Strains Of Xanthomonas Axonopodis Pv Research Articles

Characterization and variability of strains of Xanthomonas axonopodis pv. passiflorae from the state of Pará, Brazil

Bacterial spot, caused by Xanthomonas axonopodis pv. passiflorae, is a disease that has limited the cultivation of passionfruit in various orchards in Brazil. The objective of this work is to characterize and evaluate the variability of 29 strains of X. axonopodis pv. passiflorae from different municipalities producing yellow passionfruit in the state of Pará. The characterization was performed by the biochemical methods of KOH solubility, oxidase and Bactray, and the molecular methods of Xapas-F, Xapas-Ri and Xapas-Ro primers. The variability was evaluated by pathogenicity test and RAPD (randomly amplified polymorphic DNA). The strains of X. axonopodis pv. passiflorae were Gram-negative and oxidase-negative, and tests with the Bactray kit showed no relation between the collection municipality and group composition. The Xapas-F/Ri/Ro primers were specific for the strains. The primers used amplified 118 polymorphic bands in the RAPD reactions and the highest genetic similarity was between the strains PA15 and PA16. As the pathogenicity test evidenced a pathogenic variability, the strains PA2.1, PA4.5, PA14, PA4.2, PA4.1, PA4.6, PA3.4 and PA4.3 present the highest severity values for the disease. The strains of X. axonopodis pv. passiflorae show characteristics typical of the species, and genetic and pathogenic variability among them.

Morphological and biochemical characterization of strains of Xanthomonas axonopodis pv. vignicola isolated from cowpea grown in three agroecological zones in Nigeria

Cowpea bacterial blight (Xanthomonas axonopodis pv vignicola) is a devastating disease of cowpea in Nigeria. Accurate identification is the first step for an effective management option. Thus, X. axonopodis pv. vignicola isolated from cowpea (Vigna unguinculata (L.) Walp) grown in different Agro-ecological zones in Nigeria were characterized using morphological characteristics, biochemical and antibiotic sensitivity tests. Pot experiments were conducted on two susceptible lines of cowpea (IT90k – 76) and (IT84s – 2246 – 4) in the screen house. In vivo infectivity studies were carried out to determine the effect of each of theX. axonopodis pv. vignicola isolate groups on the cowpea. All isolates from each of the agroecological zones were classified into Group 1 (light yellow) and Group 2 (brownish yellow) based on their morphological presentation on nutrient agar medium. Results from four of the biochemical test showed differences between the two morphologically different groups. Group 2 isolates showed a large zone of inhibition to a Lincomycin hydrochloride while isolates from Group 1 were not affected by the action of the same antibiotic. Blight symptom was only observed when the two types of isolates were combined and inoculated simultaneously. This study showed that there are two morpho-types of X. axonopodis pv. vignicola and their synergetic infectivity results in blight symptoms expression in cowpea.

Consequences of the taxonomic revision of Xanthomonas axonopodis pv. dieffenbachiae and of the analysis of its associated pathogenicity for the recommendation of regulation (EPPO A2 List)

Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of bacterial blight of Araceae (aroids), is a regulated pest in several countries and is included in the EPPO A2 List. Reference strains of Xanthomonas axonopodis pv. dieffenbachiae have recently been reclassified into the species Xanthomonas phaseoli, Xanthomonas citri and Xanthomonas euvesicatoria on the basis of different features, including multilocus sequence analysis, average nucleotide identity and homology in DNA–DNA hybridization analyses. Based on pathogenicity tests, Constantin et al. (2017) proposed naming the pathogens on aroids as X. phaseoli pv. dieffenbachiae, X. phaseoli pv. syngonii and X. citri pv. aracearum. Recommendations are made on how to deal with these changes for the group of pathogenic bacteria for Araceae. The name Xanthomonas axonopodis pv. dieffenbachiae on the EPPO List should be adjusted to the names proposed in the taxonomic study by Constantin et al. (2016). The current EPPO Diagnostic Standard is directed at strains pathogenic on Anthurium. They mainly belong to X. phaseoli pv. dieffenbachiae, but some also to X. citri pv. aracearum that are not detected by the EPPO Diagnostic Standard. Xanthomonas phaseoli pv. syngonii strains are also aggressive, but with a host range restricted to Syngonium. The pathogenicity specific to aroids of the bacterial isolates reclassified as Xanthomonas euvesicatoria was not confirmed and no pathovar epithet has been retained for these strains.

Genetic Variability within Xanthomonas axonopodis pv. punicae, Causative Agent of Oily Spot Disease of Pomegranate

Bacterial blight is one of the most devastating diseases of pomegranate in India. It is known to be caused by the strains of Xanthomonas axonopodis pv. punicae (xap). As the control of disease varies with the use of pesticide agents, it becomes imperative to study the genetic variation among the pathogenic strains. Thirty-six strains of Xanthomonas axonopodis pv. punicae, were isolated from the diseased fruits of 3 varieties of pomegranate originating from 3 different provinces of Maharashtra, India. All the strains characterized phenotypically and genotypically were found to have diversity. The genetic diversity among the 36 Xanthomonas isolates was assessed using RAPD based techniques. A cluster dendrogram based on the random amplified polymorphic DNA (RAPD) showed that genetic diversity existed among the isolates of Xanthomonas. The genomic variation was found to be in the range of 0.55% to 0.95% among the isolates. The cluster analysis based upon band patterns formed two major clusters with 4 sub-groups. It can be concluded that this variability was genetic in nature irrespective of their location and the variety of pomegranate.

Genome sequencing reveals a new lineage associated with lablab bean and genetic exchange between Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans.

Common bacterial blight is a devastating seed-borne disease of common beans that also occurs on other legume species including lablab and Lima beans. We sequenced and analyzed the genomes of 26 strains of Xanthomonas axonopodis pv. phaseoli and X. fuscans subsp. fuscans, the causative agents of this disease, collected over four decades and six continents. This revealed considerable genetic variation within both taxa, encompassing both single-nucleotide variants and differences in gene content, that could be exploited for tracking pathogen spread. The bacterial strain from Lima bean fell within the previously described Genetic Lineage 1, along with the pathovar type strain (NCPPB 3035). The strains from lablab represent a new, previously unknown genetic lineage closely related to strains of X. axonopodis pv. glycines. Finally, we identified more than 100 genes that appear to have been recently acquired by Xanthomonas axonopodis pv. phaseoli from X. fuscans subsp. fuscans.

Pyramiding B Genes in Cotton Achieves Broader But Not Always Higher Resistance to Bacterial Blight

Near-isogenic lines of upland cotton (Gossypium hirsutum) carrying single, race-specific genes B4, BIn, and b7 for resistance to bacterial blight were used to develop a pyramid of lines with all possible combinations of two and three genes to learn whether the pyramid could achieve broad and high resistance approaching that of L. A. Brinkerhoff's exceptional line Im216. Isogenic strains of Xanthomonas axonopodis pv. malvacearum carrying single avirulence (avr) genes were used to identify plants carrying specific resistance (B) genes. Under field conditions in north-central Oklahoma, pyramid lines exhibited broader resistance to individual races and, consequently, higher resistance to a race mixture. It was predicted that lines carrying two or three B genes would also exhibit higher resistance to race 1, which possesses many avr genes. Although some enhancements were observed, they did not approach the level of resistance of Im216. In a growth chamber, bacterial populations attained by race 1 in and on leaves of the pyramid lines decreased significantly with increasing number of B genes in only one of four experiments. The older lines, Im216 and AcHR, exhibited considerably lower bacterial populations than any of the one-, two-, or three-B-gene lines. A spreading collapse of spray-inoculated AcBIn and AcBInb7 leaves appears to be a defense response (conditioned by BIn) that is out of control.

RNAseq analysis of cassava reveals similar plant responses upon infection with pathogenic and non-pathogenic strains of Xanthomonas axonopodis pv. manihotis.

An RNAseq-based analysis of the cassava plants inoculated with Xam allowed the identification oftranscriptional upregulation of genes involved in jasmonate metabolism, phenylpropanoid biosynthesis and putative targets for a TALE. Cassava bacterial blight, a disease caused by the gram-negative bacterium Xanthomonas axonopodis pv. manihotis (Xam), is a major limitation to cassava production worldwide and especially in developing countries. The molecular mechanisms underlying cassava susceptibility to Xam are currently unknown. To identify host genes and pathways leading to plant susceptibility, we analyzed the transcriptomic responses occurring in cassava plants challenged with either the non-pathogenic Xam strain ORST4, or strain ORST4(TALE1 Xam ) which is pathogenic due to the major virulence transcription activator like effector TALE1 Xam . Both strains triggered similar responses, i.e., induction of genes related to photosynthesis and phenylpropanoid biosynthesis, and repression of genes related to jasmonic acid signaling. Finally, to search for TALE1 Xam virulence targets, we scanned the list of cassava genes induced upon inoculation of ORST4(TALE1 Xam ) for candidates harboring a predicted TALE1 Xam effector binding element in their promoter. Among the six genes identified as potential candidate targets of TALE1 Xam a gene coding for a heat shock transcription factor stands out as the best candidate based on their induction in presence of TALE1 Xam and contain a sequence putatively recognized by TALE1 Xam .

High-Quality Draft Genome Sequences of Xanthomonas axonopodis pv. glycines Strains CFBP 2526 and CFBP 7119.

We report here the high-quality draft genome sequences of two strains of Xanthomonas axonopodis pv. glycines, the causal agent of bacterial pustule on soybeans. Comparison of these genomes with those of phylogenetically closely related pathovars of Xanthomonas spp. will help to understand the mechanisms involved in host specificity and adaptation to host plants.

Characterization of Strains of Xanthomonas axonopodis pv. dieffenbachiae from Bacterial Blight of Caladium and Identification of Sources of Resistance for Breeding Improved Cultivars

Bacterial blight, caused by Xanthomonas axonopodis pv. dieffenbachiae (Xad), is the most common foliar disease of caladium, an ornamental aroid grown for its colorful foliage. The disease can reduce the marketability of caladiums produced as potted plants and lower the yield of caladiums grown for tuber production. Three bacterial strains were isolated from symptomatic caladiums and identified as Xad using fatty acid analysis, carbon source use, and the sequence of the 16S-23S spacer, and tested for virulence against three susceptible cultivars. Two strains were virulent to all of the cultivars; however, one strain was differentially pathogenic, virulent against two cultivars, but not to the usually highly susceptible ‘Candidum’. In greenhouse inoculation tests of 17 cultivars and one breeding line, four cultivars were ranked as highly susceptible, nine as moderately susceptible, and five as resistant. Ten of these cultivars were also evaluated with natural infection in the field with good agreement between the results of the greenhouse and field evaluations. Cultivars White Queen, Florida Red Ruffles, Florida Sweetheart, Candidum Jr., and Mrs. Arno Nehrling have been identified as resistant to bacterial blight in greenhouse or field evaluations and can potentially be used in future breeding efforts to produce improved cultivars.

Role of rpfF in Virulence and Exoenzyme Production of Xanthomonas axonopodis pv. glycines, the Causal Agent of Bacterial Pustule of Soybean

Ten strains of Xanthomonas axonopodis pv. glycines, the causal agent of bacterial pustule of soybean, which were isolated from various soybean growing regions of Thailand, produced an extracellular diffusible factor (DSF) related to a well-characterized quorum sensing molecule produced by other Xanthomonas spp. Genomic DNA of the 10 strains of X. axonopodis pv. glycines contained rpfF, a gene encoding for the biosynthesis of the DSF in X. campestris pv. campestris. The rpfF gene from one strain of X. axonopodis pv. glycines was fully sequenced, and the 289 aa product is closely related to RpfF of other Xanthomonas spp. (95 to 98% identical). Three independently generated rpfF mutants of X. axonopodis pv. glycines strain No12-2 were defective in the production of a DSF, as expected if rpfF encodes for DSF biosynthesis in X. axonopodis pv. glycines. The rpfF mutants of X. axonopodis pv. glycines exhibited reduced virulence on soybean and produced less than wild-type levels of extracellular polysaccharide and the extracellular enzymes carboxylmethylcellulase, protease, endo-beta-1,4-mannanase, and pectate lyase. Transcripts for three genes that encode for the extracellular enzymes protease, endoglucanase, and pectate lyase were at lower abundance in an rpfF mutant than in the parental strain of X. axonopodis pv. glycines. These results indicate that X. axonopodis pv. glycines produces a diffusible signal related to the DSF of X. campestris pv. campestris, which contributes to virulence and exoenzyme production by this phytopathogenic bacterium.

Resistance to Citrus Canker Caused by Xanthomonas axonopodis pv. citri in an Accession of Citrus sp. Similar to Grapefruit.

Citrus canker is caused by Xanthomonas axonopodis pv. citri. All grapefruit (Citrus paradisi Macf.) cultivars are highly susceptible to the A-group strains of Xanthomonas axonopodis pv. citri. A type of grapefruit (Citrus sp.) called Dalan Dan (DD) has shown high resistance in the field in Misiones, Argentina where it is widely planted. The DD is Accession No. NC678 in the Campinas Citrus Germplasm Collection (Centro APTA Citrus Sylvio Moreira, Cordeiropolis, SP, Brazil) and Accession No. CCC-618 Dalan Dan in the Argentina Citrus Germplasm Collection at INTA (Concordia, Entre Ríos). Molecular studies completed in Brazil have shown high similarities among grapefruit varieties and DD (1). We report here the characterization of quantitative resistance to citrus canker in DD. Quantitative resistance in citrus is expressed as lower number of lesions per leaf area (3). Lesions per square centimeter were calculated after 3 to 4 weeks of two sets of inoculation of three inoculum levels (5 × 102, 2.5 × 103, and 5 × 103 cells per ml) in leaves of developing leaf flushes. The period of greatest susceptibility in DD was 15 to 23 days for the three inoculum levels; 1.35 ± 0.34 to 0.01 ± 0.01, 4.98 ± 0.91 to 0.91 ± 0.48, and 9.85 ± 2.17 to 1.91 ± 1.37 lesions per cm2, respectively. By contrast, the period of greatest susceptibility in grapefruit cv. Duncan was 15 to 29 days for the same three inoculum levels; 2.78 ± 0.46 to 1.22 ± 0.39, 17.29 ± 2.79 to 5.55 ± 0.93, and 22.87 ± 3.27 to 11.95 ± 3.55 lesions per cm2, respectively. The results shown here suggest that the DD citrus accession is susceptible to citrus canker for a very short time during the development of leaf flushes, which results in very high resistance in the field. This citrus germplasm material could be valuable as a source of resistance to improve other varieties or as a replacement of grapefruit due to the similar fruit and juice qualities.

Assessment of the Genetic Diversity of Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans as a Basis To Identify Putative Pathogenicity Genes and a Type III Secretion System of the SPI-1 Family by Multiple Suppression Subtractive Hybridizations

Fluorescent amplified fragment length polymorphism revealed that strains of Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans are genetically distinct and can be grouped into four genetic lineages. Four suppression subtractive hybridizations were then performed to isolate DNA fragments present in these bean pathogens and absent from closely related xanthomonads. Virulence gene candidates were identified such as homologs of hemagglutinins, TonB-dependent receptors, zinc-dependent metalloproteases, type III effectors, and type IV secretion system components. Unexpectedly, homologs of the type III secretion apparatus components (SPI-1 family), usually reported in animal pathogens and insect symbionts, were also detected.

Characterization of Xanthomonas spp. strains by bacteriocins

Twenty-five strains of Xanthomonas axonopodis pv. citri and 14 strains of Xanthomonas spp. were tested for bacteriocin production. X. axonopodis pv. passiflorae strains were sensitive to the bacteriocins produced by the 25 X. axonopodis pv. citri strains evaluated in this study while strains of X. axonopodis pv. manihotis and X. campestris pv. campestris showed variable sensitivity. Only five of the 25 X. axonopodis pv. citri strains were not inhibited by the bacteriocins produced by the two X. axonopodis pv. passiflorae strains. The bacteriocins produced by the Xanthomonas axonopodis pv. citri (FDC-806) and X. axonopodis pv. passiflorae (Mar-2850 A) strains were thermolabile, resistant to lysozyme and sensitive to DNAse. The bacteriocin produced by X. axonopodis pv. passiflorae was resistant to the action of proteinase K, trypsin and RNAse while the bacteriocin produced by X. axonopodis pv. citri was sensitive to these enzymes. The bacteriocins produced by X. axonopodis pv. passiflorae and X. axonopodis pv. citri were called passifloricin and citricin, respectively.

Agressividade de linhagens de Xanthomonas axonopodis pv. aurantifolii Tipo C em lima ácida 'Galego'

O presente trabalho objetivou gerar informações referentes à agressividade de linhagens de Xanthomonas axonopodis pv. aurantifolii Tipo C(Xaa-C), produtoras (PP) e não produtoras de pigmento (NP) escuro em meio de cultura, comparativamente a X. axonopodis pv. citri Tipo A (Xac) . Os tratamentos foram formados por 14 linhagens, sendo sete de Xaa-C PP, cinco Xaa-C NP, e duas linhagens de Xac. As linhagens foram inoculadas através de ferimentos, em folhas de lima ácida 'Galego' (Citrus aurantifolia Swingle), com agulha previamente mergulhada em uma suspensão de células bacterianas (10(7) UFC/mL). Foram realizadas dez repetições para cada tratamento, representadas por uma planta cada. As plantas foram mantidas em casa de vegetação durante todo o experimento. As linhagens diferiram entre si quanto ao período de incubação, diâmetro e populações bacterianas das lesões e, comparativamente, Xaa-C NP mostraram-se mais agressivas do que Xaa-C PP. Algumas linhagens induziram sintomas que diferiram quanto à presença e extensão de anasarca, halo amarelo e saliência do tecido necrosado.

Regulation of resistance to copper in Xanthomonas axonopodis pv. vesicatoria.

Copper-resistant strains of Xanthomonas axonopodis pv. vesicatoria were previously shown to carry plasmid-borne copper resistance genes related to the cop and pco operons of Pseudomonas syringae and Escherichia coli, respectively. However, instead of the two-component (copRS and pcoRS) systems determining copper-inducible expression of the operons in P. syringae and E. coli, a novel open reading frame, copL, was found to be required for copper-inducible expression of the downstream multicopper oxidase copA in X. axonopodis. copL encodes a predicted protein product of 122 amino acids that is rich in histidine and cysteine residues, suggesting a possible direct interaction with copper. Deletions or frameshift mutations within copL, as well as an amino acid substitution generated at the putative start codon of copL, caused a loss of copper-inducible transcriptional activation of copA. A nonpolar insertion of a kanamycin resistance gene in copL resulted in copper sensitivity in the wild-type strain. However, repeated attempts to complement copL mutations in trans failed. Analysis of the genomic sequence databases shows that there are copL homologs upstream of copAB genes in X. axonopodis pv. citri, X. campestris pv. campestris, and Xylella fastidiosa. The cloned promoter area upstream of copA in X. axonopodis pv. vesicatoria did not function in Pseudomonas syringae or in E. coli, nor did the P. syringae cop promoter function in Xanthomonas. However, a transcriptional fusion of the Xanthomonas cop promoter with the Pseudomonas copABCDRS was able to confer resistance to copper in Xanthomonas, showing divergence in the mechanisms of regulation of the resistance to copper in phytopathogenic bacteria.

Molecular characterisation of Xanthomonas strains isolated from aroids in Mauritius

Mauritius is one of the largest world producers of Anthurium cut flowers but outbreaks of bacterial blight have never been reported on the island. This work was about the characterisation and identification of bacterial strains isolated from Anthurium andreanum, Dieffenbachia maculata and Aglaonema simplex in Mauritius. Fifteen strains, that showed the morphological properties of Xanthomonas on conventional media, were tested on two semi-selective media (Esculin-trehalose and cellobiose-starch). ELISA tests using a panel of monoclonal antibodies were carried out and three out of 15 strains reacted with a Xanthomonas-specific monoclonal antibody (MAb XII). Analysis using four sets of ribosomal primers revealed that the same three Mauritius strains shared conserved PCR products with reference xanthomonads including virulent strains of Xanthomonas axonopodis pv. dieffenbachiae ( Xad). BIOLOG tests and the Sherlock Microbial Identification system (MIDI) identified these three new strains at the species level as X. axonopodis. The complementary tests that were carried out clearly confirmed that the three strains are xanthomonads and, moreover, a DNA probe which showed specificity to Xad strains suggested that the three Mauritius strains are non-virulent forms of the pathogen causing Anthurium blight.

Detection and Characterization of a New Strain of Citrus Canker Bacteria from Key/Mexican Lime and Alemow in South Florida.

In the Wellington and Lake Worth areas of Palm Beach County, FL, citrus canker appeared on Key/Mexican lime (Citrus aurantiifolia) and alemow (C. macrophylla) trees over a period of about 6 to 7 years before detection, but nearby canker-susceptible citrus, such as grapefruit (C. × paradisi) and sweet orange (C. sinensis), were unaffected. Colonies of the causal bacterium, isolated from leaf, stem, and fruit lesions, appeared similar to the Asiatic group of strains of Xanthomonas axonopodis pv. citri (Xac-A) on the nutrient agar plate, but the growth on lima bean agar slants was less mucoid. The bacterium produced erumpent, pustule-like lesions of typical Asiatic citrus canker syndrome after inoculation into Key/Mexican lime, but brownish, flat, and necrotic lesions on the leaves of Duncan grapefruit, Madame Vinous sweet orange, sour orange (C. aurantium), citron (C. medica), Orlando tangelo (C. reticulata × C. × paradisi), and trifoliate orange (Poncirus trifoliata). The bacterium did not react with the Xac-A specific monoclonal antibody A1 using enzyme-linked immunosorbent assay (ELISA) and could not be detected by polymerase chain reaction (PCR)-based assays using primers selected for Xac-A. DNA reassociation analysis confirmed that the pathogen, designated as Xac-AW, was more closely related to Xac-A and Xac-A* strains than X. axonopodis pv. aurantifolii or the citrus bacterial spot pathogen (X. axonopodis pv. citrumelo). The strain can be easily differentiated from Xac-A and Xac-A* using ELISA, PCR-based tests, fatty acid analysis, pulsed-field gel electrophoresis of genomic DNA, and host specificity.

Identification of Pathotypes of Xanthomonas axonopodis pv. manihotis in Africa and Detection of Quantitative Trait Loci and Markers for Resistance to Bacterial Blight of Cassava

ABSTRACT Cassava suffers from bacterial blight attack in all growing regions. Control by resistance is unstable due to high genotype-environment interactions. Identifying genes for resistance to African strains of Xanthomonas axonopodis pv. manihotis can support breeding efforts. Five F(1) cassava genotypes deriving from the male parent 'CM2177-2' and the female parent 'TMS30572' were used to produce 111 individuals by backcrossing to the female parent. In all, 16 genotypes among the mapping population were resistant to stem inoculation by four strains of X. axonopodis pv. manihotis from different locations in Africa, and 19 groups with differential reactions to the four strains were identified, suggesting that the strains represent different pathotypes. Four genotypes were resistant to leaf inoculation, and three were resistant to both stem and leaf inoculations. Genotypes with susceptible, moderately resistant, and resistant reactions after leaf and stem inoculation partly differed in their reactions on leaves and stems. Based on the genetic map of cassava, single-markeranalysis of disease severity after stem-puncture inoculation was performed. Eleven markers were identified, explaining between 16 and 33.3% of phenotypic variance of area under disease progress curve. Five markers on three and one linkage groups from the female- and male-derived framework of family CM8820, respectively, seem to be weakly associated with resistance to four strains of X. axonopodis pv. manihotis. Based on the segregation of alleles from the female of family CM8873, one marker was significantly associated with resistance to two X. axonopodis pv. manihotis strains, GSPB2506 and GSPB2511, whereas five markers were not linked to any linkage group. The quantitative trait loci (QTL) mapping results also suggest that the four African strains belong to four different pathotypes. The identified pathotypes should be useful for screening for resistance, and the QTL and markers will support breeding for resistance.

The leucine-responsive regulatory protein (lrp) gene for characterization of the relationship among Xanthomonas species.

Characterization of strains of Xanthomonas axonopodis pv. citri by using DNA fingerprints that were generated from primers for enterobacterial repetitive intergenic consensus (ERIC) elements led to the discovery of differential sequences for a leucine-responsive regulatory protein (lrp) gene in two subgroups of strains with different host ranges on Citrus spp. DNA hybridization and PCR-based assays that used different sets of primers were designed to detect the core sequence, as well as to obtain the entire sequence of the lrp gene for several Xanthomonas species and pathovars. Higher variability was observed at the nucleotide level than at the amino acid level among the different species and pathovars, revealing selection pressure on the lrp gene, which is presumably due to an essential role of the gene in bacterial metabolism. Moderate variability in the 3' and 5' domains was used to study relationships among different species within the genus XANTHOMONAS: Species of this genus that were isolated from citrus, as well as other pathovars of X. axonopodis, showed highly similar lrp gene sequences, whereas other Xanthomonas species, especially Xanthomonas campestris, had sequences that were more dissimilar to that of X. axonopodis. Thus, the lrp gene sequence is useful to distinguish X. axonopodis pv. citri groups and promising for polyphasic taxonomic analysis of the genus XANTHOMONAS: Data from analysis of lrp gene sequences support the current concepts for classification of xanthomonads, which are based on other approaches.

Metabolic diversity of Xanthomonas axonopodis pv. vignicola, causal agent of cowpea bacterial blight and pustule

Fifty-five strains of Xanthomonasaxonopodis pv. vignicola, isolated from blight and pustule symptoms of cowpea leaves, originating from 11 countries, were characterized for their carbon-source metabolization pattern using the Biolog GN microplate system. Great variation was found between strains according to origin. Dextrin, glycogen and succinamic acid were not used by strains from Benin, Uganda or Thailand, but by all the other strains (excluding two strains from Mozambique), whereas N-acetyl-D-glucosamine and malonic acid were used by the strains from Benin, Uganda and Thailand, but generally not by the other strains. The strains from Benin, Uganda and Thailand, as well as strains from Venezuela, Brazil and Mozambique, clustered separately from the others in multivariate analysis. Nineteen substrates were used by all the strains, 47 not by any strain and 29 only by some strains. No considerable differences were found between strains isolated from blight symptoms and from pustules. Virulence of strains was not related to the metabolic pattern. The Biolog database was not representative of the diversity of X. axonopodis pv. vignicola, since all strains were identified as Xanthomonas campestris, although belonging to eight pathovars, while only eight of nine strains from Benin and both strains from Thailand were identified as X. campestris pv. vignicola. The Biolog system appeared to be useful for characterizing the diversity of X. axonopodis pv. vignicola strains. A set of representative strains based on metabolic and molecular diversity, virulence and geographic origin is suggested for screening for resistant cowpea cultivars.