A multiplex RT-PCR assay for the simultaneous detection of prevalent viruses infecting pepper (Capsicum annuum L.)

Nemes, Katalin; Salánki, Katalin

doi:10.1016/j.jviromet.2020.113838

Cited by 13 publications

(7 citation statements)

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“…In this study, we report on the development, optimization, and validation of a multiplex PCR assay for the simultaneous detection of five RNA viruses (ChiVMV, CMV, PMMoV, CaCV, and LCCV) and a DNA virus (ChiLCV) which are of the most predominant occurrence and a major threat to chilli cultivation in India. Earlier, a multiplex RT-PCR assay for the simultaneous detection of CMV, tobacco mosaic virus, PMMoV, PVY, and TSWV in chilli was reported [ 60 ]; however, to the best of our knowledge, there is no report on multiplex PCR assay for the detection of six RNA and DNA viruses, which was attempted in the present study. We designed the primers to target the conserved regions of the viral genome in order to achieve specific amplification and avoid any chances of cross-reactivity.…”

Section: Discussionmentioning

confidence: 89%

“…The development of a multiplex PCR assay is often complex since primers must comply with several similar conditions that all primers can function under the same PCR conditions like compatibility, avoiding cross-binding, competition, and sufficient sensitivity [ 56 ]. Multiplex PCR has been successfully employed earlier to identify multiple viruses in different crops plants such as tomato [ 59 ], chilli/pepper [ 60 ], and tobacco [ 61 ]. Chilli is an important spice crop of India that is grown in almost all geographical areas.…”

Section: Discussionmentioning

confidence: 99%

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A Simplified Multiplex PCR Assay for Simultaneous Detection of Six Viruses Infecting Diverse Chilli Species in India and Its Application in Field Diagnosis

et al. 2022

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Chilli is infected by at least 65 viruses globally, with a mixed infection of multiple viruses leading to severe losses being a common occurrence. A simple diagnostic procedure that can identify multiple viruses at once is required to track their spread, initiate management measures and manage them using virus-free planting supplies. The present study, for the first time, reports a simplified and robust multiplex PCR (mPCR) assay for the simultaneous detection of five RNA viruses, capsicum chlorosis orthotospovirus (CaCV), chilli veinal mottle virus (ChiVMV), large cardamom chirke virus (LCCV), cucumber mosaic virus (CMV), and pepper mild mottle virus (PMMoV), and a DNA virus, chilli leaf curl virus (ChiLCV) infecting chilli. The developed mPCR employed six pairs of primer from the conserved coat protein (CP) region of the respective viruses. Different parameters viz., primer concentration (150–450 nM) and annealing temperature (50 °C), were optimized in order to achieve specific and sensitive amplification of the target viruses in a single reaction tube. The detection limit of the mPCR assay was 5.00 pg/µL to simultaneously detect all the target viruses in a single reaction, indicating a sufficient sensitivity of the developed assay. The developed assay showed high specificity and showed no cross-amplification. The multiplex PCR assay was validated using field samples collected across Northeast India. Interestingly, out of 61 samples collected across the northeastern states, only 22 samples (36%) were positive for single virus infection while 33 samples (54%) were positive for three or more viruses tested in mPCR, showing the widespread occurrence of mixed infection under field conditions. To the best of our knowledge, this is the first report on the development and field validation of the mPCR assay for six chilli viruses and will have application in routine virus indexing and virus management.

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Section: Discussionmentioning

confidence: 89%

Section: Discussionmentioning

confidence: 99%

A Simplified Multiplex PCR Assay for Simultaneous Detection of Six Viruses Infecting Diverse Chilli Species in India and Its Application in Field Diagnosis

et al. 2022

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“…Los áfidos Aphis gossypii Glover 1877 y Myzus persicae (Sulzer 1776) (Simbaqueba & Serna, 2021) son las únicas especies de áfidos plagas del cultivo de ají Capsicum spp. (Solanaceae) (Tálaga-Taquinas et al, 2020), que ocasionan pérdidas económicas al transmitir virus (Nemes & Salánki, 2020). Su principal método de control es la aspersión de insecticidas (Santos et al, 2017), causando perturbaciones ambientales (Guigón-Guigón & González-González, 2007) y contaminación por trazas en los frutos (Chilumuru et al, 2015), por lo que es necesario desarrollar estrategias más amigables como el control biológico.…”

Section: Introductionunclassified

Depredación, reproducción y desarrollo de la catarina Hippodamia convergens (Coleoptera: Coccinellidae)

Naranjo-Acosta¹,

Díaz-Mesa²,

Melo-Cerón³

et al. 2022

Rev. Biol. Trop.

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Introducción: Mantener poblaciones de insectos depredadores en el agroecosistema ofreciéndoles presas y recursos alimenticios provenientes de las plantas es importante para mejorar el control biológico y contribuir con la conservación. Objetivo: Determinar si el consumo de presas y recursos alimenticios florales potencia los atributos biológicos del depredado Hippodamia convergens. Métodos: Las larvas de Hippodamia convergens fueron alimentadas con dos especies de áfidos, mientras que los adultos fueron alimentados con áfidos, espigas de maíz y dos especies de plantas no cultivadas. La presencia de azúcar en el intestino del depredador se comprobó mediante la prueba de antronas frías, y la presencia de polen mediante el método de acetólisis. Los experimentos demográficos se realizaron durante los primeros 40 días después de la emergencia de la hembra bajo condiciones controladas (25 °C ± 0.5; 75 % ± 1.75 HR; L12:D12). Resultados: H. convergens no consumió fructosa de Sorghum halepense, Parthenium hysterophorus o Zea mays, sino únicamente polen. En condiciones controladas (25 °C ± 0.5, 75 % ± 1.75 HR), el tiempo de desarrollo (huevo-adulto) del depredador fue más corto (21.36 días) cuando consumió Rhopalosiphum maidis en lugar de Uroleucon nigrotibium (24.6 días), mientras que la supervivencia (L1- adulto) y la proporción sexual no cambió. La fecundidad promedio fue mayor (55.5 huevos / 40 días) al consumir U. nigrotibium con polen de P. hysterophorus, que solo U. nigrotibium (22.5 huevos / 40 días), o R. maidis con polen de Zea mays (11 huevos / 40 días). La tasa intrínseca de crecimiento natural fue mayor al consumir U. nigrotibium con polen de P. hysterophorus (0.055), que U. nigrotibium (0.034) o R. maidis con polen de Z. mays (0.019). La tasa de depredación (L1-L5) fue mayor al consumir R. maidis (0.65) que U. nigrotibium (0.51). Conclusiones: Las especies de áfidos y su combinación con polen de plantas no cultivadas afectan de manera diferente el tiempo de desarrollo, reproducción y tasa de depredación de H. convergens.

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“…XhoII digestion of the original PCR product resulted in only the two predicted restriction fragments (418/1423 bp). Thalia plant was also tested with negative results for tobamoviruses, cucumber mosaic virus, and tomato spotted wilt virus with multiplex RT-PCR (Nemes and Salánki 2020). This is the first report of SCMV infecting Thalia dealbata, a perennial herbaceous ornamental aquatic plant.…”

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confidence: 99%

Thalia dealbata, a new host of sugarcane mosaic virus

et al. 2023

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A multiplex RT-PCR assay for the simultaneous detection of prevalent viruses infecting pepper (Capsicum annuum L.)

Cited by 13 publications

References 33 publications

A Simplified Multiplex PCR Assay for Simultaneous Detection of Six Viruses Infecting Diverse Chilli Species in India and Its Application in Field Diagnosis

A Simplified Multiplex PCR Assay for Simultaneous Detection of Six Viruses Infecting Diverse Chilli Species in India and Its Application in Field Diagnosis

Depredación, reproducción y desarrollo de la catarina Hippodamia convergens (Coleoptera: Coccinellidae)

Thalia dealbata, a new host of sugarcane mosaic virus

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