PCR multiplex para a detecção de BSV e CMV em bananeiras micropropagadas

Um protocolo de PCR multiplex foi estabelecido para a detecção do Banana streak virus (BSV) e do Cucumber mosaic virus (CMV) em bananeiras micropropagadas. Estes vírus são responsáveis por perdas na produção de bananas em todo o mundo. Alguns trabalhos descrevem a integração do BSV no genoma B da bananeira. Contudo, a existência de bananeiras híbridas livres do BSV tem sido demonstrada. Ademais, determinadas estirpes do CMV não são transmitidas mecanicamente sob condições de laboratório, nem tampouco detectadas por testes sorológicos. Como conseqüência, a indexação de matrizes para cultura de tecido algumas vezes se mostra ineficiente. A metodologia apresentada neste trabalho sobrepõe esta dificuldade, pois se baseia na detecção do ácido nucléico viral presente em amostras foliares de bananeira. Na reação, foram usados os oligonucleotídeos BADNA 1A e BADNA 4, para a detecção do BSV, e "CMV senso" e "CMV antisenso" para a detecção do CMV. Após a eletroforese foi verificada a presença de dois fragmentos de DNA amplificados simultaneamente, um dos quais com 597 pb correspondente ao BSV e o outro, com 488 pb, correspondente ao CMV. Este resultado indica que o PCR multiplex pode ser utilizado como uma ferramenta adicional na indexação do BSV e do CMV em bananeiras propagadas por cultura de tecido.

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DETEKSI CMV DAN PYMoV PADA BENIH LADA (Piper nigrum) DENGAN TEKNIK ELISA

Buletin Penelitian Tanaman Rempah dan Obat ◽

10.21082/bullittro.v27n2.2016.155-162 ◽

2017 ◽

Vol 27 (2) ◽

pp. 155

Author(s):

Maya Mariana ◽

NFN Miftakhurohmah

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Mottle Virus ◽

Piper Nigrum ◽

Streak Virus ◽

Banana Streak Virus ◽

Double Antibody ◽

Yellow Mottle ◽

Das Elisa

Salah satu kendala penting pada budidaya tanaman lada yaitu penyakit kerdil yang disebabkan oleh Piper yellow mottle virus (PYMoV) dan Cucumber mosaic virus (CMV). Tindakan pencegahan penyebaran penyakit dapat dilakukan salah satunya dengan penggunaan benih sehat bebas dari infeksi PYMoV dan CMV. Tujuan penelitian adalah mendeteksi keberadaan PYMoV dan CMV pada benih lada siap tanam secara serologi menggunakan teknik ELISA. Benih lada yang dideteksi berumur 5 bulan, berasal dari penangkar benih di Sukabumi (Jawa Barat) dan Purbalingga (Jawa Tengah). Varietas lada yang diambil di Sukabumi adalah Natar 1, Petaling, dan Lampung Daun Kecil., masing-masing sebanyak 10 sampel, sedangkan di Purbalingga hanya ada Natar 1, diambil 30 sampel. Di setiap lokasi pembibitan juga dilakukan pengamatan terhadap gejala infeksi virus yang ditemukan. Deteksi virus dilakukan secara Double Antibody Sandwich (DAS)-ELISA menggunakan antiserum Banana streak virus (BSV) untuk PYMoV dan antiserum CMV untuk deteksi CMV. Hasil ELISA dibaca nilai absorbannya dengan elisa reader pada panjang gelombang 405 nm. Sampel dinilai positif, jika nilai absorbansinya 1,5 kali lebih besar daripada kontrol negatif. Gejala infeksi virus pada benih bervariasi, yaitu klorotik, belang, dan belang disertai perubahan bentuk daun. Hasil pengujian ELISA menunjukkan bahwa 66% benih lada dari Sukabumi dan 46% dari Purbalingga terinfeksi virus CMV. Namun, tidak ada satu pun yang menunjukkan reaksi positif terhadap antiserum PYMoV. Hal ini diduga karena konsentrasi PYMoV terlalu rendah sehingga tidak terdeteksi secara ELISA. Untuk mencegah penyebaran virus, deteksi virus pada benih lada penting dilakukan sebelum digunakan sebagai bahan tanaman.

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Cucumber Mosaic Virus, Tobacco Streak Virus, and Cucumber Mosaic Virus Satellite RNA Associated with Mosaic and Ringspot Symptoms in Ajuga reptans in Ohio

Plant Disease ◽

10.1094/pdis.1997.81.10.1214d ◽

1997 ◽

Vol 81 (10) ◽

pp. 1214-1214 ◽

Cited By ~ 1

Author(s):

J. R. Fisher ◽

S. T. Nameth

Keyword(s):

United States ◽

Molecular Weight ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

The United States ◽

Negative Control ◽

Enzyme Linked Immunosorbent Assay ◽

Low Molecular Weight ◽

Streak Virus ◽

Ajuga Reptans

Creeping bugleweed (Ajuga reptans L.) is a perennial ornamental commonly grown as a ground cover in temperate climates. Commercial samples of the A. reptans cultivars Royalty, var. Atropurpurea Bronze, Bronze Beauty, and Burgundy Glow showing mosaic and ringspot symptoms were tested for the presence of virus infection by direct antibody sandwich enzyme-linked immunosorbent assay (ELISA) and viral-associated double-stranded (ds) RNA analysis. Cucumber mosaic cucumovirus (CMV) was detected by ELISA and dsRNA analysis in symptomatic samples of all cultivars tested. ELISA values were considered positive if the absorbance values were twice the negative control. Negative control values were established with asymptomatic tissue of the cv. Bronze Beauty. Tobacco streak ilarvirus (TSV) was detected only by ELISA in symptomatic samples of all cultivars except Royalty. No dsRNA suggestive of TSV was detected. Alfalfa mosaic virus (AMV) was detected by ELISA and dsRNA analysis in symptomatic samples of all cultivars tested except Royalty and var. Atropurpurea Bronze. dsRNA analysis also indicated the presence of a low molecular weight, possible satellite (sat) RNA associated with all symptomatic and asymptomatic Royalty and var. Atropurpurea Bronze plants tested. Northern (RNA) blot analysis with a digoxigenin-labeled full-length clone of the (S) CARNA-5 (-) CMV satRNA (ATCC no. 45124) confirmed that the low molecular weight RNA associated with the Royalty and var. Atropurpurea Bronze cultivars was indeed CMV satRNA. Only AMV has been previously reported in A. reptans in the United States (1). This is the first report of CMV and its satRNA, as well as TSV, in A. reptans in the United States. Reference: (1) W. T. Schroeder and R. Provvidenti. Plant Dis. Rep. 56:285, 1972.

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Banana streak virus Identified for the First Time in Peru in Cavendish Banana (Musa AAA)

Plant Disease ◽

10.1094/pdis-91-7-0906b ◽

2007 ◽

Vol 91 (7) ◽

pp. 906-906 ◽

Cited By ~ 1

Author(s):

C. Pasberg-Gauhl ◽

B. E. L. Lockhart ◽

F. Castro-Mendivil Dibos ◽

J. C. Rojas Llanque

Keyword(s):

Mosaic Virus ◽

Disease Incidence ◽

Horizontal Transmission ◽

Export Market ◽

Streak Virus ◽

Degenerate Primers ◽

Specific Primers ◽

Banana Streak Virus ◽

Cavendish Banana ◽

Northern Peru

In October of 2005, a field survey was done in the province of Piura in northern Peru to determine the cause of a disease known locally as “mosaico” that was affecting organic Cavendish banana (Musa AAA) grown for the export market. Disease symptoms consisted of pronounced chlorotic and necrotic lesions on leaves of affected plants. Twenty-four farms were visited, and at each location, 10 randomly selected plants at flowering stage were evaluated for disease incidence and severity. Plants showing virus-like symptoms were observed in 18 of the 24 locations (75%). Fifty-two banana leaf samples, 27 from plants showing virus-like symptoms and 25 from asymptomatic plants, were tested for the presence of Banana streak virus (BSV), Cucumber mosaic virus (CMV), and Banana mild mosaic virus (BanMMV) by immunosorbent electron microscopy (ISEM) using partially purified leaf tissue extracts (2).The same extracts were also tested by immunocapture PCR (IC-PCR) for presence of BSV and specific BSV isolates (BSV-OL, BSV-GF, BSV-IM, and BSV-CAV) using badnavirus-specific degenerate primers and BSV isolate-specific primers, respectively (1). Seventeen of 27 leaf samples showing virus-like symptoms (63%) tested positive for BSV by ISEM and IC-PCR using badnavirus, but not isolate-specific, primers. The symptoms on the 10 samples that tested negative were not typical of BSV infection. One asymptomatic leaf sample (4%) also tested positive for BSV. To validate the PCR results, the nucleotide sequence of the amplicon from a plant showing the most prevalent foliar symptom type was determined. This sequence (GenBank Accession No. DQ674317) had ≤86% homology to the corresponding ORF III polyprotein region of BSV and other badnaviruses. Neither CMV nor BanMMV was detected in any of the 52 samples tested. From these results, it was concluded that “mosaico” disease of organic Cavendish bananas in northern Peru is associated frequently with BSV infection and that there is a high incidence of BSV infection in this area. To our knowledge, this is the first report of BSV occurrence in Peru. It was both surprising and interesting that neither BSV-OL nor BSV-GF, the two BSV isolates found most commonly in banana (Musa AAA) and plantain (Musa AAB) in South and Central America (B. E. L. Lockhart, unpublished), was detected in Cavendish banana in northern Peru. Failure to detect BSV-OL and BSV-GF suggests that field infection may be due to vertical transmission by clonal propagation rather than to horizontal transmission from local plantain and that control of “mosaico” disease could therefore be achieved by use of virus-free planting material. References: (1) A. D. W. Geering et al. Phytopathology 90:921, 2000. (2) B. E. L. Lockhart et al. Phytopathology 82:921, 1992.

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First Report of Cucumber mosaic virus in Banana from Iran

Plant Disease ◽

10.1094/pd-89-0914a ◽

2005 ◽

Vol 89 (8) ◽

pp. 914-914 ◽

Cited By ~ 2

Author(s):

T. Ghotbi ◽

K. Bananej

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Plant Viruses ◽

Economic Losses ◽

Enzyme Linked Immunosorbent Assay ◽

Streak Virus ◽

Serological Tests ◽

Tropical Fruit ◽

Test Plants ◽

Das Elisa

Banana bunchy top virus (BBTV), Banana streak virus (BSV), and Cucumber mosaic virus (CMV) (genus Cucumovirus, family Bromoviridae [2]) cause widespread economic losses on banana (Musa sp.) throughout the world and have been reported on banana in different countries including Pakistan along its southeastern border with Iran (1). A survey was conducted from 2004–2005 to identify viruses infecting banana in greenhouses in different growing areas in northern Iran, Mazandaran Province (Sari, Babol, Behshahr, and Ghaemshahr cities). A total of 180 samples from seven banana-growing greenhouses with symptoms of mosaic, chlorosis, stunting, and fruit malformation were collected. All samples were tested for CMV with polyclonal antibodies using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) (CMV strain D subgroup I; gifted by H. Lecoq, INRA, Avignion, France). For sap inoculation onto indicator test plants, selected ELISA-positive leaf samples were ground in chilled 0.01 M phosphate buffer, pH 7.0, containing 0.15% 2-mercaptoethanol. Chlorotic and necrotic local lesions developed on Chenopodium amaranticolor and Vigna unguiculata (cv. Mashad local) 10 and 12 days postinoculation, respectively. Cucumis sativus and Nicotiana rustica also developed systemic mosaic symptoms (3). All indicator test plants were rechecked for the presence of CMV using DAS-ELISA. On the basis of serological tests and indicator host plants reactions, CMV was identified in 32% of samples including Sari (13.8%), Babol (2.7%), Behshahr (10%) and Ghahemshahr (5%), respectively. Fifty-five samples did not react with CMV antiserum but the presence of symptoms resembling BBTV and BSV (4) emphasizes the need for further investigations to confirm the presence and identities of other viruses. References: (1) J. Bird and F. L. Wellman. Phytopathology 52:286, 1962. (2) S. K. Choi et al. J. Virol. Methods 83:67, 1999. (3) A. J. Gibbs and B. D. Harrison. Descriptions of Plant Viruses. No.1. CMI/AAB, Surrey, England, 1970. (4) R. C. Ploetz et al., eds. Compendium of Tropical Fruit Diseases. The American Phytopathological Society, St. Paul, MN, 1994.

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Identificación y caracterización molecular del genoma completo de tres virus en cultivos de lulo (Solanum quitoense) de Antioquia (Colombia)

Revista Colombiana de Ciencias Hortícolas ◽

10.17584/rcch.2018v12i2.7692 ◽

2018 ◽

Vol 12 (2) ◽

pp. 281-292 ◽

Cited By ~ 8

Author(s):

Yuliana Gallo ◽

Luisa Fernanda Toro ◽

Helena Jaramillo ◽

Pablo Andrés Gutiérrez ◽

Mauricio Marín

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Yellow Vein ◽

Streak Virus ◽

Rt Pcr ◽

Potato Yellow Vein Virus

El lulo es uno de los renglones agrícolas más promisorios para la región andina de Colombia, gracias a sus excelentes características organolépticas y a su potencial para el procesamiento industrial. En Antioquia, se ha observado en los últimos años la presencia de plantas con síntomas de enfermedades virales, que incluyen amarillamientos intervenales, mosaicos y deformación de brotes. En este trabajo, se evaluó dicha situación en un grupo (bulk) de muestras foliares de plantas sintomáticas de lulo obtenidas en municipios del Oriente Antioqueño, utilizando la metodología molecular de secuenciación de nueva generación (NGS), y la confirmación posterior por RT-PCR. Los análisis bioinformáticos de las secuencias obtenidas (10.777.822 de reads) indicaron la presencia de tres virus de RNA en el transcriptoma evaluado; el Cucumber mosaic virus (CMV) se encontró en mayores niveles de infección (40,4% del total de reads), mientras que los otros virus correspondieron al Potato yellow vein virus (PYVV) (0,09%) y Alstroemeria necrotic streak virus (ANSV) (0,06%), siendo posible obtener sus genomas completos. La ocurrencia de los tres virus fue confirmada en plantas individuales de lulo mediante pruebas de RT-PCR/secuenciación Sanger con cebadores específicos diseñados a partir de los datos de NGS (ANSV y CMV), o con cebadores reportados previamente (PYVV). En el futuro será necesario evaluar los efectos de estos virus sobre los rendimientos, longevidad de plantas y calidad de semilla en los cultivos de lulo en el país, así como sus métodos de transmisión, rango de hospedantes y sintomatología específica.

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Infecção viral simples e dupla em Alstroemeria sp.

Revista Brasileira de Horticultura Ornamental ◽

10.14295/rbho.v5i1.38 ◽

1999 ◽

Vol 5 (1) ◽

Cited By ~ 1

Author(s):

Ligia M. L. Duarte ◽

Patrícia V. Seabra ◽

Eliana B. Rivas ◽

Sílvia R. Galleti ◽

Amélia V. Alexandre

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Tomato Spotted Wilt Virus ◽

Sao Paulo ◽

Tobacco Streak Virus ◽

Streak Virus ◽

São Paulo ◽

Tomato Spotted Wilt ◽

Wilt Virus

Amostras de Alstroemeria sp., provenientes de 2 regiões produtoras do Estado de São Paulo (Brasil) e da Itália (material quarentenado), foram submetidas a testes de transmissão mecânica, serológicos (DASELISA e decoração) e observações ao microscópio eletrônico. Das 33 amostras analisadas, 1 1 estavam infectadas por vírus, tendo sido identificados Cucumber mosaic virus (CMV), Tobacco streak virus (TSV) e Tomato spotted wilt virus (TSWV) tanto em infecções simples como mistas (TSV + CMV, TSV + TSWV e TSV + CMV + Potyviridae). Em 3 amostras infectadas foram observadas, ao microscópio eletrônico, partículas alongadas, porém não foi possível a identificação do vírus ao nível de espécie. Os dados obtidos mostram que a ocorrência de infecções mistas em Alstroemeria é um fato comum, envolvendo principalmente o TSV.

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