Comparison of the starch-iodine reaction technique with the Recombinase Polymerase Amplification for citrus Huanglongbing (HLB) diagnosis

Comparison of the starch-iodine reaction technique with the Recombinase Polymerase Amplification for citrus Huanglongbing (HLB) diagnosis

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Huanglongbing (HLB) is one of the most important citrus diseases worldwide. A field level diagnosis is difficult when it is based on visual symptoms, since they can be confused with similar symptoms of other diseases or nutritional deficiencies of the plant. Molecular methods are used for the diagnosis of the disease, which are laborious, time-consuming and costly, in addition to not allowing rapid detection in the field. In relation to this problem, the method of starch-iodine reaction (RAI) was proposed as a cheap alternative for the diagnosis of HLB by various authors, however, the method has not been widely used, possibly due to lack of diffusion. In the present work the efficacy of the RAI diagnostic method was demonstrated, by comparison with the molecular method of Amplification by recombinase and polymerase (ARP); a molecular method that allows the detection of HLB in the field, but with higher cost. A total of 825 leaf samples were taken from 33 sweet orange (Citrus sinensis L.) plants from orchards of the Department of Caazapá, Paraguay. The samples were analyzed by RAI method, staining and intensity of coloration were observed by optical microscopy. Among these, 33 samples were selected with positive and negative reactions to be analyzed by the ARP method. Results revealed a correlation of 93.6% between both methods. The RAI method, in addition to differentiating HLB symptoms from other diseases and nutritional deficiencies in the plant, is a simple, fast, effective, and economical method for diagnosis if the disease.

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Main Authors: González Segnana, Luis Roberto, Acosta Aveiro, Darío Rafael, Pino Quintana, Cesar Darío, González Espínola, Diego Dionisio
Format: Digital revista
Language:spa
Published: Facultad de Ciencias Agrarias de la Universidad Nacional de Asunción 2020
Online Access:https://www.agr.una.py/revista/index.php/ria/article/view/586
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institution UNA PY
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country Paraguay
countrycode PY
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databasecode rev-ria
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libraryname Biblioteca de la Facultad de Ciencias Agrarias de la UNA PY
language spa
format Digital
author González Segnana, Luis Roberto
Acosta Aveiro, Darío Rafael
Pino Quintana, Cesar Darío
González Espínola, Diego Dionisio
spellingShingle González Segnana, Luis Roberto
Acosta Aveiro, Darío Rafael
Pino Quintana, Cesar Darío
González Espínola, Diego Dionisio
Comparison of the starch-iodine reaction technique with the Recombinase Polymerase Amplification for citrus Huanglongbing (HLB) diagnosis
author_facet González Segnana, Luis Roberto
Acosta Aveiro, Darío Rafael
Pino Quintana, Cesar Darío
González Espínola, Diego Dionisio
author_sort González Segnana, Luis Roberto
title Comparison of the starch-iodine reaction technique with the Recombinase Polymerase Amplification for citrus Huanglongbing (HLB) diagnosis
title_short Comparison of the starch-iodine reaction technique with the Recombinase Polymerase Amplification for citrus Huanglongbing (HLB) diagnosis
title_full Comparison of the starch-iodine reaction technique with the Recombinase Polymerase Amplification for citrus Huanglongbing (HLB) diagnosis
title_fullStr Comparison of the starch-iodine reaction technique with the Recombinase Polymerase Amplification for citrus Huanglongbing (HLB) diagnosis
title_full_unstemmed Comparison of the starch-iodine reaction technique with the Recombinase Polymerase Amplification for citrus Huanglongbing (HLB) diagnosis
title_sort comparison of the starch-iodine reaction technique with the recombinase polymerase amplification for citrus huanglongbing (hlb) diagnosis
description Huanglongbing (HLB) is one of the most important citrus diseases worldwide. A field level diagnosis is difficult when it is based on visual symptoms, since they can be confused with similar symptoms of other diseases or nutritional deficiencies of the plant. Molecular methods are used for the diagnosis of the disease, which are laborious, time-consuming and costly, in addition to not allowing rapid detection in the field. In relation to this problem, the method of starch-iodine reaction (RAI) was proposed as a cheap alternative for the diagnosis of HLB by various authors, however, the method has not been widely used, possibly due to lack of diffusion. In the present work the efficacy of the RAI diagnostic method was demonstrated, by comparison with the molecular method of Amplification by recombinase and polymerase (ARP); a molecular method that allows the detection of HLB in the field, but with higher cost. A total of 825 leaf samples were taken from 33 sweet orange (Citrus sinensis L.) plants from orchards of the Department of Caazapá, Paraguay. The samples were analyzed by RAI method, staining and intensity of coloration were observed by optical microscopy. Among these, 33 samples were selected with positive and negative reactions to be analyzed by the ARP method. Results revealed a correlation of 93.6% between both methods. The RAI method, in addition to differentiating HLB symptoms from other diseases and nutritional deficiencies in the plant, is a simple, fast, effective, and economical method for diagnosis if the disease.
publisher Facultad de Ciencias Agrarias de la Universidad Nacional de Asunción
publishDate 2020
url https://www.agr.una.py/revista/index.php/ria/article/view/586
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spelling oai:ojs.www.agr.una.py:article5862020-11-05T21:02:09Z Comparison of the starch-iodine reaction technique with the Recombinase Polymerase Amplification for citrus Huanglongbing (HLB) diagnosis Comparación del método de reacción almidón – iodo con la Amplificación por Polimerasa y Recombinasa para el diagnóstico del Huanglongbing (HLB) de los cítricos González Segnana, Luis Roberto Acosta Aveiro, Darío Rafael Pino Quintana, Cesar Darío González Espínola, Diego Dionisio HLB citrus detection HLB cítricos detección. Huanglongbing (HLB) is one of the most important citrus diseases worldwide. A field level diagnosis is difficult when it is based on visual symptoms, since they can be confused with similar symptoms of other diseases or nutritional deficiencies of the plant. Molecular methods are used for the diagnosis of the disease, which are laborious, time-consuming and costly, in addition to not allowing rapid detection in the field. In relation to this problem, the method of starch-iodine reaction (RAI) was proposed as a cheap alternative for the diagnosis of HLB by various authors, however, the method has not been widely used, possibly due to lack of diffusion. In the present work the efficacy of the RAI diagnostic method was demonstrated, by comparison with the molecular method of Amplification by recombinase and polymerase (ARP); a molecular method that allows the detection of HLB in the field, but with higher cost. A total of 825 leaf samples were taken from 33 sweet orange (Citrus sinensis L.) plants from orchards of the Department of Caazapá, Paraguay. The samples were analyzed by RAI method, staining and intensity of coloration were observed by optical microscopy. Among these, 33 samples were selected with positive and negative reactions to be analyzed by the ARP method. Results revealed a correlation of 93.6% between both methods. The RAI method, in addition to differentiating HLB symptoms from other diseases and nutritional deficiencies in the plant, is a simple, fast, effective, and economical method for diagnosis if the disease. El Huanglongbing (HLB) es una de las enfermedades más importantes de los cítricos a nivel mundial. Su diagnóstico a nivel de campo es difícil cuando se basa en los síntomas visuales ya que pueden ser confundidos con síntomas similares de otras enfermedades o deficiencias nutricionales de la planta. Métodos moleculares son utilizados para el diagnóstico de la enfermedad, los cuales son laboriosos, requieren tiempo y tienen un costo elevado, además de no permitir una rápida detección en el campo. En relación a esta problemática, el método de diagnóstico de reacción almidón-iodo (RAI) fue propuesto por diversos autores como una alternativa barata para la diagnosis de HLB, sin embargo, el método no ha sido ampliamente utilizado, posiblemente por falta de difusión. En el presente trabajo se demostró la eficacia del método de diagnóstico por RAI, mediante su comparación con el método molecular de Amplificación por recombinasa y polimerasa(ARP), siendo éste un método molecular que permite la detección de HLB a nivel de campo, pero que tiene un costo superior. Fueron extraídas un total de 825 muestras de hojas de 33 plantas de naranjo dulce (Citrus sinensisL.) de plantíos del Departamento de Caazapá, Paraguay. Las muestras fueron analizadas por el método de RAI, la tinción e intensidad de la coloración fueron observadas por microscopia óptica. Entre estas se seleccionaron 33 muestras con reacciones positivas y negativas para ser analizadas por el método de ARP. Resultados revelaron una correlación del 93,6% entre ambos métodos. El método de RAI, además de permitir diferenciar los síntomas de HLB de otras enfermedades y deficiencias nutricionales en la planta, es un método sencillo, rápido, eficaz, y económico para el diagnóstico de la enfermedad. Facultad de Ciencias Agrarias de la Universidad Nacional de Asunción 2020-11-05 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf application/epub+zip https://www.agr.una.py/revista/index.php/ria/article/view/586 10.18004/investig.agrar.2020.diciembre.2202586 Investigación Agraria; Vol. 22 No. 2 (2020): July-December; 100-105 Investigación Agraria; Vol. 22 Núm. 2 (2020): Julio-Diciembre; 100-105 Investigación Agraria; v. 22 n. 2 (2020): Julho-Dezembro; 100-105 2305-0683 1684-9086 spa https://www.agr.una.py/revista/index.php/ria/article/view/586/451 https://www.agr.una.py/revista/index.php/ria/article/view/586/461 https://creativecommons.org/licenses/by/4.0

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