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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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Development and evaluation of Taqman assays for the differentiation of Dickeya (sub)species - van der Wolf et al. 2014

Development and evaluation of Taqman assays for the differentiation of Dickeya (sub)species - van der Wolf et al. 2014 | Diagnostic activities for plant pests | Scoop.it

TaqMan assays were developed for the detection of seven Dickeya species, namely D. dianthicola, D. dadantii, D. paradisiaca, D. chrysanthemi, D. zeae, D. dieffenbachiae and D. solani. 

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The organization of proficiency testing in plant pathology (qualitative methods of analysis) according to the ISO/IEC 17043: example of the French national reference laboratory - Chabirand et al 2014

The organization of proficiency testing in plant pathology (qualitative methods of analysis) according to the ISO/IEC 17043: example of the French national reference laboratory - Chabirand et al 2014 | Diagnostic activities for plant pests | Scoop.it

The article presents the experience of the French National Reference laboratories with proficiency testing for qualitative tests. 

Petter Françoise's insight:

EPPO is about to approve a new standard Guidelines for the organization of interlaboratory comparisons by plant pest diagnostic laboratories whyihc includes both test performance studies and proficiency testing.

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Survival and Transmission of Potato Virus Y, Pepino Mosaic Virus, and Potato Spindle Tuber Viroid in Water Mehle et al 2014

Hydroponic systems and intensive irrigation are used widely in horticulture and thus have the potential for rapid spread of water-transmissible plant pathogens.

This artcile presents the role of water as a source of plant infection for three mechanically transmissible plant pathogens that constitute a serious threat to tomato and potato production: pepino mosaic virus (PepMV), potato virus Y (PVY), and potato spindle tuber viroid (PSTVd). It concludes that water should be taken into account in future epidemiology and risk assessment studies.

Petter Françoise's insight:

 

Article on the margin of diagnostics

Pepino mosaic virus (PepMV),and potato spindle tuber viroid (PSTVd) are pest recommended for regulation by EPO and diagnostic protcols have been developped

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Improved PCR for identification of members of the genus Xanthomonas Adriko et al. 2014

Improved PCR for identification of members of the genus Xanthomonas Adriko et al. 2014 | Diagnostic activities for plant pests | Scoop.it

This article presents a PCR based approach that can reliably and effectively differentiate xanthomonads from non-xanthomonads as well as separating the strains belonging to the two described groups of the genus Xanthomonas. The study thus offers valuable tools for disease surveillance and management. It can effectively be applied in rapid assessment of new disease occurrences, for which no specific detection tools could be in place.

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Sensitive detection of Tomato ringspot virus by real-time TaqMan RT-PCR targeting the highly conserved 3′-UTR region

A real-time TaqMan RT-PCR assay was developed for the rapid and sensitive detection of Tomato ringspot virus (ToRSV), an important plant virus which infects a wide range of fruit and ornamental crops. This TaqMan real-time assay was tested using four different reagent kits and was shown to be robust and stable, with no significant differences in sensitivity between kits. It is expected that the implementation of this TaqMan real-time RT-PCR assay will facilitate efficient phytosanitary certification of nursery stock requiring testing for ToRSV by regulatory agencies, and will also have wider uses for the general detection of ToRSV in a range of hosts.

Petter Françoise's insight:

An EPPO Diagnostic protocol PM 7/49 was adopted for this pest in 2004

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Development of reverse transcription loop-mediated isothermal amplification assay for rapid detection of an emerging potyvirus: Tomato necrotic stunt virus

Development of reverse transcription loop-mediated isothermal amplification assay for rapid detection of an emerging potyvirus: Tomato necrotic stunt virus | Diagnostic activities for plant pests | Scoop.it

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for sensitive detection of ToNStV. The sensitivity of RT-LAMP was comparable to that of conventional RT-PCR, with detection of ToNStV in a reaction containing only 8 pg of total tomato RNA or with 1:20,000 dilution of crude tissue extract. This assay was able to detect ToNStV in a broad range of solanaceous plant species. The RT-LAMP for ToNStV was specific with no cross-reactivity to other potyviruses (i.e. Potato virus Y and Tobacco etch virus), as well as several other common tomato viruses. RT-LAMP should complement RT-PCR and real-time RT-PCR assays reported previously, with a potential to provide a simple, rapid, and sensitive field diagnostic method for ToNStV.

Petter Françoise's insight:

Tomato necrotic stunt virus (ToNStV) is an emerging potyvirus that causes severe stunting to infected tomato plants.

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A Multiplex-PCR Method for Strain Identification and Detailed Phylogenetic Analysis of AY-Group Phytoplasmas

A Multiplex-PCR Method for Strain Identification and Detailed Phylogenetic Analysis of AY-Group Phytoplasmas | Diagnostic activities for plant pests | Scoop.it

this multiplex-PCR system is useful for rapid strain identification and detailed phylogenetic analysis of phytoplasmas.

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Aster yellows (AY) group (16SrI) phytoplasmas are associated with over 100 economically important diseases worldwide and represent the most diverse and widespread phytoplasma group.

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Rapid detection and quantification of viable Potato Cyst Nematodes using qPCR in combination with propidium monoazide - Christoforou et al.

Rapid detection and quantification of viable Potato Cyst Nematodes using qPCR in combination with propidium monoazide - Christoforou et al. | Diagnostic activities for plant pests | Scoop.it

A real-time PCR combined with propidium monoazide (PMA) was developed for the quantification of viable PCN. This dye renders exposed DNA of dead cells unable to amplify and thus, only DNA from viable / intact PCN juveniles is PCR amplified and detected. The novelty of the present method lies in the simultaneous quantitative and qualitative estimation of viable PCN inoculums using species specific primers and Taqman probes. 

Petter Françoise's insight:

The EPPO protocol on G. pallida and G. rostochiensis is under revision. The Panel on Diagnostics in Nematology will be invited to consider this new method. 

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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 HBL disease: ‘Candidatus Liberibacter asiaticus’, ‘Ca. L. africanus’ or ‘Ca. L. americanus’ is essential for the preventive control of the disease. Real-time PCR is a useful tool for bacterial detection. Nucleic acids purification steps, however, limits the number of samples that can be processed by PCR. Universal detection of ‘Ca. Liberibacter’ species was achieved by a direct tissue-printing and spotting of plant leaf petiole extract or squashing of individual psyllids onto paper or nylon membranes. 

Petter Françoise's insight:

These tests are included in the EPPO Protocol on HLB which is now ready to be presented for adoption. 

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Microlepidoptera on Solanaceae

Microlepidoptera on Solanaceae | Diagnostic activities for plant pests | Scoop.it
Microleptidoptera on Solanaceae contains an interactive key, fact sheets, descriptions, and images to aid in the identification of microlepidoptera on solanaceous plants.
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Development of the simultaneous detection of Ralstonia solanacearum race 3 and Clavibacter michiganensis subsp. sepedonicus in potato tubers by a multiplex real-time PCR assay

Development of the simultaneous detection of Ralstonia solanacearum race 3 and Clavibacter michiganensis subsp. sepedonicus in potato tubers by a multiplex real-time PCR assay | Diagnostic activities for plant pests | Scoop.it

Clavibacter michiganensis subsp. sepedonicus and Ralstonia solanacearum (Smith) Yabuuchi et al. race 3 are the causal agents of ring-rot and brown-rot of potato respectively. These diseases represent a serious threat to potato production in temperate climates. Both bacteria are listed as A2 pests in the EPPO region and as zero-tolerance quarantine organisms in the European Union. All the detection tests developed so far were only focused on the detection of a single pathogen while the absence of both bacteria has to be certified in the seed tubers. We have therefore developed a new multiplex real-time PCR assay to simultaneously detect both bacteria in a single assay. Additionally, the reliability of this molecular diagnostic test has been improved by the simultaneous amplification of an internal control, corresponding to a potato gene co-extracted from the sample. The polyvalence and the specificity of each set of bacterial primers and probes were evaluated on more than 90 bacterial strains. The limit of detection of this triplex real-time protocol was similar to those observed with other molecular protocols previously developed for the individual detection of one of these bacteria. A concordance of 100 % was obtained in a blind test mimicking the routine application of the technology. In conclusion, this new protocol represents a straightforward and convenient method potentially adapted to primary screening of potato tubers.

Petter Françoise's insight:

EPPO has adopted  diagnostic protocols for both pests

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A bead-based suspension array for the multiplexed detection of begomoviruses and their whitefly vectors

A bead-based suspension array for the multiplexed detection of begomoviruses and their whitefly vectors | Diagnostic activities for plant pests | Scoop.it

A bead-based array that detects plant viruses and their insect vectors is described.

The multiplexed array was shown to be highly specific, sensitive and reproducible.

Comparison with qPCR showed equal sensitivity of both methods.

The array may facilitate the improved management of invasive pests and diseases.

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Application of a modified EDTA-mediated exudation technique and guttation fluid analysis for Potato spindle tuber viroid RNA detection in tomato plants (Solanum lycopersicum)

Application of a modified EDTA-mediated exudation technique and guttation fluid analysis for Potato spindle tuber viroid RNA detection in tomato plants (Solanum lycopersicum) | Diagnostic activities for plant pests | Scoop.it

 In this study, a modified method based on an EDTA-mediated phloem exudation technique was applied for detection of PSTVd in the phloem of infected tomato plants. RT-PCR, nucleic acid sequencing, and Southern blot analyses of RT-PCR products verified the presence of viroid RNA in phloem exudates. In addition, the guttation fluid collected from the leaves of PSTVd-infected tomato plants was analyzed revealing the absence of viroid RNA in the xylem sap. To our knowledge, this is the first report of PSTVd RNA detection in phloem exudates obtained by the EDTA-mediated exudation technique.

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Molecular Detection of the Laurel Wilt Fungus, Raffaelea lauricola

Molecular Detection of the Laurel Wilt Fungus, Raffaelea lauricola | Diagnostic activities for plant pests | Scoop.it
Plant Disease, Volume 98, Issue 4, Page 559-564, April 2014.
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Q-collect an EU funded FP 7 framework project on collections

Q-collect an EU funded FP 7 framework project on collections | Diagnostic activities for plant pests | Scoop.it
Q-collect
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Learn more about the Q-collect project by visiting it's website.

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Molecular characterisation and diagnostics of some Longidorus species (Nematoda: Longidoridae) from Russia and other countries using rRNA genes - Subbotin et al. 2014

Molecular characterisation and diagnostics of some Longidorus species (Nematoda: Longidoridae) from Russia and other countries using rRNA genes - Subbotin et al. 2014 | Diagnostic activities for plant pests | Scoop.it

The needle nematodes of the genus Longidorus can cause diseases of various crops and trees, and are comprised of more than 150 valid species. Eleven valid and six unidentified species of the genusLongidorus collected in different regions of Russia, two states of USA, Germany, New Zealand and Ukraine were molecularly characterized using analysis of the partial 18S rRNA and the D2–D3 expansion segments of the 28S rRNA gene sequences. 

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International Plant Protection Convention Expert Consultation on Draft Diagnostic Protocols (ECDP)

The IPPC has initiated an Expert Consultation on several draft  Diagnostic Protocols

Please contribute!

Petter Françoise's insight:

The expert consultation  aims at ensuring improvement on quality for the development of a draft diagnostic protocol (DP), through inputs and feedback, in a scientific basis, from a wider number of experts worldwide not part of the DP drafting group. A wider consultation of experts on draft protocols in earlier stages of development is crucial to ensure the quality of the protocol and to facilitate the adoption process. 

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Longicorn ID: Tool for Diagnosing Cerambycoid Families, Subfamilies, and Tribes

Longicorn ID: Tool for Diagnosing Cerambycoid Families, Subfamilies, and Tribes | Diagnostic activities for plant pests | Scoop.it
Check out APHIS’ new online tool to help non-experts identify large, tree-killing beetles (Cerambycids)! http://t.co/SgUT7DGgU4
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Detection of Tomato yellow leaf curl virus in imported tomato fruit in northern Europe

Imported tomato fruits infected with Tomato yellow leaf curl virus (TYLCV) were identified on the market in northern Europe using paper-based FTA Classic Cards (Whatman), polymerase chain reaction (PCR) and partial DNA sequence analyses.

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An EPPO diagnostic protocol PM 7/50(1) Tomato yellow leaf curl and Tomato mottle begomoviruses was adopted in 

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4th Meeting of the Panel on Diagnostics and Quality Assurance, Paris, 2013-11-13/15

4th Meeting of the Panel on Diagnostics and Quality Assurance, Paris, 2013-11-13/15 | Diagnostic activities for plant pests | Scoop.it
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Read the the short report of the last Panel on Diagnostics and Quaity Assurance

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Fera is recruiting a bacteriologist

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Fera asked me to disseminate this announcement

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Methods in virus diagnostics: From ELISA to next generation sequencing Boonham et al 2014

Methods in virus diagnostics: From ELISA to next generation sequencing Boonham et al 2014 | Diagnostic activities for plant pests | Scoop.it
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A review article involving some  of our regional experts!

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Development of a Multiplexed Bead-Based Suspension Array for the Detection and Discrimination of Pospiviroid Plant Pathogens

Development of a Multiplexed Bead-Based Suspension Array for the Detection and Discrimination of Pospiviroid Plant Pathogens | Diagnostic activities for plant pests | Scoop.it

Efficient and reliable diagnostic tools for the routine indexing and certification of clean propagating material are essential for the management of pospiviroid diseases in horticultural crops. This study describes the development of a true multiplexed diagnostic method for the detection and identification of all nine currently recognized pospiviroid species in one assay using Luminex bead-based suspension array technology.

Petter Françoise's insight:

The EPPO Panel on Diagnostics in Virology and Phytoplasmology agreed at its last meeting that a  generic Standard on the detection of pospiviroids should be prepared for the next meeting of the Panel

http://www.eppo.int/MEETINGS/2013_meetings/virology.htm

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Internship position with the IPPC Secretariat on the Diagnostic Protocol programme

Internship position with the IPPC Secretariat on the Diagnostic Protocol programme | Diagnostic activities for plant pests | Scoop.it
Petter Françoise's insight:

The intern will work with the Secretariat lead for diagnostic protocols and will assist in the organization of the Technical Panel on Diagnostic Protocols (TPDP) meetings, the development and formatting of technical discussion papers and meeting reports, and in editing draft diagnostic protocols.

 

If you want to know more please visit the IPPC Website

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Knapco's curator insight, January 31, 6:07 AM

IPPC aims to protect world plants resources from pests and plant diseases.

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The Complete Genome Sequence of ‘Candidatus Liberibacter americanus’, Associated with Citrus Huanglongbing

The Complete Genome Sequence of ‘Candidatus Liberibacter americanus’, Associated with Citrus Huanglongbing | Diagnostic activities for plant pests | Scoop.it

Liberibacter spp. form a Rhizobiaceae clade of phloem-limited pathogens of limited host range. Two obligately parasitic species have been sequenced: ‘Candidatus Liberibacter asiaticus’, which causes citrus huanglongbing (HLB) worldwide, and ‘Ca. L. solanacearum’, which causes potato “zebra chip” disease. 

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