8 research outputs found

    First Report of Citrus variegation virus in Palestine Sweet Lime, as Coffee Shade in Costa Rica

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    Symptoms resembling those of Citrus variegation virus (CVV) were observed in old Palestine sweet lime trees (Citrus limettioides Tan.) used as coffee shade in the Central Valley in Costa Rica. The symptoms include leaf flecking, mosaic, malformation, and dwarfing. This disease was transmitted by grafting to “Valencia” sweet orange (C. sinensis L.), Etrog Citron (C. medica L.), and sweet lime under greenhouse conditions. Symptoms were severe in Palestine sweet lime all year long, but sweet orange and Etrog citron were asymptomatic under high temperature. Reverse transcription polymerase chain reaction (RT-PCR) was used to confirm the presence of CVV in young leaves from sweet lime trees collected from field, and other Citrus spp. grafted and maintained in the greenhouse at University of Costa Rica. Fresh tissue from trees infected with CVV obtained from the USDA (Riverside, California), was used as positive control. Furthermore, the sequence obtained (605 bp) was analyzed by Blastn algorithm which showed 97% homology with CVV RNA 3 (GenBank Accession No. AF434912). Additionally, small scale virus purification was carried out and isometric particles (26 to 32 nm) were observed under the electron microscope. To our knowledge, this is the first report of the presence of CVV in Costa Rica infecting Palestine sweet limes. Also, we remark on the potential of Palestine sweet lime as a host plant indicator in biological indexing of CVVMinisterio de Ciencia y Tecnología, Fondo Reinserción/[222-2004]/MICIT- CONICIT/Costa RicaUniversidad de Costa Rica/[801-A5-506]/UCR/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM)UCR::Vicerrectoría de Docencia::Ciencias Agroalimentarias::Facultad de Ciencias Agroalimentarias::Escuela de AgronomíaUCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Biologí

    Memoria del II Coloquio Internacional sobre Diversidad Cultural y Estudios Regionales

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    Desde la Sede de Occidente y, específicamente, en el seno de la Coordinación de Investigación y con el apoyo de la Dirección de la Sede y las Coordinaciones de Docencia, Administración y Acción Social, así como de la Vicerrectoría de Investigación, se han celebrado en el 2011 y en el 2012 dos coloquios internacionales sobre diversidad cultural y estudios regionales. El propósito de ambos consistió principalmente en motivar a investigadores e investigadoras de las distintas unidades académicas de la Universidad de Costa Rica y de otras instituciones de Educación Superior, así como a representantes de Centros e Institutos de Investigación nacionales e internacionales, a presentar resultados de investigaciones que contribuyeran a un mayor conocimiento de los procesos culturales y que dieran a conocer resultados que permitieran la comprensión de las realidades de distintos sectores y regiones. En el 2011 se presentaron cuarenta y seis ponencias y se dictaron tres conferencias magistrales, y en el 2012 se expusieron cincuenta ponencias y se impartieron tres conferencias magistrales.UCR::Sedes Regionales::Sede de Occidente::Recinto San Ramón::Centro de Investigaciones sobre Diversidad Cultural y Estudios Regionales (CIDICER

    Citrus leprosis pathosystem: virus-vector-host relationship

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    A leprose dos citros é considerada uma das doenças mais destrutivas da indústria citrícola. O patossistema dessa doença envolve o agente causal o Citrus leprosis virus C, o vetor Brevipalpus phoenicis Geijskes (Acari: Tenuipalpidae) e as plantas hospedeiras. Este vírus com genoma conhecido é membro tipo do gênero Cilevirus. Causa sintomas localizados nas folhas, frutos e caules e está restrito ao continente Americano. Apesar de muitos esforços para se conhecer o patossistema, ainda existem muitas questões pendentes sobre as interações. O presente trabalho teve como objetivo obter informações detalhadas sobre as relações vetorvírus- hospedeiro da leprose citoplasmática, estando dividido em três objetivos: 1. Procurou-se uma planta indicadora padrão que fosse de facil obtenção, manejo e baixo custo, e que expressase em curto tempo os sintomas de CiLV-C e outros vírus transmitidos por Brevipalpus (VTBr); 2. Determinar parâmetros como o período de acesso para aquisição e inoculação do CiLV-C pelo ácaro vetor, o período de retenção do vírus pelo ácaro, avaliação da capacidade das diferentes fases do ácaro de transmitir o vírus e a % de indivíduos de uma população de ácaros colonizando plantas afetadas pela leprose capazes de transmitir o vírus; 3. Avaliar um grande número de espécies de plantas, de diferentes famílias botânicas, quanto à suscetibilidade experimental à infecção pelo CiLV-C, pelo ácaro vetor. A presença de CiLV-C nestes ensaios foi confirmada por testes de ELISA, RT-PCR, microscopia eletrônica de transmissão e imunofluorescência. O feijoeiro comum (Phaseolus vulgaris L.) reage com lesões necróticas locais em apenas cinco dias à inoculação com ácaros virulíferos, podendo-se reduzir para dois dias se as plantas forem incubadas a 28ºC. O cv. ,,Una\" foi selecionado como planta-teste padrão dentre 113 cvs. avaliados. Outros vírus transmitidos por Brevipalpus do tipo citoplasmático (VTBr) como vírus da pinta verde do maracujá (Passion fruit green spot virus PFGSV), vírus da mancha anular de Ligustrum (Ligustrum ringspot virus LigRSV) e mancha verde do hibisco (Hibiscus green spot vírus HGSV) também causaram lesões locais necróticas em feijoeiro. Usando feijão como planta teste e ácaros virulíferos, se determinou o período de acesso à aquisição do vírus-4 h; período de acesso à inoculação do vírus-4h; período de retenção do vírus no ácaro de 12 dias, porcentagem de ácaros virulíferos, colonizando tecidos infectados, transmitirem CiLV-C em até 45%. Os experimentos também confirmaram que todas as etapas de desenvolvimento do ácaro (larvas, proto-deutoninfa e adultos) são capazes de transmitir CiLV-C, inclusive confirmou-se que o macho tem a capacidade de transmissão e que o vírus pode ser adquirido a partir de lesões de folhas, frutos e caules. Não se constatou transmissão transovariana do CiLV-C. Foram testadas 140 espécies, de 45 famílias, dentro das quais 62 espécies de 26 famílias apresentaram lesões localizadas nas folhas. Destas, 46 espécies sintomáticas, o CiLV-C foi detectado em pelo menos um dos testes para sua detecção, confirmando a transmissão. O conhecimento de parâmetros de alimentação, retenção do vírus e porcentagem de ácaros viruliferos na população, além do genoma de hospedeiras suscetíveis ao vírus tem importantes implicações no entendimento da epidemiologia, na quarentena e podem oferecer indícios da origem do CiLV-C.Citrus leprosis (CL), caused by the Citrus leprosis virus C (CiLV-C), is reported only in the American continent. The pathosystem of the CL involves the causal agent, the main vector Brevipalpus phoenicis Geijeskes (Acari: Tenuipalpidae) and the susceptible hosts. For long time only Citrus spp. were considered the sole susceptible host. The entire genome of the CiLV-C was sequenced and a new genus, Cilevirus, was assigned for this virus. CL is characterized by the induction of localized symptoms on the leaves, fruits and stems. Important advances were made recently for the understanding of CL pathosystem, but despite these efforts little is known about details of the virus/vector/host relationship. The present work aimed to cover such deficiencies. In the first place a search of suitable indicator plant was made and the common bean (Phaseolus vulgaris L.) was found to respond with localized necrotic lesions after infestation with viruliferous B. phoenicis in five days and when infested leaves are incubated at 28°C. Furthermore, bean plants are easy and cheap to produce and handle. The black bean cv. ,,IAC Una\" was adopted as a standard test variety, among 113 assayed cultivars of various genetic backgrounds. Common bean plants mite-inoculated with other cytoplasmic-type Brevipalpus-transmitted viruses (BrTVs) [Passion fruit green spot virus (PFGSV), Solanum violaefolium ringspot virus (SvRSV), Ligustrum ringspot virus (LigRSV) and Hibiscus green spot virus (HGSV)] also responded with necrotic local lesions and could serve as test plants for these viruses. Detecion of these viruses were made by RTPCR and/or transmission electron microscopy. Using common bean as test plant, some parameters of the vector/virus relationship were determined: virus acquisition feeding period- 4 h; virus inoculation feeding period- 4h; period of retention of the virus by a single viruliferous mite- at least 12 days; percentage of viruliferous mites from a mites colonizing infected tissues- 45%. The experiments also confirmed that all the developmental stages of the mite (larvae, proto- and deutonymph, adult) as well as males are able to transmit CiLV-C. No transovarial passage of the CiLV-C was registered. The virus can be acquired from lesions of leaves, fruits and stems. To assess the experimental host range of CiLV-C, a large number of plant species were inoculated with B. phoenicis, viruliferous to CiLV-C, under experimental conditions. Of the140 species tested, belonging to 45 families, 62 (of 26 families) produced localized lesions on inoculated leaves. Of these 62 plants producing local lesions, 45 had the presence of CiLV-C confirmed by at least one of the assays to detect the virus (RT-PCR, ELISA, transmission electron microscopy and immunofluorescence). These assays were also used to confirm the presence of CiLV-C in transmission experiments. Although only few non Citrus species were found naturally infected by CiLV-C, present results show that a large number of plant species are susceptible to the virus with implications on the epidemiology, quarantine and the evolution of the citrus leprosis pathosystem

    Detection of Plantago asiatica mosaic virus in lily hybrid plants(Lilium spp.) in Costa Rica grown from imported bulbs

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    Plantago asiatica mosaic virus (PlAMV) was detected in Costa Rica infecting lily plants showing foliar chlorotic and necrotic streaking. Virus identity was established by TEM, sequencing and ELISA. Pathogenicity was confirmed on Nicotiana benthamiana by mechanical inoculation, and PlAMV was detected by ELISA and RT-PCR.Universidad de Costa Rica/[801-B3-126]/UCR/Costa RicaUniversidad de Costa Rica/[801-A1-801]/UCR/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM)UCR::Vicerrectoría de Docencia::Ciencias Agroalimentarias::Facultad de Ciencias Agroalimentarias::Escuela de Agronomí

    First report of Iris yellow spot virus in Costa Rica

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    Copyright © 2017, Australasian Plant Pathology Society Inc.Iris yellow spot virus (IYSV), an emerging disease of onion crops, was identified by transmission electron microscopy, enzyme linked immunosorbent assay, and reverse transcription polymerase chain reaction in Costa Rica. Onion plants had straw-colored, elongated lesions and tip dieback. Costa Rican IYSV nucleocapsid partial sequences (15 isolates) grouped with isolates from North and Central America and from New Zealand.Universidad de Costa Rica/[801-B3-126]/UCR/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM)UCR::Vicerrectoría de Docencia::Ciencias Agroalimentarias::Facultad de Ciencias Agroalimentarias::Escuela de Agronomí

    First Report of Xylella fastidiosa in Avocado in Costa Rica

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    Copyright 2008 American Phytopathology Society Journals.Since the late 1990s, chlorotic mottling, marginal scorch, deformation of leaves, defoliation, shortening of internodes, and branch dieback have been observed in avocado trees (Persea americana Mill.) in Costa Rica. The symptoms are not uniformly distributed in the tree, so some branches are symptomatic while others are not. These symptoms are similar to several leaf scorch diseases caused by the bacterium Xylella fastidiosa Wells (2,4). This bacterium has been detected in coffee and citrus plants in Costa Rica. Of 227 avocado trees tested by double-antibody sandwich (DAS)-ELISA with X. fastidiosa specific antiserum (Agdia Inc., Elkhart, IN) from 2000–2004, 188 were positive. Results of ELISA tests of individual trees varied with the season and branches tested. Fifteen greenhouse-grown, ELISA-negative avocado seedlings were grafted with budwood from an ELISA-positive tree. Eight of these developed scorch symptoms and one also showed chlorotic mottling and deformation, showing that the disease is graft transmitted. All of these features are characteristic of diseases caused by X. fastidiosa (2,4). Transmission electron microscopy of leaf petioles from three field trees positive by ELISA, revealed rod-shaped bacilli approximately 1.6 to 2.0 μm long and 0.3 μm in diameter with a rippled cell wall inside xylem vessels and embedded in a matrix; morphology and measurements that are consistent with those reported for X. fastidiosa (2). DNA extraction and PCR attempts have been limited by mucilaginous sap from avocado. Positive PCR results (approximately 472-bp band) were obtained from two of the grafted seedlings and seven field trees from two distinct geographical locations (Alajuela and San José provinces) with DNA extractions from the plant sap using DNeasy Plant Mini Kit (Qiagen GmbH, Hilden, Germany) following a modified protocol (1) and nested PCR (3). Four of the PCR products, including one from the grafted seedlings, were cloned and sequenced in duplicate. GenBank sequences EU021997 to EU022000 present 99 to 100% sequence identity to a Pierce's disease strain from California (Temecula1) and 94 to 95% to a citrus variegated chlorosis strain from Brazil (Found-5). Several attempts have been made to isolate the bacterium in ‘periwinkle wilt’ and buffered cysteine-yeast extract media with negative results, probably because of the rapid production of mucilaginous sap when the avocado tissues were sampled. To our knowledge, this is the first report of X. fastidiosa in avocado trees.Fundación CRUSAUniversidad de Costa Rica/[801-A2-528]/UCR/Costa RicaUniversidad de Costa Rica/[801-A1-801]/UCR/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM

    Catharanthus roseus (Apocynaceae) naturalmente infectada con diversos fitoplasmas en Costa Rica

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    Phytoplasmas (class Mollicutes) are causal agents of plant diseases with an economic impact on crops or threatening local biodiversity. A survey was conducted from 2012 to 2016 on infected Catharanthus roseus plants that exhibited symptoms reminiscent of phytoplasma infection throughout Costa Rica. A total of 73 plants were collected exhibiting symptoms such as virescence, phyllody, axillary proliferation, little leaf, leaf malformation, chlorosis, or yellowing. All samples were tested by nested PCR using phytoplasma universal and specific primer pairs. Phytoplasma infection was detected in 52 (71.2 %) of the plants collected. Phytoplasmas of six subgroups belonging to 16Sr groups I, III, IX, XIII and XV were identified based on sequencing and in silico RFLP analyses. ´Candidatus Phytoplasma asteris´ (16SrI) was the predominant group among the positive samples (n = 30) showing variety of symptoms and wide distribution from sea level to ca. 1 400 masl in six of the seven Costa Rican provinces. Group 16SrIII was the second most abundant (14 samples); and the remaining three groups were seldom found in C. roseus (8 samples). Moreover, group 16SrXIII phytoplasma was detected for the first time in the country. To the best of our knowledge, this is the first report of natural infection of C. roseus with phytoplasma subgroups 16SrI-B, 16SrI-P, 16SrIII-F, 16SrIX-F, 16SrXIII-A, and 16SrXV-B in Costa Rica and Central America.Catharanthus roseus (Apocynaceae) naturalmente infectada con diversos fitoplasmas en Costa Rica. Los fitoplasmas (clase Mollicutes) son agentes causales de enfermedades de plantas que provocan pérdidas económicas o amenazan la biodiversidad local. Una recolecta de plantas de Catharanthus roseus que mostraban síntomas de posible infección con fitoplasmas se realizó en diferentes lugares de Costa Rica desde 2012 a 2016. Un total de 73 plantas fueron recolectadas con síntomas tales como viriscencia, filodia, brotación axilar múltiple, reducción foliar, deformación foliar, clorosis, y amarillamiento. Todas las muestras fueron evaluadas mediante PCR anidado usando los pares de imprimadores universales y específicos para fitoplasmas. Infección por fitoplasmas se detectó en 52 (71.2 %) de las muestras. Fitoplasmas de seis subgrupos dentro de los grupos 16Sr I, III, IX, XIII y XV fueron identificados basados en secuenciación del ADN y análisis de polimorfismos de restricción (RFLP) in silico. El grupo predominante encontrado en las muestras positivas (n = 30) fue el 16SrI (‘Candidatus Phytoplasma asteris’), éste mostró variedad de síntomas y amplia distribución desde el nivel del mar hasta casi los 1 400 m.s.n.m. en seis de las siete provincias de Costa Rica. El grupo 16SrIII fue el segundo más abundante (14 muestras); y los restantes tres grupos se encontraron en pocas muestras de C. roseus (8 muestras). Además, fitoplasmas del grupo 16SrXIII se detectaron por primera vez en el país. De acuerdo a nuestro conocimiento, este es el primer informe de infección natural de C. roseus con fitoplasmas de los subgrupos 16SrI-B, 16SrI-P, 16SrIII-F, 16SrIX-F, 16SrXIII-A y 16SrXV-B en Costa Rica y Centroamérica.Universidad de Costa Rica/[801-A1-801]/UCR/Costa RicaUniversidad de Costa Rica/[801-B3-091]/UCR/Costa RicaUniversidad de Costa Rica/[801-B7-138]/UCR/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM

    Virus-vector relationship in the Citrus leprosis pathosystem

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    Citrus leprosis has been one of the most destructive diseases of citrus in the Americas. In the last decade important progress has been achieved such as the complete genome sequencing of its main causal agent, Citrus leprosis virus C (CiLV-C), belonging to a new genus Cilevirus. It is transmitted by Brevipalpus yothersi Baker (Acari: Tenuipalpidae), and is characterized by the localized symptoms it induces on the leaves, fruits and stems. It occurs in the American continents from Mexico to Argentina. The virus was until recently considered restricted to Citrus spp. However, it was found naturally infecting other plants species as Swinglea glutinosa Merrill and Commelina benghalensis L., and has been experimentally transmitted by B. yothersi to a large number of plant species. Despite these advances little is known about the virus-vector relationship that is a key to understanding the epidemiology of the disease. Some components of the CiLV-C/B. yothersi relationship were determined using the common bean (Phaseolus vulgaris L. cv. ‘IAC Una’) as a test plant. They included: (a) the virus acquisition access period was 4 h; (b) the virus inoculation access period was 2 h; (c) the latent period between acquisition and inoculation was 7 h; (d) the period of retention of the virus by a single viruliferous mite was at least 12 days; (d) the percentage of viruliferous individuals from mite colonies on infected tissues ranged from 25 to 60%. The experiments confirmed previous data that all developmental stages of B. yothersi (larva, protonymph and deutonymph, adult female and male) were able to transmit CiLV-C and that transovarial transmission of the virus did not occur. CiLV-C can be acquired from lesions on leaves, fruits and stems by B. yothersi. Based on the distribution of lesions produced by single viruliferous B. yothersi on bean leaves, it is concluded that they tend to feed in restricted areas, usually near the veins. The short latent and transmission periods during the larval stage suggest that the CiLV-C/B. yothersi relationship is of the persistent circulative type.Fundación de apoyo a la investigación estatal/[2014/08458-9]/Fapesp/BrasilConsejo Nacional de desarrollo científico y tecnológico/[47.2425/2013-7]/CNPq/BrasilFundación de apoyo a la investigación estatal/[2008/57477-5]/Fapesp/BrasilFundación de apoyo a la investigación estatal/[2013/25713-0]/Fapesp/BrasilUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM
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