The Potential Use of Isothermal Amplification Assays for In-Field Diagnostics of Plant Pathogens

Plants, Journal Year: 2021, Volume and Issue: 10(11), P. 2424 - 2424

Published: Nov. 10, 2021

Rapid, sensitive, and timely diagnostics are essential for protecting plants from pathogens. Commonly, PCR techniques used in laboratories highly sensitive detection of DNA/RNA viral, viroid, bacterial, fungal pathogens plants. However, using PCR-based methods in-field is a challenge sometimes nearly impossible. With the advent isothermal amplification methods, which provide nucleic acids at certain temperature do not require thermocyclic equipment, going beyond laboratory has become reality molecular diagnostics. The stage ceases to be limited by time instruments. Challenges solve involve finding suitable approaches rapid user-friendly plant preparation amplicons after amplification. Here, we summarize phytopathogens based on different types discuss their advantages disadvantages. In this review, consider combination with extraction compatible phytodiagnostics. Molecular out-of-lab conditions particular importance against order quickly prevent control spread disease. We believe that development rapid, equipment-free acid future phytodiagnostics, its benefits already visible.

Language: Английский

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Contribution of the EU Reference Laboratory for Insects and Mites to EFSA's pest categorization of non‐EU Scolytinae on non‐coniferous hosts

EPPO Bulletin, Journal Year: 2025, Volume and Issue: unknown

Published: March 27, 2025

Abstract The European Food Safety Authority (EFSA) was mandated by the Union (EU) to perform a group pest categorization of exotic Scolytinae (Coleoptera: Curculionidae) whose establishment in EU territory would present significant risk terms plant health. This should be used reconsider listing ‘Non‐European Scolytinae’ as quarantine pests current Regulation. In first stage, EFSA short‐listed 88 species developing on broadleaf hosts and known for their mobility, having impact levels documented literature either absent or with limited presence EU. then asked Reference Laboratory (EURL) provide comments related its expertise diagnosis insects, assess practical technical implications including these list submitted future regulation? EURL proceeded case‐by‐case examination ranked overall feasibility identification, from A (both morphological molecular tools available) D (none available). Several issues were also raised. particular, identification using sequences hosted NCBI GenBank Bold databases proved highly hazardous. case Euwallacea fornicatus complex especially problematic because multiple taxonomy available distinction constitutive species. As result, we proposed recommendations consideration modifying EFSA's selected developed next working programme.

Language: Английский

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A first inference of the phylogeography of the worldwide invader Xylosandrus compactus

Journal of Pest Science, Journal Year: 2021, Volume and Issue: 95(3), P. 1217 - 1231

Published: Oct. 20, 2021

Language: Английский

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Rapid identification of Ips sexdentatus (Boerner, 1766) (Curculionidae) from adults and frass with real-time PCR based on probe technology

Journal of Plant Diseases and Protection, Journal Year: 2024, Volume and Issue: 131(5), P. 1473 - 1481

Published: April 27, 2024

Abstract A molecular tool has been developed for the identification of Ips sexdentatus (Börner 1776) (Coleoptera Curculionidae Scolytidae), well-known six thooted bark beetle, widely distributed in Eurasia, where it infests several species genus Pinus and occasionally a few conifer genera Abies , Larix Picea . The test can be useful both countries I. is handled as quarantine and, to greater reason, Europe discriminate biological traces this commonly found beetle from those produced by regulated pests. protocol based on real-time PCR with TaqMan probe technology whole insect bodies (adults) well artificial frass contaminated DNA beetle. here direct indirect proven effective terms analytical specificity, sensitivity, reliability reproducibility. recommended practical diagnostic allowing rapid toothed presence any trace other xylophagous pests collected at points entry during phytosanitary surveys.

Language: Английский

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Pest categorisation of non‐EU Scolytinae on non‐coniferous hosts

EFSA Journal, Journal Year: 2024, Volume and Issue: 22(9)

Published: Sept. 1, 2024

Abstract The EFSA Panel on Plant Health performed a group pest categorisation for the EU territory of non‐EU Scolytinae (Coleoptera: Curculionidae) non‐coniferous hosts, which total 6495 known species. Most species attack apparently healthy, weakened or dead trees, either feeding phloem (‘bark beetles’ subgroup) fungi inoculated into sapwood (‘ambrosia subgroup). Smaller subgroups feed and reproduce in seeds fruits, herbaceous plants. Some are polygynous, males initiate gallery chamber new host attract females. Others monogamous, females galleries. Many respond to primary volatile attractants emitted by some produce aggregation pheromones that conspecifics both sexes. attacking living hosts often associated with contribute weakening defences provide nutrients insects. inbreeding; offspring mate their sisters rarely leave natal tree. larvae all develop pupate within hosts. Based catalogues other published data, database was constructed providing information reproductive habits, geographic distribution Köppen–Geiger climate types countries where occur. were screened exclude following categories: (i) 708 conifers; (ii) 127 present at least four Member States (iii) 440 occurring areas climatic conditions not EU. Among remaining 5220 species, 88 mobility, occupying two landmasses separated geographical barriers had impact levels documented literature, extracted. They grouped subcategories: 12 high plant health; 16 low doubtful impact; 48 no (iv) never been recorded as ‘introduced’ consulted but landmasses. All could enter wood products, plants planting, establish because available, is suitable parts Control measures inhibit introduction available. There considerable uncertainty regarding potential many Methods reliable identification lacking. For criteria assessed consideration quarantine met. Nevertheless, able method discriminate confidently between clearly meet status those do not.

Language: Английский

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Detection of invasive and native beetle species within trees by chemical analysis of frass

Scientific Reports, Journal Year: 2023, Volume and Issue: 13(1)

Published: July 22, 2023

Abstract In recent years, several invasive woodborers (Coleoptera: Cerambycidae) have been found in Japan. Aromia bungii is a worldwide important pest of fruits and ornamental species the genus Prunus . It invaded Japan early 2010s now causes heavy damage to stone fruit trees. Anoplophora glabripennis Apriona swainsoni are destructive pests street, horticultural The first step intercepting these beetles detect their presence infestation, as accurate identification crucial for management. Ejected frass major sign infestation likely holds information on insect. We focused chemicals both larvae frass, conducted GC–MS analysis three native malasiaca all four species, 4 or 5 species-specific hydrocarbons were detected frass. These results indicate that could allow definitive detection wood-boring pests.

Language: Английский

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FUNGI OBTAINED FROM OLIVE TWIG DIEBACK AND ADULTS OF THE ALIEN PEST XYLOSANDRUS COMPACTUS (EICHHOFF) (COLEOPTERA CURCULIONIDAE SCOLYTINAE)

Redia, Journal Year: 2022, Volume and Issue: 105, P. 197 - 204

Published: Dec. 12, 2022

Olive (Olea europaea L.) is a traditional crop of great socioeconomic importance in the Mediterranean Basin. In 2021, survey was performed an Italian nursery located Central Italy where unusual symptoms and widespread dieback had been observed on young potted olive trees. Twigs branches all plants affected by dryness showed presence alien bark beetle Xylosandrus compactus. Woody tissues colonized X. compactus were transferred into laboratory several fungi such as Cladosporium perangustum Neofusicoccum parvum then identified. addition to these species, Geosmithia pallida also reported for first time this plant. Studies its associated are currently progress further evaluate potential risks cultivation. Key Words: Olea europea, fungi, beetles.

Language: Английский

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An accurate, efficient, and economical identification technology for black twig borer based on species-specific cytochrome C oxidase subunit I PCR assay

Journal of Economic Entomology, Journal Year: 2023, Volume and Issue: 116(4), P. 1372 - 1378

Published: June 27, 2023

Abstract Xylosandrus compactus (Eichhoff) (Coleoptera: Curculionidae, Scolytinae) is a worldwide invasive species that causes huge economic loss and environmental damage in many countries. Traditional morphological characteristics make it hard to identify scolytines due their tiny size. Besides, the intercepted insect samples are incomplete, limitation of (larvae pupae) morphology makes identification more difficult. The majority caused by adults fungi serve as nutrition for larvae. They destroy plant trunks, branches, twigs, affecting transport tissues both weak healthy plants. An accurate, efficient, economical molecular technique X. not restricted professional taxonomic knowledge necessary. In present study, tool based on mitochondrial DNA gene, cytochrome C oxidase subunit I (COI) was developed. A species-specific COI (SS-COI) PCR assay designed regardless developmental stage. Twelve commonly found eastern China, namely compactus, crassiusculus, discolor, germanus, borealis, amputates, eupatorii, mancus, Xyleborinus saxesenii, Euwallacea interjectus, E. fornicatus, Acanthotomicus suncei, were included study. Additionally, specimens from 17 different areas well specimen collected United Stated, also analyzed. Results demonstrated accuracy high efficiency assay, stage or type specimen. These features provide good application prospect fundamental departments can be used prevent harmful consequences spread compactus.

Language: Английский

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Rapid identification of Ripersiella (= Rhizoecus) hibisci (Kawai & Takagi, 1971) (Hemiptera: Rhizoecidae) with TaqMan probe‐based real‐time PCR

EPPO Bulletin, Journal Year: 2024, Volume and Issue: 54(1), P. 41 - 48

Published: April 1, 2024

Abstract Ripersiella hibisci (Kawai & Takagi, 1971) is a root mealybug included in the EPPO A1 list of pests recommended for regulation and an EU Quarantine pest (Annex IIA). This causes damage to several species monocotyledonous dicotyledonous ornamental foliage potted plants, especially bonsais. The underground location R. makes its detection difficult during phytosanitary controls, moreover, it can be confused with other non‐regulated mealybugs (e.g. Planococcus citri , P. ficus Rhizoecus falcifer dianthi ). Thus, molecular test based on real‐time PCR was designed rapid unambiguous identification this mealybug. Analytical sensitivity, analytical specificity, repeatability reproducibility were assessed as part validation. showed 100% specificity (exclusivity inclusivity), allowing target distinguished from all non‐target tested (exclusivity); inclusivity demonstrated, validating different populations pest. efficient tool (in various life stages), which particularly useful when screening samples needed limit or avoid introduction

Language: Английский

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Identification of the ambrosia beetle Anisandrus dispar (Fabricius) (Coleoptera Curculionidae Scolytinae) using TaqMan™ probe assay on biological samples

iForest - Biogeosciences and Forestry, Journal Year: 2023, Volume and Issue: 16(3), P. 182 - 187

Published: June 30, 2023

The European shot-hole borer Anisandrus dispar (Fabricius) (Coleoptera Curculionidae Scolytinae) is a well-known ambrosia beetle living on shrubs and several ornamental fruiting trees where it can cause heavy damages. Like other harmful xyleborine species, A. represent potential threat outside its native region. Molecular diagnostic tools lead to accurate identification of xylophagous species hidden in wooden matrix phytosanitary surveys at entry points. A molecular assay based qPCR TaqMan™ Probes was developed for the from different matrices. To setup perform test, DNA extraction carried out adults, larvae, artificial samples wood chips oak healthy plants whose lysates were contaminated with known amount adults. has proven inclusive dispar, exclusive towards non-target organisms, showing 100% analytical specificity. limit detection 0.32 pg µL-1 insect adult 0.8 containing Quercus spp. 0.1 ng DNA. Repeatability reproducibility showed low values independently used extraction, confirming possible use diagnostics biological even if not directly related presence developmental stages. presented approach may be adjusted applied purposes quarantine pests rapidly detect infestations vegetal matrices globally traded.

Language: Английский

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