scholarly journals EFICIÊNCIA NA DETECÇÃO DOS VÍRUS PLRV, PVX E PVY EM TECIDOS VEGETAIS DE GEMA APICAL DORMENTE MAIS ESTOLÃO, BROTOS E FOLHAS DA BATATA, PELO MÉTODO SOROLÓGICO ENZYME-LINKED IMMUNOSORBENT ASSAY-ELISA

2000 ◽  
Vol 1 (1) ◽  
pp. 80 ◽  
Author(s):  
Milton Vasconcelos GUEDES
Keyword(s):  
Solanum Tuberosum ◽  
Immunosorbent Assay ◽  
Solanum Tuberosum L ◽  
Enzyme Linked Immunosorbent Assay ◽  
De Se ◽  
Das Elisa

A propagação vegetativa da batata (Solanum tuberosum L.) é a causa da degenerescência de variedades uma vez que o tubérculo-semente utilizado no plantio pode perpetuar os vírus incidentes na planta-mãe. Daí a necessidade de renovação constante de estoques de batata-semente, bem como, de se obter técnicas eficientes de diagnóstico. Os objetivos gerais deste trabalho foram (1) comparar a eficiência na detecção dos vírus PLRV, PVX e PVY por meio do teste sorológico DAS-ELISA em tecidos vegetais de diferentes origens:- da gema apical + estolão, brotos e folhas, após quebra natural de dormência, das variedades Baraka, Contenda e Elvira; (2) verificar a confiabilidade da detecção de vírus em tecidos vegetais dormentes ou récem-brotados, visando reduzir o tempo de obtenção dos resultados em relação ao uso de folhas. Os objetivos específicos foram: avaliar a incidência dos diferentes vírus em amostras de campos de batata-consumo, na segunda geração de multiplicação; comparar os resultados obtidos com o equipamento de leitura de ELISA e os da leitura visual. A homogeneidade das variâncias dos tratamentos foi testada pelo teste de Bartlett e a comparação das médias dos tratamentos foi feita utilizando-se o teste de Tukey, ambos ao nível de 5% de probabilidade. Os resultados foram analisados utilizando-se o programa estatístico MSTATC, versão 2.11. A incidência média das diferentes viroses nas amostras das variedades Baraka, Contenda e Elvira, foi maior para PVY e PLRV, e muito baixa para PVX. O PVY foi o vírus de maior incidência nas três variedades: 14,1% em Baraka, 10,9% em Elvira e 2,9% em Contenda. O PLRV ocorreu com incidência de 3,6% na variedade Elvira, 2,3% em Baraka e 0,7% em Contenda. O PVX foi detectado em níveis inferiores a 0,5% nas três variedades; a variedade Contenda foi a que apresentou os menores índices médios de incidência para os três vírus, quando comparados às demais variedades estudadas; o teste DAS-ELISA pode ser utilizado na detecção dos vírus PLRV e PVX, tanto em gema apical dormente + estolão, como em brotos ou folhas, após quebra natural de dormência nas variedades Baraka, Contenda e Elvira; a eficiência do teste DAS ELISA pode variar de acordo com a origem do tecido vegetal utilizado; o vírus PVY, não pode ser detectado com segurança na gema apical dormente + estolão, para as três variedades testadas. Tendo em vista os resultados apresentados pode-se concluir que a detecção das viroses em brotos foi mais eficiente para todas as variedades estudadas antecipando-se os resultados e proporcionando um ganho de no mínimo 30 dias.

scholarly journals Variations of Leek yellow stripe virus Concentration in Garlic and Its Incidence in Argentina

Plant Disease ◽  
2002 ◽  
Vol 86 (10) ◽  
pp. 1085-1088 ◽  
Author(s):  
Vilma C. Conci ◽  
Pablo Lunello ◽  
Diana Buraschi ◽  
Rusell R. Italia ◽  
Sergio F. Nome
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Concentration ◽  
Santa Cruz ◽  
Leek Yellow Stripe Virus ◽  
Random Samples ◽  
Crop Cycle ◽  
White Type ◽  
Yellow Stripe ◽  
Das Elisa

The purpose of this work was to determine variations in titer of Leek yellow stripe virus (LYSV) throughout the crop cycle and bulb storage, and to evaluate the incidence of infected plants in the main garlic-production regions of Argentina. One hundred plants with LYSV from each of five cultivars were analyzed by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) in six different vegetative stages in White- and Red-type garlic cultivars and seven stages in cv. Rosado Paraguayo, throughout the year. In two White-type garlic cultivars, LYSV showed peaks of viral concentration in May, at the beginning of the crop cycle, and in November, just before harvest. In two Red-type garlic selections, an increase was detected in November (period of bulbing). The highest virus titers for these four garlic cultivars were detected in devernalized clove. In Rosado Paraguayo, the peak virus concentration occurred in September prior to harvesting. In a survey at 14 different localities in Argentina, 3,066 random samples were analyzed. LYSV was found in 80 to 98% of the plants from all regions, except in Santa Cruz, where 34% of plants were infected. The importance of this study is that it allows us to recommend the most suitable moment of the year to make the analysis with DAS-ELISA.

scholarly journals Detección serológica y molecular del Potato virus X (PVX) en tubérculos-semilla de papa (Solanum tuberosum L. y Solanum phureja Juz. Bukasov) en Antioquia, Colombia

2016 ◽  
Vol 18 (1) ◽  
Author(s):  
Daniela García Ruíz ◽  
Manuel Andrés Olarte Quintero ◽  
Pablo Andrés Gutiérrez Sánchez ◽  
Mauricio Alejandro Marín Montoya
Keyword(s):  
Solanum Tuberosum ◽  
Potato Virus ◽  
Solanum Tuberosum L ◽  
Potato Virus X ◽  
Solanum Phureja ◽  
Das Elisa

<p>El Potato virus (PVX) es uno de los virus más limitantes del cultivo de la papa en el mundo. Es transmitido solamente por contacto y por tubérculo-semilla. Su control se fundamenta en la siembra de tubérculos certificados por su sanidad viral y en la disponibilidad de metodologías de diagnóstico altamente sensibles. En este trabajo se evaluó la prevalencia del PVX en cuatro diferentes tejidos de tubérculos de Solanum tuberosum subsp. andigena var. Diacol-Capiro y S. phureja var. Criolla Colombia utilizando pruebas de DAS-ELISA para 128 submuestras y de RT-qPCR para 32 grupos de submuestras (4 submuestras/grupo). Los resultados de las pruebas serológicas  indicaron la presencia de PVX en el 6,25 y 50% de las submuestras analizadas para la variedad Diacol-Capiro y Criolla Colombia, respectivamente; mientras que los niveles de prevalencia del PVX utilizando la detección por RT-qPCR fueron del 93,75%, independientemente de la variedad de papa y del tejido evaluado. Los valores promedio del ciclo umbral (Ct) en las RT-qPCR fueron de 25,6 (Ct=18,02 a 34,49) y el análisis de las curvas de desnaturalización permitió identificar dos variantes del virus con valores de Tm de 79,5±1°C y 83,7±1°C. La secuenciación de los amplicones obtenidos por RT-qPCR para los controles positivos y para dos de las muestras, confirmó su naturaleza viral. Estos resultados señalan unos muy altos niveles de prevalencia de PVX en el material de siembra de papa en Antioquia y la necesidad de fortalecer los programas de certificación de semilla con pruebas de detección como RT-qPCR.</p>

scholarly journals DETECTION AND ELIMINATION VIRUS ON POTATO (Solanum tuberosum L.)

2018 ◽  
Vol 5 (2) ◽  
pp. 92
Author(s):  
Debora Margareth ◽  
Ida Ayu Astarini ◽  
I Gede Rai Maya Temaja
Keyword(s):  
Solanum Tuberosum ◽  
Serological Test ◽  
Solanum Tuberosum L ◽  
Sampling Technique ◽  
Purposive Sampling ◽  
Potato Plants ◽  
Potato Crops ◽  
Das Elisa ◽  
Random Program

This study aims to determine the symptoms of virus attacks on potato crops, identification of viruses that attack potato plants in Pancasari village, Bali, efectivity of thermotherapy technique to eliminate in vivo potato. The research was taken using purposive sampling technique and complete random program. Virus symptoms were observed on two potato fields. Serological test employed DAS-ELISA with PVX, PVS, PVY and PLRV antiserum. In vivo elimination virus were done by thermoteraphy in 37°C±1°C (4 days), 34°C±1°C (3 days) on 3 weeks and 33°C±1°C (4 days), 30°C±1°C (3 days) on 2 weeks. Results of the study at Pancasari Village Bali shown varied symptoms on potato leaves, such as mottle, mosaic, malformation, leafroll, witchesbroom. Through serological test, had identified viruses were 11.11% PVX, 55.56% PVS, 80% PVY and 11.11% PLRV which were infected farmer’s potato. In vivo thermotherapy was succeed decreasing PVS and PVY level during 2 weeks on 33°C±1°C (4 days) and 30°C±1°C (3 days).

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 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 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.

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