealed that both CsBAK1 (C. sinensis BAK1) or CmBAK1(C. medica BAK1) could inhibit the growth of Xcc to some extent when transiently expressed in the susceptible 'Bingtang' genotype of sweet orange. Critical regions of the CmBAK1 promoter sequence were identified by creating downstream deletions and exposing mutants to Xcc to determine effects on the resistance phenotype; a 426 bp region (−2000~–1574) was identified as a key functional region responsible for eliciting the hypersensitive response in plants. Through screening arrayed Citron C-05 cDNA libraries by yeast one-hybrid assays, a basic APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF) transcription factor of CmRAP2-13 that binds directly to the 426 bp key sequence and activates expression of CmBAK1 was identified. Moreover, transcriptional analysis revealed an obvious increase in transcript levels of CsRAP2-13 in Citron C-05, American citron, and Finger citron. In this study, we present the identification of transcriptional activators that are found to interact with BAK1 proteins in response to Xcc. These results reveal a coordinated regulatory mechanism of RAP2-13, which may be involved in defence responses through the regulation of BAK1. [ABSTRACT FROM AUTHOR]

Xanthomonas axonopodis pv. citri (Xac) belongs to the Gram-negative species, causing citrus canker that seriously affects the fruit yield and quality of many rutaceae plants. Herein, we found that compound 2-(butyldisulfanyl) quinazolin-4(3H)-one exhibited remarkable anti-Xac activity in vitro with a half effective concentration (EC50) of 2.6 μg/mL, while the positive controls thiodiazole-copper with 57 μg/mL and bismerthiazol with 68 μg/mL and this compound showed great anti-citrus canker activity in vivo. This active compound also was confirmed to reduce biofilm formation, increase the level of reactive oxygen species, damage the morphological structure of the bacteria, and cause bacterial death. Proteomics and RT-qPCR analysis results indicated that this compound down-regulated the expression of enzymes in the MEP (2-methyl-D-erythritol 4-phosphate) pathway and might achieve destructive ability of Xac. Overall, this study indicates that such derivatives could be a promising scaffold to develop novel bactericides to control citrus canker. [ABSTRACT FROM AUTHOR]

Bacterial spot (BS) of tomato (S. lycopersicum), caused by Xanthomonas spp., namely X. euvesicatoria (Xeu), is one of the major threats for the production of this crop worldwide. Developing new biocontrol solutions against this disease will allow disease management strategies to be less based on Cu compounds. Nanoclays, such as montmorillonite (NMT), have been under investigation for their antimicrobial activity, or as delivery tools/stabilizers for organic compounds, such as essential oils (EOs), that also possess antimicrobial activity against plant pathogens. This work aims to assess how the application of NMT alone or incorporating S. montana EO on Xeu-infected hosts (var. Oxheart) affects the shoots' redox status and antioxidant defense mechanisms. In vitro shoots, grown on Murashige and Skoog medium, were divided in two groups, Xeu-infected and uninfected (control) shoots. Shoots of each group were then treated with NMT, S. montana EO, EO-NMT. Results show that the NMT was able to reduce Xeu bacterial amount, while reducing ROS production and keeping the transcript levels of the defense-related genes close to those of the control. When applied to uninfected shoots, the treatments triggered the production of ROS and upregulated the phenylpropanoid and hormone pathway, which suggest that they act as defense elicitors. Globally, the results indicate that NMT has the potential to integrate BS management strategies, due to its antimicrobial activity, and that EO and/or nanoclays could be successfully employed as new disease preventive strategies, since they enhance the healthy shoots' defense, thus potentially limiting the pathogen establishment. [ABSTRACT FROM AUTHOR]

Bacterial Leaf Spot (BLS) is a serious bacterial disease of chilli (Capsicum spp.) caused by at least four different Xanthomonas biotypes: X. euvesicatoria pv. euvesicatoria, X. euvesicatoria pv. perforans, X. hortorum pv. gardneri, and X. vesicatoria. Symptoms include black lesions and yellow halos on the leaves and fruits, resulting in reports of up to 66% losses due to unsalable and damaged fruits. BLS pathogens are widely distributed in tropical and subtropical regions. Xanthomonas is able to survive in seeds and crop residues for short periods, leading to the infections in subsequent crops. The pathogen can be detected using several techniques, but largely via a combination of traditional and molecular approaches. Conventional detection is based on microscopic and culture observations, while a suite of Polymerase Chain Reaction (PCR) and Loop-Mediated Isothermal Amplification (LAMP) assays are available. Management of BLS is challenging due to the broad genetic diversity of the pathogens, a lack of resilient host resistance, and poor efficacy of chemical control. Some biological control agents have been reported, including bacteriophage deployment. Incorporating stable host resistance is a critical component in ongoing integrated management for BLS. This paper reviews the current status of BLS of chilli, including its distribution, pathogen profiles, diagnostic options, disease management, and the pursuit of plant resistance. [ABSTRACT FROM AUTHOR]

Plants grow together with microbes that have both negative and positive impacts on the host, while prokaryotes are in turn also hosts for viruses, co-evolving together in a complex interrelationship. Most research focuses on the interaction of either bacterial pathogens interacting with the plant host, or the impact on viruses on their pathogenic bacterial hosts. Few studies have investigated the co-evolution of bacterial pathogens with their host plants as well as with their bacterial viruses. In this work, we aimed to identify the genes that were associated with both phage sensitivity and host pathogenicity of the bacterium Xanthomonas oryzae pv. oryzae (Xoo), which is the most important bacterial rice pathogen. Using the Tn5 transposon mutation technology, we created a library of Xoo strain C2 comprising 4524 mutants, which were subsequently tested for phage infectability. The phage infection tests showed that less than 1% of the mutants (n = 36) were resistant to phage infection, which was attributed to the Tn5 insertion in 19 genes. Interestingly, three out of 19 genes that conveyed resistance to the phage resulted in reduced pathogenicity to rice seedlings compared to the wild type. We identified three genes involved in both phage infection and bacterial virulence, which were studied by knockout mutants and complementation experiments. All of the three knockout mutants were resistant to infection by phage X2, while the complemented strains restored the susceptibility to the bacterial virus. Surprisingly, the genes are also essential for pathogenicity, which we confirmed by single knockout mutants corresponding to the Tn5 mutants. All three genes are involved in lipopolysaccharide synthesis, thus changing the cell envelope surface molecule composition. Our work shows a possible balance in terms of the connection between bacterial virulence and phage resistance, supporting the deployment of phages for the biocontrol of plant pathogens. [ABSTRACT FROM AUTHOR]

The genus of Xanthomonas contains many well-known plant pathogens with the ability to infect some of the most important crop plants, thereby causing significant economic damage. Unfortunately, classical pest-control strategies are neither particularly efficient nor sustainable and we are, therefore, in demand of alternatives. Here, we present the isolation and characterization of seven novel phages infecting the plant-pathogenic species Xanthomonas translucens and Xanthomonas campestris. Transmission electron microscopy revealed that all phages show a siphovirion morphology. The analysis of genome sequences and plaque morphologies are in agreement with a lytic lifestyle of the phages making them suitable candidates for biocontrol. Moreover, three of the isolated phages form the new genus "Shirevirus". All seven phages belong to four distinct clusters underpinning their phylogenetic diversity. Altogether, this study presents the first characterized isolates for the plant pathogen X. translucens and expands the number of available phages for plant biocontrol. [ABSTRACT FROM AUTHOR]

Xanthomonas arboricola pv. juglandis (hereafter X. juglandis) is the etiological agent of walnut blight, the most important bacterial disease affecting walnut production worldwide. Currently, the disease is treated mainly with copper-derived compounds (e.g., CuSO4) despite the evidence of genetic resistance in these strains. Regarding the effectiveness and sustainability, the use of a bacteriophage appears to be a biocontrol alternative to reduce X. juglandis load and symptomatology of walnut blight. Here, the phages f20-Xaj, f29-Xaj, and f30-Xaj were characterized, and their effectiveness in walnut orchards against walnut blight was determined. These bacteriophages showed a specific lytic infection in X. juglandis strains isolated from Chile and France. Phylogenetic analysis of the complete genome of f20-Xaj and f30-Xaj indicates that these phages belong to the Pradovirus genus. In the field, the cocktail of these bacteriophages showed similar effectivity to CuSO4 in the reduction of incidence and severity in walnut tissue. Moreover, the bacterial load of X. juglandis was significantly reduced in the presence of bacteriophages in contrast to a CuSO4 treatment. These results show that the use of bacteriophages can be an alternative to combat the symptoms of walnut blight caused by X. juglandis. [ABSTRACT FROM AUTHOR]

The plant-specific ASR (abscisic acid, stress and ripening) transcription factors are pivotal regulators of plant responses to abiotic stresses. However, their functions in plant disease resistance remain largely unknown. In this study, we revealed the role of OsASR6 in rice plants' resistance to two important bacterial diseases caused by Xanthomonas oryzae pv. oryzae (Xoo) and X. oryzae pv. oryzicola (Xoc) and elucidated the mechanisms underlying OsASR6-regulated resistance. The expression of OsASR6 was strongly elevated in response to both Xoo and Xoc challenges. Silencing of OsASR6 in OsASR6-RNAi transgenic plants markedly enhanced rice resistance to the two bacterial pathogens. Moreover, comparative transcriptome analyses for OsASR6-RNAi and wild-type plants inoculated and uninoculated with Xoc demonstrated that OsASR6 suppressed rice resistance to Xoc by comprehensively fine-tuning CIPK15- and WRKY45-1-mediated immunity, SA signaling and redox homeostasis. Further luciferase reporter assays confirmed that OsASR6 negatively regulated CIPK15 but not WRKY45-1 expression in planta. Overexpression of OsCIPK15 strongly enhanced rice resistance to Xoo and Xoc. Collectively, these results reveal that OsASR6 alleviates rice resistance through the transcriptional suppression of OsCIPK15, and thus links calcium signaling to rice resistance against X. oryzae. Our findings provide insight into the mechanisms underlying OsASR6-mediated regulation of rice resistance to X. oryzae. [ABSTRACT FROM AUTHOR]