RESUMO O Potato vírus Y (PVY) é reconhecido como um dos patógenos mais comuns e destrutivos que ocorre nas áreas produtoras de batata em todo o mundo. Recentemente a PVYNTN e a PVYN:O/N-Wi emergiram como as principais estirpes presentes nos campos de batata brasileiros. Neste estudo, amostras de batata provenientes do Sul de Minas Gerais foram testadas inicialmente por DAS-ELISA e RT-PCR multiplex para discriminar as estirpes de PVY. Posteriormente, parte deste material foi testada por RT-qPCR para confirmar e quantificar os vírus nos tecidos infectados. Foi investigada a sensibilidade das técnicas para detecção de isolados de PVY presentes nos locais amostrados, bem como a ocorrência de infecções mistas, visando entender o quadro epidemiológico geral deste patógeno nos campos produtores de batata. No teste RT-PCR multiplex, as amostras com sorotipos O e N foram identificadas como infectadas pelas estirpes PVYNTN e PVYN:O/N-Wi. Quando testadas por RT-PCR para amplificação de PVYE, 41 amostras (67,2%) foram positivas, mostrando o perfil eletroforético característico para esta estirpe recombinante, e 9 amostras mostraram padrões atípicos para PVYE recombinante, mostrando a variabilidade dos isolados de PVY que ocorrem no campo. A melhor técnica para detectar infecção mista foi RT-qPCR, sendo que a concentração de PVYNTN foi muito maior que a de PVYN:O/N-Wi. Esses resultados mostram a importância da utilização de método mais adequado para o diagnóstico e levantamento de estirpes de PVY em lavouras de batata e revelam, pela primeira vez, a disseminação do recombinante de PVYE para diversos campos de batata brasileiros. (PVY mundo PVYNO/NWi PVYNONWi PVYN O/N Wi brasileiros estudo DASELISA DAS ELISA RTPCR RT PCR Posteriormente RTqPCR qPCR infectados amostrados mistas PVYNO/NWi. Wi. 4 67,2% 672 67 2 (67,2% positivas campo RTqPCR, qPCR, revelam vez PVYNO NWi ON 67,2 6 (67,2 67, (67, (67 (6 (

ABSTRACT Potato virus Y (PVY) is recognized as one of the most common and destructive pathogens seriously affecting potato producing areas worldwide. More recently PVYNTN and PVYN:O/N-Wi, have emerged as the main strains present in the PVY infected plants detected in Brazilian potato fields. In this study, samples of potato collected in south part of Minas Gerais - Brazil were first tested by DAS-ELISA and then by RT-PCR multiplex in order to discriminate the PVY strains. Afterward, part of them was tested by RT-qPCR to confirm and quantify the viruses in infected tissues. The sensitivity of the techniques for detecting PVY isolates present in the sampled locations was investigated, as well as the occurrence of mixed infections, aiming to understand the general epidemiological picture of this pathogen in potato producing fields. In the multiplex RT-PCR test, the samples with O and N serotypes were identified as infected with PVYNTN and PVYN:O/N-Wi strains. When tested by RT-PCR for amplification of PVYE, 41 samples (67,2%) were positive, having a characteristic electrophoretic profile for this recombinant strain, and 9 isolates were also observed with atypical patterns for recombinant PVYE. The best technique to detect mixed infection was RT-qPCR, with the concentration of PVYNTN being much higher than that of PVYN:O/N-Wi. These results show the importance of using the most suitable method for the diagnosis and surveying of PVY strains in crop fields and reveal, for the first time, the dissemination of PVYE recombinants to several Brazilian potato fields. (PVY worldwide PVYNO/NWi, PVYNONWi PVYN O/N Wi, Wi study DASELISA DAS ELISA RTPCR RT PCR Afterward RTqPCR qPCR tissues investigated infections test PVYNO/NWi 4 67,2% 672 67 2 (67,2% positive strain RTqPCR, qPCR, PVYNO/NWi. Wi. reveal time PVYNO NWi ON 67,2 6 (67,2 67, (67, (67 (6 (

RESUMO A diagnose das bacterioses causadoras de manchas foliares na cultura do cafeeiro (Coffea arabica), mancha aureolada (Pseudomonas syringae pv. garcae), mancha bacteriana (P. syringae pv. tabaci), crestamento bacteriano (P. cichorii) e mancha escura (Robbsia andropogonis) é considerada um desafio para os fitopatologistas. A identificação incorreta pode ocorrer quando o diagnóstico é baseado exclusivamente em sintomas e propriedades bioquímicas. Assim, o objetivo com este estudo foi identificar e diferenciar as espécies e patovares de Pseudomonas patogênicos ao cafeeiro, possibilitando um levantamento da ocorrência dessas bactérias nas principais regiões produtoras de Minas Gerais, Brasil. Primeiramente, a patogenicidade dos isolados foi confirmada por inoculação em C. arabica cv. Catuaí Vermelho IAC 99. Em seguida, análises bioquímicas, combinadas com PCR convencional, reação em cadeia de polimerase de elemento repetitivo (rep-PCR) e filogenia baseada em sequências do gene rpoD, foram usadas para caracterizar 84 isolados de Pseudomonas de lavouras de café e viveiros em Minas Gerais. Com base na filogenia do gene rpoD, 73 isolados foram identificados como P. syringae pv. garcae, cinco como P. syringae pv. tabaci e seis como P. cichorii. Os resultados de rep-PCR sugerem alta variabilidade genérica nas populações de Pseudomonas syringae pv. garcae e P. cichorii. Este é o primeiro relato da ocorrência de mancha foliar bacteriana (P. syringae pv. tabaci) em campo de produção de café no Estado de Minas Gerais. Os resultados confirmaram a prevalência de P. syringae pv. garcae em lavouras de café no Estado e o conhecimento gerado contribuirá para o desenvolvimento de primers espécie-específicos para identificação e detecção deste patógeno.

ABSTRACT The diagnosis of foliar bacterial diseases in coffee (Coffea arabica), such as halo blight (Pseudomonas syringae pv. garcae), bacterial leaf spot (P. syringae pv. tabaci), bacterial blight (P. cichorii), and dark leaf spot (Robbsia andropogonis), is considered a challenge for plant pathologists. The misidentification has been occurring when the diagnosis is solely based on symptoms and biochemical properties. Thus, the objective of this study was to identify and differentiate species and pathovars of Pseudomonas pathogenic to coffee plants, enabling a survey of the occurrence of these bacteria in the main producing regions of Minas Gerais state, Brazil. Firstly, the pathogenicity of the isolates was confirmed by inoculation in C. arabica cv. Catuaí Vermelho IAC 99. Then, biochemical analyses, combined with, repetitive element-polymerase chain reaction (rep-PCR) and phylogeny based on rpoD gene sequences were used to characterize 84 Pseudomonas isolates from coffee crops and nurseries. Based on rpoD-phylogeny, 73 isolates were identified as P. syringae pv. garcae, five as P. syringae pv. tabaci and six as P. cichorii. The rep-PCR results suggest a high genetic variability in populations of Pseudomonas syringae pv. garcae and P. cichorii. This is the first report of the occurrence of bacterial leaf spot (P. syringae pv. tabaci) in the coffee-producing filed in Minas Gerais State. The findings confirmed the prevalence of P. syringae pv. garcae in coffee production fields in the State and the generated knowledge will contribute for the development of species-specific primers for the identification and detection of this pathogen.

ABSTRACT The transmission of tobamovirus by tomato and pepper seeds is an important mean of virus introduction in crops. Therefore, detecting its presence in the seed becomes essential for the preventive control of virus diseases. In this study, a method was proposed for the detection of Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) in tomatoes (Solanum lycopersicum) and Pepper mild mottle virus (PMMoV) in pepper (Capsicum annum) seeds. Seed lots with different levels of incidence were analyzed by biological, serological, and molecular methods. Using DAS-ELISA technique, it was possible to detect TMV up to the limit rate of 1:170 (1 contaminated seed: total seeds) and the ToMV up to 1:200 in tomato seeds; PMMoV in pepper seeds was detected up to 1:140. The IC-RT-PCR detected the TMV and ToMV up to the limit of 1:400 and PMMoV up to the limit of 1:300. The assembled lots containing only 1 contaminated seed in 1000 (1:1000) were combined into 30 sub lots for DAS-ELISA analysis and 10 sub lots for IC-RT-PCR analysis. Both techniques, DAS-ELISA and IC-RT-PCR, were efficient to detect the three viruses in all analyzed samples, but the detection of tobamoviruses with RT-PCR and biological tests was not reliable. Based on the results of this study, in which a combination of seeds in sub lots was made to reduce the number of tests performed, it is possible to make significant savings in the cost of the diagnostic methods routinely conducted in official laboratories, with high efficiency and reliability.

O alelo Ry adg é muito utilizado pelos melhoristas, por conferir resistência extrema a todas as estirpes do PVY. Entretanto, a estirpe necrótica possui grande capacidade de recombinação, de modo que, novos variantes genéticos capazes de provocar perdas consideráveis na produtividade, vêm surgindo recentemente. Até o momento, nem todas as estirpes recombinantes de PVY necrótico foram testados quanto à sua reação com o gene Ry. Neste estudo, objetivou-se identificar clones de batata, portadores do alelo de resistência e avaliar a sua reação aos seguintes isolados recombinantes: NTN (PVY NTN), Wilga (PVY N-Wi) e "top crespo" (PVY E). Clones avançados de batata, do programa de melhoramento da Universidade Federal de Lavras, foram avaliados por meio de um marcador molecular específico para o alelo Ry adg. Os clones portadores do alelo de resistência foram enxertados em plantas de fumo infectadas com o PVY N e três isolados recombinantes. Os clones portadores do alelo de resistência não foram infectados com nenhum dos recombinantes e nem com o PVY N no teste de enxertia. Esses resultados confirmam que o alelo Ry adg confere resistência também aos variantes testados.

The Ry adg allele is widely used by breeders to confer extreme resistance to all strains of PVY. However, the necrotic strain has increased recombination resulting in recent considerable losses in productivity. Thus far, not all necrotic recombinant strains of PVY have been tested for their reaction to the Ry gene. The objective of this study was to identify potato clones carrying the resistant allele and to assess their reaction to the following recombinant strains: NTN (PVY NTN), Wilga (PVY N-Wi), and "curly top" (PVY E). Advanced clones from the potato breeding program at Universidade Federal de Lavras were evaluated through a specific molecular marker for the Ry adg allele. The clones carrying the resistance allele were grafted on tobacco plants infected with necrotic recombinant strains of PVY. The clones carrying the allele for resistance to PVY were not infected with any of the recombinants during the grafting test. These results confirm that resistance to necrotic recombinant strains has not yet been overcome and that the Ry adg allele also confers resistance to the three recombinant strains tested.

Com o objetivo de identificar o tipo de herança, o número de genes e os parâmetros genéticos e fenotípicos da resistência do acesso PI 595201 a um isolado de PRSV-W da região Norte do Brasil foi conduzido um experimento na área experimental do Departamento de Produção Vegetal da Universidade Federal do Tocantins, em condições de casa de vegetação. A partir do cruzamento entre a cultivar suscetível Crimson Sweet (P1) e o acesso resistente PI 595201 (P2) foi obtida a geração F1 e as gerações segregantes, cujas plantas foram inoculadas e avaliadas quanto à resistência com base nos sintomas nas folhas. Foram avaliadas 48, 48, 80, 200, 100 e 100 plantas das gerações P1, P2, F1, F2, RC11 e RC12, respectivamente, em delineamento inteiramente casualizado com quatro repetições. A partir da análise de componentes de médias no controle da resistência determinaram-se dois genes, predominando os efeitos gênicos aditivos, seguido de efeitos de dominância incompleta no sentido de aumentar a resistência. As herdabilidades para a resistência, no sentido amplo e restrito, foram de 0,48 e 0,38, respectivamente.

Aiming to identify the type of inheritance, the number of genes and genetic and phenotypic parameters of PI 595201 access resistance to an isolate of PRSV-W in the North of Brazil, was conducted an experiment in the experimental area of the Department of Plant Production of the Universidade Federal do Tocantins, in a greenhouse. From the cross between the susceptible cultivar 'Crimson Sweet' (P1) and resistant access 'PI 595201' (P2) was obtained F1 and segregating generations, whose plants were inoculated and evaluated for resistance based on leaf symptoms. Was evaluated 48, 48, 80, 200, 100 and 100 plants of P1, P2, F1, F2, RC11 and RC12 generations, respectively, in a completely randomized design with four replications. From the analysis of medium components in the control of resistance were determined two genes, predominantly additive gene effects, followed by effects of incomplete dominance to increase the resistance. The heritability for resistance in broad and narrow sense were 0.48 and 0.38, respectively.

O crestamento bacteriano comum, causado por Xanthomonas axonopodis pv. phaseoli (Xap), é a principal bacteriose do feijoeiro no Brasil, sendo transmitida principalmente por sementes. O presente trabalho teve como objetivo aperfeiçoar uma técnica, por meio de diferentes métodos de preparação do extrato, para a detecção de Xap, bem como sua detecção simultânea com Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff) nos extratos de sementes de feijão,via PCR. A partir de amostras de sementes de feijão inoculadas artificialmente com Xap e lotes comerciais, foram avaliados: extrato bruto obtido diretamente das sementes, extrato concentrado por filtração em membrana de 0,22µM de diâmetro, extrato concentrado por centrifugação e extrato plaqueado em meio semi-seletivo XCP1 com e sem antibióticos (BIO-PCR). Avaliou-se a presença simultânea de Xap e Cff em 10 lotes comerciais de sementes de feijão através da reação multiplex, utilizandos-se os primers X4c, X4e, CffFOR2 e CffREV4. A partir do extrato bruto, do extrato concentrado por centrifugação e por filtragem em membrana Millipore® não foi possível a detecção de Xap nas sementes de feijão artificialmente contaminadas nem nos 47 lotes comerciais de sementes/ grãos de feijão. A técnica de BIO-PCR permitiu a detecção de Xap a partir de extratos de sementes de feijão artificialmente contaminadas e em 18 dos 47 lotes comerciais. A técnica de detecção simultânea de Xap e Cff no mesmo gel é viável, por amplificar fragmentos de DNA típicos de cada fitobactéria. O uso do meio de cultura XCP1 sem adição de antibióticos permitiu detectar Xap com período de incubação menor em um dia em comparação à detecção utilizando-se o meio de cultura com antibióticos

Common bacterial blight, caused by Xanthomonas axonopodis pv. phaseoli (Xap), is the major bacteriosis affecting bean plants in Brazil and is mainly transmitted by seeds. This study aimed to improve a technique, using different methods of extract preparation, for the detection of Xap alone and simultaneously with Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff) in bean seed extracts, via PCR. Samples of bean seeds artificially inoculated with Xap and commercial batches were used to evaluate: crude extract obtained directly from the seeds, extract concentrated by filtration in 0.22ìM-diameter membrane, extract concentrated by centrifugation and extract plated on semiselective XCP1 medium with and without antibiotics (BIO-PCR). The simultaneous presence of Xap and Cff in 10 commercial batches of bean seeds was assessed through multiplex reaction, using the primers X4c, X4e, and CffFOR2 CffREV4. By using crude extract, extract concentrated by centrifugation and by filtration in millipore® membrane, detection of Xap in bean seeds artificially contaminated or in 47 commercial batches of bean seeds/grains was not possible. BIO-PCR technique allowed the detection of Xap in extracts from artificially contaminated bean seeds and in 18 of the 47 commercials batches. The technique of simultaneous detection of Xap and Cff on the same gel is feasible, for amplifying DNA fragments typical of each phytobacterium. The use of XCP1 culture medium without adding antibiotics allowed the detection of Xap with shorter incubation period in one day, compared to detection using the culture medium with antibiotics.

Blister spot (Colletotrichum gloeosporioides) is now widespread in most coffee producing states of Brazil, becoming a limiting factor for production. The lack of data relating to the reproduction of typical symptoms (light green, oily patches) leaves a gap within the pathosystem, forcing the search for new methodologies for monitoring the disease. Monitoring of genetically modified organisms has proven to be an effective tool in understanding the host x pathogen interactions. Thus, the present study was carried out to evaluate the effectiveness of two systems of genetic transformation in obtaining mutants using the gfp reporter gene. Using the two transformation systems (PEG and electroporation) revealed the efficiency of both, confirmed by fluorescence microscopy and resistance to the antibiotic hygromycin-B, when incorporated into the culture medium. The fungus maintained its cultural and morphological characteristics when compared to wild strains. When inoculated on coffee seedlings, it was found that the pathogenicity of the processed isolates had not changed.

O objetivo deste trabalho foi avaliar a flutuação populacional do ácaro Brevipalpus phoenicis, vetor do Coffee ringspot virus (CoRSV) - agente causal da mancha-anular do cafeeiro -, bem como a sua distribuição em plantas de café e o efeito de acaricidas no seu controle em lavouras cafeeiras. O experimento foi realizado no Município de Coromandel, na região do Alto do Paranaíba, MG, durante dois anos. No segundo ano, avaliou-se o progresso da doença, e determinou-se a relação entre o crescimento populacional do ácaro e a incidência da doença. Observou-se que a mancha-anular e o ácaro ocorreram durante todo o ano, com maiores populações e incidência da doença nos meses com menores índices pluviais e temperaturas amenas. As análises de Pearson mostraram correlação negativa entre os parâmetros climáticos e a população do ácaro. Na análise da incidência da doença em diferentes épocas, verificou-se correlação positiva entre a primeira avaliação e as avaliações subsequentes. Quanto à distribuição espacial, foi observada a presença do ácaro em toda a planta; porém, as folhas são preferidas pelos adultos, enquanto os frutos são preferidos para oviposição. Os acaricidas são eficientes para manter a população do ácaro em baixos níveis, com consequente diminuição da incidência da doença.

The objective of this work was to evaluate the population fluctuation of the mite Brevipalpus phoenicis, vector of the Coffee ringspot virus (CoRSV) - the causal agent of the coffee ringspot disease -, as well as its distribution in coffee plants, and the effect of acaricides in its control in coffee crops. The experiment was carried out at the municipality of Coromandel, in the region of Alto do Paranaíba, MG, Brazil, during two years. In the second year, the progress of the disease was evaluated, and the relationship between the mite population and disease incidence was determined. Ringspot disease and mites were detected throughout the year, with larger populations and disease incidence in the months with lower rainfall and mild temperatures. Pearson analyses showed negative correlation between the climatic parameters and the mite population. In the analysis of the disease incidence in different periods, a positive correlation was found among the first and subsequent evaluations. Regarding spatial distribution, mite presence was observed in the whole plant; however, leaves are preferred by the adults, whereas the fruit are preferred for oviposition. The acaricides are effective in keeping the mite population at low levels, with consequent reduction of disease incidence.

O objetivo deste trabalho foi determinar a presença do alelo Pvr4, que confere resistência contra o PepYMV (Pepper yellow mosaic virus), em genótipos de pimentão comunmente encontrados no mercado brasileiro, com uso de um marcador molecular codominante tipo CAPS. A resistência ao PepYMV, nos genótipos CM-334-INRA, Myr-29 e em genótipos derivados do híbrido comercial Mônica-R, foi detectada como associada à banda de 444 pb, ligada ao alelo de resistência Pvr4. As plantas resistentes homozigotas (pvr4/pvr4) mostraram uma banda de 444 pb, as suscetíveis (Pvr4+/Pvr4+) uma banda de 458 pb e as resistentes heterozigotas (Pvr4+/Pvr4) mostraram as duas bandas. No entanto, no acesso resistente CM-334-UFV, nos híbridos Magali-R e Martha-R, assim como em populações derivadas desse acesso e desses híbridos, a resistência ao PepYMV não esteve associada ao marcador CAPS. O acesso CM-334-UFV ('Criollo de Morelos-334', de Viçosa, MG) distinguiu-se do CM-334-INRA ('Criollo de Morelos-334', da França); embora ambos os acessos tenham sido resistentes ao PepYMV, apenas em CM-334-INRA foi encontrada a associação da resistência com a banda de 444 pb.

The objective of this work was to determine the presence of the Pvr4 allele, which controls the resistance to the PepYMV (Pepper yellow mosaic virus), in sweet pepper genotypes commonly available in the Brazilian market, using a CAPS codominant molecular marker. The resistance to PepYMV, in the genotypes CM-334-INRA, Myr-29, and in genotypes derived from the hybrid Mônica-R, was found to be associated with the 444 bp band linked to the resistance allele Pvr4. Homozygous resistant plants (pvr4/pvr4) showed a single band of 444 bp, the susceptible ones (Pvr4+/Pvr4+) showed a band of 458 bp, and the heterozygous resistant plants (Pvr4+/Pvr4) showed both bands. However, in the resistant accession CM-334-UFV, and in the hybrids Magali-R and Martha-R, as well as in populations derived from this accession and these hybrids, the resistance to PepYMV was not associated to the CAPS marker. The accession CM-334-UFV ('Criollo de Morelos-334' from Viçosa, MG, Brazil) was distinct from CM-334-INRA ('Criollo de Morelos-334', from France); although both accessions were resistant to PepYMV, the association of resistance with the 444 bp band was found only in CM-334-INRA.

Atualmente, existe a necessidade de se desenvolver métodos sensíveis, baratos, reproduzíveis e rápidos para a detecção de fitobactérias em sementes. O objetivo deste trabalho foi otimizar um método de obtenção das células bacterianas e a técnica de PCR para detecção de Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), em sementes de feijão. Foi utilizado o primer CffFOR2-REV4, desenhado a partir do fragmento amplificado via PCR baseado na seqüência repetitiva (Rep-PCR). Avaliaram-se também quatro métodos de preparação dos extratos de sementes de feijão para a obtenção células de Cff : 1) extrato bruto de sementes; 2 ) extrato concentra do por filtração em membra na milipore (0,22Mu de diâmetro) e ressuspensão em água; 3) extrato concentrado por centrifugação a 10 .00 0 xg por 15 minutos no volume de 20 ou de 80 mL e 4) Bio-PCR. Dentre esses métodos, tanto a Bio-PCR quanto a concentração do extrato por centrifugação, seja no volume de 20 ou de 80 mL, possibilitaram amplificar o segmento de DNA de 306 pb, característico de Cff. Essas duas técnicas, além de detectar a bactéria, apresentam alta sensibilidade, detectando até 1 semente inoculada artificialmente artificialmente com Cff em 999 sementes sadi as. Analisaram-se dezessete lotes comerciais de sementes de feijão pelo método de concentração de extrato por centrifugação, sendo qu e em doze detectou-se a bactéria Cff, observado pela presença de bandas com 306 pb. Portanto, foi possível otimizar um método de obtenção das células bacterianas e técnica de PCR para detecção de Cff em sementes de feijão, que seja sensível, reproduzível e de fácil execução, que poderá ser utilizado rotineiramente em laboratórios de análises de sementes.

Currently, there is a need to develop sensitive methods, inexpensive, reproducible and rapid detection of phytobacteria seeds. The objective of this study was to optimize a method for obtaining bacterial cells and the PCR technique to detect Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), in bean seeds using PCR. It was used one pairs of primers CffFOR2-REV4, designed from an PCR amplified fragment of the conserved repetitive sequence (Rep-PCR). The primer pair CffFOR2-REV4 was selected in this study, because demonstrated higher reproducibility and efficacy in the detection of Cff strains in bean seeds. Four methods were also evaluated in the seed extract preparation for PCR: 1) rough seed extract; 2) millipore membrane filter (0.22Mu diameter) concentrated seed extract and subsequently ressuspended in water; 3) centrifuge concentrated seed extract 10.000 xg for 15 minutes for 20 and 80 mL; 4) Bio-PCR. Among those methods, either Bio-PCR or centrifuge concentrated extract, in a total 20 or 80 mL suspension volume, produced a 306bp DNA fragment, diagnostic for Cff. Those two techniques detected the bacterium and presented high sensitivity, detecting up to 1 Cff physiological conditioning artificially contaminated seed in a total 999 healthy ones. A total of seventeen commercial bean seed lots were analyzed by the method of concentration of the extract by centrifugation, and twelve detected in the bacterium Cff, observed by the presence of bands with 306 bp. Therefore, it was possible to optimize a method of obtaining the bacterial cells and PCR for detection of Cff in bean seeds, that is sensitive, reproducible and easy to perform, which can be used routinely in laboratories for seed.

O objetivo deste trabalho foi associar um marcador microssatélite ao alelo Ty-1 de resistência a Begomovirus em tomateiro, e avaliar a eficiência desta associação na seleção de linhagens resistentes ao vírus. Os marcadores SSR-47 e SSR-48 foram testados em linhagens isogênicas contrastantes quanto à presença do alelo Ty-1 (LA-3473, LA-3474, LA-3475). O marcador SSR-47, por ter detectado polimorfismo nas linhagens, foi o único utilizado nas etapas subsequentes da pesquisa. Detectada a associação entre o SSR-47 e o alelo Ty-1, testou-se sua eficiência na seleção de genótipos avançados de tomateiro. Para confirmar a eficiência da seleção, foi realizada a avaliação fenotípica das plantas com padrões contrastantes de bandas para SSR-47, quanto à resistência a Begomovirus. Plantas que apresentaram banda única de 191 pb foram resistentes ao Begomovirus, pelo teste de inoculação por enxertia; aquelas com banda única de 180 pb foram suscetíveis; e as plantas com bandas de 191 e 180 pb foram resistentes. A distância máxima entre o Ty-1 e o marcador SSR-47 foi de 2,7 cM. Este marcador foi efetivo em caracterizar genótipos portadores do alelo Ty-1. As respostas das plantas à infecção pelo Begomovirus, induzida via enxertia, são consistentes com as reações previstas com o uso do marcador molecular SSR-47.

The objective of this work was to associate a microsatellite marker with the Ty-1 allele that controls resistance to Begomovirus in tomatoes, and to assess the eficiency of this association in the selection of virus-resistant lines. Microsatellite markers SSR-47 and SSR-48 were tested in near isogenic tomato lines (LA-3473, LA-3474, LA-3475) with contrasting genotypes for the Ty-1 allele. The marker SSR-47 detected polymorphism in the lines, and was the only one used in the subsequent research phases. An association between the SSR-47 marker and the Ty-1 locus was detected, and its efficiency for selection of begomovirus-resistant genotypes in tomato was assessed. To confirm the selection efficiency, plants with contrasting banding patterns for the SSR-47 marker were tested for their reaction to Begomovirus. All plants with a single 191-bp band were resistant to Begomovirus, through inoculation test by grafting. Plants with a single 180-bp band were susceptible, and plants with both 191bp and 180 bp bands were resistant. The maximum distance between Ty-1 and the SSR-47 marker was 2.7 cM. This marker was efficient to identify genotypes bearing the Ty-1 allele. Plant responses to infection by Begomovirus inoculated through grafting were consistent with reactions predicted with the use of the SSR-47 marker.

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.

O objetivo do trabalho foi verificar o efeito de tratamentos inseticidas, na produção e no controle da disseminação de vírus por afídeos vetores e o teor de resíduos observados nos tubérculos submetidos a esses tratamentos. Foram empregadas as cultivares, Ágata e Emeraude com Potato vírus Y. O modelo utilizado foi o de blocos inteiramente casualizados, com 12 tratamentos e 4 repetições, sendo: 1) pulverização semanal (PS), com a última aos 55 dias após o plantio (DAP); 2) PS com a última aos 69 DAP; 3) PS, com a última aos 69 DAP + aplicação de Cabrio Top; 4) pulverização a cada cinco dias, sendo a última aos 69 DAP; 5) PS sendo a última aos 80 DAP e 6) controle, sem pulverização, todos combinados com dois tratamentos de sulco (Imidacloprid e Phorate). Observou-se que a maioria dos tratamentos não impediu a disseminação dos vírus.

The objective the work was to investigate the effect of insecticide treatments on virus spread and in potato yield, as well as to evaluate the residue value of the insecticides employed in the tubers subjected to these treatments. The experiment was carried out with two cultivars in randomized blocks with 40 tubers, 4 replications and 12 treatments, as follows: 1) insecticide spraying (SI) on leaves once a week, with the last application at 55 days after planting (DAP); 2) SI on leaves once a week, with the last application at 69 DAP; 3) SI on leaves once a week, with the last application at 69 DAP + application of Cabrio Top; 4) SI on leaves every five days, with the last application at 69 DAP; 5) SI on leaves once a week, with the last application at 80 DAP; and 6) control, with no insecticides after plant emergence; and all of them combined with 2 soil treatments (Imidacloprid and Phorate). Those results show that the application of insecticides is useless when the potato seed has a high PVY incidence.

O marcador molecular SCAR 'Sw-421' é localizado a 1,0 cM do alelo Sw-5, proveniente de Lycopersicon peruvianum (L.), que confere resistência ao vira-çabeça em tomateiro. Contudo, o mesmo ainda não havia sido testado em populações avançadas de tomateiro. O objetivo do trabalho foi distinguir plantas resistentes homozigotas (Sw-5/Sw-5), resistentes heterozigotas (Sw-5/Sw-5+) e suscetíveis (Sw-5+/Sw-5+) pelo marcador SCAR 'Sw-421'. As amplificações de bandas de 940 pb e 900 pb caracterizaram os genótipos resistentes homozigotos e suscetíveis, respectivamente. Duas bandas (900 pb e 940 pb) foram observadas nos genótipos heterozigotos, confirmando a herança codominante do marcador. De 57 plantas das progênies isogênicas avaliadas 18 (31,6%) plantas foram caracterizadas como resistentes, 8 (14,0%) como heterozigotas e 31 (54,4%) plantas suscetíveis. O marcador molecular SCAR 'Sw-421' constitui importante ferramenta para seleção e piramidação de alelos de resistência, especialmente quando se utilizam outras fontes de resistência ao vira-cabeça, como por exemplo, a fonte Rey de los Tempranos.

The SCAR (Sequence Characterized Amplified Region) 'Sw-421' molecular marker is located at 1.0 cM from the Sw-5 allele, originated from Lycopersicon peruvianum (L.), which confers resistance to the tomato spotted wilt virus (TSWV). However, it had not been tested yet in advanced tomato populations. The goal of this study was to distinguish resistant homozygotes (Sw-5/Sw-5) and heterozygotes (Sw-5/Sw-5+) from susceptible (Sw-5+/Sw-5+) plants in crossing populations with the Stevens cultivar and advanced backcrossing populations by using 'Sw421' SCAR marker. The amplification of 940 bp and 900 bp bands characterized the resistant homozygotes and susceptible controls, respectively. A two band pattern (900 bp and 940 bp) was observed in heterozygote genotypes (Sw-5/Sw-5+), which confirmed the co-dominant inheritance mechanism of the marker. Fifty seven plants from the isogenic progenies were characterized based on bands pattern: 18 plants (31.6%) were identified as resistant homozygotes, 8 plants (14.0%) as resistant heterozygotes and 31 plants (54.4%) were characterized as susceptible. The SCAR 'Sw-421' marker is an important tool for selection and pyramid resistance alleles, mainly when other resistance sources to the TSWV are available, such as the Rey de los Tempranos source.

O objetivo deste trabalho foi avaliar genótipos de tomateiro, quanto à resistência a begomovírus, e caracterizar, por meio do marcador molecular SSR-47, híbridos de tomate de mesa portadores do alelo de resistência ao begomovírus Ty-1, com potencial comercial. Os 24 híbridos experimentais, heterozigotos no loco Ty-1, depois de infectados via enxertia, apresentaram sintomas intermediários, em comparação aos identificados pelas linhagens homozigotas Ty-1/Ty-1 e pelos genótipos suscetíveis Ty-1+/Ty-1+, o que indica a dominância incompleta do alelo Ty-1. Esses híbridos foram considerados como parcialmente tolerantes a begomovírus. Os híbridos experimentais TEX-246, TEX-261, TEX-253, TEX-256, TEX-262, TEX-252, TEX-251 e TEX-268 aliaram médias elevadas de produção total e de massa média dos frutos; e os híbridos TEX-246, TEX-253, TEX-256, TEX-262 e TEX-252 apresentaram valores elevados também para meia-vida da firmeza e foram, portanto, considerados competitivos em comparação aos padrões comerciais usados como testemunhas. O marcador molecular SSR-47 foi eficiente em caracterizar genótipos portadores do alelo Ty-1. A infecção do begomovírus, induzida via enxertia, manifestou sintomas, nos genótipos testados, condizentes com os resultados obtidos com o marcador molecular SSR-47.

The objectives of this study were to evaluate tomato genotypes for begomovirus resistance and to assess SSR-47 fingerprinting patterns associated with alleles Ty-1, in hybrids of fresh-market tomato with commercial potential. Twenty-four experimental genotypes, heterozygous at Ty-1 locus, were infected with begomovirus via grafting, and showed intermediate symptoms compared to the ones identified by homozygous Ty-1/Ty-1 lines and by susceptible genotypes Ty-1+/Ty-1+, which indicates the incomplete dominance of the Ty-1 allele. These hybrids were considered to possess partial resistance to begomovirus. The experimental hybrids TEX-246, TEX-261, TEX-253, TEX-256, TEX-262, TEX-252, TEX-251 and TEX-268 combined high total yields with high mean fruit mass. The hybrids TEX-246, TEX-253, TEX-256, TEX-262 and TEX-252 showed also longer half-life for fruit firmness and were thus considered competitive with current commercial hybrids used as checks. The molecular marker SSR-47 was effective for fingerprinting of tomato genotypes bearing the resistance allele Ty-1. The graft-induced infection of begomovirus showed symptoms in the genotypes tested that are in agreement with the results of fingerprinting with the marker SSR-47.