Development of a simplified NASBA protocol for detecting viable cells of the citrus pathogen <i>Xanthomonas citri</i> subsp. c<i>itri</i> under different treatments

Scuderi, G; Golmohammadi, M.; Cubero, Jaime; López, Marı́a M.; Cirvilleri, G.; Llop, Pablo

doi:10.1111/j.1365-3059.2010.02305.x

Cited by 18 publications

(23 citation statements)

References 38 publications

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“…Several PCR-based diagnostic tools were developed with the aim of specifically detecting X. citri pv. citri, has been developed (Scuderi et al, 2010). pvs citri and aurantifolii : primers 2/3 (Hartung et al, 1993), XCF/R (Miyoshi et al, 1998), 4/7 (Hartung et al, 1996, J-pth1/2 (Cubero and Graham, 2002) and VM3/4 (Mavrodieva et al, 2004).…”

Section: Detection and Identificationmentioning

confidence: 99%

Scientific Opinion on the risk to plant health of Xanthomonas citri pv. citri and Xanthomonas citri pv. aurantifolii for the EU territory

2014

EFS2

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The Panel conducted a pest risk assessment for Xanthomonas campestris (all strains pathogenic to Citrus) for the EU territory and an assessment of the effectiveness of present EU requirements against Xanthomonas strains pathogenic to citrus. The risk assessment was conducted under the scenario of absence of the current specific EU plant health legislation and the assumption that citrus-exporting countries apply measures to reduce yield and quality losses. Risk reduction options were systematically identified and evaluated. The strains of X. campestris pathogenic to citrus have been reclassified as four distinct infraspecific taxa within two species: X. citri and X. alfalfae. Only two pathovars (X. citri pv. citri and X. citri pv. aurantifolii) are responsible for the citrus bacterial canker that presents a major risk for the citrus industry in the EU. Seven entry pathways have been identified and evaluated. The likelihood of entry was rated unlikely for fruit, very likely for fruit plants for planting, moderately likely for ornamental plants for planting and unlikely for leaves and twigs. The uncertainty of probability of entry was rated as high. The probability of establishment was rated as moderately likely to likely with a medium uncertainty because host plants are widely present in EU areas where environmental conditions are suitable. Once established, spread would be likely with a low uncertainty. The impact of the disease, even if control measures are applied, was rated as moderate to major with a medium uncertainty. The disease would cause yield losses in areas where citrus is the main crop, increase the need for control measures and create environmental problems.The combined EU regulations have been shown to be effective in preventing the introduction of X. citri pv. citri or X. citri pv. aurantifolii in the EU, as no outbreaks of citrus canker in the EU territory have been reported. Vernière for the preparatory work on this Scientific Opinion and EFSA staff: Svetla Kozelska, Tilemachos Goumperis and Olaf Mosbach Schulz for the support provided to this Scientific Opinion. Xanthomonas citri pv. citri and Xanthomonas citri pv. aurantifolii pest risk assessment EFSA Journal 2014;12(2):3556 4 Xanthomonas citri pv. citri and Xanthomonas citri pv. aurantifolii pest risk assessment EFSA Journal 2014;12(2):3556 6importance of maintaining the absence of X. citri pv. citri and X. citri pv. aurantifolii in citrusproducing areas and of the risk reduction options to maintain this absence.The effectiveness of current EU phytosanitary measures to reduce the risk of introduction of X. citri pv. citri and X. citri pv. aurantifolii ranges from moderate to high. However, the requirements for buffer zones of pest free production sites in areas infested by X. citri pv. citri or X. citri pv. aurantifolii are insufficiently detailed.The probability of entry of X. citri pv. citri and X. citri pv. aurantifolii via import of plants for planting for citrus production and of ornamental rutaceous plants (species listed in section 3.1...

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Section: Detection and Identificationmentioning

confidence: 99%

Scientific Opinion on the risk to plant health of Xanthomonas citri pv. citri and Xanthomonas citri pv. aurantifolii for the EU territory

2014

EFS2

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“…; Scuderi et al . ). Despite the economic interest in Xcc, most of the currently available PCR‐based diagnostic tools are prone to false‐positive reactions, as documented by Delcourt et al .…”

Section: Introductionmentioning

confidence: 97%

Rapid and accurate identification of Xanthomonas citri subspecies citri by fluorescence in situ hybridization

Waite

Griffin

Taylor

et al. 2016

Letters in Applied Microbiology

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Xanthomonas citri subsp. citri (Xcc) is an aggressive and hardy pathogen of citrus plants worldwide. Outbreaks are difficult and costly to contain and the establishment of citrus canker results in restricted trade. In order to extend the existing toolkit for identification of Xcc we developed a novel diagnostic approach based on fluorescence in situ hybridization. Our approach is of comparable specificity and sensitivity to existing methods but can be performed directly on infected tissue making it significantly faster than existing PCRs, and requiring fewer laboratory resources.

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“…However, only recently have molecular methods been developed to detect and quantify only the viable Xcc (Scuderi et al. ; Golmohammadi et al. ).…”

Section: Introductionmentioning

confidence: 99%

“…; Scuderi et al. ; Songwattana and Ketudat‐Cairns ; Peyrou et al. ), and depending on the PCR method, detection is possible from 10 to 10 3 colony‐forming units (CFU) per ml, while limits of bioassay in some tests have been 10 2 , but as little as 1 bacterium injection–infiltrated into a grapefruit leaf has been shown to cause disease (Gottwald and Graham ).…”

Section: Introductionmentioning

confidence: 99%

“…Important techniques used to detect or quantify Xcc include molecular PCR-based methods (Hartung et al 1993;Cubero et al 2001;Graham 2002, 2005;Park et al 2006;Shiotani et al 2009;Golmohammadi et al 2012), immunological antibody assays (Alvarez et al 1991), injection-infiltration bioassay or needle-prick pathogenicity tests (Koizumi 1971;Stall et al 1982;Schubert et al 2001;Graham and Leite 2004;Shiotani et al 2009;Gottwald et al 2009) and culture-based methods (Goto 1970;Stall et al 1982;Timmer 1988;Gottwald et al 2009;Bock et al 2005Bock et al , 2010. However, only recently have molecular methods been developed to detect and quantify only the viable Xcc (Scuderi et al 2010;Golmohammadi et al 2012). The available techniques complement each other for our understanding of the pathogen ecology and epidemiology.…”

Section: Introductionmentioning

confidence: 99%

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A Comparison of Pathogen Isolation in Culture and Injection–infiltration Bioassay of Citrus Leaves for DetectingXanthomonas citrisubsp.citri

Bock¹,

Gottwald²,

Graham

2013

Journal of Phytopathology

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Citrus canker [caused by Xanthomonas citri subsp. citri (Xcc)] can cause yield loss of susceptible citrus and result in trade restrictions of fresh fruit. For both regulatory purposes and epidemiological studies, accurate detection and quantification of viable inoculum are critical. Two accepted methods used to detect and quantify Xcc are injection–infiltration bioassay and culture, but these two methods have not been directly compared using field‐obtained samples. The two methods were compared using washates of lesions taken from fruit, leaves and shoots in a commercial orchard in Florida in 2009–2010 and 2010–2011, with bioassay being the assumed standard. Despite some misclassifications, true positives (sensitivity) and true negatives (specificity) were the dominant classes using culture. False positives for lesions from shoots ranged from 13.1 to 21.4% in 2009–2010 and 2010–2011, respectively, and false positives for lesions from fruit and leaves ranged from 4.3 to 15.7%, in the two seasons, respectively. The false positive rate for culture compared with injection–infiltration bioassay was highest (0.16–0.55), due to more frequent recovery of Xcc by culture at ≤103 colony‐forming units (CFU) Xcc per ml. The false negative rate was consistently lower (0.02–0.21), confirming that in only a few cases did culture fail to detect Xcc when it was present. The area under the curve for receiver operator characteristic analysis ranged from 0.80 to 0.97, confirming that culture provided an accurate diagnosis in most cases. There was a higher frequency of lesions from shoots with a CFU ≤103 Xcc compared with lesions from fruit or leaves, making culture more effective at detecting these. The data demonstrate that culture is a reliable way to detect and quantify Xcc compared with injection–infiltration bioassay, particularly when the CFU is ≤103 Xcc per ml.

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Development of a simplified NASBA protocol for detecting viable cells of the citrus pathogen Xanthomonas citri subsp. citri under different treatments

Cited by 18 publications

References 38 publications

Scientific Opinion on the risk to plant health of Xanthomonas citri pv. citri and Xanthomonas citri pv. aurantifolii for the EU territory

Scientific Opinion on the risk to plant health of Xanthomonas citri pv. citri and Xanthomonas citri pv. aurantifolii for the EU territory

Rapid and accurate identification of Xanthomonas citri subspecies citri by fluorescence in situ hybridization

A Comparison of Pathogen Isolation in Culture and Injection–infiltration Bioassay of Citrus Leaves for DetectingXanthomonas citrisubsp.citri

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