ABSTRACT Zinnia sp. and hairy beggartick (Bidens pilosa) plants exhibiting symptoms of possible virus infection were found in the municipality of Santa Bárbara d’Oeste, São Paulo State, Brazil. Flexuous filamentous particles and cytoplasmatic inclusions typical of potyvirus infection were observed by transmission electron microscopy, respectively, in leaf extracts and cells of symptomatic leaves. Infection of both plants with bidens mottle virus (BiMoV) was confirmed by RT-PCR using potyvirus universal primers, followed by nucleotide sequencing of the amplicons. The nearly complete genome sequence of the Brazilian isolate, named BiMoV-BR, is 9700 nucleotides long and shares 95.6 % identity with the corresponding nucleotide sequence of a BiMoV isolate from the United States. BiMoV-BR was mechanically transmitted and caused systemic infection on plants of Zinnia sp., hairy beggarstick, sunflower (Helianthus annuus), and lettuce (Lactuca sativa). Myzus persicae transmitted the virus to Zinnia sp. plants with efficacy of 8 % and 42 %, using one and ten aphids per plant, respectively. This is the first detection of BiMoV in Brazil. Further studies are necessary to evaluate the distribution of this potyvirus in the country. sp Bidens pilosa dOeste, dOeste d Oeste, Oeste d’Oeste State Brazil microscopy respectively leaves (BiMoV RTPCR RT PCR primers amplicons BiMoVBR, BiMoVBR BR, BR 970 956 95 6 95. States beggarstick Helianthus annuus, annuus , annuus) Lactuca sativa. sativa . sativa) 4 plant country 97 9

ABSTRACT Symptoms of severe vein clearing, leaf rolling, and blistering were observed in cassava plants in southwestern Bahia State, Brazil. The plants were collected, vegetatively propagated, and kept in a greenhouse. Leaf samples tested positive for begomovirus infection by Polymerase chain reaction (PCR). Complete nucleotide sequences of DNA-A and DNA-B of isolate PSLDV-Man04 revealed 99.89 % and 96.2 % identity, respectively, with the bipartite begomovirus passionfruit severe leaf distortion virus (PSLDV) genome. This PSLDV isolate was experimentally transmitted to two and four of the five cassava plants and ten passion fruit plants, respectively, inoculated with Bemisia tabaci Middle East-Asia Minor 1 (MEAM1). This is the first report of the natural infection of cassava plants by the PSLDV begomovirus in Brazil. clearing rolling State Brazil collected propagated greenhouse PCR. PCR . (PCR) DNAA DNA A DNAB B PSLDVMan04 PSLDVMan Man04 Man PSLDV-Man0 9989 99 89 99.8 962 96 2 96. identity respectively (PSLDV genome EastAsia East Asia MEAM1. MEAM1 MEAM (MEAM1) (PCR PSLDVMan0 Man0 PSLDV-Man 998 9 8 99. (MEAM1 (MEAM

ABSTRACT Sunflower (Helianthus annuus) plants showing symptoms of chlorosis, mosaic, chlorotic ringspot, and necrosis on younger leaves were found in a small experimental plot in Piracicaba, in the state of São Paulo, Brazil. Preliminary examinations by transmission electron microscopy of symptomatic leaf tissue revealed flexuous filamentous particles 13-15 nm wide and 700-750 nm long, and cytoplasmatic cylindrical inclusions typical of those found in plant cells infected by members of the Potyvirus genus. Total RNA extracted from symptomatic leaves and subjected to RT-PCR followed by partial nucleotide sequencing confirmed the presence of a potyvirus in the affected plants, which was identified as sunflower chlorotic mottle virus (SuCMoV), a member of the Sunflower chlorotic mottle virus (genus Potyvirus, family Potyviridae) species. Mechanical transmission assays with extracts of symptomatic sunflower leaves reproduced the original symptoms in sunflowers, mosaic symptoms in Zinnia elegans, and chlorotic local lesions in Chenopodium amaranticolor and C. quinoa. Sunflower and zinnia plants became infected after aphid transmission experiments with Myzus persicae. RT-PCR tests using specific primers for SuCMoV confirmed the presence of this virus in experimentally infected plants, meeting the criteria of Koch’s postulate. This is the first report of SuCMoV infecting sunflower plants in Brazil.

ABSTRACT: In the last decades, the high incidence of viruses transmitted by whiteflies has become a problem in the tomato fields, threatening, more recently, the potato crops. The present study carried out a survey of begomoviruses and criniviruses in tomato and potato crops, from 2015 to 2018, in the municipalities of Araucária, Campo do Tenente, Campo Largo, Contenda, Lapa, Faxinal, Morretes, Reserva, Castro, Palmeira and São Mateus do Sul, in Paraná State, Brazil. Total DNA and RNA from leaves were extracted and used as templates to detect, respectively, begomoviruses by PCR and criniviruses by RT-PCR. Out of 215 tomato samples, 14 from Faxinal were infected by crinivirus. The other tomato samples and 243 potato samples showed negative results for begomovirus and crinivirus. Results indicated a low incidence (6.5%) of crinivirus infecting tomato crops in Paraná State, and the nucleotide sequence of one amplified fragment shared 99.71% identity with tomato chlorotic virus (ToCV). ABSTRACT decades fields threatening recently 201 2018 Araucária Tenente Largo Contenda Lapa Morretes Reserva Castro Sul State Brazil detect respectively RTPCR. RTPCR RT PCR. RT-PCR 21 1 24 6.5% 65 6 5 (6.5% 9971 99 71 99.71 ToCV. ToCV . (ToCV) 20 2 6.5 (6.5 997 9 7 99.7 (ToCV 6. (6. 99. (6 (

RESUMO: Nas últimas décadas, a alta incidência de vírus transmitidos por mosca-branca tornou-se um problema nos campos de tomateiros, ameaçando, mais recentemente, a cultura da batateira. O presente trabalho teve como objetivo realizar um levantamento de begomovírus e crinivírus em lavouras de tomateiro e batateira nos municípios de Araucária, Campo do Tenente, Campo Largo, Contenda, Lapa, Faxinal, Morretes, Reserva, Castro, Palmeira e São Mateus do Sul, no Estado do Paraná, Brasil, de 2015 à 2018. DNA e RNA totais de folhas foram extraídos e utilizados como molde para detectar begomovírus por PCR e crinivírus por RT-PCR. Das 215 amostras de tomateiros coletadas, 14 provenientes de Faxinal estavam infectadas por crinivírus. As demais amostras de tomateiro e as 243 amostras de batateira analisadas apresentaram resultados negativo para begomovírus e crinivírus. Os resultados indicaram baixa incidência (6,5%) de crinivírus infectando lavouras de tomateiros no Estado do Paraná e a sequência de nucleotídeos de um amplicon apresentou 99,71% de identidade com o crinivírus tomato chlorotic virus (ToCV). RESUMO décadas moscabranca mosca branca tornouse tornou se ameaçando recentemente Araucária Tenente Largo Contenda Lapa Morretes Reserva Castro Sul Brasil 201 2018 RTPCR. RTPCR RT PCR. RT-PCR 21 coletadas 1 24 6,5% 65 6 5 (6,5% 9971 99 71 99,71 ToCV. ToCV . (ToCV) 20 2 6,5 (6,5 997 9 7 99,7 (ToCV 6, (6, 99, (6 (

RESUMO A pimenta-de-cheiro, Capsicum chinense é uma cultura importante na Bacia Amazônica, cultivada principalmente por pequenos produtores. Plantas de C. chinense em um campo experimental localizado no norte do estado brasileiro do Amazonas, foram encontradas apresentando sintomas característicos de infecção viral. Extratos de amostras de folhas sintomáticas examinados ao microscópio eletrônico de transmissão revelaram a presença de partículas alongadas e flexuosas e de partículas isométricas. Análises moleculares permitiram identificar a presença do pepper yellow mosaic virus (PepYMV) e do cucumber mosaic virus (CMV). Pulgões, identificados como Aphis gossypii foram encontrados colonizando pimenteiras-de-cheiro neste campo experimental e podem representar o provável vetor de PepYMV e CMV. Este trabalho relata a primeira ocorrência desses vírus infectando C. chinense no estado do Amazonas.

ABSTRACT The habanero chilli pepper, Capsicum chinense is an important crop in the Amazon Basin, mainly grown by small-scale producers. Capsicum chinense plants in an experimental field in the northern Brazilian state of Amazonas were found exhibiting characteristic symptoms of viral infection. Leaf sap from symptomatic plants examined under a transmission electron microscope revealed the presence of elongated flexuous particles and isometric particles. Using molecular assays, the viruses were identified as pepper yellow mosaic virus (PepYMV) and cucumber mosaic virus (CMV). Aphids, identified as Aphis gossypii, were found colonizing the C. chinense plants in the field and may be the vector for both PepYMV and CMV. We report the first occurrence of these viruses infecting C. chinense in the state of Amazonas.

ABSTRACT: Severe mosaic symptoms, accompanied by yellow spots, abnormally small leaves, fruit malformation and cracking, reduced plant growth, and high levels of whitefly (Bemisia tabaci MEAM1) infestation were observed in passionflower (Passiflora edulis) orchards in southwestern Bahia, Brazil. The aim of this work was to identify the species of begomovirus infecting the passionflowers, its prevalence in southwestern Bahia, and the spatial and temporal dynamics of the disease. Leaf samples from symptomatic plants collected at 57 orchards located in ten counties were evaluated by PCR for begomovirus infection. Complete nucleotide sequences of DNA-A for two isolates revealed 97 % identity with Passionfruit severe leaf distortion virus (PSLDV). The occurrence of PSLDV in 57 orchards was evaluated based on the presence of characteristic disease symptoms. Approximately 235,000 visually assessed plants exhibited symptoms characteristic of begomovirus infection. Epidemiological studies, conducted in two orchards in Dom Basílio County, showed that disease progress was relatively slow until 121 days after transplanting (DAT), but more rapid in the following 35 days, reaching 100 % infected plants by 156 DAT. The exponential model was fitted to the temporal dynamic of the disease for both areas. An aggregated pattern of diseased plants was predominant for almost all evaluations. It is possible that the primary and secondary spread of the pathogen occurred concurrently during the epidemic progression in both areas, especially late in the season. Containment measures to prevent the virus and the vector from spreading to other passionfruit producing areas in Brazil should be implemented.

Zucchini yellow mosaic virus (ZYMV) causes substantial economic losses in cucurbit crops. Although ZYMV has been present in Brazil for more than 20 years, there is little information about the biological and molecular characteristics of the isolates found in the country. This study aimed to characterize the experimental hosts, pathotypes and genetic diversity of a collection of eleven Brazilian ZYMV isolates within the coat protein gene. For biological analysis, plant species from Amaranthaceae, Chenopodiaceae, Cucurbitaceae, Fabaceae, Solanaceae, and Pedaliaceae were mechanically inoculated and pathotypes were identified based on the reaction of a resistant Cucumis melo, accession PI414723. All of the cucurbit species/varieties and Sesamum indicum were systemically infected with all isolates. The nucleotide sequence variability of the coat protein gene ranged from 82 % to 99 % compared to the corresponding sequences of ZYMV isolates from different geographical locations. No recombination event was detected in the coat protein gene of the isolates.

In the present work, the susceptibility of wild and domesticated plants of Momordica charantia to viruses from the genus Potyvirus that cause diseases in papaya (Carica papaya) and cucurbit crops was evaluated. The plants were subjected to experimental and natural infection with Papaya ringspot virus types P and W (PRSV-P and PRSV-W) and Zucchini yellow mosaic virus (ZYMV). None of the potyviruses infected the wild type of M. charantia through mechanical or aphid inoculation or under field exposition, whereas the domesticated type was only infected by isolates of ZYMV. In addition, both wild and domesticated types of M. charantia were not infected in natural conditions by an isolate of Zucchini lethal chlorosis virus, genus Tospovirus, transmitted by Frankliniella zucchini. These data clearly indicated that this wild type of M. charantia does not seem to have a role in the epidemiology of the diseases caused by these three potyviruses in Brazil.

Catharanthus roseus is a perennial, evergreen herb in the family Apocynaceae, which is used as ornamental and for popular medicine to treat a wide assortment of human diseases. This paper describes a new potyvirus found causing mosaic symptom, foliar malformation and flower variegation in C. roseus. Of 28 test-plants inoculated mechanically with this potyvirus, only C. roseus and Nicotiana benthamiana developed systemic mosaic, whereas Chenopodium amaranticolor and C. quinoa exhibited chlorotic local lesions. The virus was transmitted by Aphis gossypii and Myzus nicotianae. When the nucleotide sequence of the CP gene (768nt) was compared with other members of the Potyviridae family, the highest identities varied from 67 to 76 %. For the 3' UTR (286nt), identities varied from 16.8 to 28.6 %. The name Catharanthus mosaic virus (CatMV) is proposed for this new potyvirus.

Plantas de campânula (Campanula medium) exibindo mosaico e necrose foliar e anéis em flores foram coletadas em uma estufa comercial de flores na região de Atibaia, SP. Suspeitando de possível etiologia viral, amostras de tecido lesionado foram analisadas por ensaios de transmissão mecânica, microscopia eletrônica e sorologia. Todos os resultados apontaram para a presença do Tomato spotted wilt virus (TSWV) como o responsável pelos sintomas. Esse é o primeiro relato deste patógeno em campânula no Brasil.

Plants of bellflower (Campanula medium) exhibiting symptoms of mosaic, leaf necrosis and flower ring spot were found in a commercial crop in a greenhouse in Atibaia, São Paulo State. Electron microscopy, serology and biological assays indicated the presence of Tomato spotted wilt virus (TSWV). This is the first report of this pathogen in bellflower plants in Brazil.

O Tomato yellow vein streak virus (ToYVSV) é uma espécie putativa de begomovirus que infecta o tomateiro (Solanum lycopersicon) em diversas regiões do Brasil onde se cultiva essa solanácea, sendo a espécie prevalente no estado de São Paulo até 2005. Estudou-se a interação do ToYVSV com a Bemisia tabaci biótipo B e identificaram-se hospedeiras alternativas deste vírus. Os períodos de acesso mínimo de aquisição (PAA) e de inoculação (PAI) foram de 30 min e 10 min, respectivamente. A porcentagem de plantas infectadas chegou até cerca de 75% após um PAA e PAI de 24 h. O período de latência do vírus no vetor foi de 16 horas. O ToYVSV foi retido pela B. tabaci até 20 dias após a aquisição do vírus. Não foi detectada transmissão do vírus para progênie da B. tabaci biótipo B oriundas de insetos virulíferos. De 34 espécies de plantas testadas como hospedeiras somente Capsicum annuum, Chenopodium amaranticolor, C. quinoa, Datura stramonium, Gomphrena globosa, Nicotiana clevelandii e N. tabacum cv. TNN foram suscetíveis à infecção com o ToYVSV, por meio de inoculação com a B. tabaci. As espécies susceptíveis ao ToYVSV serviram de fonte de inóculo para a transmissão do vírus para tomateiros por meio de B. tabaci biótipo B.

The Tomato yellow vein streak virus (ToYVSV) is a putative species of begomovirus, which was prevalent on tomato crops in São Paulo State, Brazil, until 2005. The objectives of this study were to evaluate the interaction between ToYVSV and its vector Bemisia tabaci biotype B and to identify alternative hosts for the virus. The minimum acquisition and inoculation access periods of ToYVSV by B. tabaci were 30 min and 10 min, respectively. Seventy five percent of tomato-test plants were infected when the acquisition and inoculation access periods were 24 h. The latent period of the virus in the insect was 16 h. The ToYVSV was retained by B. tabaci until 20 days after acquisition. First generation of adult whiteflies obtained from viruliferous females were virus free as shown by PCR analysis and did not transmit the virus to tomato plants. Out of 34 species of test-plants inoculated with ToYVSV only Capsicum annuum, Chenopodium amaranticolor, C. quinoa, Datura stramonium, Gomphrena globosa, Nicotiana clevelandii and N. tabacum cv. TNN were susceptible to infection. B. tabaci biotype B was able to acquire the virus from all these susceptible species, transmitting it to tomato plants.

O ZLCV é um tospovírus encontrado com freqüência causando severos danos em cucurbitáceas. Nesse trabalho avaliaram-se os danos causados pelo ZLCV em abobrinha de moita 'Caserta', em campo na ESALQ/USP, Piracicaba-SP, onde esse vírus é freqüente. Plantas obtidas pela semeadura direta foram monitoradas periodicamente quanto à infecção pelo ZLCV por meio dos sintomas e por PTA-ELISA. Monitorou-se ainda a contaminação com Papaya ringspot virus - type W e Zucchini yellow mosaic virus, desconsiderando a produção dessas plantas. As plantas foram agrupadas em função da época de aparecimento dos sintomas do ZLCV, avaliando a produção de frutos comerciais (FC) e não comerciais (FNC) de cada grupo e comparando com a de plantas que permaneceram sem sintomas até o final do experimento. As plantas que apresentaram sintomas até os 23 dias após a emergência (DAE) não produziram qualquer tipo de frutos. FC foram colhidos de plantas que apresentaram sintomas a partir dos 42 DAE. Mesmo assim, houve redução de 78,5 % na produção de FC. Plantas que mostraram sintomas por ocasião da última colheita (55 DAE) apresentaram redução na produção de FC de 9,6 %. A infecção com o ZLCV até o início da frutificação inviabiliza a produção de FC de abobrinha de moita 'Caserta'.

Zucchini lethal chlorosis virus (ZLCV) is a tospovirus frequently associated with severe yield loss on cucurbit crops. The purpose of this work was to evaluate the damage caused by this virus on zucchini squash (Cucurbita pepo cv. Caserta) under field condition. The experiment was carried out at the Campus of the College of Agriculture, University of São Paulo, Piracicaba, SP, Brazil, were ZLCV is prevalent. After germination, zucchini squash plants were periodically monitored for the presence of characteristic symptoms induced by ZLCV and PTA-ELISA for virus indexing. Infection by Papaya ringspot virus - type W and Zucchini yellow mosaic virus were also monitored by PTA-ELISA, and plants infected by these viruses were disregarded for yield evaluation. Plants were grouped based on the time the symptoms were first seen. Fruits harvested from each plant within each group were classified as marketable (M) and non-marketable (NM) based on the phenotype. Plants that did not show symptoms by the end of the crop were considered healthy and their yield was used as control. Zucchini squash plants that showed symptoms of ZLCV infection up to 23 after emergency did not yield any fruit. Marketable fruits were first harvested only from plants that showed symptoms 42 days after emergency. However, the yield of marketable fruits was reduced by 78.5 %, as compared to that from asymptomatic plants. Plants that showed symptoms 55 days after emergency showed a reduction on the yield of marketable fruit of 9.6%.

O Papaya ringspot virus - type W (PRSV-W) e o Zucchini yellow mosaic virus (ZYMV) são os vírus predominantes em culturas de cucurbitáceas no Brasil, onde geralmente causam danos significativos na produção. As doenças causadas por ambos os vírus são de difícil controle. O objetivo desse trabalho foi avaliar o efeito das estirpes fracas PRSV-W-1 e ZYMV-M na produção de abobrinha de moita (Cucurbita pepo L. cvs. Samira, Novita Plus, AF 2847, and Yasmin) em condições de estufa plástica e de campo. Plantas infectadas com a estirpe ZYMV-M sob condições de estufa plástica não exibiram sintomas foliares típicos da doença e alterações na quantidade e qualidade dos frutos produzidos. No entanto, quando infectadas com a estirpe PRSV-W-1, ou PRSV-W-1 + ZYMV-M, as plantas mostraram sintomas severos de mosaico foliar e redução na qualidade dos frutos. Não houve alteração no número e no peso médio dos frutos colhidos dessas plantas. Quando as plantas dessas variedades foram infectadas com a estirpe PRSV-W-1 e avaliadas simultaneamente em campo e em estufa plástica, constatou-se que a intensificação de sintomas foi mais pronunciada sob condições de estufa plástica. A produção quantitativa não foi alterada. Fatores ambientais parecem ter influenciado a intensificação dos sintomas induzidos pela estirpe PRSV-W-1.

Papaya ringspot virus - type W (PRSV-W) and Zucchini yellow mosaic virus (ZYMV) are the most prevalent viruses in cucurbit crops in Brazil and responsible for frequent yield losses. Diseases caused by these viruses are difficult to control. The objective of this work was to evaluate the effects of mild strains PRSV-W-1 and ZYMV-M on the yield of Cucurbita pepo L. cvs. Samira, Novita Plus, AF 2847, and Yasmin, under plastic greenhouse and field conditions. Plants infected with ZYMV-M and grown in a plastic greenhouse did not exhibit typical leaf symptoms or significant alterations in quantitative and qualitative fruit yield. However, when infected with PRSV-W-1, or PRSV-W-1 + ZYMV-M, the plants exhibited severe leaf mosaic symptoms and reduced fruit quality, although there were no changes in the number and mean fruit weight harvested from these plants. When these plants were infected with PRSV-W-1 and studied simultaneously in the field and plastic greenhouse, intensification of symptoms in the fruits and leaves was more pronounced under the greenhouse conditions. Quantitative yield did not change. Environmental factors seem to influence symptoms induced by PRSV-W-1.

O vírus do mosaico do caupi (Cowpea aphid-borne mosaic virus - CABMV) é um potyvirus que causa uma das mais importantes doenças do maracujazeiro no Brasil. O vírus é transmitido por diversas espécies de afídeos de maneira não persistente, não circulativa. A reação de 16 espécies de Passiflora à infecção com quatro isolados brasileiros do CABMV, por meio de inoculação mecânica foi avaliada em condições de casa-de-vegetação. Somente a espécie selvagem P. suberosa foi resistente à infecção com todos os isolados do CABMV, em dois ensaios independentes. Plantas de P. suberosa enxertadas em plantas de P. edulis f. flavicarpa infectadas com o CABMV também não desenvolveram sintomas da doença. O vírus também não foi detectado por RT-PCR nas folhas superiores das plantas, sugerindo que essa espécie é imune ao CABMV.

Cowpea aphid-borne mosaic virus (CABMV) is a potyvirus that causes the most serious virus disease of passion fruit crops in Brazil. It is transmitted by several species of aphids in a non-persistent, non-circulative manner. The reaction of 16 species of Passiflora to infection by mechanical inoculation with four Brazilian isolates of CABMV was evaluated under greenhouse conditions. Only P. suberosa, a wild species, was resistant to infection by all virus isolates, in two independent assays. P. suberosa grafted onto infected P. edulis f. flavicarpa did not develop symptoms; neither was the virus detected by RT-PCR in the upper leaves, suggesting that this species is immune to CABMV.

Plantas de Capsicum annuum cv. Magali R, resistentes ao Pepper yellow mosaic virus (PepYMV), exibindo sintomas severos de mosaico amarelo, malformação foliar e subdesenvolvimento foram encontradas em plantios na região de Lins, SP, Brasil, em 2003/04. Partículas semelhantes àquelas do gênero Potyvirus foram observadas em extrato foliar de planta infectada examinado em microscópio eletrônico de transmissão. O extrato foliar também reagiu com anti-soro contra o PepYMV em PTA-ELISA. Além de C. annuum cv. Magali R, esse potyvirus também infectou sistemicamente C. annuum cv. Rubia R, que é resistente ao PepYMV. A seqüência de nucleotídeos de parte do gene da proteína capsidial (CP) desse potyvirus apresentou 96-98% de identidade com a de outros isolados do PepYMV. A seqüência parcial de nucleotídeos da região 3' não traduzida (3' NTR) apresentou 94-96% de identidade com a do PepYMV. Esses resultados são indicativos de que o potyvirus que quebrou a resistência em pimentão é um isolado do PepYMV.

Plants of Capsicum annuum cv. Magali R, resistant to Pepper yellow mosaic virus (PepYMV), which showed severe yellow mosaic, leaf malformation and stunting were observed during the 2003/04 growing season in Lins, São Paulo State, Brazil. Potyvirus-like particles observed in leaf sap from infected plants under the electron microscope reacted with an antiserum against PepYMV in PTA-ELISA. In addition to C. annuum cv. Magali R, this potyvirus also infected systemically the resistant C. annuum cv. Rubia R. The nucleotide sequence of part of the CP gene of this potyvirus shared 96-98% identity with that of other PepYMV isolates. The partial nucleotide sequence of the 3' NTR showed 94-96% identity with that of PepYMV. These data indicate that this potyvirus is a resistance-breaking isolate of PepYMV.