Detection of Bemisia tabaci Meam1 and Med cryptic species in Oklahoma

The agricultural importance of Oklahoma combined with the sheer efficiency of insect pests for virus transmission makes whiteflies and the viruses they transmit a recipe for economic losses in the agriculture sector. Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) are small pests that quickly develop insecticide-resistance. Therefore, chemical control of infestations is difficult in affected crops i.e., pepper, sweet potatoes, cucumber, tomatoes, beans, hibiscus, etc. Two main cryptic species of Bemisia tabaci are reported to cause damage in the U.S, B. tabaci MEAM1 (Middle East Asia Minor 1, also called biotype B) and B. tabaci MED (Mediterranean, biotype Q). The two cryptic species transmit different viruses. Since minimal documentation is available for whiteflies in Oklahoma, insects were collected from different host plants in Stillwater, Oklahoma from August to October 2022. DNA was extracted and PCR was performed with primers that discriminate between MEAM1 and MED cryptic species. The partial mtCOI gene was also sequenced and a phylogenetic tree constructed. The results indicate the presence of MEAM 1 and MED cryptic species in Oklahoma. This is the first report of MED in Oklahoma. The correct identification of B. tabaci cryptic species contributes to structure management strategies, identifying the ecological niches, and pest movement.Entomology and Plant Patholog

Morphological and molecular characterization of powdery mildew on watermelon plants in São Paulo state

Powdery mildew is one of the most commom foliar diseases of cultivated and wild cucurbits worldwide. Six different fungal species have been reported to be associated with powdery mildew of cucurbits, such as Podosphaera xanthii and Golovinomyces cichoracearum. The aim of the study was to identify the causal agent of the disease in watermelon plants (Citrullus lanatus) cultivated in greenhouses of the State of São Paulo. DNA total was extracted using Chelex method and the ITS region was amplified via standard PCR reaction using the specific primers PMITS1 and PMITS2. A fragment of 700pb was amplified and the sequence showed 99% of identity with P. xanthii (accession number JQ728480). Morphological observation showed that the conidia were ellipsoidal-ovoid, had fibrosin bodies and a bifurcated germinative tube in the lateral position. These characteristics and the molecular analyses confirmed P. xanthi as the etiological agent of powdery mildew in watermelon plants in São Paulo State. Morphological and molecular characterization of powdery mildew on watermelon plants in São Paulo statePowdery mildew is one of the most commom foliar diseases of cultivated and wild cucurbits worldwide. Six different fungal species have been reported to be associated with powdery mildew of cucurbits, such as Podosphaera xanthii and Golovinomyces cichoracearum. The aim of the study was to identify the causal agent of the disease in watermelon plants (Citrullus lanatus) cultivated in greenhouses of the State of São Paulo. DNA total was extracted using Chelex method and the ITS region was amplified via standard PCR reaction using the specific primers PMITS1 and PMITS2. A fragment of 700pb was amplified and the sequence showed 99% of identity with P. xanthii (accession number JQ728480). Morphological observation showed that the conidia were ellipsoidal-ovoid, had fibrosin bodies and a bifurcated germinative tube in the lateral position. These characteristics and the molecular analyses confirmed P. xanthi as the etiological agent of powdery mildew in watermelon plants in São Paulo State

Reaction of lettuce genotypes to Lettuce mosaic virus-Most (LMV-Most) and characterization of the translation factor eIF4E

O objetivo deste trabalho foi avaliar genótipos de alface quanto à reação ao Lettuce mosaic virus (LMV; Most-type, isolado AF-199) e variações no fator eucariótico de tradução eIF4E. Todos os genótipos inoculados foram suscetíveis ao LMV, que foi detectado por RT-PCR com primers específicos. Porém, os acessos 169501, 169501C, 172918A e 162499, apresentaram sintomas tardios e somente nas folhas inoculadas. O sequenciamento da porção codificadora para eIF4E mostrou que estes genótipos apresentam padrão eIF4E0 (mol0), típico para suscetibilidade ao LMV, o que indica que o fenótipo encontrado não está correlacionado com as variações de nucleotídeos neste fator de tradução.The objective of this work was to evaluate lettuce genotypes for their reaction to Lettuce mosaic virus (LMV; Most-type, isolate AF-199) and variations of the eukaryotic translation initiation factor eIF4E. All inoculated genotypes were susceptible to LMV, which was detected by RT-PCR using specific primer pairs. However, the accessions 169501, 169501C, 172918A, and 162499 showed late development of symptoms that appeared only on the inoculated leaves. Sequencing of the coding region of eIF4E showed that these genotypes have an eIF4E0 (mol0) standard typical for their susceptibility to LMV, indicating that the phenotype found is not correlated to nucleotide variations in this translation factor

Only the B biotype of Bemisia tabaci is present on vegetables in São Paulo State, Brazil

Bemisia tabaci (Genn.) is one of the most important pests in cultivated areas of vegetables and ornamental crops around the world. Based on the mitochondrial cytochrome oxidize I (mtCOI) sequence, there is evidence that B. tabaci should be considered a cryptic species complex of 11 groups containing 24 species. Two of the groups, Middle East-Asia Minor 1 and Mediterranean include biotypes B and Q, respectively. In this study we evaluated the mtCOI sequence of B. tabaci populations collected in sites of the state of São Paulo, Brazil. Using PCR-RFLP with Taq I, a typical biotype B profile was obtained for all specimens. Based on the comparison with mtCOI reference sequences we found four haplotypes all belonging to the Middle East-Asia Minor 1. They occurred in the hosts pepper (Capsicum annuum L.), tomato (Solanum lycopersicum L.), eggplant (Solanum melongena L.) and cucurbitaceae plants.Bemisia tabaci (Genn.) é considerada uma das mais importantes pragas em cultivos de hortaliças e ornamentais em todo o mundo. Baseado na análise da seqüência mitocondrial (citocromo oxidase I - mtCOI) foi proposto recentemente que B. tabaci deva ser considerado um complexo críptico de espécies, contendo 11 grupos e 24 espécies. Dois destes grupos: Middle East-Asia Minor e Mediterranean englobam os biótipos B e Q, respectivamente. Avaliou-se a sequência mtCOI de espécimes de B. tabaci coletados em regiões do estado de São Paulo, Brasil. Por PCR-RFLP utilizando-se a enzima Taq I, pôde-se observar somente o padrão típico de clivagem para o biótipo B. Comparando-se com sequências consenso, todas as moscas brancas foram classificadas no grupo Middle East-Asia Minor e puderam ser separadas em quatro haplótipos, indicando prevalência do biótipo B em áreas de pimentão (Capsicum annuum L.), tomate (Solanum lycopersicum L.), cucurbitáceas e berinjela (Solanum melongena L.) do Estado de São Paulo

A aplicação de fungicidas pode incrementar a produção de tomateiro coinfectado por Begomovirus e Crinivirus

The objective of this work was to evaluate the effect of fungicide application on the concentration of Tomato severe rugose virus (ToSRV, Begomovirus) in the 'Mariana' hybrid tomato coinfected with ToSRV and Tomato chlorosis virus (ToCV, Crinivirus) and the progression of viral concentration by qPCR, as well as to quantify fruit yield and quality. Experiment I consisted in the application of fungicides after sowing (pretreatment): pyraclostrobin+metiram (P+M) (at 3 g L-1) + boscalid (B) (at 0.3 g L-1), followed by biweekly sprayings with P+M (4 g L-1); in experiment II, there was no application at sowing (control treatment), only 4 g L-1 P+M biweekly. ToSRV and ToCV transmissions were performed using sweetpotato whiteflies (Bemisia tabaci, biotype B), at 15, 30, 45, 60, and 70 days after transplanting (DAT). There was an increase in yield, better fruit quality, and a reduction in the ToSRV concentration in the plants, when the viruses were transmitted late and pretreatment was performed. Tray pretreatment in sowing with P+M (3 g L-1) and B (0.3 g L-1), followed by biweekly sprayings with P+M (4 g L-1), increases fruit yield and quality in the 'Mariana' hybrid tomato coinfected at 45, 60, and 75 DAT by ToSRV and ToCV, and there is a reduction in the concentration of ToSRV.O objetivo deste trabalho foi avaliar o efeito da aplicação de fungicidas na concentração de Tomato severe rugose virus (ToSRV, Begomovirus) no híbrido de tomateiro 'Mariana' coinfectado por ToSRV e Tomato chlorosis virus (ToCV, Crinivirus) e a progressão da concentração viral por qPCR, bem como quantificar a produção e a qualidade de frutos. O experimento I consistiu na aplicação dos fungicidas após a semeadura (pré-tratamento): piraclostrobina+metiram (P+M) (a 3 g L-1) + boscalida (B) (a 0,3 g L-1), seguida de pulverizações quinzenais com P+M (4 g L-1); no experimento II, não houve aplicação na semeadura (controle), apenas 4 g L-1 de P+M quinzenalmente. As transmissões de ToSRV e ToCV foram realizadas com uso de moscas-brancas (Bemisia tabaci, biótipo B), aos 15, 30, 45, 60 e 70 dias após o transplante (DAT). Houve aumento da produção, melhor qualidade dos frutos e redução da concentração de ToSRV nas plantas, quando os vírus foram transmitidos tardiamente e o pré-tratamento foi realizado. O pré-tratamento na bandeja, na semeadura, com P+M (3 g L-1) e B (0,3 g L-1), seguido de pulverizações quinzenais com P+M (4 g L-1), aumenta a produção e a qualidade dos frutos, em tomateiro híbrido 'Mariana' coinfectado aos 45, 60 e 75 DAT por ToSRV e ToCV, e há redução da concentração de ToSRV

Quebra da resistência em pimentão contra o Pepper yellow mosaic virus

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.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

Variabilidade genética em isolados de Curtobacterium flaccumfaciens

A cultura do feijoeiro está sujeita à incidência de várias doenças que acarretam perdas significativas na produção, dentre as quais encontra-se a murcha-de-curtobacterium ou murcha bacteriana, causada por Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff). Atualmente a murcha-de-curtobacterium tem se constituído em um novo problema para a cultura do feijoeiro em várias regiões brasileiras. A resistência genética tem sido o meio mais eficiente no controle da doença, porém a possibilidade da existência de variabilidade genética presente em isolados de Cff, pode ser uma conseqüência maléfica ao melhoramento visando a obtenção de cultivares de feijoeiro resistentes, especialmente na estabilidade e durabilidade da resistência. Neste sentido, o presente trabalho teve como objetivo o estudo da variabilidade genética de 26 isolados de Curtobacterium flaccumfaciens, 20 dos quais provenientes de feijoeiro (Cff), coletados em diferentes regiões do Brasil, quatro provenientes de coleções internacionais (Cff) e dois endofíticos de citros (C. flaccumfaciens). Foram utilizados dois pares de oligonucleotídeos, CffFOR2-CffREV4 e CF4-CF5, avaliando-se na especificidade em reação de PCR, para a caracterização dos 26 isolados. No estudo da variabilidade genética, utilizou-se da técnica rep-PCR e os iniciadores REP, ERIC e BOX. A partir do padrão eletroforético gerado pela amplificação dessas seqüências repetitivas no DNA genômico dos 26 isolados bacterianos, foram realizadas análises pertinentes (UPGMA SM) e obtenção de um dendograma. Considerando-se um índice de similaridade de 75%, os isolados foram distribuídos em quatro grupos distintos. Os isolados de Cff provenientes do Paraná e DF foram separados em grupos diferentes, enquanto que isolados endofíticos de citros não formaram um grupamento distinto dos de feijoeiro. Os isolados procedentes do Estados de São Paulo mostraram-se geneticamente heterogêneos, alguns se agruparam com o isolado de USA, Santa Catarina e de citros, enquanto outros agruparam-se com isolados provenientes da França e Paraná. A avaliação da especificidade dos dois pares de oligonucleotídeos, mostrou que os oligonucleotídeos CffFOR2-CffREV4 detectaram todos os 26 isolados. Os oligonucleotídeos CF4-CF5 apresentaram menor especificidade não detectando dois isolados de Cff de feijoeiro (2928) e (2936) e os isolados endofíticos de citros. Portanto como ferramenta para detecção de Cff em feijoeiro os oligonucleotídeos CffFOR2-CffREV4 revelaram ser os mais indicados.The bean crop is exposed to many diseases that lead to significant losses, such as the bacterial wilt of common bean caused by Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff). Currently, the bacterial wilt of common bean has been a new problem for the bean crops in several Brazilian regions. The genetic resistance has been the most efficient approach for disease control; however, the possible existence of genetic variability in Cff isolates may be a harmful consequence to the plant improvement especially in relation to the resistance stability and durability. For this reason, this study aimed the evaluation of the genetic variability of 26 Curtobacterium flaccumfaciens isolates, 20 isolates are from bean plants (Cff) collected in different Brazilian regions, four isolates are from international collections (Cff) and two isolates are from citrus endophytic community (C. flaccumfaciens). Two pairs of primers (CffFOR2-CffREV4 and CF4-CF5) were evaluated for its specificity in PCR reaction in the characterization of the 26 isolates studied. In the genetic variability evaluation, the rep-PCR technique with the REP, ERIC and BOX primers was used. From the electrophoresis pattern generated by the amplification of these repetitive sequences in the genomic DNA of the 26 bacterial isolates, a pertinent analysis (UPGMA SM) and a dendogram were performed. Considering a 75% similarity index, the isolates were distributed into four distinct groups. The Cff isolates from Paraná and Distrito Federal were separated in distinct groups, while citrus plant endophytic isolates did not form a distinct group from the bean plant ones. The isolates from São Paulo were genetically heterogeneous; and some of them were grouped with the USA, Santa Catarina and citrus plant isolates, while others were grouped with the ones from France and Paraná. The specificity evaluation of the two pairs of primers (CffFOR2-CffREV4 and CF4-CF5) used in the identification of Cff isolates in bean plant and citrus endophytic Curtobacterium flaccumfaciens isolates showed that the CffFOR2-CffREV4 combination was highly specific in the identification for the 26 isolates. The CF4-CF5 primers showed less specificity for two Cff isolateds in bean plant (2928) and (2936) and for the citrus endophytic isolates. Therefore, as an auxiliary tool in the Cff identification in bean plant, the primers CffFOR2-CffREV4 are the best indication