scholarly journals PEWARISAN KETAHANAN MELON (Cucumis melo L.) KULTIVAR MELODI GAMA 3 TERHADAP Kyuri green mottle mosaic virus (RESISTANCE’S INHERITANCE TO Kyuri green mottle mosaic virus IN MELON (Cucumis melo L.) MELODI GAMA 3 CULTIVAR)

2017 ◽  
Vol 20 (2) ◽  
pp. 59 ◽  
Author(s):  
Budi Setiadi Daryono ◽  
Faizatul Fitriyah
Keyword(s):  
Mosaic Virus ◽  
Optical Density ◽  
Immunosorbent Assay ◽  
Cucumis Melo ◽  
Mosaic Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Financial Loss ◽  
Double Antibody ◽  
Cucumis Melo L ◽  
Das Elisa

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).

scholarly journals Development of a triple antibody sandwich enzyme-linked immunosorbent assay for cassava mosaic disease detection using a monoclonal antibody to Sri Lankan cassava mosaic virus

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Saengsoon Charoenvilaisiri ◽  
Channarong Seepiban ◽  
Mallika Kumpoosiri ◽  
Sombat Rukpratanporn ◽  
Nuchnard Warin ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Mosaic Disease ◽  
Cassava Mosaic Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sri Lankan ◽  
Stem Cuttings ◽  
Field Samples ◽  
Cassava Stem ◽  
Cassava Mosaic Virus

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.

scholarly journals Deteksi Squash mosaic virus pada Lima Varietas Mentimun (Cucumis sativus L.)

2017 ◽  
Vol 8 (2) ◽  
pp. 104 ◽  
Author(s):  
Erika Rosminim Purba ◽  
Susanti Mugi Lestari ◽  
Yudia Nurhaelena ◽  
Sri Hendrastuti Hidayat
Keyword(s):  
Mosaic Virus ◽  
Disease Severity ◽  
Immunosorbent Assay ◽  
Disease Incidence ◽  
Virus Titer ◽  
Seed Transmission ◽  
Mosaic Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cucumis Sativus L ◽  
Generative Phase

<p align="center"><strong><em>ABSTRACT</em></strong></p><p><em><span style="text-decoration: underline;">Squash</span></em><em> <span style="text-decoration: underline;">mosaic</span> <span style="text-decoration: underline;">virus</span> </em><em>(SqMV) is a seed-borne pathogen which infect many <span style="text-decoration: underline;">Cucurbitaceae </span>crops.  Infection of SqMV has been reported from several vegetable growing areas in Indonesia. The objective of this research was to determine the percentage of seed-borne SqMV on five cucumber varieties i.e. ‘Jupiter’, ‘Venus’, ‘Japan File’, ‘Vario’, and ‘Calista’ and the effect of SqMV infection on mosaic disease development. Five cucumber varieties were mechanically inoculated with SqMV, followed by observation on symptom development, incubation period, and disease incidence. Seed-borne virus was detected by Dot Immunobinding Assay (DIBA) and indirect Enzyme-linked immunosorbent assay (ELISA) methods following growing-on test. The plants showed varied symptoms including green mosaic, green-yellow mosaic, vein clearing, and fruit malformation. Disease severity and virus titer showed general trend, i.e. low during inflorescence period and increasing on fruiting period; with the exception on ‘Japan File’ which showed decreasing of disease severity since generative phase. All commercial seeds (F1) tested evidently infected by SqMV with high incidence (100%), whereas infection of SqMV on F2 seeds of ‘Venus’ reached 60.87%.</em></p><p><em>Keywords: DIBA, disease incidence, ELISA, seed transmission, virus titer</em></p><p align="center"><strong> </strong></p><p align="center"><strong> </strong></p><p align="center"><strong>ABSTRAK</strong><strong> </strong></p><p><em>Squash mosaic virus </em>(SqMV) adalah patogen terbawa benih yang banyak menginfeksi tanaman <em>Cucurbitaceae</em>, dan keberadaannya di Indonesia sudah meluas. Tujuan penelitian ialah mengetahui persentase SqMV terbawa benih pada lima varietas mentimun yaitu ‘Yupiter’, ‘Venus’, ‘Japan File’, ‘Vario’, dan ‘Calista’ dan mengetahui pengaruh infeksi SqMV terhadap perkembangan penyakit mosaik. Lima varietas mentimun diinokulasi dengan SqMV secara mekanis kemudian diamati gejala yang muncul, periode inkubasi, dan insidensi penyakit. Pengujian virus terbawa benih dilakukan dengan menumbuhkan benih, selanjutnya deteksi virus dilakukan menggunakan metode <em>Dot Immunobinding Assay </em>(DIBA)<em> </em>dan<em> indirect Enzyme-linked immunosorbent assay </em>(<em>ELISA</em>). Tanaman mentimun menunjukkan gejala infeksi SqMV yang bervariasi yaitu mosaik hijau, mosaik kuning hijau, pemucatan tulang daun, dan malformasi pada buah. Pengamatan keparahan penyakit dan titer virus menunjukkan pola perkembangan penyakit mosaik yaitu menurun pada fase berbunga dan meningkat lagi pada fase berbuah, kecuali varietas Japan File memberikan respons yang berbeda karena penurunan keparahan penyakit berlanjut sejak fase generatif. Benih komersial (F1) yang banyak digunakan petani terbukti membawa SqMV dengan infeksi mencapai 100% dan tanaman varietas ‘Venus’ yang terinfeksi SqMV menghasilkan benih keturunan (F2) yang membawa SqMV dengan efisiensi mencapai 60.87%.</p><p>Kata kunci: DIBA, ELISA, insidensi penyakit, titer virus, tular benih</p>

scholarly journals Insiden Penyakit Virus Tular Umbi pada Tigabelas Varietas Bawang Merah Asal Jawa Barat dan Jawa Tengah

2016 ◽  
Vol 21 (2) ◽  
pp. 164
Author(s):  
Neni Gunaeni ◽  
Astri W Wulandari ◽  
Ati Srie Duriat ◽  
Agus Muharam
Keyword(s):  
Immunosorbent Assay ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Onion Yellow Dwarf Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Diseases ◽  
Double Antibody ◽  
Yellow Dwarf Virus ◽  
Yellow Stripe ◽  
Das Elisa

Penyakit virus tular umbi merupakan salah satu kendala dalam meningkatkan produksi bawang merah. Hal ini disebabkan oleh virus yang infeksinya bersifat sistemik. Apabila partikel virus berada dalam jaringan benih umbi, maka akan sulit untuk dikendalikan dan dapat membawa masalah baru pada pertanaman berikutnya. Penelitian bertujuan mengetahui insiden penyakit virus tular umbi pada 13 varietas bawang merah yang berasal dari Jawa Barat dan Jawa Tengah. Penelitian dilaksanakan di Rancaekek (elevasi 650 m dpl.) dan di Laboratorium Balai Penelitian Tanaman Sayuran Lembang (elevasi 1.250 m dpl.), sejak bulan Agustus sampai November 2004. Perlakuan terdiri atas 13 varietas bawang merah, yaitu: Lodra, Sumenep, Batu, Merah Maja, Merah Cigugur, Ciniru, Bima, Bima Curut, Bima Timor, Bima Arjuna, Kuning Tablet, Kuning Gombong, dan Philipina. Rancangan yang digunakan ialah acak kelompok dengan tiga ulangan. Hasil penelitian menunjukkan bahwa (1) insiden penyakit virus tular umbi pada masing-masing varietas bawang merah asal Jawa Barat dan Jawa Tengah berturut-turut yaitu varietas Lodra 84,67%, Sumenep 82,56-100%, Batu 39,86-78,67%, Merah Maja 95,25%, Merah Cigugur 100%, Ciniru 66,27%, Bima Curut 78,57%, Bima 100%, Bima Timor 57,98%, Bima Arjuna 47,96%, Kuning Tablet 57,48%, Kuning Gombong 97,92%, dan Philipina 97,92 %, (2) gejala infeksi virus pada daun umumnya berupa  klorosis, mosaik bergaris kuning vertikal terputus-putus, garis-garis hijau vertikal, dan ukuran daun menjadi kecil, (3) gejala-gejala tersebut bereaksi positif dengan OYDV(onion yellow dwarf virus) dan SYSV (shallot yellow stripe virus) berdasarkan uji DAS-ELISA (double antibody sandwich-enzyme linked immunosorbent assay). Informasi mengenai insiden virus tular umbi pada bawang merah ini sangat penting dalam rangka mengembangkan metode perbenihan bawang merah bebas virus. <br /><br />Virus disease is one of major problems in increasing shallots production, because its infection has a systemic character. If it is already in shallots bulb tissues, the virus is difficult to be controlled and will cause new problems to the next planting. The experiment was aimed to determine incidence of bulb-borne virus diseases on  thirteen varieties of shallots (Allium cepa var. ascalonicum) originated from West  and Central Java. The experiment was carried out at Indonesian Vegetable Research Institute Lembang (1,250 m asl.) and Rancaekek (650 m asl.), from  August to November 2004. The shallot varieties tested were Lodra, Sumenep, Batu, Merah Maja, Merah Cigugur, Ciniru, Bima, Bima Curut, Bima Timor, Bima Arjuna, Kuning Tablet, Kuning Gombong, and Philipina. A randomized complete block design with three replications were used in this experiment. The results of the experiment showed that  (1) incidence of virus diseases in shallots bulb on variety Lodra was 84.67%, Sumenep 82.56-100%, Batu 39.86-78.67%, Merah Maja 95.25%, Merah Cigugur 100%, Ciniru 66.27%, Bima Curut 78.57%, Bima 100%, Bima Timor 57.98%, Bima Arjuna 47.96%, Kuning Tablet 57.48%, Kuning Gombong 97.92%,  and Philipina 97.92 %, (2) the virus symptoms exhibited on infected shallots were  yellow stripe mosaic, chlorosis,  green stripe leaf,  and leaves became small, and (3) the symptoms were associated with OYDV (onion yellow dwarf virus) and SYSV (shallots yellow stripe virus) base on DAS-ELISA (double antibody sandwich-enzyme linked immunosorbent assay). Information on the incidence of viral diseases on shallots bulb is very important to develop the production technology of virus-free shallots bulb.<br /><br /><br />

scholarly journals Differentiation between Human Coronaviruses NL63 and 229E Using a Novel Double-Antibody Sandwich Enzyme-Linked Immunosorbent Assay Based on Specific Monoclonal Antibodies

2010 ◽  
Vol 18 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Patricia Sastre ◽  
Ronald Dijkman ◽  
Ana Camuñas ◽  
Tamara Ruiz ◽  
Maarten F. Jebbink ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Respiratory Tract ◽  
Immunosorbent Assay ◽  
Respiratory Tract Infections ◽  
Enzyme Linked Immunosorbent Assay ◽  
Double Antibody ◽  
Human Coronaviruses ◽  
Tract Infections ◽  
Das Elisa ◽  
Potential Tool

ABSTRACTHuman coronaviruses (HCoVs) are responsible for respiratory tract infections ranging from common colds to severe acute respiratory syndrome. HCoV-NL63 and HCoV-229E are two of the four HCoVs that circulate worldwide and are close phylogenetic relatives. HCoV infections can lead to hospitalization of children, elderly individuals, and immunocompromised patients. Globally, approximately 5% of all upper and lower respiratory tract infections in hospitalized children are caused by HCoV-229E and HCoV-NL63. The latter virus has recently been associated with the childhood disease croup. Thus, differentiation between the two viruses is relevant for epidemiology studies. The aim of this study was to develop a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) as a potential tool for identification and differentiation between HCoV-NL63 and HCoV-229E. The nucleocapsid (N) proteins of HCoV-NL63 and HCoV-229E were expressed in anEscherichia colisystem and used to immunize mice in order to obtain monoclonal antibodies (MAbs) specific for each virus. Three specific MAbs to HCoV-NL63, one MAb specific to HCoV-229E, and four MAbs that recognized both viruses were obtained. After their characterization, three MAbs were selected in order to develop a differential DAS-ELISA. The described assay could detect up to 3 ng/ml of N protein and 50 50% tissue culture infective doses/ml of virus stock. No cross-reactivity with other human coronaviruses or closely related animal coronaviruses was found. The newly developed DAS-ELISA was species specific, and therefore, it could be considered a potential tool for detection and differentiation of HCoV-NL63 and HCoV-229E infections.

scholarly journals Antalya İli Kumluca İlçesindeki Biber Yetiştirilen Seralarda Domates Lekeli Solgunluk Virüsü (Tomato spotted wilt tospovirus-TSWV)’ nün Yaygınlığı ve Konukçu Yabancı Ot Türlerinin Belirlenmesi

2022 ◽  
Vol 9 (sp) ◽  
pp. 2565-2570
Author(s):  
Serkan Yeşil ◽  
Özder Gömlekli
Keyword(s):  
Plant Species ◽  
Immunosorbent Assay ◽  
Virus Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Double Antibody ◽  
Tomato Spotted Wilt ◽  
Plant Families ◽  
Das Elisa ◽  
Pepper Plants

There are many viruses that infect pepper and limit its production. Among these viruses, Tomato spotted wilt tospovirus (TSWV) infects crops in 35 plant families that are economically important, including pepper. In the present study; totaly 156 leaf samples were collected, including 57 from pepper plants showing virus-like symptoms and 99 from weeds and/or plants other than peppers in and around the greenhouse, through surveys carried out in pepper greenhouses in Kumluca district of Antalya province, from September to December 2020. Then, the plant leaf samples were tested to determine TSWV infections by the Double Antibody Sandwich Enzyme-linked Immunosorbent Assay (DAS-ELISA) method. According to result of the tests, it was determined that 55.76% of the tested leaf samples were infected with TSWV, while this rate was determined as 96.49% for pepper samples and 32.32% for other plant samples. During the survey studies, it was revealed that the leaf samples of 13 out of 31 weed and different plant species except pepper were infected with at least one of the viruses. In addition, pepper plants showing symptoms TSWV-like symptoms in pepper greenhouses were counted during the survey, and the prevalence of this virus disease was calculated on the basis of Kumluca district and neighborhoods. As a result of these calculations, the prevalences of TSWV; for Kumluca, Mavikent, Beykonak, Salur, Hacıveliler, Adrasan, Merkez, and Kavakköy were determined as 26.93%, 26.92%, 32.27%, 20.66%, 21.13%, 17.66%, 13%, and 25%, respectively.

scholarly journals Inheritance of Resistance to Watermelon Mosaic Virus in Cucumis melo L.

HortScience ◽  
1994 ◽  
Vol 29 (2) ◽  
pp. 107-110 ◽  
Author(s):  
Raphael Z. Gilbert ◽  
Molly M. Kyle ◽  
Henry M. Munger ◽  
Stewart M. Gray
Keyword(s):  
Mosaic Virus ◽  
Cucumis Melo ◽  
Dominant Gene ◽  
Zucchini Yellow Mosaic Virus ◽  
Leaf Tissue ◽  
Yellow Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Watermelon Mosaic Virus ◽  
Papaya Ringspot Virus ◽  
Cucumis Melo L

Resistance to watermelon mosaic virus (WMV) was transferred by successive backcrossing with selection from Cucumis melo PI 414723 to three melon varieties. Levels of resistance to virus accumulation in leaf tissue were evaluated using enzyme-linked immunosorbent assay, and procedures are described to select resistant individuals efficiently and accurately in segregating populations. Resistance is controlled by a single dominant. gene designated Wmr. Plants that carry this gene initially develop mosaic symptoms on inoculated leaves, but eventually recover from symptoms, and low or no virus can be detected in the youngest leaves. In contrast, susceptible plants show similar symptoms initially, but remain stunted and symptomatic with reduced fruit yield and fruit quality. Co-infection with other cucurbit viruses, specifically cucumber mosaic virus, papaya ringspot virus, and zucchini yellow mosaic virus, did not overcome resistance to WMV conferred by Wmr.

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