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Diagnostic activities for plant pests
Information on diagnostic activities on pests and diseases of plants follow the activities of EPPO Panels on diagnostics
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EPPO and Diagnostics for plant pests of quarantine significance

EPPO and Diagnostics for plant pests of quarantine significance | Diagnostic activities for plant pests | Scoop.it

 

'Diagnostic activities for plant pests' is maintained by the Secretariat of the European and Mediterranean Plant Protection Organization (EPPO) and its aim is to share information on diagnostics.

 

EPPO is an intergovernmental organization created in 1951 which currently has 50 member countries. EPPO is responsible for harmonization and cooperation among the National Plant Protection Organizations (official authorities) of its member countries. EPPO helps its members in their efforts to protect plant health in agriculture, forestry and the uncultivated environment (standard-setting activities and exchange of information).

 

EPPO has established a programme on diagnostics since 1992

view the approved diagnostic protocols on the EPPO official website:

http://archives.eppo.int/EPPOStandards/diagnostics.htm

 

On this website, a page dedicated to EPPO activities on diagnostics is also available at: http://www.eppo.int/QUARANTINE/diagnostic_activities.htm

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The EPPO Secretariat maintains other scoop.it magazines onPest Risk Analysis (http://www.scoop.it/t/pest-risk-analysis) Pest Alerts (http://www.scoop.it/t/pest-alerts), Invasive Alien Plants (http://www.scoop.it/t/invasive-alien-plants), Video of plant pests (http://www.scoop.it/t/pests-on-videos), Communication on pests (http://www.scoop.it/t/communication-and-citizen-sciences-on-pests-and-invasive-alien-species)

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The Fitness for Purpose of Analytical Methods (2014)

The Fitness for Purpose of Analytical Methods (2014) | Diagnostic activities for plant pests | Scoop.it

This guide was first issued in 1998. Since the release of the first edition, however, there have been many changes in terminology, working practices, reference documents and requirements. This second edition, produced by the Eurachem Method Validation Working Group, forms a thorough revision of the 1998 edition.

This second edition accommodates the main changes in international standards and practice. The new edition also includes notes on some aspects of validation that are specific to qualitative test methods.

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EPPO has developed Standards on Quality Assurance including  PM 7/98(2)Specific requirements for laboratories preparing accreditation for a plant pest diagnostic activity

http://onlinelibrary.wiley.com/doi/10.1111/epp.12118/pdf

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Q-collect Workshop for collections and users of biological materia Kleinmachnow, DE, 2014-11-27/2

Q-collect Workshop for collections and users of biological materia  Kleinmachnow, DE, 2014-11-27/2 | Diagnostic activities for plant pests | Scoop.it
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Read the web-report of the Q-collect Workshop!

Q-collect is a EU FP7 funded project which started on the 1st of October 2013 and aims to improve the status of reference collections important to plant health

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Detection and quantification of airborne inoculum of Hymenoscyphus pseudoalbidus using real-time PCR assays - Chandelier - et al. 2014

Detection and quantification of airborne inoculum of Hymenoscyphus pseudoalbidus using real-time PCR assays - Chandelier - et al. 2014 | Diagnostic activities for plant pests | Scoop.it

 A method based on real-time polymerase chain reaction (PCR) and the use of rotating-arm spore traps was developed for quantifying airborne Hymenoscyphus pseudoalbidus ascospores. The method was sensitive and reproducible, and the collection efficiency was 10% of the spores present in the air. The temporal ascospore dispersal pattern was studied over 3 years by collecting spores every 15 days for a 24 h air-sampling period during the ash-growing season. The highest production was detected from the end of June to the beginning of September. The overall ascospore production did not differ significantly among stands within a specific year but there were differences from year to year. There was a positive correlation between air temperature and the number of ascospores trapped, with most of the positive samples being observed at temperatures above 12°C. The vertical profile of ascospore dispersal showed a strong decrease in ascospore density within a height of 3 m, regardless of date of collection. An analysis of the spore traps installed at increasing distances from an infected stand showed that most of the ascospores were deposited downwind within 50 m of the stand. These data are discussed in context of the epidemiology of the disease.

Petter Françoise's insight:

A Diagnostic protocol on Hymenoscyphus pseudoalbidus was adopted in 2013

http://onlinelibrary.wiley.com/doi/10.1111/epp.12061/pdf 

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Detection of living Bursaphelenchus xylophilus in wood, using reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) - Leal et al - 2014

Detection of living Bursaphelenchus xylophilus in wood, using reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) - Leal et al - 2014 | Diagnostic activities for plant pests | Scoop.it

Pinewood nematode (PWN), Bursaphelenchus xylophilus, the causal agent of pine wilt disease, is an inhabitant of native pine species of North America, where its presence has minor impact. In contrast, the introduction of this nematode to forests in Asia and Europe has devastated some pine stands and is recognized as a pest of significant phytosanitary concern by the National Plant Protection Organizations of several countries. The ability to detect PWN in internationally traded wood products is crucial to reduce the spread of this organism. Currently, the majority of molecular techniques for the detection of PWN rely on the presence of genomic DNA and thus fail to differentiate between living and dead PWN. The detection of dead nematodes could lead to unnecessary trade disruption. Therefore, accurate techniques for the detection of and differentiation between living and dead PWN are critical. We have developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay, which specifically identifies living PWN in wood by detecting the presence of mRNA encoding an expansin gene as a viability marker. This diagnostic method was found to be more sensitive, faster and less dependent on expensive laboratory equipment than PCR. In addition, unlike PCR, it allows for simple colour detection of amplification products. This method will help resolve disputes over the detection of PWN by clarifying whether it originates from live or dead organisms. Where approved treatments are implemented, unnecessary trade disruption will be avoided, thus protecting market access of wood products from PWN-infested areas.

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This test will be evaluated for inclusion in the EPPO Diagnostic Protocol on PWN

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Affordable and reliable plant sap-mediated template preparation for the detection of various phytopathogens by PCR assay - Sahana et al. 2014

Affordable and reliable plant sap-mediated template preparation for the detection of various phytopathogens by PCR assay - Sahana et al. 2014 | Diagnostic activities for plant pests | Scoop.it

Plant pathogens like ‘Ca. Liberibacter’, phytoplasma, viruses and viroids cause diseases to almost all economically important plants. A simple and fast sap-mediated polymerase chain reaction (PCR) method for the detection of these pathogens infecting various plant hosts is described in the present study. Sap from selected plants was drawn aseptically on parafilm, from the mid-rib of young leaves. Depending on the type of host plant, sap was diluted to optimal concentration before PCR analysis. ‘Ca. Liberibacter’, Citrus tristeza virus (CTV) and Citrus exocortis viroid (CEVd) infecting citrus, and ‘Ca. Phytoplasma’ infecting pepper and sandal trees were tested by sap-mediated PCR. The reliability of this procedure was evaluated by comparing the findings with previously described protocols. The sap-mediated nucleic acid template preparation for PCR assay is devoid of laborious nucleic acid extraction and expensive chemicals. Hence the present method is rapid, economical and so can be employed for diagnosis of large number of plant samples

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Development and Validation of Real-Time PCR Tests for the Identification of Four Spodoptera Species: S. eridania, S. frugiperda, S.littoralis, and S. litura (Lepidoptera: Noctuidae) B.T.L.H. Van De...

Development and Validation of Real-Time PCR Tests for the Identification of Four Spodoptera Species: S. eridania, S. frugiperda, S.littoralis, and S. litura (Lepidoptera: Noctuidae) B.T.L.H. Van De... | Diagnostic activities for plant pests | Scoop.it

The genus Spodoptera comprises 31 species, 4 of which are listed as quarantine pests for the European Union: Spodoptera eridania (Cramer), Spodoptera frugiperda (Smith), Spodoptera littoralis (Boisduval), and Spodoptera litura (F.). In international trade, the earlier life stages (eggs and larvae) are being intercepted at point of inspection most frequently, challenging the possibilities of morphological identification. To realize a rapid and reliable identification for all stages, we developed and validated four simplex real-time polymerase chain reaction identification tests based on the mitochondrial cytochrome b gene using dual-labeled hydrolysis probes. Method validation on dilutions of extracted DNA of the target organisms showed that low levels of template (up to 0.2–100 pg) can reliably be identified. No cross-reactivity was observed with 14 nontarget Spodoptera and 5 non-Spodoptera species in the specific Spodoptera tests. The tests showed to be repeatable, reproducible (both 100%), and robust. The new Spodoptera tests have proven to be suitable tools for routine identification of all life stages of S. eridania, S. frugiperda, S. littoralis, and S. litura.

Petter Françoise's insight:

A diagnostic protocol on Spodoptera species is in development in EPPO

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New adopted Diagnostic Protocols on Phyllosticta citricarpa (McAlpine) Aa on fruit and Xanthomonas citri subsp. citri | International Plant Protection Convention

New adopted Diagnostic Protocols on Phyllosticta citricarpa (McAlpine) Aa on fruit and Xanthomonas citri subsp. citri | International Plant Protection Convention | Diagnostic activities for plant pests | Scoop.it

Two new IPPC protocols have been adopted. 

EPPO will adjust its dignostic protocols for these pests. 

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Quantitative analysis of “flavescence doreé” phytoplasma with droplet digital PCR Mehle et al 2014

Quantitative analysis of “flavescence doreé” phytoplasma with droplet digital PCR Mehle et al 2014 | Diagnostic activities for plant pests | Scoop.it

The first droplet-digital-PCR-based absolute quantification of “flavescence dorée” phytoplasma, agent of a quarantine phytoplasma yellows in grapevine was performed. Quantitative PCR that targets the secY gene was transferred to droplet digital PCR and used for absolute quantification of “flavescence dorée” phytoplasma, without the need for calibration curves. The sensitivity of the two assays compares well and was shown that it could be used for quantification and quality control of DNA based on in-house reference materials typically used in diagnostics and metrological laboratories. This new tool has great potential for monitoring phytoplasma kinetics, such as there relationship with the progress of an infection, and variations of the phytoplasma titer through the season and screening plants for resistance.

Petter Françoise's insight:

The EPPO standards PM 7/79(1) Grapevine flavescence dorée phytoplasma is under revision. 

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Association of ‘Candidatus Liberibacter solanacearum’ with a Vegetative Disorder of Celery in Spain and Development of a Real-Time PCR Method for Its Detection Teresani et al. 2014

Association of ‘Candidatus Liberibacter solanacearum’ with a Vegetative Disorder of Celery in Spain and Development of a Real-Time PCR Method for Its Detection Teresani et al. 2014 | Diagnostic activities for plant pests | Scoop.it

A new symptomatology was observed in celery (Apium graveolens) in Villena, Spain in 2008. Symptomatology included an abnormal amount of shoots per plant and curled stems. These vegetative disorders were associated with ‘Candidatus Liberibacter solanacearum’ and not with phytoplasmas. Samples from plant sap were immobilized on membranes based on the spot procedure and tested using a newly developed real-time polymerase chain reaction assay to detect ‘Ca. L. solanacearum’. Then, a test kit was developed and validated by intralaboratory assays with an accuracy of 100%. Bacterial-like cells with typical morphology of ‘Ca. Liberibacter’ were observed using electron microscopy in celery plant tissues. A fifth haplotype of ‘Ca. L. solanacearum’, named E, was identified in celery and in carrot after analyzing partial sequences of 16S and 50S ribosomal RNA genes. From our results, celery (family Apiaceae) can be listed as a new natural host of this emerging bacterium.

Petter Françoise's insight:

An EPPO Diagnostic protocol is in preparation for ‘Ca. L. solanacearum’. 

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Four draft Diagnostic protocols sent for Member consultation (MC) by the International Plant Protection Convention

Four draft Diagnostic protocols sent for Member consultation (MC) by the  International Plant Protection Convention | Diagnostic activities for plant pests | Scoop.it

Following clearance by the Standards Committee (SC), four draft diagnostic protocols are sent for member consultation by the IPPC Secretariat to contracting parties, national plant protection organizations (NPPOs), regional plant protection organizations (RPPOs) and international organizations.

Comments must be submitted through the IPPC contact point in the IPPC 

 

Petter Françoise's insight:

The protocols are:

Erwinia amylovora (Burrill)

Genus Anastrepha

Ditylenchus dipsaci and Ditylenchus destructor

Phytoplasmas

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EPPO Bulletin - Early View - Wiley Online Library

EPPO Bulletin - Early View - Wiley Online Library | Diagnostic activities for plant pests | Scoop.it

Read some article relevant for diagnostics


The MeloTuber Test: a real-time TaqMan® PCR-based assay to detect the root-knot nematodes Meloidogyne chitwoodi and M. fallax directly in potato tubers

Short report of the Workshop on Ct cut off values

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and more.....

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LAMP assay and rapid sample preparation method for on-site detection of Flavescence dorée phytoplasma in grapevine - Kogovšek et al. 2014

LAMP assay and rapid sample preparation method for on-site detection of Flavescence dorée phytoplasma in grapevine - Kogovšek et al. 2014 | Diagnostic activities for plant pests | Scoop.it

In Europe the most devastating phytoplasma associated with grapevine yellows (GY) diseases is a quarantine pest flavescence dorée (FDp) from the 16SrV taxonomic group. The on-site detection of FDp with an affordable device would contribute to faster and more efficient decisions on the control measures for FDp. Therefore, a real-time isothermal LAMP assay for detection of FDp was validated according to the EPPO standards and MIQE guidelines. The FD LAMP assay was shown to be specific and extremely sensitive, since it detected FDp in all leaf samples that were determined to be FDp infected using quantitative real-time PCR. The whole procedure of sample preparation and testing was designed and optimised for on-site detection and can be completed in one hour. The homogenisation procedure of the grapevine samples (leaf vein, flower or berry) was optimised to allow direct testing of the crude homogenates with the LAMP assay, without the need for a DNA extraction, and was shown to be extremely sensitive.

Petter Françoise's insight:

The EPPO protocol for FDp is under revision.   

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The Top 10 oomycete pathogens in molecular plant pathology - Kamoun et al. 2014

The Top 10 oomycete pathogens in molecular plant pathology - Kamoun et al. 2014 | Diagnostic activities for plant pests | Scoop.it

Oomycetes form a deep lineage of eukaryotic organisms that includes a large number of plant pathogens which threaten natural and managed ecosystems. We undertook a survey to query the community for their ranking of plant-pathogenic oomycete species based on scientific and economic importance. In total, we received 263 votes from 62 scientists in 15 countries for a total of 33 species. The Top 10 species and their ranking are: (1) Phytophthora infestans; (2, tied) Hyaloperonospora arabidopsidis; (2, tied) Phytophthora ramorum; (4)Phytophthora sojae; (5) Phytophthora capsici; (6) Plasmopara viticola; (7) Phytophthora cinnamomi; (8, tied) Phytophthora parasitica; (8, tied) Pythium ultimum; and (10) Albugo candida. This article provides an introduction to these 10 taxa and a snapshot of current research. We hope that the list will serve as a benchmark for future trends in oomycete research.

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After viruses (2011), fungi (2012), bacteria (2012),  nematodes (2013), now the oomycetes 

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Development and Validation of Real-Time PCR Tests for the Identification of Four Spodoptera Species Van De Vossenberg, & Van Der Straten (2014)

Development and Validation of Real-Time PCR Tests for the Identification of Four Spodoptera Species  Van De Vossenberg, & Van Der Straten (2014) | Diagnostic activities for plant pests | Scoop.it
The genus Spodoptera comprises 31 species, 4 of which are listed as quarantine pests for the European Union: Spodoptera eridania (Cramer), Spodoptera frugiperda (Smith), Spodoptera littoralis (Boisduval), and Spodoptera litura (F.). In international trade, the earlier life stages (eggs and larvae) are being intercepted at point of inspection most frequently, challenging the possibilities of morphological identification. To realize a rapid and reliable identification for all stages, we developed and validated four simplex real-time polymerase chain reaction identification tests based on the mitochondrial cytochrome b gene using dual-labeled hydrolysis probes. Method validation on dilutions of extracted DNA of the target organisms showed that low levels of template (up to 0.2‐100 pg) can reliably be identified. No cross-reactivity was observed with 14 nontarget Spodoptera and 5 non-Spodoptera species in the specific Spodoptera tests. The tests showed to be repeatable, reproducible (both 100%), and robust. The new Spodoptera tests have proven to be suitable tools for routine identification of all life stages of S. eridania, S. frugiperda, S. littoralis, and S. litura.
Petter Françoise's insight:

An EPPO diagnostic protocol on Spodoptera species is in development and includes this real-time PCR test. It will be sent for EPPO member country consultation shortly.

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Detection, Diversity, and Population Dynamics of Waterborne Phytophthora ramorum Populations Eyre & Garbelotto 2014

Detection, Diversity, and Population Dynamics of Waterborne Phytophthora ramorum Populations Eyre & Garbelotto 2014 | Diagnostic activities for plant pests | Scoop.it

Information on detection of P. ramorum 

Petter Françoise's insight:

An EPPO diagnostic protocol exist on P. ramorum and is considered for revision

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3rd Meeting of the Panel on Diagnostics in Virology and Phytoplasmology

3rd Meeting of the Panel on Diagnostics in Virology and Phytoplasmology | Diagnostic activities for plant pests | Scoop.it

read a short report of the last Panel! 

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Tissue-print and squash real-time PCR for direct detection of ‘Candidatus Liberibacter’ species in citrus plants and psyllid vectors - Bertolini et al- 2014

Tissue-print and squash real-time PCR for direct detection of ‘Candidatus Liberibacter’ species in citrus plants and psyllid vectors - Bertolini et al- 2014 | Diagnostic activities for plant pests | Scoop.it

Huanglongbing (HLB) disease is seriously threatening and/or damaging the citrus industry worldwide. Accurate detection of the three species associated with HLB disease, ‘Candidatus Liberibacter asiaticus’, ‘Candidatus Liberibacter africanus’ and ‘Candidatus Liberibacter americanus’, is essential for the preventive control of the disease. Real-time PCR is a useful tool for bacterial detection. However, nucleic acid purification steps limit the number of samples that can be processed by PCR. Universal detection of ‘Ca. Liberibacter’ species was achieved by direct tissue-printing and spotting of plant leaf petiole extracts or squashing of individual psyllids onto paper or nylon membranes. Primers were designed and used with TaqMan chemistry for accurate detection of the bacterium in immobilized targets (prints of 10 overlapping leaf pedicels per tree, or squashed single vectors), by extraction with water and direct use for real-time PCR. This simplified method was validated and could detect HLB-liberibacters in 100% of leaves with symptoms and 59% of symptomless leaves collected from HLB-infected trees. The use of direct assays as template showed good agreement with use of purified DNA (κ = 0·76 ± 0·052). The squash assay allowed detection of the bacterium in 40% of mature Diaphorina citri that fed on leaves of HLB-infected trees with or without symptoms. A commercial ready-made kit based on this technology showed 96% accuracy in intra-laboratory performance studies. The simplified direct methods of sample preparation presented herein can be effectively adopted for use in rapid screening of HLB agents in extensive surveys, certification schemes or for epidemiological and research studies.

Petter Françoise's insight:

An EPPO Diagnostic Protocol on HLB is about to be adopted and includes these tests!

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Optimising droplet digital PCR analysis approaches for detection and quantification of bacteria: a case study of fire blight and potato brown rot -Dreo et al 2014

Optimising droplet digital PCR analysis approaches for detection and quantification of bacteria: a case study of fire blight and potato brown rot -Dreo et al 2014 | Diagnostic activities for plant pests | Scoop.it

Here we report on the first assessment of droplet digital PCR (ddPCR) for detection and absolute quantification of two quarantine plant pathogenic bacteria that infect many species of the Rosaceae and Solanaceae families: Erwinia amylovora and Ralstonia solanacearum. An open-source R script was written for the ddPCR data analysis. Analysis of a set of samples with known health status aided the assessment and selection of different threshold settings (QuantaSoft analysis, definetherain pipeline and manual threshold), which led to optimal diagnostic specificity. The interpretation of theE. amylovora ddPCR was straightforward, and the analysis approach had little influence on the final results and the concentrations determined. The sensitivity and linear range were similar to those for real-time PCR (qPCR), for the analysis of both bacterial suspensions and plant material, making ddPCR a viable choice when both detection and quantification are desired. With the R. solanacearum ddPCR, the use of a high global threshold was necessary to exclude false-positive reactions that are sometimes observed in healthy plant material. ddPCR significantly improved the analytical sensitivity over that of qPCR, and improved the detection of low concentrations of R. solanacearum in potato tuber samples. Accurate and rapid absolute quantification of both of these bacteria in pure culture was achieved by direct ddPCR. Our data confirm the suitability of these ddPCR assays for routine detection and quantification of plant pathogens and for preparation of defined in-house reference materials with known target concentrations.

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Rapid diagnostic detection of plum pox virus in Prunus plants by isothermal AmplifyRP® using reverse transcription-recombinase polymerase amplification Zhang et al 2014

Rapid diagnostic detection of plum pox virus in Prunus plants by isothermal AmplifyRP® using reverse transcription-recombinase polymerase amplification Zhang et al 2014 | Diagnostic activities for plant pests | Scoop.it

In this study, the innovative development of two AmplifyRP® tests is reported for a rapid isothermal detection of PPV using reverse transcription-recombinase polymerase amplification. In an AmplifyRP® test, all specific recombination and amplification reactions occur at a constant temperature without thermal cycling and the test results are either recorded in real-time with a portable fluorescence reader or displayed using a lateral flow strip contained inside an amplicon detection chamber. The major improvement of this assay is that the entire test from sample preparation to result can be completed in as little as 20 min and can be performed easily both in laboratories and in the field. The results from this study demonstrated the ability of the AmplifyRP® technique to detect all nine PPV strains (An, C, CR, D, EA, M, Rec, T, or W). Among the economic benefits to pathogen surveys is the higher sensitivity of the AmplifyRP® to detect PPV when compared to the conventional ELISA and ImmunoStrip® assays. This is the first report describing the use of such an innovative technique to detect rapidly plant viruses affecting perennial crops.

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Coupling scanning electron microscopy with DNA bar coding: a novel approach for thrips identification - Kumar et al. 2014

Coupling scanning electron microscopy with DNA bar coding: a novel approach for thrips identification - Kumar et al. 2014 | Diagnostic activities for plant pests | Scoop.it

The specific objective of this pilot study was to assess if the thrips specimens processed to be observed under scanning electron microscopy (SEM) can be further utilized for DNA based assays. Larvae and adults of S. dorsalis were subjected to traditional morphological identification using high resolution SEM prior to their DNA extraction. Sequence results of both mtCO1 and ITS rDNA of individual larva and adult S. dorsalis were found to be in agreement with the taxonomic identification conducted using SEM and each result confirmed the other technique. Our results suggest that steps involved during specimen preparation and observation under SEM does not impact DNA analysis of the sample. The two techniques together could be used for the correct identification of various thrips species using the same specimen. The proof of concept was further tested on three other important thrips species Frankliniella occidentalis (Pergande), F. schultzei(Trybom) and Thrips palmi Karny, which confirmed the robustness of the technique. 

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2014 July meeting of the TPDP - Moving forward. | International Plant Protection Convention

2014 July meeting of the TPDP - Moving forward. | International Plant Protection Convention | Diagnostic activities for plant pests | Scoop.it

Message from the IPPC Secretariat

 

The Technical Panel on Diagnostic Protocol (TPDP) met from 07 to 12 July 2014 at the European and Mediterranean Plant Protection Organization (EPPO) Headquarters in Paris, France.

The meeting’s agenda was designed to cover the revision of 7 draft diagnostic protocols (DPs) which were submitted to the expert consultation on IPP this year, discussion of the status of all 31 DPs on the work programme and the TPDP working procedures and related documents, such as the Instruction to Authors. The TPDP also started to develop a gap and a SWOT analysis.

The panel thanked all the DP authors involved in the drafting groups. During the revision of the DPs, the meeting had the presence of two authors: Mr Dominique COLLINS (UK) for draft DP on Genus Liriomyza (2006-017) and Mr Thomas PRIOR (UK) for draft DP on Xiphinema americanumm (2004-025).

The meeting achieved several important outcomes and specific plans for this and the upcoming year. In looking toward 2015, the TPDP expects that 8 draft DPs will be sent out for member consultation, divided in the two consultation periods, and 5 draft DPs to the SC for approval for adoption. It was noted that 4 draft DPs are currently under member consultation and 2 draft DPs under theDP notification period.

More information on the TPDP activities can be found here.

https://www.ippc.int/core-activities/standards-setting/expert-drafting-groups/technical-panels/technical-panel-diagnostic-protocols

Petter Françoise's insight:

EPPO has been very honoured of hosting the IPPC technical Panel on Diagnostic Protocols from 2012 to 2014

We will welcome the PAnel again anytime!

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13th Workshop on EPPO Diagnostic Protocols for fungi held jointly with the 17th Annual Meeting of the European Mycological Network Budapest, 2014-05-06/0

13th Workshop on EPPO Diagnostic Protocols for fungi held jointly with the 17th Annual Meeting of the European Mycological Network  Budapest, 2014-05-06/0 | Diagnostic activities for plant pests | Scoop.it

read a short report of the workshop.

Petter Françoise's insight:

From 2015, EPPO will establish  a Panel on DIagnostics in Mycology. 

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PM 7/98 (2) Specific requirements for laboratories preparing accreditation for a plant pest diagnostic activity

PM 7/98 (2) Specific requirements for laboratories preparing accreditation for a plant pest diagnostic activity | Diagnostic activities for plant pests | Scoop.it

This revised version of PM 7/098 was prepared following a survey on its use in EPPO plant pest diagnostic laboratories. 

 

Petter Françoise's insight:

PM 7/098 is one of the most consulted EPPO Standard in recent years. 

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Development of a real-time PCR method for the detection of the dagger nematodes Xiphinema index, X. diversicaudatum, X. vuittenezi and X. italiae, and for the quantification of X. index numbers Van...

Development of a real-time PCR method for the detection of the dagger nematodes Xiphinema index, X. diversicaudatum, X. vuittenezi and X. italiae, and for the quantification of X. index numbers Van... | Diagnostic activities for plant pests | Scoop.it

The ectoparasitic dagger nematodes Xiphinema index and X. diversicaudatum, often at low numbers in the soil, are vectors of grapevine nepoviruses, which cause huge agronomical problems for the vineyard industry. This study reports a method, based on real-time PCR, for the specific detection of these species and of the closely related non-vector species X. vuittenezi and X. italiae. Specific primers and TaqMan probes were designed from the ribosomal DNA internal transcribed spacer 1 (ITS1), enabling the specific detection of single individuals of each of the X. index, X. diversicaudatum, X. italiae and X. vuittenezi species from the general nematode population. This specificity of detection and absence of false positive reaction was confirmed in samples of each species mixed with the three otherXiphinema species or mixed with nematodes representative from other genera (non-plant-parasitic Dorylaimida, Longidorus sp.,Meloidogyne spp., Globodera spp. and Pratylenchus sp.). The method was shown to be valid for the relative quantification of X. indexnumbers through its use, from crude nematode extracts of soil samples, in a greenhouse assay of grapevine accessions ranging from highly susceptible to resistant. As an alternative to time-consuming microscopic identification and counting, this real-time PCR method will provide a fast, sensitive and reliable diagnostic and relative quantification technique for X. index nematodes extracted from fields or controlled conditions.

Petter Françoise's insight:

EPPO has a diagnostic protocol PM 7/95(1): Xiphinema americanum sensu lato   http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2338.2009.02326.x/abstract ;

This protocol is under revision. An IPPC protocol is also in preparation. 

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18th Meeting of the EPPO Panel on Diagnostics in Bacteriology Wageningen (NL), 2014-03-18/20

18th Meeting of the EPPO Panel on Diagnostics in Bacteriology Wageningen (NL), 2014-03-18/20 | Diagnostic activities for plant pests | Scoop.it

Read a short report of the last Panel on Diagnostics in Bacteriology. 

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