A duplex-PCR method for species- and pathovar-level identification and detection of the quarantine plant pathogen Xanthomonas arboricola pv. pruni

Pothier, Joël F.; Pagani, M.C.; Pelludat, Cosima; Ritchie, D. F.; Duffy, Brion

doi:10.1016/j.mimet.2011.03.019

Cited by 51 publications

(58 citation statements)

References 23 publications

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“…Strains were identified as X. arboricola based upon phenotypic characteristics and biochemical tests as described by Schaad (40). and amplification of a specific PCR fragment using the PCR test with XarbQ-F and XarbQ-R primers developed by Pothier et al (41). A set of 27 strains formed convex, yellow-pigmented colonies, which were characterized as Gramnegative rods able to perform oxidative metabolism of glucose, galactose, mannose, cellobiose, trehalose, and arabinose and hydrolysis of gelatin, esculin, starch, and Tween 20 (except for strains CFBP 7645 and CFBP 7636, which did not hydrolyze Tween 20).…”

Section: Resultsmentioning

confidence: 99%

“…A set of 27 strains formed convex, yellow-pigmented colonies, which were characterized as Gramnegative rods able to perform oxidative metabolism of glucose, galactose, mannose, cellobiose, trehalose, and arabinose and hydrolysis of gelatin, esculin, starch, and Tween 20 (except for strains CFBP 7645 and CFBP 7636, which did not hydrolyze Tween 20). A single amplicon with the correct size of 432 bp was obtained for each of these 27 strains using the PCR test (41). None of the 27 strains induced symptoms on walnut (Juglans regia), the plant from which they were isolated, after inoculation on walnut seedlings.…”

Section: Resultsmentioning

confidence: 99%

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Phylogenetic and Variable-Number Tandem-Repeat Analyses Identify Nonpathogenic Xanthomonas arboricola Lineages Lacking the Canonical Type III Secretion System

Essakhi

Cesbron

Saux

et al. 2015

Appl Environ Microbiol

118

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bXanthomonas arboricola is conventionally known as a taxon of plant-pathogenic bacteria that includes seven pathovars. This study showed that X. arboricola also encompasses nonpathogenic bacteria that cause no apparent disease symptoms on their hosts. The aim of this study was to assess the X. arboricola population structure associated with walnut, including nonpathogenic strains, in order to gain a better understanding of the role of nonpathogenic xanthomonads in walnut microbiota. A multilocus sequence analysis (MLSA) was performed on a collection of 100 X. arboricola strains, including 27 nonpathogenic strains isolated from walnut. Nonpathogenic strains grouped outside clusters defined by pathovars and formed separate genetic lineages. A multilocus variable-number tandem-repeat analysis (MLVA) conducted on a collection of X. arboricola strains isolated from walnut showed that nonpathogenic strains clustered separately from clonal complexes containing Xanthomonas arboricola pv. juglandis strains. Some nonpathogenic strains of X. arboricola did not contain the canonical type III secretion system (T3SS) and harbored only one to three type III effector (T3E) genes. In the nonpathogenic strains CFBP 7640 and CFBP 7653, neither T3SS genes nor any of the analyzed T3E genes were detected. This finding raises a question about the origin of nonpathogenic strains and the evolution of plant pathogenicity in X. arboricola. T3E genes that were not detected in any nonpathogenic isolates studied represent excellent candidates to be those responsible for pathogenicity in X. arboricola. E ubacteria constitute a major component of the commensal microbiota, and the nature of their interaction with plants is still unknown (1). Xanthomonas strains living in close association with plants but causing no apparent disease symptoms on their host have been reported (2, 3). On the basis of amplified fragment length polymorphism (AFLP) analysis, Gonzalez et al. (2) showed that nonpathogenic Xanthomonas strains colonizing cassava were clearly distinguished from Xanthomonas axonopodis pv. manihotis strains that cause cassava bacterial blight. Although some of these nonpathogenic strains have been characterized genetically and phenotypically, little is known about their epidemiological or ecological importance.In previous studies, the genetic diversity and population structure of Xanthomonas have been investigated using DNA-DNA hybridization (4, 5), repetitive-sequence PCR (rep-PCR) (4-7), AFLP (4, 8), and fluorescent AFLP (9, 10). As an alternative to these methods, the comparative sequence analysis of protein-encoding genes has also been widely explored. For example, Parkinson et al. (11) used the gyrB gene, which encodes the subunit B protein of DNA gyrase, for establishing a phylogenetic relationship among 203 Xanthomonas pathotype strains. Young et al. (12, 13) used a multilocus sequence analysis (MLSA) based on four genes (dnaK, encoding the chaperone protein; fyuA, encoding one tonB-dependent transporter; gyrB and rpoD, encoding the R...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Phylogenetic and Variable-Number Tandem-Repeat Analyses Identify Nonpathogenic Xanthomonas arboricola Lineages Lacking the Canonical Type III Secretion System

Essakhi

Cesbron

Saux

et al. 2015

Appl Environ Microbiol

118

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“…At species level: PCR amplifications were performed using Xanthomonas specie-specific primers XarbQF and XarbQR for qumA gene, a quinate dehydrogenase [23]. Reaction contained a total volume of 40 µl with 50 ng of DNA template, 1X PCR buffer, 0.2 µM of each primer (GBT Oligos, Buenos Aires), 0.2 µM of each dNTP, 2.5 mM MgCl 2 , 1%, DMSO and 1.2 units Taq DNA polymerase (Thermo Fischer Scientific, Invitrogen Argentina S.A.).…”

Section: Resultsmentioning

confidence: 99%

First Report of Apical Necrosis in Walnut Cultivars from Northern Argentinean Patagonia

Temperini¹,

Pardo²,

Pose³

2017

J Plant Pathol Microbiol

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Walnut blight caused by Xanthomonas arborícola pv. juglandis and apical necrosis are severe diseases that cause premature drop of walnuts all around the world. Evidence from previous studies proposes Xanthomonas arborícola pv. juglandis together with Fusarium spp. and Alternaria spp. as causal agents of apical necrosis. While walnut blight is a well-known pathology in Argentina, apical necrosis has not been reported yet in this country. However, during 2013 and 2014 seasons, serious damages were registered in the nut producing region of Río Negro Middle Valley in northern Argentinean Patagonia. Diseased fruits exhibited symptoms that matched those described for walnut blight and apical necrosis. Therefore, the aim of this study was to determine the incidence of these pathologies and the microorganisms involved in their etiology. As a result, it was possible to identify morphologically and molecularly the bacteria isolated from damaged fruits, confirming their identity as Xanthomonas arborícola pv. juglandis. Moreover, A. tenuissima was found in a high prevalence in damaged as well as in healthy fruits, whereas Fusarium spp. was isolated with a low frequency in both. When pathogenicity tests were performed, it was demonstrated the ability of the three microorganisms to produce typical lesions in healthy nuts. Thus, we can presume that both diseases occurred during this couple of years in this region. This is the first report of apical necrosis in Argentina

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“…In neglected peach orchards, X. arboricola pv. pruni can damage 25 % to 75 % of the fruits (Dunegan 1932;Pothier et al 2011b).…”

Section: Communicated By E Dirlewangermentioning

confidence: 99%

Identification of a major QTL for Xanthomonas arboricola pv. pruni resistance in apricot

Socquet-Juglard

Duffy

Pothier

et al. 2012

Tree Genetics & Genomes

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Xanthomonas arboricola pv. pruni causes bacterial spot of stone fruit resulting in severe yield losses in apricot production systems. Present on all continents, the pathogen is regulated in Europe as a quarantine organism. Host resistance is an important component of integrated pest management; however, little work has been done describing resistance against X. arboricola pv. pruni. In this study, an apricot population derived from the cross "Harostar" × "Rouge de Mauves" was used to construct two parental genetic maps and to perform a quantitative trait locus analysis of resistance to X. arboricola pv. pruni. A population of 101 F1 individuals was inoculated twice for two consecutive years in a quarantine greenhouse with a mixture of bacterial strains, and disease incidence and resistance index data were collected. A major QTL for disease incidence and resistance index accounting respectively for 53 % (LOD score of 15.43) and 46 % (LOD score of 12.26) of the phenotypic variation was identified at the same position on linkage group 5 of "Rouge de Mauves." Microsatellite marker UDAp-452 co-segregated with the resistance, and two flanking microsatellites, namely BPPCT037 and BPPCT038A, were identified. When dividing the population according to the alleles of UDAp-452, the subgroup with unfavorable allele had a disease incidence of 32.6 % whereas the group with favorable allele had a disease incidence of 21 %, leading to a reduction of 35.6 % in disease incidence. This study is a first step towards the markerassisted breeding of new apricot varieties with an increased tolerance to X. arboricola pv. pruni.

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A duplex-PCR method for species- and pathovar-level identification and detection of the quarantine plant pathogen Xanthomonas arboricola pv. pruni

Cited by 51 publications

References 23 publications

Phylogenetic and Variable-Number Tandem-Repeat Analyses Identify Nonpathogenic Xanthomonas arboricola Lineages Lacking the Canonical Type III Secretion System

Phylogenetic and Variable-Number Tandem-Repeat Analyses Identify Nonpathogenic Xanthomonas arboricola Lineages Lacking the Canonical Type III Secretion System

First Report of Apical Necrosis in Walnut Cultivars from Northern Argentinean Patagonia

Identification of a major QTL for Xanthomonas arboricola pv. pruni resistance in apricot

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