Next‐generation methods for early disease detection in crops

Pest Management Science, Journal Year: 2023, Volume and Issue: 80(2), P. 245 - 261

Published: Aug. 21, 2023

Plant pathogens are commonly identified in the field by typical disease symptoms that they can cause. The efficient early detection and identification of essential procedures to adopt effective management practices reduce or prevent their spread order mitigate negative impacts disease. In this review, traditional innovative methods for plant highlighting major advantages limitations presented discussed. Traditional techniques diagnosis used pathogen focused typically on DNA, RNA (when molecular methods), proteins peptides serological methods) pathogens. Serological based mainly enzyme-linked immunosorbent assay (ELISA) most common method due high-throughput potential low cost. This technique is not particularly reliable sufficiently sensitive many during asymptomatic stage infection. For non-cultivable laboratory, nucleic acid-based technology best choice consistent identification. Lateral flow systems tools allow fast accurate results even conditions, but have sensitivity issues be overcome. PCR assays performed last-generation portable thermocyclers may provide rapid situ. advent instruments speed detection, commercial costs, potentially revolutionize pathology. review provides information current methodologies different © 2023 Authors. Pest Management Science published John Wiley & Sons Ltd behalf Society Chemical Industry.

Language: Английский

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Rapid on-site differentiation of two invasive whitefly cryptic species using LAMP

Scientific Reports, Journal Year: 2025, Volume and Issue: 15(1)

Published: Feb. 12, 2025

Loop-mediated isothermal amplification (LAMP) is a promising technique for detecting pest species used on the spot due to its simplicity and rapidity. The whitefly regarded as complex encompassing over 44 cryptic species. These are morphologically indistinguishable but exhibit notable differences in their biological traits. Among them, Bemisia tabaci MEAM1 MED most invasive devastating members. Accurately quickly distinguishing between these two whiteflies field directly affects implementation of control measures. However, it often difficult distinguish when they occur together require extensive specialist knowledge or instrumentation. one limitations current LAMP that reaction mixture requires cold chain, which not ideal detection spot. To promote actual application spot, methods differentiate B. were developed laboratory, Trialeurodes vaporariorum was negative control. Then effect preservation condition (room temperature low temperature) efficiency compared. study showed MED- MEAM1- can MEAM1, MED, T. after low-temperature more than 24 h. Finally, we validated method with temperature-controlled hot-water cup proved effectiveness applied field. results demonstrated provides available technical support

Language: Английский

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Molecular Identification of Agrilus anxius (Coleoptera: Buprestidae) Using a qPCR Assay With Locked Nucleic Acid (LNA) Probe

Journal of Applied Entomology, Journal Year: 2025, Volume and Issue: unknown

Published: March 6, 2025

ABSTRACT In the present study, a biomolecular diagnostic assay based on qPCR‐LNA (Locked Nucleic Acid) probes was developed for identification of Agrilus anxius (Coleoptera: Buprestidae) from adult insects. The performance new protocol also evaluated indirect diagnosis insect's presence artificially contaminated frass. designed primer and probe were able to distinguish in silico all A. samples nontarget species, with 100% match homologous sequences found GenBank databases. molecular sensitive, specific repeatable. analytical sensitivity (limit detection—LoD) adults frass 6.4 fg/μL 0.08 pg/μL, respectively. This assay, by analysing eDNA samples, will allow early detection an area before it has caused major impact. analysis is becoming increasingly used tool spatial survey programs phytosanitary services could play decisive role pest surveillance.

Language: Английский

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Specificity and Sensitivity of a Rapid LAMP Assay for Early Detection of Emerald Ash Borer (Agrilus planipennis) in Europe

Forests, Journal Year: 2023, Volume and Issue: 14(2), P. 436 - 436

Published: Feb. 20, 2023

Buprestids are an emerging threat to broadleaf forests across the world. Species such as emerald ash borer (EAB, Agrilus planipennis) seriously threaten (Fraxinus spp.) in North America and Europe. As it continues spreading west from European Russia, native populations will suffer dramatic losses. Due their cryptic lifestyle of egg larval stages on developing bark vascular tissue, buprestids other wood borers can be difficult detect. Early detection tools vital implement fast eradication measures, prevent establishment invasive species populations. Detection methods using polymerase chain reaction (PCR) assays target specific taxa extremely timely obtain results especially since samples need transported laboratory first. However, loop-mediated isothermal amplification (LAMP) eDNA highly sensitive providing within 30 min after sample extraction. In this study, we investigated specificity sensitivity EAB LAMP assay early tool The was when tested against 12 spp., five buprestids, two Scolytinae, cerambycids (n = 24). amplified DNA a concentration low 0.02 pg/µL. These demonstrate that is specific, used detect monitor urban settings.

Language: Английский

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eDNA-based detection of the invasive crayfish Pacifastacus leniusculus in streams with a LAMP assay using dependent replicates to gain higher sensitivity

Scientific Reports, Journal Year: 2022, Volume and Issue: 12(1)

Published: April 21, 2022

Abstract LAMP assays are becoming increasingly popular in the field of invasive species detection but still underused eDNA-based monitoring. Here, we propose a assay designed to detect North American crayfish Pacifastacus leniusculus water samples from streams. The presence P. was detected through this new all one nine sites sampled. No correlation found between ddPCR absolute concentration measurements and number LAMP-positive technical replicates. However, showed that using dependent replicates could significantly enhance sensitivity assay. Applied other assays, it improve thus allow for more efficient use aquatic ecosystems.

Language: Английский

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Development of a loop-mediated isothermal amplification (LAMP) assay for the identification of the invasive wood borer Aromia bungii (Coleoptera: Cerambycidae) from frass

3 Biotech, Journal Year: 2021, Volume and Issue: 11(2)

Published: Jan. 19, 2021

Abstract The red-necked longhorn beetle Aromia bungii (Faldermann, 1835) (Coleoptera: Cerambycidae) is native to east Asia, where it a major pest of cultivated and ornamental species the genus Prunus . Morphological or molecular discrimination adults larval specimens required identify this invasive wood borer. However, recovering stages from trunks branches causes extensive damage plants timewasting. An alternative approach consists in applying non-invasive diagnostic tools biological traces (i.e., fecal pellets, frass). In way, infestations host can be detected without destructive methods. This paper presents protocol based on both real-time visual loop-mediated isothermal amplification (LAMP), using DNA A. extracted particles frass. Laboratory validations demonstrated robustness protocols adopted their reliability was confirmed performing an inter-lab blind panel. LAMP assay qPCR SYBR Green method F3/B3 external primers were equally sensitive, more sensitive than conventional PCR (sensitivity > 10 3 same starting matrix). protocol, due relatively easy performance method, could useful tool apply rapid monitoring management its outbreaks.

Language: Английский

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Identification of the ambrosia beetle Xyleborinus saxesenii (Ratzeburg, 1837) (Coleoptera: Curculionidae: Scolytinae) from frass and adult DNA by TaqMan Probe real‐time‐PCR

EPPO Bulletin, Journal Year: 2024, Volume and Issue: 54(1), P. 84 - 94

Published: Jan. 22, 2024

Abstract A molecular tool has been set up for the unambiguous identification of ambrosia beetle, Xyleborinus saxesenii (Ratzeburg, 1837) (Coleoptera Curculionidae Scolytinae), cosmopolitan fruit‐tree pinhole borer, widely distributed in temperate regions five continents, where it infests a wide range hardwoods and softwoods. The test was based on real‐time PCR with TaqMan probe technology developed whole insect bodies (adults) as well frass produced by beetle. shown to meet criteria established EPPO harmonization diagnostic methods. In particular, gave good results terms analytical specificity (inclusivity exclusivity) sensitivity, fully repeatable reproducible. Since X. is one most commonly intercepted beetles during phytosanitary controls worldwide, this practical will be useful its rapid beetle biological material (frass, body fragments). can countries quarantine species, region.

Language: Английский

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Rapid Detection of Pityophthorus juglandis (Blackman) (Coleoptera, Curculionidae) with the Loop-Mediated Isothermal Amplification (LAMP) Method

Plants, Journal Year: 2021, Volume and Issue: 10(6), P. 1048 - 1048

Published: May 22, 2021

The walnut twig beetle Pityophthorus juglandis is a phloem-boring bark responsible, in association with the ascomycete Geosmithia morbida, for Thousand Cankers Disease (TCD) of trees. recent finding TCD Europe prompted development effective diagnostic protocols early detection members this insect/fungus complex. Here we report highly efficient, low-cost, and rapid method detecting beetle, or even just its biological traces, from environmental samples: loop-mediated isothermal amplification (LAMP) assay. method, designed on 28S ribosomal RNA gene, showed high specificity sensitivity, no cross reactivity to other beetles wood-boring insects. test was successful very small amounts target insect’s nucleic acid, limit values 0.64 pg/µL 3.2 WTB adults frass, respectively. A comparison (both real time visual) conventional PCR did not display significant differences terms LoD. This LAMP protocol will enable quick, P. areas new infestations phytosanitary inspections at vulnerable sites (e.g., seaports, airports, loading stations, storage facilities, wood processing companies).

Language: Английский

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Colorimetric loop-mediated isothermal amplification assay for detection and ecological monitoring of Sarocladium oryzae, an important seed-borne pathogen of rice

Frontiers in Plant Science, Journal Year: 2022, Volume and Issue: 13

Published: Aug. 18, 2022

Accurate and timely disease detection plays a critical role in achieving sustainable crop protection. Globally, rice has been staple for centuries plagued by the diseases that greatly hamper its productivity. Sheath rot, an emerging of caused seed-borne pathogen Sarocladium oryzae , reportedly heavy losses to agricultural produce recent years. Our study led development validation LAMP assay early S. causal agent sheath rot from live-infected tissues, seeds, weeds, environmental samples. The could detect as low 1.6 fg/μl 15 min. was implemented bio-surveil presence this testing it on three weed species ( Echinochloa colona crus-galli Cyperus teneriffae ) growing around fields. results showed two viz . E. used test 13 different varieties seeds. In total, did not show their seeds while rest were found harbor pathogen. developed can effectively be screen live samples including field soil.

Language: Английский

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Development of novel LAMP and qPCR assays for rapid and specific identification of Bronze birch borer (Agrilus anxius)

Environmental DNA, Journal Year: 2023, Volume and Issue: 5(6), P. 1177 - 1190

Published: Nov. 1, 2023

Abstract Buprestids are an emerging threat to broadleaf forests across the world. Bronze birch borer ( Agrilus anxius , BBB) poses a serious European species if insect were be introduced. Due their cryptic lifestyle feeding on vascular tissue of host plants, buprestids and other wood borers can difficult observe or detect. Early detection tools vital swiftly implement eradication measures prevent establishment introduced species. In this study, we developed novel qPCR LAMP assays for BBB investigated specificity sensitivity use as early in forests. Plant chemicals may limit these assays, so conducted testing with extracted foliage plant tissues determine potential inhibition effects DNA amplification. Both specific target when tested against 17 /buprestid species, two Scolytinae, five Cerambycids N = 24). varied assay amplifying at concentration low 20 fg/μL, whereas amplified 3.2 pg/μL. extracts from leaves did not impact either assay, reaching similar levels. contrast, reduced 0.04 ng/μL compared 0.008 control. These results demonstrate that both highly sensitive used detect frass identify larvae well monitor spread A. . resulted more than overall. Thus, needed quickly make fast management decisions initial screening samples, method is optimal. However, fine critical, then preferential.

Language: Английский

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