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PURPOSE: To evaluate long terms outcomes of botulinum toxin in infantile esotropia by measuring the amount of microtropia 24 months after injection. Secondary purpose was to identify predictive factors of microtropia. METHODS: A retrospective, single-center study was performed at the university medical center in Bordeaux between 2001 and 2018, including all patients with infantile esotropia greater than 20 D. All patients received 5 or 7,5 IU of botulinum toxin A in each medial rectus, once or twice depending on the angle of deviation after the first injection and after wearing full optical correction at least two months. We noted the angle at 1, 6, 12 and 24 months, the occurrence of any complications and the need for later strabismus surgery. The primary endpoint was the achievement of a microtropia less than 8 diopters (D) at 24 months post-injection. We evaluated the predictive factors for microtropia with a Fischer's test. RESULTS: We included 30 patients with esotropia greater than 20 D. The mean follow-up after injection was 48 months ±30. The mean age was 16.24 months (7-29 months) with a female predominance in the population (SR=0.43). The mean pre-injection deviation was 41.25±12.17 D. The majority of patients were mildly (40%) or moderately (40%) hyperopic. At 24 months, 46.7% microtropias were obtained (95% CI: 28.9%-64.5%). The change in mean angle at 1, 6, 12 and 24 months post-injection was -8.57±25.21 D; 14.48±13.40 D; 18.38±12.07 D and 21.23±14.97 D, respectively. No factors were predictive of microtropia. Of the 30 children, 3 had transient ptosis requiring strips and 12 showed an exotropia at 1 month. All complications were self-limited and without consequences. 3 children had a second injection of botulinum toxin, which in 2/3 of the cases resulted in a long-lasting microtropia. 26.7% (n=8) of the children underwent secondary surgery. Obtaining a microtropia 24 months after injection statistically significantly reduced the need for secondary strabismus surgery: 92.9% P=0.039% CI 95% (0.002; 1.0606). CONCLUSION: Botulinum toxin appears to be a less invasive and more conservative alternative to surgery in children with infantile esotropia. In 46.7% of cases, microtropia is achieved. An improvement was noted in 90% (n=27) of the children with a reduction of half (21.23 D) of the mean post-injection angle at 24 months. When effective, it significantly reduces the need for secondary surgery.
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Toxinas Botulínicas Tipo A , Esotropia , Fármacos Neuromusculares , Toxinas Botulínicas Tipo A/efeitos adversos , Criança , Esotropia/tratamento farmacológico , Esotropia/cirurgia , Feminino , Humanos , Lactente , Masculino , Fármacos Neuromusculares/efeitos adversos , Músculos Oculomotores , Procedimentos Cirúrgicos Oftalmológicos , Estudos Retrospectivos , Resultado do Tratamento , Visão BinocularRESUMO
Bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae represents a severe threat to rice cultivation in Mali. Characterizing the pathotypic diversity of bacterial populations is key to the management of pathogen-resistant varieties. Forty-one X. oryzae pv. oryzae isolates were collected between 2010 and 2013 in the major rice growing regions in Mali. All isolates were virulent on the susceptible rice variety Azucena; evaluation of the isolates on 12 near isogenic rice lines, each carrying a single resistance gene, identified six new races (A4 to A9) and confirmed race A3 that was previously reported in Mali. Races A5 and A6, isolated in Office du Niger and Sélingué, were the most prevalent races in Mali. Race A9 was the most virulent, circumventing all of the resistance genes tested. Xa3 controlled six of seven races (i.e., 89% of the isolates tested). The expansion of race A9 represents a major risk to rice cultivation and highlights the urgent need to identify a local source of resistance. We selected 14 isolates of X. oryzae pv. oryzae representative of the most prevalent races to evaluate 29 rice varieties grown by farmers in Mali. Six isolates showed a high level of resistance to X. oryzae pv. oryzae and were then screened with a larger collection of isolates. Based on the interactions among the six varieties and the X. oryzae pv. oryzae isolates, we characterized eight different pathotypes (P1 to P8). Two rice varieties, SK20-28 and Gigante, effectively controlled all of the isolates tested. The low association observed among races and pathotypes of X. oryzae pv. oryzae suggests that the resistance observed in the local rice varieties does not simply rely on single known Xa genes. X. oryzae pv. oryzae is pathogenically and geographically diverse. Both the races of X. oryzae pv. oryzae characterized in this study and the identification of sources of resistance in local rice varieties provide useful information to inform the design of effective breeding programs for resistance to bacterial leaf blight in Mali.
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Oryza , Xanthomonas , Mali , Doenças das PlantasAssuntos
Glaucoma/congênito , Hemorragia Pós-Operatória/etiologia , Hemorragia Retiniana/etiologia , Criança , Feminino , Glaucoma/tratamento farmacológico , Glaucoma/cirurgia , Humanos , Pressão Intraocular , Macula Lutea , Mitomicina/administração & dosagem , Hemorragia Pós-Operatória/diagnóstico por imagem , Hemorragia Pós-Operatória/cirurgia , Hemorragia Retiniana/diagnóstico por imagem , Hemorragia Retiniana/cirurgia , Tomografia de Coerência Óptica , Trabeculectomia/efeitos adversos , Acuidade Visual , VitrectomiaAssuntos
Inibidores da Angiogênese/administração & dosagem , Bevacizumab/administração & dosagem , Neovascularização de Coroide/diagnóstico por imagem , Neovascularização de Coroide/tratamento farmacológico , Adolescente , Neovascularização de Coroide/etiologia , Feminino , Humanos , Injeções Intravítreas , Distrofia Macular Viteliforme/complicaçõesRESUMO
Multilocus variable-number tandem-repeat analysis (MLVA) is efficient for routine typing and for investigating the genetic structures of natural microbial populations. Two distinct pathovars of Xanthomonas oryzae can cause significant crop losses in tropical and temperate rice-growing countries. Bacterial leaf streak is caused by X. oryzae pv. oryzicola, and bacterial leaf blight is caused by X. oryzae pv. oryzae. For the latter, two genetic lineages have been described in the literature. We developed a universal MLVA typing tool both for the identification of the three X. oryzae genetic lineages and for epidemiological analyses. Sixteen candidate variable-number tandem-repeat (VNTR) loci were selected according to their presence and polymorphism in 10 draft or complete genome sequences of the three X. oryzae lineages and by VNTR sequencing of a subset of loci of interest in 20 strains per lineage. The MLVA-16 scheme was then applied to 338 strains of X. oryzae representing different pathovars and geographical locations. Linkage disequilibrium between MLVA loci was calculated by index association on different scales, and the 16 loci showed linear Mantel correlation with MLSA data on 56 X. oryzae strains, suggesting that they provide a good phylogenetic signal. Furthermore, analyses of sets of strains for different lineages indicated the possibility of using the scheme for deeper epidemiological investigation on small spatial scales.
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Repetições Minissatélites , Tipagem Molecular , Oryza/microbiologia , Doenças das Plantas/microbiologia , Xanthomonas/classificação , Xanthomonas/genética , Monitoramento Epidemiológico , Epidemiologia Molecular/métodosRESUMO
BACKGROUND: Statin therapy is a cornerstone therapy for secondary prevention after acute coronary syndrome (ACS). However, the use of these drugs can be limited by side effects, mainly muscular pain. Ezetimibe is a newer lipid-lowering agent, with fewer side effects. AIMS: The present study was designed to compare a commercially available association of ezetimibe and simvastatin (E-S) to high dose Rosuvastatin on cholesterol and muscular enzyme levels and occurrence of muscular pain. METHODS: All consecutive ACS statin-naïve patients with LDL cholesterol (LDL-C)>100mg/dL randomly received either high dose statin (Rosuvastatin 20mg) or E-S 10/40-mg. All patients had one-month follow-up with biological testing and clinical examination. We compared the two groups on the biological efficiency and incidence of muscular pain. RESULTS: One hundred and twenty-eight patients were randomized; 64 received E-S and 64 Rosuvastatin. In the two groups, the lowering of LDL-C level (Δ=51%) at one month was significant (P<0.01) without any difference in the rate of lowering on LDL-C or HDL-C suggesting that E-S is as effective as high dose Rosuvastatin (P=0.77 and P=0.99). The rate of patients reaching the objective of LDL-C<100mg/dL (45%) and LDL-C<70mg/dL (51%) was not different in the two clusters (P=0.65). Incidence of muscular pain was 15% higher in patients treated with Rosuvastatin (P=0.01) without any difference on CPK level (P=0.6). CONCLUSION: Using an association of E-S in an effective alternative strategy to high dose Rosuvastatin with a lower incidence of muscular pain, which might impact adherence to medication after ACS.
Assuntos
Síndrome Coronariana Aguda/tratamento farmacológico , Anticolesterolemiantes/administração & dosagem , Azetidinas/administração & dosagem , Fluorbenzenos/administração & dosagem , Inibidores de Hidroximetilglutaril-CoA Redutases/administração & dosagem , Pirimidinas/administração & dosagem , Sulfonamidas/administração & dosagem , Anticolesterolemiantes/efeitos adversos , Azetidinas/efeitos adversos , Quimioterapia Combinada , Ezetimiba , Feminino , Fluorbenzenos/efeitos adversos , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/efeitos adversos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Pirimidinas/efeitos adversos , Rosuvastatina Cálcica , Sulfonamidas/efeitos adversosRESUMO
Bacterial leaf streak (BLS) caused by Xanthomonas oryzae pv. oryzicola was first reported in Africa in the 1980s. Recently, a substantial reemergence of this disease was observed in West Africa. Samples were collected at various sites in five and three different rice-growing regions of Burkina Faso and Mali, respectively. Sixty-seven X. oryzae pv. oryzicola strains were isolated from cultivated and wild rice varieties and from weeds showing BLS symptoms. X. oryzae pv. oryzicola strains were evaluated for virulence on rice and showed high variation in lesion length on a susceptible cultivar. X. oryzae pv. oryzicola strains were further characterized by multilocus sequence analysis (MLSA) using six housekeeping genes. Inferred dendrograms clearly indicated different groups among X. oryzae pv. oryzicola strains. Restriction fragment length polymorphism analysis using the transcriptional activator like effector avrXa7 as probe resulted in the identification of 18 haplotypes. Polymerase chain reaction-based analyses of two conserved type III effector (T3E) genes (xopAJ and xopW) differentiated the strains into distinct groups, with xopAJ not detected in most African X. oryzae pv. oryzicola strains. XopAJ functionality was confirmed by leaf infiltration on 'Kitaake' rice Rxo1 lines. Sequence analysis of xopW revealed four groups among X. oryzae pv. oryzicola strains. Distribution of 43 T3E genes shows variation in a subset of X. oryzae pv. oryzicola strains. Together, our results show that African X. oryzae pv. oryzicola strains are diverse and rapidly evolving, with a group endemic to Africa and another one that may have evolved from an Asian strain.
Assuntos
Variação Genética , Oryza/microbiologia , Doenças das Plantas/microbiologia , Xanthomonas/genética , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Sequência de Bases , Burkina Faso , DNA Bacteriano/química , DNA Bacteriano/genética , Genética Populacional , Haplótipos , Mali , Dados de Sequência Molecular , Tipagem de Sequências Multilocus , Filogenia , Folhas de Planta/microbiologia , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNA , Virulência , Xanthomonas/isolamento & purificação , Xanthomonas/patogenicidadeRESUMO
The species Xanthomonas oryzae is comprised of two designated pathovars, both of which cause economically significant diseases of rice in Asia and Africa. Although X. oryzae is not considered endemic in the United States, an X. oryzae-like bacterium was isolated from U.S. rice and southern cutgrass in the late 1980s. The U.S. strains were weakly pathogenic and genetically distinct from characterized X. oryzae pathovars. In the current study, a draft genome sequence from two U.S. Xanthomonas strains revealed that the U.S. strains form a novel clade within the X. oryzae species, distinct from all strains known to cause significant yield loss. Comparative genome analysis revealed several putative gene clusters specific to the U.S. strains and supported previous reports that the U.S. strains lack transcriptional activator-like (TAL) effectors. In addition to phylogenetic and comparative analyses, the genome sequence was used for designing robust U.S. strain-specific primers, demonstrating the usefulness of a draft genome sequence in the rapid development of diagnostic tools.
Assuntos
Variação Genética , Genoma Bacteriano , Oryza/microbiologia , Xanthomonas/classificação , Xanthomonas/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , Genótipo , Dados de Sequência Molecular , Filogenia , Poaceae/microbiologia , Análise de Sequência de DNA , Estados Unidos , Xanthomonas/isolamento & purificaçãoRESUMO
Bacterial leaf streak (BLS) caused by Xanthomonas oryzae pv. oryzicola is prevalent in Asia where it can decrease yield by as much as 30%. In Africa, BLS has been reported in Madagascar, Nigeria, Senegal, and recently in Mali (1). The pathogen is seed transmitted and rice seeds can be a source of primary inoculum (3). In October 2009, leaf streak symptoms were observed on 3-month-old field rice grown in three regions of Burkina Faso (Haut-Bassin, Cascades, and East Center). Disease was found on cultivated Oryza sativa (varieties TS2, FKR19, and FKR56N), wild rice species (O. longistaminata and O. barthii), and weeds. Symptoms consisted of water-soaked lesions that developed into translucent, yellow streaks with visible exudates at the leaf surface. Yellow-pigmented Xanthomonas-like colonies were isolated on PSA semiselective medium (peptone 10 g, sucrose 10 g, bacto agar 16 g, distilled water 1,000 ml, actidione 50 mg liter-1, cephalexin 40 mg liter-1, and kasugamycin 20 mg liter-1). A multiplex PCR developed for the identification of Xanthomonas oryzae pathovars (2) was used to check the identity of Xanthomonas-like isolates. X. oryzae pv. oryzicola strains BLS256 from the Philippines and CFBP 7331 from Mali were used as positive controls. Three expected DNA fragments (331, 691, and 945 bp) corresponding to X. oryzae pv. oryzicola were obtained from all isolates using the multiplex PCR. No fragment was observed for negative controls (distilled water as the template). Five X. oryzae pv. oryzicola isolates were further analyzed by sequence analysis using portions of the gyrB housekeeping gene together with reference strains. Two sequence types were identified among Burkinabe isolates differing by only one nucleotide. When compared with the nucleotide database with BLAST, three isolates (BAI6, BAI15, and BAI19) were 100% identical to the type culture strain X. oryzae pv. oryzicola BLS256 (gyrB sequence was obtained from GenBank AAQN01000001.1) while the other two (BAI5 and BAI20) demonstrated 99% sequence similarity. The nucleotide sequence of isolate BAI5 was submitted to GenBank (HQ112342). Pathogenicity tests were performed on greenhouse-grown 3-week-old rice plants cv. Nipponbare. Cultures were grown overnight in PSA medium and adjusted in sterile water to 1 × 108 CFU/ml and inoculated into rice leaves with the blunt end of a 1-ml syringe. Four infiltrations were done per isolate per leaf and two leaves were inoculated per plant. Control plants were inoculated with sterile water. After 15 days of incubation in the greenhouse at 27 ± 1°C with a 12-h photoperiod, inoculated leaves exhibited water-soaked lesions with yellow exudates that were identical to symptoms seen in the field. Control plants remained symptomless. Colonies with morphology typical of Xanthomonas were recovered from the symptomatic leaves and typed using multiplex PCR to fulfill Koch's postulates. Three isolates have been deposited in the Collection Française de Bactéries Phytopathogènes (CFBP) and identified as X. oryzae pv. oryzicola strains CFBP7341-43. To our knowledge, this is the first report of X. oryzae pv. oryzicola in Burkina Faso. Further surveys and strain collection will be necessary to evaluate the geographic distribution and prevalence of BLS in Burkina Faso and neighboring countries. References: (1) C. Gonzalez et al. Mol. Plant-Microbe Interact. 20:534, 2007. (2) J. Lang et al. Plant Dis. 94:311, 2010. (3) G. Xie and T. Mew. Plant Dis. 82:1007, 1998.
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Single nucleotide polymorphisms (SNP) are the most abundant type of DNA polymorphism found in animal and plant genomes. They provide an important new source of molecular markers that are useful in genetic mapping, map-based positional cloning, quantitative trait locus mapping and the assessment of genetic distances between individuals. Very little is known on the frequency of SNPs in cassava. We have exploited the recently-developed collection of cassava expressed sequence tags (ESTs) to detect SNPs in the five cultivars of cassava used to generate the sequences. The frequency of intra-cultivar and inter-cultivar SNPs after analysis of 111 contigs was one polymorphism per 905 and one per 1,032 bp, respectively; totaling 1 each 509 bp. We have obtained further information on the frequency of SNPs in six cassava cultivars by analysis of 33 amplicons obtained from 3' EST and BAC end sequences. Overall, about 11 kb of DNA sequence was obtained for each cultivar. A total of 186 SNPs (136 and 50 from ESTs and BAC ends, respectively) were identified. Among these, 146 were intra-cultivar polymorphisms, while 80 were inter-cultivar polymorphisms. Thus the total frequency of SNPs was one per 62 bp. This information will help to develop new strategies for EST mapping as well as their association with phenotypic characteristics.
Assuntos
Etiquetas de Sequências Expressas , Manihot/genética , Polimorfismo de Nucleotídeo Único , Biologia Computacional , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
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.
RESUMO
Plant disease resistance genes (R genes) show significant similarity amongst themselves in terms of both their DNA sequences and structural motifs present in their protein products. Oligonucleotide primers designed from NBS (Nucleotide Binding Site) domains encoded by several R-genes have been used to amplify NBS sequences from the genomic DNA of various plant species, which have been called Resistance Gene Analogues (RGAs) or Resistance Gene Candidates (RGCs). Using specific primers from the NBS and TIR (Toll/Interleukin-1 Receptor) regions, we identified twelve classes of RGCs in cassava (Manihot esculenta Crantz). Two classes were obtained from the PCR-amplification of the TIR domain. The other 10 classes correspond to the NBS sequences and were grouped into two subfamilies. Classes RCa1 to RCa5 are part of the first subfamily and were linked to a TIR domain in the N terminus. Classes RCa6 to RCa10 corresponded to non-TIR NBS-LRR encoding sequences. BAC library screening with the 12 RGC classes as probes allowed the identification of 42 BAC clones that were assembled into 10 contigs and 19 singletons. Members of the two TIR and non-TIR NBS-LRR subfamilies occurred together within individual BAC clones. The BAC screening and Southern hybridization analyses showed that all RGCs were single copy sequences except RCa6 that represented a large and diverse gene family. One BAC contained five NBS sequences and sequence analysis allowed the identification of two complete RGCs encoding two highly similar proteins. This BAC was located on linkage group J with three other RGC-containing BACs. At least one of these genes, RGC2, is expressed constitutively in cassava tissues.
Assuntos
Genes de Plantas , Imunidade Inata/genética , Manihot/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Proteínas de Ligação a DNA , Dados de Sequência Molecular , Filogenia , Doenças das Plantas , Receptores de Interleucina-1 , Alinhamento de SequênciaRESUMO
ABSTRACT Cassava bacterial blight, caused by Xanthomonas axonopodis pv. manihotis, is a widespread disease that affects cassava (Manihot esculenta). We collected 238 X. axonopodis pv. manihotis strains by intensively sampling single fields in four edaphoclimatic zones (ECZs) in Colombia. DNA polymorphism of different X. axonopodis pv. manihotis populations was assessed by restriction fragment length polymorphism (RFLP) analyses, repetitive sequence-based polymerase chain reaction (rep-PCR), and amplified fragment length polymorphism (AFLP) assays. Genetic diversity, phenetic relationships among strains, and the coefficient of genetic differentiation were determined. All strains were tested for aggressiveness on the susceptible cassava cv. MCOL 1522. Strains were also tested for virulence on cassava differentials adapted to the strains' respective ECZs. Our study showed that the Colombian X. axonopodis pv. manihotis population has a high degree of genetic diversity. The hierarchical analysis of diversity showed genotypic differentiation at all levels, among ECZs, among fields within ECZs, and among strains within fields planted to several cassava genotypes. New RFLP haplotypes were detected, leading to the characterization of a new pathotype. Dendrograms from AFLP were more robust than those from RFLP data. A close association between the strains' geographical origin and DNA polymorphism was obtained using RFLP and AFLP data. We suggest that the host played a role in causing pathogen differentiation.
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Cassava bacterial blight (CBB) is caused by Xanthomonas axonopodis pv. manihotis (Xam). Resistance is found in Manihot esculenta and, in addition, has been introgressed from a wild relative, M. glaziovii. The resistance is thought to be polygenic and additively inherited. Ninety-three varieties of M. esculenta (Crantz) were assessed by AFLPs for genetic diversity and for resistance to CBB. AFLP analysis was performed using two primer combinations and a 79.2% level of polymorphism was found. The phenogram obtained showed between 74% and 96% genetic similarity among all cassava accessions analysed. The analysis permitted the unique identification of each individual. Two Xam strains were used for resistance screening. Variation in the reaction of cassava varieties to Xam strains was observed for all plant accessions. The correlation of resistance to both strains, had a coefficient of 0.53, suggesting the independence of resistance to each strain. Multiple correspondence analysis showed a random distribution of the resistance/susceptibility response with respect to overall genetic diversity as measured by AFLP analysis. A total heterozygosity index was calculated to determine the diversity within clusters as well as among them. Our results demonstrate that resistance to CBB is broadly distributed in cassava germplasm and that AFLP analysis is an effective and efficient means of providing quantitative estimates of genetic similarities among cassava accessions.
Assuntos
Variação Genética , Manihot/genética , Doenças das Plantas/genética , Polimorfismo de Fragmento de Restrição , Manihot/classificação , Manihot/imunologia , Manihot/microbiologiaRESUMO
[This corrects the article on p. 4430 in vol. 63.].
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Cassava bacterial blight, caused by Xanthomonas axonopodis pv. manihotis, is of significant concern wherever cassava is grown. The movement of infected, asymptomatic stems is a major means of pathogen dispersal. A reliable and sensitive diagnostic procedure is necessary for the safe movement of cassava planting material. We used a cloned and sequenced pathogenicity gene of X. axonopodis pv. manihotis to develop a polymerase chain reaction (PCR) test for this pathogen. A set of primers directed the amplification of an 898-bp fragment in all 107 pathogenic strains of X. axonopodis pv. manihotis tested. PCR products were not observed when genomic DNA was tested for 27 strains of other xanthomonads, for saprophytic bacteria, or for five nonpathogenic strains of X. axonopodis pv. manihotis. The primers worked well for pathogen detection in direct PCR assays of X. axonopodis pv. manihotis colonies grown on liquid medium and in PCR assays of extracts from leaf and stem lesions. The minimum number of cells that could be detected from cassava stem and leaf lesions was 3 × 102 to 104 CFU/ml. The PCR assays proved to be relativyel sensitive and could become very useful in detecting the pathogen in cassava planting material.
RESUMO
Analyses of DNA polymorphism and virulence variation were used to evaluate the population structure of Xanthomonas axonopodis pv. manihotis, the pathogen causing cassava bacterial blight in Colombia. We collected strains from the major cassava-growing regions which can be grouped into different edaphoclimatic zones (ECZs) according to environmental conditions, production constraints, and economic parameters. DNA polymorphism was assessed by a restriction fragment length polymorphism analysis, using an X. axonopodis pv. manihotis plasmid DNA sequence (pthB) as a probe to evaluate the genetic relatedness among 189 Colombian strains. The sampling intensity permitted the estimation of genetic differentiation within and among ECZs, sites, and fields and even within an individual plant. A multiple correspondence analysis indicated that the Colombian X. axonopodis pv. manihotis population showed a high degree of diversity relative to X. axonopodis pv. manihotis populations studied previously, and the entire collection was grouped into seven clusters. A general correlation was observed between the clusters and the geographical origin of the strains, as each cluster was largely composed of strains from the same ECZ. Representative strains, identified with pthB, were further characterized by ribotyping, hybridization to two repetitive genomic probes (pBS6 and pBS8), and restriction analysis of plasmid contents to evaluate the complementarity of these markers. Virulence variation was observed within the Colombian collection. Strains of different aggressiveness were found in all ecological zones, but no correlation between virulence variation and DNA polymorphism was observed. The genetic and virulence analyses contribute to understanding the X. axonopodis pv. manihotis population structure in Colombia.
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A collection of 51 Xanthomonas campestris strains from throughout the world was studied to detect and assess genetic diversity among pathogens of small grains. Isolates from barley, bread wheat, bromegrass, canary grass, cassava, maize, orchard grass, rice, rough-stalked meadow grass, rye, timothy, and triticale were analyzed by pathogenicity tests on bread wheat cv. Alondra and barley cv. Corona, indirect immunofluorescence, and restriction fragment length polymorphism (RFLP). Three probes were used for the RFLP analysis. They were an acetylaminofluorene-labelled 16S+23S rRNA probe from Escherichia coli and two (sup32)P-labelled restriction fragments from either plasmidic (pBSF2) or chromosomal (pBS8) DNA of X. campestris pv. manihotis. Strains clustered in 9 and 20 groups with the rRNA probe and the pBSF2 DNA probe, respectively. Strains of X. campestris pv. graminis, X. campestris pv. phleipratensis, and X. campestris pv. poae are shown to be related but are also distinguishable by RFLP patterns, serology, and pathogenicity on bread wheat. Strains pathogenic only for barley and not for wheat grouped together. Another group is temporarily designated deviant X. campestris pv. undulosa. These South American isolates from bread wheat did not react by indirect immunofluorescence and produced atypical lesions in pathogenicity tests. The results stress the need to perform pathogenicity tests before strains are named at the pathovar level. The importance of the different probes used for epidemiological studies or phylogenetic studies of closely related strains is underlined.
RESUMO
Fifty-one strains representing Xanthomonas campestris pv. manihotis and cassavae and different pathovars occurring on plants of the family Euphorbiaceae were characterized by ribotyping with a 16S+23S rRNA probe of Escherichia coli and by restriction fragment length polymorphism analysis with a plasmid probe from X. campestris pv. manihotis. Pathogenicity tests were performed on cassava (Manihot esculenta). Histological comparative studies were conducted on strains of two pathovars of X. campestris (vascular and mesophyllic) that attack cassava. Our results indicated that X. campestris pv. manihotis and cassavae have different modes of action in the host and supplemented the taxonomic data on restriction fragment length polymorphism that clearly separate the two pathovars. The plasmid probe could detect multiple restriction fragment length polymorphisms among strains of the pathovar studied. Ribotyping provides a useful tool for rapid identification of X. campestris pathovars on cassava.