Economic relevance and adoption of monitoring and management ofthe fruit fly in citrus orchards of western Santa Catarina / Relevância econômica e adoçãodo monitoramento e manejo da mosca-das-frutas em pomares de citros do oeste catarinense

The west of Santa Catarina is the main citrus producing region in the state. The objective of this workwas to carry out a surveyabout the citriculture economic relevance and the monitoring and management of fruit flies by citrus growers in the west of Santa Catarina. A questionnaire with 13 questions was applied in locoto 58 citrus growers and the responses were organized for results analysis. In the west of Santa Catarina, for most of the respondents(67.8%), the citriculture is a complementary activity of another main activities practiced in the farm. Most citrus growers (91.38%) consider the fruit fly one of the mainphytosanitary problems of the crop and only 12.07% report a low level of incidence of this pest in their orchards. However, most farmers do not monitor the insect (41.4%) or are unaware of the subject (13.8%). Of those who monitor, 56.25% use traps, while43.75% monitor by visual observation of the fruits. Chemical control is carried out by almost half of citrus growers, erroneously based on the presence of fallen fruit or even by calendar applications. These presented data are worrying and reveal the negligence of citrus growers with the practice of monitoring this important pest.(AU)

A mesorregião do Oeste catarinense é a principal região produtora de citros do estado. O objetivo deste trabalho foi realizar um levantamento acerca da relevância econômica da atividade citrícola e do monitoramento e manejo de mosca-das-frutas adotado pelos citricultores do oeste catarinense. Um questionário com 13 perguntas foiaplicado in locoa 58 citricultores e as respostas foram organizadas para análise dos resultados. No oeste catarinense, para grande parte dos entrevistados (67,8%), a citricultura é uma atividade complementar de outras principais. A maioria dos citricultores (91,38%) considera a mosca-das-frutas um dos principais problemas fitossanitários da cultura e apenas 12,07% relatam como baixo o grau de incidência desta praga em seus pomares. No entanto, boa parte dos agricultores não faz monitoramento do inseto (41,4%) ou desconhece o assunto (13,8%). Dos que realizam monitoramento, 56,25% o fazem com uso de armadilhas, enquanto 43,75% monitoram por meio da observação visual dos frutos. O controle químico é realizado por praticamente metade dos citricultores, tendo,erroneamente,como base de decisão a presença de frutos caídos ou ainda por meio de aplicações por calendário. Os dados apresentados são preocupantes e revelam a negligência de boa parte dos citricultores com o monitoramento desta importante praga.(AU)

Lime sulfur to control citrus flat mite and its interactions with the entomopathogenic fungus Lecanicillium muscarium

Lime sulfur is one of the few products indicated to control Brevipalpus yothersi in Brazilian organic citrus orchards. Other strategies, such as the use of entomopathogenic fungi should be evaluated, and Lecanicillium muscarium is one of the basic choices for pest management. Knowledge of the interactions between lime sulfur and this entomopathogen is critical for developing control strategies. With this goal, it was conducted the toxicological characterization of lime sulfur to B. yothersi and the compatibility evaluation with L. muscarium. Finally, the effects of L. muscarium and lime sulfur mixtures on B. yothersi control were evaluated. Product evaluation for B. yothersi was done through direct and residual contact bioassay, and different concentrations of lime sulfur mixed in potato dextrose agar culture medium were used to evaluate compatibility with L. muscarium. Lime sulfur was effective against adults of B. yothersi and caused eggs unviability of up to 71.0%, at a dose of 80 L per 2,000 L of H2O. The lethal concentration (LC50 and LC99) of lime sulfur estimated for mite adults were 246.62 and 858.5 µg of sulfur per mL of H2O (ppm a.i.). Lime sulfur concentrations of 180 to 560 ppm a.i. showed promise for use in combination with L. muscarium. However, concentrations of 1,000 and 5,600 ppm significantly reduced colony size and the number of spores/colony. The mixture of 100 and 180 ppm a.i. of lime sulfur with L. muscarium (108 conidia·mL­1) was not able to reduce the lethal time of entomopathogen on B. yothersi.

Morphological and molecular identification of Cladosporium sphaerospermum isolates collected from tomato plant residues / Identificação morfológica e molecular de isolados de Cladosporium sphaerospermum coletados de resíduos de plantas de tomate

This study was conducted at the Agriculture College University of Karbala, Iraq to isolate and morphologically and molecularly diagnose thirteen Cladosporium isolates collected from tomato plant residues present in desert regions of Najaf and Karbala provinces, Iraq. We diagnosed the obtained isolates by PCR amplification using the ITS1 and ITS4 universal primer pair followed by sequencing. PCR amplification and analysis of nucleotide sequences using the BLAST program showed that all isolated fungi belong to Cladosporium sphaerospermum. Analysis of the nucleotide sequences of the identified C. sphaerospermum isolates 2, 6, 9, and 10 showed a genetic similarity reached 99%, 98%, 99%, and 99%, respectively, with those previously registered at the National Center for Biotechnology Information (NCBl). By comparing the nucleotide sequences of the identified C. sphaerospermum isolates with the sequences belong to the same fungi and available at NCBI, it was revealed that the identified C. sphaerospermum isolates 2, 6, 9, and 10 have a genetic variation with those previously recorded at the National Center for Biotechnology Information (NCBl); therefore, the identified sequences of C. sphaerospermum isolates have been registered in GenBank database (NCBI) under the accession numbers MN896004, MN896107, MN896963, and MN896971, respectively.

Este estudo foi conduzido na Agriculture College University of Karbala, Iraque, para isolar e diagnosticar morfológica e molecularmente treze isolados de Cladosporium coletados de resíduos de plantas de tomate presentes nas regiões desérticas das províncias de Najaf e Karbala, no Iraque. Diagnosticamos os isolados obtidos por amplificação por PCR usando o par de primers universais ITS1 e ITS4 seguido de sequenciamento. A amplificação por PCR e a análise de sequências de nucleotídeos usando o programa BLAST mostraram que todos os fungos isolados pertencem a Cladosporium sphaerospermum. A análise das sequências de nucleotídeos dos isolados 2, 6, 9 e 10 de C. sphaerospermum identificados mostrou similaridade genética de 99%, 98%, 99% e 99%, respectivamente, com aqueles previamente registrados no National Center for Biotechnology Informações (NCBl). Ao comparar as sequências de nucleotídeos dos isolados de C. sphaerospermum identificados com as sequências pertencentes aos mesmos fungos e disponíveis no NCBI, foi revelado que os isolados 2, 6, 9 e 10 de C. sphaerospermum identificados têm variação genética com aqueles anteriormente registrados no National Center for Biotechnology Information (NCBl). Portanto, as sequências identificadas de isolados de C. sphaerospermum foram registradas no banco de dados GenBank (NCBI) sob os números de acesso MN896004, MN896107, MN896963 e MN896971, respectivamente.

Morphological and molecular identification of Cladosporium sphaerospermum isolates collected from tomato plant residues / Identificação morfológica e molecular de isolados de Cladosporium sphaerospermum coletados de resíduos de plantas de tomate

This study was conducted at the Agriculture College University of Karbala, Iraq to isolate and morphologically and molecularly diagnose thirteen Cladosporium isolates collected from tomato plant residues present in desert regions of Najaf and Karbala provinces, Iraq. We diagnosed the obtained isolates by PCR amplification using the ITS1 and ITS4 universal primer pair followed by sequencing. PCR amplification and analysis of nucleotide sequences using the BLAST program showed that all isolated fungi belong to Cladosporium sphaerospermum. Analysis of the nucleotide sequences of the identified C. sphaerospermum isolates 2, 6, 9, and 10 showed a genetic similarity reached 99%, 98%, 99%, and 99%, respectively, with those previously registered at the National Center for Biotechnology Information (NCBl). By comparing the nucleotide sequences of the identified C. sphaerospermum isolates with the sequences belong to the same fungi and available at NCBI, it was revealed that the identified C. sphaerospermum isolates 2, 6, 9, and 10 have a genetic variation with those previously recorded at the National Center for Biotechnology Information (NCBl); therefore, the identified sequences of C. sphaerospermum isolates have been registered in GenBank database (NCBI) under the accession numbers MN896004, MN896107, MN896963, and MN896971, respectively.(AU)

Este estudo foi conduzido na Agriculture College University of Karbala, Iraque, para isolar e diagnosticar morfológica e molecularmente treze isolados de Cladosporium coletados de resíduos de plantas de tomate presentes nas regiões desérticas das províncias de Najaf e Karbala, no Iraque. Diagnosticamos os isolados obtidos por amplificação por PCR usando o par de primers universais ITS1 e ITS4 seguido de sequenciamento. A amplificação por PCR e a análise de sequências de nucleotídeos usando o programa BLAST mostraram que todos os fungos isolados pertencem a Cladosporium sphaerospermum. A análise das sequências de nucleotídeos dos isolados 2, 6, 9 e 10 de C. sphaerospermum identificados mostrou similaridade genética de 99%, 98%, 99% e 99%, respectivamente, com aqueles previamente registrados no National Center for Biotechnology Informações (NCBl). Ao comparar as sequências de nucleotídeos dos isolados de C. sphaerospermum identificados com as sequências pertencentes aos mesmos fungos e disponíveis no NCBI, foi revelado que os isolados 2, 6, 9 e 10 de C. sphaerospermum identificados têm variação genética com aqueles anteriormente registrados no National Center for Biotechnology Information (NCBl). Portanto, as sequências identificadas de isolados de C. sphaerospermum foram registradas no banco de dados GenBank (NCBI) sob os números de acesso MN896004, MN896107, MN896963 e MN896971, respectivamente.(AU)

First record of Anastrepha obliqua (Diptera: Tephritidae) and a tritrophic relation with parasitoids in a citrus orchard in Pará state, Brazil / Primeiro registro de Anastrepha obliqua (Diptera: Tephritidae) e de uma relação tritrófica com parasitoides em um pomar de citros no estado do Pará, Brasil

Citriculture is a growing industry in Pará state, Brazil, but information regarding fruit flies and their associated parasitoids in this region is lacking. To address this gap in knowledge, we collected oranges (Citrus sinensis), lime oranges (C. sinensis), common sweet limes (C. limettioides), citrons (C. medica) and mandarins (C. reticulata). We recorded field infestation by Anastrepha obliqua in C. sinensis under natural conditions for the first time in the study region, and a tritrophic relationship between C. sinensis, A. obliqua, and the parasitoids Opius bellus and Asobara anastrephae was identified.(AU)

A citricultura está crescendo no Estado do Pará, Brasil, mas faltam informações sobre as pragas conhecidas como moscas-das-frutas e os parasitoides associados na região. Visando abordar esta lacuna no conhecimento, foram coletados frutos de laranja (Citrus sinensis), laranja var. lima (C. sinensis), lima-da-pérsia (C. limettioides), limão-cidra (C. medica) e tangerina (C. reticulata). Registramos, pela primeira vez, infestação em condições naturais de campo por Anastrepha obliqua em C. sinensis no Pará e identificamos uma tritrófica entre C. sinensis, A. obliqua e os parasitoides Opius bellus e Asobara anastrephae.(AU)

Interactions of adjuvants on adhesion and germination of Isaria fumosorosea on adults of Diaphorina citri

Asian citrus psyllid, Diaphorina citri, is considered the most important citrus pest worldwide, as it transmits Huanglongbing – serious citrus disease. New efficient and sustainable strategies to control this pest have been investigated and the use of entomopathogenic fungi has become a promising alternative. The objective of this study was to evaluate the effects of adjuvants a) Tween 80 at 0.01 % (v/v); b) Silwet L77 at 0.025 % (v/v) and c) KBRAdj at 0.075 % (v/v) on adhesion, germination and pathogenicity of Isaria fumosorosea ESALQ-1296(5 × 106 conidia mL–1). Female adults of D. citri used in this experiment were sprayed on Citrus limonia seedlings. The sprayed insects were analyzed through scanning electron microscopy (SEM) to identify the most susceptible integument regions for fungus attachment and the effect of adjuvants used. In the pathogenicity test, adjuvants Silwet L77 and KBRAdj presented a higher efficiency than Tween 80. Fungi adhered predominantly to the ventral posterior (abdomen) region in comparison with the dorsal anterior (thorax) region. In addition, adjuvants Silwet L77 and KBRAdj presented faster germination ( 72 h). Conidial germination in the dorsal part of the thorax of the insects was observed only with adjuvant KBRAdj 72 h post inoculation.

Interactions of adjuvants on adhesion and germination of Isaria fumosorosea on adults of Diaphorina citri

Asian citrus psyllid, Diaphorina citri, is considered the most important citrus pest worldwide, as it transmits Huanglongbing serious citrus disease. New efficient and sustainable strategies to control this pest have been investigated and the use of entomopathogenic fungi has become a promising alternative. The objective of this study was to evaluate the effects of adjuvants a) Tween 80 at 0.01 % (v/v); b) Silwet L77 at 0.025 % (v/v) and c) KBRAdj at 0.075 % (v/v) on adhesion, germination and pathogenicity of Isaria fumosorosea ESALQ-1296(5 × 106 conidia mL–1). Female adults of D. citri used in this experiment were sprayed on Citrus limonia seedlings. The sprayed insects were analyzed through scanning electron microscopy (SEM) to identify the most susceptible integument regions for fungus attachment and the effect of adjuvants used. In the pathogenicity test, adjuvants Silwet L77 and KBRAdj presented a higher efficiency than Tween 80. Fungi adhered predominantly to the ventral posterior (abdomen) region in comparison with the dorsal anterior (thorax) region. In addition, adjuvants Silwet L77 and KBRAdj presented faster germination (< 48 h) of I. fumosorosea spores when compared to Tween 80 (> 72 h). Conidial germination in the dorsal part of the thorax of the insects was observed only with adjuvant KBRAdj 72 h post inoculation.(AU)

Closely-related Xanthomonas citri subsp. citri isolates trigger distinct histological and transcriptional responses in Citrus limon

Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), has an important economic impact on the citrus industry. Extensive information is available about the disease but, nevertheless, the study of plant-pathogen interactions could provide new information in the understanding of citrus canker disease. A new isolate has been identified, Xcc AT, which has a high genetic similarity (> 90 %) to the virulent Xcc T strain based on genetic clustering analyses of the rep-PCR fingerprinting patterns, but it does not produce cankerous lesions in Citrus limon. In this study, we compared C. limon responses to Xcc AT and to the virulent Xcc T strain at both histological and transcriptional levels. Histologically, leaves inoculated with Xcc AT exhibited neither a typical disordering of the spongy mesophyll, nor a swelling of epidermis. A particular content (undetermined) was also found in mesophyll cells near the stomata, together with increased starch accumulation. The transcriptomic profiles were compared by cDNA-AFLP technique. A total of 121 fragments derived from transcript (TDF) were either specifically induced or repressed by the isolates, and 62 were sequenced. Analysis of global expression identified different classes of genes known to be involved in plant-pathogen interactions. This study constitutes the first approach of the specific interaction between the avirulent Xcc AT isolate and C. limon.

Closely-related Xanthomonas citri subsp. citri isolates trigger distinct histological and transcriptional responses in Citrus limon

Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), has an important economic impact on the citrus industry. Extensive information is available about the disease but, nevertheless, the study of plant-pathogen interactions could provide new information in the understanding of citrus canker disease. A new isolate has been identified, Xcc AT, which has a high genetic similarity (> 90 %) to the virulent Xcc T strain based on genetic clustering analyses of the rep-PCR fingerprinting patterns, but it does not produce cankerous lesions in Citrus limon. In this study, we compared C. limon responses to Xcc AT and to the virulent Xcc T strain at both histological and transcriptional levels. Histologically, leaves inoculated with Xcc AT exhibited neither a typical disordering of the spongy mesophyll, nor a swelling of epidermis. A particular content (undetermined) was also found in mesophyll cells near the stomata, together with increased starch accumulation. The transcriptomic profiles were compared by cDNA-AFLP technique. A total of 121 fragments derived from transcript (TDF) were either specifically induced or repressed by the isolates, and 62 were sequenced. Analysis of global expression identified different classes of genes known to be involved in plant-pathogen interactions. This study constitutes the first approach of the specific interaction between the avirulent Xcc AT isolate and C. limon.(AU)

Dissemination of Aleurocanthus woglumi in citrus plants, its natural enemies and new host plants in the state of Rio de Janeiro, Brazil / Disseminação de Aleurocanthus woglumi em citros, seus inimigos naturais e novas plantas hospedeiras no estado do Rio de Janeiro, Brasil

Aleurocanthus woglumi is an exotic pest, widely disseminated in Brazil, with a high preference for citrus plants, but with a polyphagous feeding habit. The contribution of the state of Rio de Janeiro in the Brazilian production of citrus fruits is small; however, there are still public policies that encourage citrus production in the state. In 2010, the appearance of this pest in Rio de Janeiro was confirmed in the municipality of Cachoeiras de Macacu. The objectives of this research were to evaluate the dissemination of A. woglumi in this state, conduct a survey of new species of host plants, identify and evaluate the population of natural enemies present at two orchards that grow 'Tahiti' limes (Citrus latifolia) infested by the pest in Cachoeiras de Macacu, RJ, Brazil. In 19 municipalities, leaves of citrus and other species of plants presenting aleyrodid nymphs were collected, and yellow sticky traps were installed to capture adults for subsequent identification. At the 'Tahiti' lime orchards, the leaf collections were done to confirm the species of Aleyrodidae , and regarding A. woglumi , the natural enemies associated with this pest were collected directly from the infested plants. The results showed that A. woglumi is wide-spread in 12 municipalities. Three new host plants for A. woglumi were identified: Artocarpus heterophyllus (Moraceae) , Pouteria caimito (Sapotaceae) and Struthanthus flexicaulis (Loranthaceae). In Cachoeiras de Macacu, a new species of parasitoid of A. woglumi nymphs was named: Encarsia pergandiella (Hymenoptera: Aphelinidae), while the insects in the Coccinellidae (Coleoptera) family have stood out as predators of this pest.(AU)

Aleurocanthus woglumi é uma praga exótica, amplamente disseminada no Brasil, com grande preferência pelos citros, mas com hábito alimentar polífago. A participação do estado do Rio de Janeiro na produção brasileira de citros é pequena; todavia, ainda ocorrem políticas públicas de incentivo à citricultura no Estado. Em 2010, a ocorrência dessa praga no estado do Rio de Janeiro foi confirmada no município de Cachoeiras de Macacu. Os objetivos deste trabalho foram avaliar a disseminação de A. woglumi nesse Estado, levantar novas espécies de plantas hospedeiras, identificar e avaliar a população de seus inimigos naturais presentes em dois pomares de lima-ácida (Citrus latifolia) 'Tahiti', infestados pela praga em Cachoeiras de Macacu, RJ, Brasil. Foram realizadas coletas de folhas de citros e de outras espécies de plantas com presença de ninfas de aleirodídeos em 19 municípios, e armadilhas adesivas amarelas foram instaladas para a captura de adultos, para posterior identificação. Nos pomares de lima-ácida, coletas de folhas foram feitas para confirmar a espécie de aleirodídeo e, em se tratando de A. woglumi , coletaram-se inimigos naturais associados a essa praga diretamente nas plantas infestadas. Os resultados mostraram que A. woglumi está disseminada em 12 municípios. Três novas espécies de plantas hospedeiras de A. woglumi foram identificadas: Artocarpus heterophyllus (Moraceae), Pouteria caimito (Sapotaceae) e Struthanthus flexicaulis (Loranthaceae). Em Cachoeiras de Macacu, uma nova espécie de parasitoide de ninfas de A. woglumi foi assinalada: Encarsia pergandiella (Hymenoptera: Aphelinidae), enquanto que os insetos representantes da família Coccinellidae (Coleoptera) se destacaram como predadores dessa praga.(AU)

Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing

Huanglongbing (HLB), a devastating citrus disease caused by the bacterium Candidatus Liberibacter spp., is now responsible for significant economic losses worldwide. Yet, no effective disease control has been found, and the non-cultivability of the bacterium has severely hampered studies on the pathogen. The 16S rDNA gene is a well-characterized sequence, essential for cell survival, and is used for bacterial identification or assignment of close relationships at the genus and species levels. Quantitative Real-Time PCR (qPCR) assays based on 16S rDNA genes are widely used in the detection of Ca. Liberibacter spp. in multiplex reactions. We have developed for the first time a set of qPCR primers based on the conserved 16S rDNA gene, which specifically and simultaneously detects in a singleplex reaction, all three bacterial species associated with HLB, and can differentiateCa.Liberibacter asiaticus or africanus from americanus by their characteristic melting curves. The assay is very sensitive, and it was possible to amplify expected DNA fragments with an efficiency of 98 % using the Syber Green system and a Ct value lower than tested methods for HLB diagnosis. The application of this fast, simple and efficient detection methodology could also be important in the detection of all species of HLB-associated Liberibacters and could contribute to early pathogen detection, a crucial step in the development of preventive strategies aimed at avoiding the dissemination of this devastating disease in HLB-free areas.

Novel set of real-time PCR primers for simultaneous detection of Liberibacter species associated with citrus Huanglongbing

Huanglongbing (HLB), a devastating citrus disease caused by the bacterium Candidatus Liberibacter spp., is now responsible for significant economic losses worldwide. Yet, no effective disease control has been found, and the non-cultivability of the bacterium has severely hampered studies on the pathogen. The 16S rDNA gene is a well-characterized sequence, essential for cell survival, and is used for bacterial identification or assignment of close relationships at the genus and species levels. Quantitative Real-Time PCR (qPCR) assays based on 16S rDNA genes are widely used in the detection of Ca. Liberibacter spp. in multiplex reactions. We have developed for the first time a set of qPCR primers based on the conserved 16S rDNA gene, which specifically and simultaneously detects in a singleplex reaction, all three bacterial species associated with HLB, and can differentiateCa.Liberibacter asiaticus or africanus from americanus by their characteristic melting curves. The assay is very sensitive, and it was possible to amplify expected DNA fragments with an efficiency of 98 % using the Syber Green system and a Ct value lower than tested methods for HLB diagnosis. The application of this fast, simple and efficient detection methodology could also be important in the detection of all species of HLB-associated Liberibacters and could contribute to early pathogen detection, a crucial step in the development of preventive strategies aimed at avoiding the dissemination of this devastating disease in HLB-free areas.(AU)

Monitoramento do ácido chiquímico no manejo de plantas daninhas com glifosato em pomar comercial de citros / Shikimic acid monitoring in weed control with glyphosate in a commercial citrus orchard

Com o objetivo de verificar o acúmulo de ácido chiquímico em plantas de laranja pêra (Citrus sinensis) num pomar comercial manejado com glifosato, um herbicida sistêmico de amplo espectro, foram coletadas amostras na Fazenda Jequitibá, tradicional no cultivo de citros, situada no Município de Santo Antônio de Posse, SP. O produtor aplicou de forma convencional Roundup® Original a 1.440 g.ha-1 de equivalente ácido (e.a.) do sal de isopropilamino de glifosato em 19/12/ 2006 na entrelinha de 15 plantas, deixando outras cinco como testemunha. A reaplicação de glifosato a 1.260 g.ha-1 de e.a. foi realizada em 2/4/2007. Em ambos os casos, imediatamente antes da aplicação e aos 3, 7, 10, 15, 20 e 35 dias após, foram coletadas 20 folhas de cada planta tanto da área tratada como da não tratada, analisando-se o teor de ácido chiquímico por cromatografia líquida de alta eficiência (CLAE) de forma isocrática após extração por micro-ondas. Os resultados mostraram não ocorrer acúmulo do ácido chiquímico nas plantas de laranja pêra, não havendo diferenças significativas nos teores deste composto entre o material proveniente da área tratada com glifosato e o daquela capinada manualmente.

In order to check the accumulation of shikimic acid in a traditional commercial grove of citrus "Pêra" cultivar (Citrus sinensis) managed for weed control with glyphosate, a systemic herbicide with wide spectrum, samples were collected at Fazenda Jequitibá, in Santo Antonio de Posse County, São Paulo State, Brazil. The producer applied the following treatments of Roundup Original® glyphosate at 1,440 g.ha-1 a.e. of the isopropylamine salt on 19 December 2006 between rows of 15 plants, leaving five others as control. The reapplication of glyphosate at 1,260 g ha-1 was done on 2 April 2007. In both cases, immediately before application and at 3, 7, 10, 15, 20 and 35 days thereafter, 20 leaves from each treated and untreated plants were collected for analysis of the content of shikimic acid by isocratic high performance liquid chromatography (HPLC) assisted with microwave. The results showed no significant differences in levels of shikimic acid between the material from the area treated with glyphosate and that weeded manually.