Detection of squash mosaic virus in seeds of melon (Cucumis melo) by enzyme-linked immunosorbent assay (ELISA)

1990 ◽  
Vol 96 (2) ◽  
pp. 91-102 ◽  
Author(s):  
A. A. J. M. Franken ◽  
D. Z. Maat ◽  
G. C. Kamminga
2017 ◽  
Vol 20 (2) ◽  
pp. 59 ◽  
Author(s):  
Budi Setiadi Daryono ◽  
Faizatul Fitriyah

Melon (Cucumis melo L.) belongs to Cucurbitaceae. Melon has high potential to be developed as main horticultural product in Indonesia. Melon is one of important foreign exchange and is the fifth biggest horticulture commodity in Indonesia. One of the problems in melon farming is mosaic disease caused by Kyuri green mottle mosaic virus (KGMMV). KGMMV infection reduces the quality and the amount of melon production. Melon farmers suffered a significant financial loss. Melodi Gama 3 (MG3) is a high yielding melon cultivar from the Genetics Laboratory, Faculty of Biology, Universitas Gadjah Mada. The use of genetically resistant melon cultivar has beneficial outcome for agriculture sector. The aim of this research was to study the resistance’s inherintance to KGMMV in MG3 melon cultivar. Two cultivars of MG3, MG3|5and MG3|8, were cultivated in the greenhouse. MAI, Glamour, Ladika, and Action melon cultivars were used as references. Resistance of KGMMV was analyzed by symptom observation and serological detection using Double Antibody Sandwich Enzyme Linked Immunosorbent Assay (DAS-ELISA). DAS-ELISA result analyzed further to establish resistance category. Description to melon cultivar phenotype variation was done. The result of this research indicates that MG3 melon cultivar is tolerant to KGMMV. The decrease of MG3 optical density was directly related with the lowering of KGMMV symptoms. The character of tolerance to KGMMV was inherited from Melodi Gama 1 (MG1) cultivar. Melon (Cucumis melo L.) merupakan tanaman buah yang tergolong dalam familia Cucurbitaceae. Tanaman melon berpotensi untuk dikembangkan sebagai produk unggulan hortikultura di Indonesia. Tanaman melon juga merupakan salah satu penghasil devisa penting Indonesia dan menempati urutan ke-5 dari kelompok hortikultura. Salah satu kendala yang sering dihadapi oleh petani melon adalah penyakit mosaik yang disebabkan oleh Kyuri green mottle mosaic virus (KGMMV). Infeksi KGMMV pada pertanian melon mengakibatkan penurunan kualitas dan kuantitas hasil, sehingga petani mengalami kerugian ekonomi yang cukup berarti. Melodi Gama 3 (MG3) merupakan kultivar melon unggul hasil rakitan Laboratorium Genetika, Fakultas Biologi, Universitas Gadjah Mada. Penggunaan kultivar melon yang tahan terhadap infeksi KGMMV secara genetis merupakan alternatif yang sangat bermanfaat dalam bidang pertanian. Penelitian ini dilakukan untuk mengetahui pewarisan ketahanan MG3 terhadap infeksi KGMMV. Melon kultivar MG3, ditumbuhkan di greenhouse. Sebagai pembanding digunakan melon kultivar yang umum ditanam petani, yaitu MAI, Glamour, Ladika, dan Action. Kelima kultivar melon tersebut diinokulasi dengan KGMMV. Parameter ketahanan KGMMV yang digunakan adalah segregasi gejala dan uji serologis dengan Double Antibody Sandwich Enzyme Linked Immunosorbent Assay (DAS-ELISA). Hasil DAS-ELISA selanjutnya dianalisis untuk mengetahui kategori ketahanannya. Dilakukan pula deskripsi pada variasi fenotip kultivar melon yang ditanam. Hasil penelitian ini menunjukkan bahwa tanaman melon kultivar Melodi Gama 3 memiliki sifat toleransi terhadap infeksi KGMMV. Toleransi ditunjukkan dengan nilai optical density (OD) yang menurun seiring dengan penurunan gejala infeksi KGMMV. Sifat ketahanan terhadap KGMMV diwariskan dari kultivar Melodi Gama 1 (MG1).


2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Saengsoon Charoenvilaisiri ◽  
Channarong Seepiban ◽  
Mallika Kumpoosiri ◽  
Sombat Rukpratanporn ◽  
Nuchnard Warin ◽  
...  

Abstract Background Cassava mosaic disease (CMD) is one of the most devastating viral diseases for cassava production in Africa and Asia. Accurate yet affordable diagnostics are one of the fundamental tools supporting successful CMD management, especially in developing countries. This study aimed to develop an antibody-based immunoassay for the detection of Sri Lankan cassava mosaic virus (SLCMV), the only cassava mosaic begomovirus currently causing CMD outbreaks in Southeast Asia (SEA). Methods Monoclonal antibodies (MAbs) against the recombinant coat protein of SLCMV were generated using hybridoma technology. MAbs were characterized and used to develop a triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) for SLCMV detection in cassava leaves and stems. Assay specificity, sensitivity and efficiency for SLCMV detection was investigated and compared to those of a commercial ELISA test kit and PCR, the gold standard. Results A TAS-ELISA for SLCMV detection was successfully developed using the newly established MAb 29B3 and an in-house polyclonal antibody (PAb) against begomoviruses, PAb PK. The assay was able to detect SLCMV in leaves, green bark from cassava stem tips, and young leaf sprouts from stem cuttings of SLCMV-infected cassava plants without cross-reactivity to those derived from healthy cassava controls. Sensitivity comparison using serial dilutions of SLCMV-infected cassava sap extracts revealed that the assay was 256-fold more sensitive than a commercial TAS-ELISA kit and 64-fold less sensitive than PCR using previously published SLCMV-specific primers. In terms of DNA content, our assay demonstrated a limit of detection of 2.21 to 4.08 × 106 virus copies as determined by quantitative real-time PCR (qPCR). When applied to field samples (n = 490), the TAS-ELISA showed high accuracy (99.6%), specificity (100%), and sensitivity (98.2%) relative to the results obtained by the reference PCR. SLCMV infecting chaya (Cnidoscolus aconitifolius) and coral plant (Jatropha multifida) was also reported for the first time in SEA. Conclusions Our findings suggest that the TAS-ELISA for SLCMV detection developed in this study can serve as an attractive tool for efficient, inexpensive and high-throughput detection of SLCMV and can be applied to CMD screening of cassava stem cuttings, large-scale surveillance, and screening for resistance.


Bragantia ◽  
2007 ◽  
Vol 66 (1) ◽  
pp. 61-68 ◽  
Author(s):  
Rosa Maria Chung ◽  
Joaquim Adelino de Azevedo Filho ◽  
Addolorata Colariccio

O trabalho teve como meta avaliar a reação de 18 linhagens superiores do programa de melhoramento de alface (Lactuca sativa L.) do IAC e de seis cultivares comerciais, ao Lettuce mosaic virus (LMV). Em condições de campo, na região de Atibaia (SP), foram observados sintomas de mosaico, nanismo e necrose em plantas das cultivares Rider, 'Karla H25' e Hortência. O vírus presente nos isolados foi identificado por meio de inoculação mecânica em plantas indicadoras e diferenciadoras e de testes sorológicos de Plate Trapped Antigen-Enzyme linked-immunosorbent assay (PTA-ELISA). Nas amostras avaliadas, identificou-se a espécie LMV pelo PTA-ELISA e do patotipo IV pela reação nas hospedeiras diferenciais. Para a avaliação do comportamento dos genótipos de alface, foi empregado o LMV isolado 'Karla H25'. Foram submetidos à inoculação 24 genótipos de alface empregando-se, como controle positivo, a alface 'White Boston' por sua suscetibilidade ao LMV. O delineamento experimental foi inteiramente ao acaso e analisado pelo teste do qui-quadrado. Detectaram-se genótipos com comportamento de suscetibilidade e de tolerância. Nos genótipos 3 e 4, foram observadas plantas com comportamento de tolerância ao LMV isolado 'Karla H25', enquanto nos demais genótipos, constataram-se plantas com comportamento suscetível. O plantio de cultivares tolerantes pode ser uma alternativa aos prejuízos causados pela infecção pelo LMV com conseqüente diminuição do uso de produtos químicos para o controle dos afídeos vetores.


1980 ◽  
Vol 10 (3) ◽  
pp. 278-283 ◽  
Author(s):  
T. Hardcastle ◽  
A. R. Gotlieb

An enzyme-linked immunosorbent assay (ELISA) method was developed to detect apple mosaic virus (ApMV) in Betulaalleghaniensis Britton. Tests for virus using bark and bud tissues of dormant trees were successful. ApMV was also detectable in old leaf tissue in August and September, as well as in newly emerging leaf tissue forced in a greenhouse in March. Whole crude antiserum used to coat ELISA plates in tests with bud tissues was a reliable substitute for purified immunoglobulin without loss of sensitivity or specificity. An attempt was made to use ELISA for quantifying virus concentrations in field samples of ApMV-infected birch leaves.


1986 ◽  
Vol 32 (2) ◽  
pp. 189-192 ◽  
Author(s):  
R. Vincent Miller ◽  
Thomas W. Carroll ◽  
David C. Sands

A protocol was developed for testing chemical treatments for activity against seed-borne barley stripe mosaic virus. Vantage barley seeds infected with the MI-3 strain of the virus were allowed to imbibe dimethylsulfoxide solutions of test compounds. Of 49 compounds tested, alone or in combination with butylated hydroxyanisol, 18 reduced the number of plants expressing viral symptoms by up to 50%. Using an enzyme-linked immunosorbent assay to detect barley stripe mosaic virus, less than 2% of the asymptomatic plants arising from five chemical treatments were found to contain detectable virus antigen. In some treatments, reductions in the number of emerged plants with virus symptoms were correlated with reduced emergence.


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