Evaluation of the Chilli veinal mottle virus CP gene expressing transgenic Nicotiana benthamiana plants for disease resistance against the virus / Avaliação do gene CP do vírus Chilli veinal mottle que expressa plantas transgênicas Nicotiana benthamiana quanto à resistência a doenças contra o vírus

Vegetables are an important source of income and high-value crops for small farmers. Chilli (Capsicum spp.) is one of the most economically important vegetables of Pakistan and it is grown throughout the country. It is a rich source of nutrition especially vitamins A, B, C and E along with minerals as folic acid, manganese (Mn), potassium (K) and molybdenum (Mo). Chilli possesses seven times more amount of vitamin C than an orange. Vitamin A, C and betacarotenoids are strong antioxidants to scavenge the free radicals. Chilli production is restricted due to various biotic factors. Among these viruses, Chilli veinal mottle virus (ChiVMV) is one of the most destructive and menacing agents that inflicts heavy and colossal losses that accounted for 50% yield loss both in quality and quantity. Pathogen-Derived Resistance (PDR) approach is considered one of the effective approaches to manage plant viruses. In this study, ChiVMV was characterized on a molecular level, the coat protein (CP) gene of the virus was stably transformed into Nicotiana benthamiana plants using Agrobacterium tumefaciens. The transgenic plants were challenged with the virus to evaluate the level of resistance of plants against the virus. It was observed that the plants expressing CP gene have partial resistance against the virus in terms of symptoms' development and virus accumulation. Translation of this technique into elite chilli varieties will be resulted to mitigate the ChiVMV in the crop as well as an economic benefit to the farmers.

Vegetais são uma importante fonte de renda e culturas de alto valor para os pequenos agricultores. A pimenta-malagueta (Capsicum spp.) é uma das hortaliças mais importantes economicamente do Paquistão e é cultivada em todo o país. É uma rica fonte de nutrição, especialmente vitaminas A, B, C e E com minerais como ácido fólico, manganês (Mn), potássio (K) e molibdênio (Mo). O pimentão possui sete vezes mais vitamina C do que a laranja. Vitaminas A e C e betacarotenoides são antioxidantes fortes para eliminar os radicais livres. A produção de pimenta é restrita devido a vários fatores bióticos. Entre esses vírus, o ChiVMV é o agente mais destrutivo e ameaçador que inflige perdas pesadas e colossais que representam 50% da perda de rendimento, tanto em qualidade quanto em quantidade. A abordagem de resistência derivada de patógenos (PDR) é considerada uma das abordagens eficazes para gerenciar os vírus de plantas. Neste estudo, ChiVMV foi caracterizado em nível molecular e o gene CP do vírus foi transformado de forma estável em plantas Nicotiana benthamiana usando Agrobacterium tumefaciens. As plantas transgênicas foram desafiadas com o vírus para avaliar seu nível de resistência contra o vírus. Observou-se que as plantas que expressam o gene CP apresentam resistência parcial ao vírus em termos de desenvolvimento de sintomas e acúmulo de vírus. A tradução dessa técnica em variedades de pimenta de elite resultará na mitigação do ChiVMV na safra, bem como em benefícios econômicos para os agricultores em termos de melhor rendimento e baixo custo de produção.

Characterization of Cowpea aphid-borne mosaic virus in Passiflora edulis 'Catarina' in the southern coast of Santa Catarina State, Brazil / Caracterização do Cowpea aphid-borne mosaic virus em Passiflora edulis cv. Catarina no litoral sul de Santa Catarina, Brasil

CABMV is a limiting virus for passion fruit crop in Brazil, its main producing country. This virus has been reported in all producing states of the country, with the state of Santa Catarina (SC) in 2017 standing as the third largest passion fruit producer. In 2017, it reached 8.4% of the national production. The southern coast is the main responsible for the increase in production, which has been supplying the domestic market. However, in that same year, this region recorded the first symptom expressions in plants and fruits. The evaluation of the samples collected in the municipalities of Sombrio, Praia Grande and São João do Sul, southern coast of SC, was performed by using a mechanical transmission to indicator plants, PTA-ELISA and RT-PCR, and by sequencing. The evaluation results were positive for CABMV and negative for CMV in PTA-ELISA. In RT-PCR, there was the formation of a 700bp ca band, expected size for Potyvirus, whose sequence comparison with those deposited in GenBank reveled 98% identity with the isolates from São Paulo State. The occurrence of the virus in the southern coast of SC did not reach a serious decrease in passion fruit production due to the union of producers, who adopted preventive management measures to control the virus, whose effect led to a consolidation of the passion fruit production chain in the region.(AU)

O CABMV é um vírus limitante para a cultura do maracujá no Brasil, principal país produtor mundial, cuja ocorrência já foi relatada em todos os estados produtores. Em 2017, o estado de Santa Catarina (SC) foi o terceiro maior produtor de maracujá no Brasil, responsável por 8,4% da produção nacional, sendo o litoral sul o principal responsável pelo aumento da produção, garantindo o abastecimento do mercado interno. Entretanto, nesse mesmo ano, essa região registrou as primeiras expressões de sintomas em plantas e frutos. Uma avaliação das amostras coletadas nos municípios de Sombrio, Praia Grande e São João do Sul, litoral sul de Santa Catarina, foi realizada por transmissão mecânica para plantas indicadoras, PTA-ELISA, RT-PCR e sequenciamento. Os resultados foram positivos para o CABMV e negativos para o CMV, tanto em PTA-ELISA quanto RT-PCR. Na RT-PCR, houve a amplificação de bandas com ca de 700pb, tamanho esperado para o Potyvirus cuja comparação de sequências com as depositadas no GenBank revelaram 98% de similaridade com os isolados do estado de São Paulo. A ocorrência do vírus na região do litoral sul de Santa Catarina não causou quebra na produção de maracujá devido à adoção conjunta de medidas preventivas de manejo pelos produtores, fato que consolidou a cadeia produtiva do maracujá na região.(AU)

Degenerate primers facilitate the detection and identification of potyviruses from the northwest region of Iran

Potyviruses are accounted for 40% of viral diseases of various crops including vegetables and legumes. Potyviruses can be transmitted through plant sap, seeds and many aphid species. Due to ease of spread of these viruses detection of such viruses are crucial as to the control of the incited diseases. This study compared the efficiencies of two couples of primers in their detection of potyviruses infecting vegetables in North-West of Iran. To identify potyviruses infecting vegetables, total RNA preparations from leaves of diseased plants were subjected to reverse transcription [RT] with oligo d[T]15, M4T or a potyvirus-specific primer, followed by polymerase chain reactions [PCR] with two pairs of degenerate primers including Sprimer/M4 or NIb2F/NIb3R that would yield [tilde] 1700 or 350 bp fragments, respectively. Amplification was achieved from 7 samples when Sprimer and M4 were used, but non- specific fragments were also amplified. However, specific amplifications from 31 samples were achieved with NIb2F2 and NIb3R. PCR products resulting from the use of NIb2F/NIb3R were subjected for cloning and sequencing. BLAST analysis of the sequenced data revealed that the PCR-amplified fragments belonged to Bean common mosaic virus [green bean], Bean yellow mosaic virus [broad bean], Potato virus Y [potato], Watermelon mosaic virus [squash, muskmelon or melon], Zucchini yellow mosaic virus [squash], and Soybean mosaic virus [bean]. This study demonstrated the efficiency of NIb2F2/NIb3R in detection of potyviruses and revealed the extent of infections with diverse potyviruses species in the northwest part of Iran.

Molecular modeling and conformational analysis of native and refolded viral genome-linked protein of Cardamom mosaic virus.

The viral genome-linked protein (VPg) of Potyviruses is covalently attached to the 5’ end of the genomic RNA. Towards biophysical characterization, the VPg coding region of Cardamom mosaic virus (CdMV) was amplified from the cDNA and expressed in E. coli. Most of the expressed VPg aggregated as inclusion bodies that were solubilized with urea and refolded with L-arginine hydrochloride. The various forms of CdMV VPg (native, denatured and refolded) were purified and the conformational variations between these forms were observed with fluorescence spectroscopy. Native and refolded CdMV VPg showed unordered secondary structure in the circular dichroism (CD) spectrum. The model of CdMV VPg was built based on the crystal structure of phosphotriesterase (from Pseudomonas diminuta), which had the maximum sequence homology with VPg to identify the arrangement of conserved amino acids in the protein to study the functional diversity of VPg. This is the first report on the VPg of CdMV, which is classified as a new member of the Macluravirus genus of the Potyviridae family.

Incidencia de potyvirus y caracterización molecular de PVY en regiones productoras de papa (Solanum tuberosum L) de Colombia / Incidence of potyvirus and molecular characterization of PVY in potato (Solanum tuberosum L) growing regions of Colombia

Los problemas virales reducen los rendimientos y la calidad del tubérculo semilla en cultivos de papa de todo el mundo. Esta investigación se planteó con el fin de evaluar los niveles de incidencia de potyvirus en diez de las principales regiones cultivadoras de papa de los departamentos de Antioquia, Boyacá, Cundinamarca y Nariño (Colombia), y las características genotípicas del virus Y de la papa (Potato virus Y, PVY), seleccionado por ser el potyvirus más limitante de este cultivo. Para la evaluación de la incidencia se utilizaron pruebas de Elisa con anticuerpos que reconocen epítopes comunes a los potyvirus, mientras que las pruebas moleculares incluyeron el análisis filogenético de secuencias parciales del gen de la cápside viral de 33 aislamientos, así como la secuenciación de una porción de los extremos 5´ y 3´del genoma de dos cepas colombianas de este virus. Los resultados confirmaron la presencia de potyvirus en los cultivos de los cuatro departamentos evaluados, con una incidencia promedio del 72%, siendo este nivel superior al 56% en todas las zonas evaluadas. Los análisis moleculares del PVY, permitieron asociar las cepas colombianas estudiadas con las razas PVYN y la variante PVYNTN, esta última responsable de la enfermedad conocida en el mundo como PTNRD (Potato tuber necrotic ringspot disease).

Potato viruses are responsible for significant reductions in seed quality and crop yields around the world. In this study, we evaluate the levels of incidence of potyvirus in ten potato growing regions of Colombia from the provinces of Antioquia, Boyacá, Cundinamarca and Nariño. As PVY is the most limiting potyvirus in potato farming, a molecular characterization of Colombian PVY strains was also performed. Incidence was evaluated by ELISA using general potyvirus antibodies. Phylogenetic analysis were made on the partial sequence of the capsid gene from 33 isolates. A portion of the 5´ and 3' genome ends was obtained from two Colombian strains. Results confirmed the presence of potyvirus in the four provinces with an average incidence of 72%. The lowest incidence value was 56%. Molecular analysis clustered all Colombian isolates with strains PVYN and PVYNTN, the latter responsible for the disease known as PTNRD (Potato tuber necrotic ringspot disease).

Combining ability of summer-squash lines with different degrees of parthenocarpy and PRSV-W resistance

The aim was to assess heterosis in a set of 16 summer-squash hybrids, and evaluate the combining capacity of the respective parental lines, which differed as to the degree of parthenocarpy and resistance to PRSV-W (Papaya Ringspot Virus-Watermelon strain). The hybrids were obtained using a partial diallel cross design (4 x 4). The lines of parental group I were 1 = ABX-037G-77-03-05-01-01-bulk, 2 = ABX-037G-77-03-05-03-10-bulk, 3 = ABX-037G77-03-05-01-04-bulk and 4 = ABX-037G-77-03-05-05-01-bulk, and of group II, 1' = ABX-037G-77-03-05-04-08-bulk, 2' = ABX-037G-77-03-05-02-11-bulk, 3' = Clarice and 4' = Caserta. The 16 hybrids and eight parental lines were evaluated for PRSV-W resistance, parthenocarpic expression and yield in randomized complete-block designs, with three replications. Parthenocarpy and the resistance to PRSV-W were rated by means of a scale from 1 to 5, where 1 = non-parthenocarpic or high resistance to PRSV-W, and 5 = parthenocarpic or high susceptibility to PRSV-W. Both additive and non-additive gene effects were important in the expression of parthenocarpy and resistance to PRSV-W. Whereas estimates of heterosis in parthenocarpy usually tended towards a higher degree, resistance to PRSV-W was towards higher susceptibility. At least one F1 hybrid was identified with a satisfactory degree of parthenocarpy, resistance to PRSV-W and high fruit-yield.

Association of cucumovirus and potyvirus with betelvine (Piper betle L.) as evidenced by ELISA and RT-PCR.

An attempt was made to detect various viruses of Piper betle grown at Mahoba and Banthara in India. DAC-ELISA and RT-PCR tests were performed in leaf sap samples of betelvine for detection of a cucumovirus (Cucumber mosaic virus) and potyvirus (Bean yellow mosaic virus) using specific antibodies and universal primers of respective viruses. DAC-ELISA could detect only CMV. However, RT-PCR detected both cucumovirus and potyvirus infection in betelvine samples. Association of CMV with betelvine was observed for the first time in the present study.