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Leafhoppers That Carry Begomoviruses on Roselle Crop (Hibiscus Sabdariffa L.)

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Leafhoppers That Carry Begomoviruses on Roselle Crop (Hibiscus Sabdariffa L.) Leafhoppers that carry begomoviruses on roselle crop (Hibiscus sabdariffaL.) Cicadélidos portadores de begomovirus en el cultivo de jamaica (Hibiscus sabdariffaL.) Julieta Martínez-Cruz, Daniel Leobardo Ochoa-Martínez*, Javier Hernández-Morales, Posgrado en Fitosanidad-Fitopatología. Colegio de Postgraduados Km. 36.5 Carretera México-Texcoco, CP. 56230, Mon- tecillo, Texcoco, Estado de México; Erika Janet Zamora-Macorra, Universidad Autónoma Chapingo Km. 38.5 Carretera México-Texcoco, CP. 56230, Texcoco, Estado de México; Sergio Ramírez-Rojas, Campo Ex- perimental Zacatepec: CEZACA-INIFAP Km. 0.5 Carretera Zacatepec-Galeana; CP. 62780, Colonia Centro; Zacatepec, Morelos. * Autor para correspondencia: [email protected] Recibido: 25 de Enero, 2018. Aceptado: 24 de Abril, 2018. Martínez-Cruz J, Ochoa-Martínez DL, Hernández-Mo- Abstract. The viruses of the Begomovirus genus rales J, Zamora-Macorra EJ, Ramírez-Rojas S. 2018. have a worldwide distribution and to date it is Leafhoppers that carry begomoviruses on roselle crop known that they are transmitted exclusively by (Hibiscus sabdariffa L.). Revista Mexicana de Fitopato- Bemisia tabaci. In Mexico, in 2016, a complex of logía 36(2): 321-330. begomoviruses associated with the yellowing of DOI: 10.18781/R.MEX.FIT.1801-4 roselle (Hibiscus sabdariffa) was reported, in which Okra yellow mosaic Mexico virus (OYMMV) is Primera publicación DOI: 03 de Mayo, 2018. present. The objective of this study was to know the First DOI publication: May 03, 2018. carrier insects of begomoviruses associated with roselle. Insects were collected from plants with Resumen. Los virus del género Begomovirus tie- yellowing, vein clearing and mosaic and analyzed nen una distribución mundial y a la fecha se sabe by PCR. Three species of leafhoppers that carry que son transmitidos exclusivamente por Bemisia OYMMV were identified: Trypanalebra maculata, tabaci. En 2016, se reportó un complejo de bego- Kunzeana scimetara and Agallia excavata. In T. movirus asociados al amarillamiento de la jamaica maculata and A. modesta, Sida golden mosaic (Hibiscus sabdariffa L.) en México,entre los que Buckup virus (SiGMBuV) and Melon chlorotic leaf se encuentra el Okra yellow mosaic Mexico virus curl virus (MCLCuV) were detected, respectively. (OYMMV). Con el propósito de conocer la ento- This is the first report of leafhoppers as carriers of mofauna asociada al cultivo de jamaica portadora begomoviruses. de begomovirus, se colectaron insectos en plantas con amarillamiento, aclaramiento de nervadu- Key words: virus vector insects, Agallia, ras y mosaico y se analizaron mediante PCR. Se Trypanalebra, Kunzeana. Publicación en línea, mayo 2018 321 Revista Mexicana de FITOPATOLOGÍA Mexican Journal of Phytopathology Fully Bilingual identificaron tres especies de cicádelidos portado- The Begomovirus genus belongs to the ras de OYMMV: Trypanalebra maculata, Kunzea- Geminiviridae family, which includes pathogens na scimetara y Agallia excavata. Asimismo, se en- that have circular genomes of single-stranded contró que T. maculata y A. modesta son portado- DNA with one or two components of 2700- ras de los begomovirus Sida golden mosaic Buckup 3000 pb within incomplete icosahedral particles virus (SiGMBuV) y Melon chlorotic leaf curl virus (geminated). They are responsible for several (MCLCuV), respectivamente. Este es el primer re- diseases that affect economically important crops porte de cicadélidos portadores de begomovirus. in tropical and subtropical regions worldwide (Moffat, 1999). Based on its host range, insect Palabras clave: insectos vectores de virus, Aga- vector, genomic composition and sequence llia, Trypanalebra, Kunzeana. similarity, the Geminiviridae family is divided into seven genera (Varsani et al., 2009; Varsani et al., 2014). One of them is the Begomovirus genus that El género Begomovirus pertenece a la familia includes over 60 species exclusively transmitted by Geminiviridae, el cual comprende patógenos que a Bemisia tabaci species complex (Markham et al., poseen genomas circulares de DNA monocatenario 1994). In Mexico, roselle (Hibiscus sabdariffa L.) compuesto por uno o dos componentes de 2700- is a crop of great economic importance, and Ayutla 3000 pb, contenidos dentro de partículas icosaédri- and Tecoanapa, Guerrero, are the municipalities cas (geminadas) incompletas. Son responsables de with the greatest cultivated area at the national level diversas enfermedades en cultivos de importancia (SIAP, 2015). Worldwide, four viruses associated económica en regiones tropicales y subtropicales with this crop have been reported: Cotton leaf del mundo (Moffat, 1999). Con base en su gama curl virus (CLCuV), Malva vein clearing virus de hospedantes, insecto vector, organización de su (MVCV) (Brunt et al., 1996), Okra mosaic virus genoma y similitud de secuencias, la familia Gemi- (OkMV) (Stephan et al., 2008) and Mesta yellow niviridae se divide en siete géneros (Varsani et al., vein mosaic virus (MYVMV) (Chatterjee et al., 2009; Varsani et al., 2014). Uno de éstos es el gé- 2008). In Mexico, a begomovirus complex was nero Begomovirus que agrupa más de 60 especies found associated with roselle yellowing, including transmitidas exclusivamente por un complejo de Okra yellow mosaic Mexico virus (OYMMV) especies de Bemisia tabaci (Markham et al., 1994). (Velázquez et al., 2016). In the 2015 cycle, the En México, la jamaica (Hibiscus sabdariffa L.) es disease was detected in two plots in Tecoanapa un cultivo de gran importancia económica, Ayut- with 100% incidence. Whitefly populations are la y Tecoanapa, Guerrero, son los municipios con very low on crops in that production area. Based on mayor superficie cultivada a nivel nacional (SIAP, this, the objective of the present study was to find 2015). En el ámbito mundial se reportan cuatro out if there are other insects that carry OYMMV or virus asociados al cultivo, Cotton leaf curl virus other begomoviruses. (CLCuV), Malva vein clearing virus (MVCV) In 2016, two samplings of the municipalities (Brunt et al., 1996), Okra mosaic virus (OkMV) of Tecoanapa and Ayutla, Guerrero, were done (Stephan et al., 2008) y Mesta yellow vein mosaic (Table 1). The first sampling took place in the 2015 virus (MYVMV) (Chatterjee et al., 2008). En México, cycle, from August 3 to 5, when the crop was in se consignó un complejo de begomovirus asociados its vegetative stage, in locations where there was Publicación en línea, mayo 2018 322 Revista Mexicana de FITOPATOLOGÍA Fully Bilingual Mexican Journal of Phytopathology al amarillamiento de la jamaica, entre los cuales a high incidence of plants showing yellowing. The se encuentra Okra yellow mosaic Mexico virus second sampling took place during the 2016 cycle, (OYMMV) (Velázquez et al., 2016). En el ciclo from November 26 to 28, when calyces were being 2015 esta enfermedad se presentó en dos parcelas harvested, in plots where the incidence of yellowing de Tecoanapa con una incidencia de 100%. Sien- was high. In all cases, insects were collected with do además muy bajas las poblaciones de mosquita a sweep net from roselle plants and surrounding blanca en el cultivo en esa zona productora. Con weeds showing yellowing, vein clearing and base en lo anterior, el objetivo del presente estudio mosaic, and placed in plastic containers containing fue conocer si existen otros insectos portadores de 96% ethanol. In the laboratory, the insects were OYMMV u otros begomovirus. separated according to their morphological Durante 2016 realizaron dos recorridos en los similarities and kept at -20 °C. Total DNA was municipios de Tecoanapa y Ayutla, Guerrero (Cua- extracted from 2 or 3 individuals in each insect dro 1). El primero del 3 al 5 de agosto cuando el group using CTAB (Sambrook and Rusell, 2001). cultivo se encontraba en etapa vegetativa en locali- The remaining insects in each group were kept in dades donde se tuvo una alta incidencia de plantas ethanol to be identified later, in case they tested con amarillamiento en el ciclo 2015. El segundo positive for begomoviruses through PCR using recorrido se hizo del 26 al 28 de noviembre en la AV494/AC1048 universal primers and under the época de cosecha de cálices en sitios donde la inci- amplification conditions reported by Wyatt and dencia de amarillamiento fue alta en el ciclo 2016. Brown (1996), which amplify a 550 pb fragment. En todos los casos se colectaron insectos de plantas The amplified products were sequenced and de jamaica y maleza adyacente a ellas que mostra- compared to those in the GenBank database. The ban amarillamiento, aclaramiento de nervaduras y insects that tested positive for begomoviruses were mosaico con una red de golpeo y se colocaron en mounted, identified using taxonomic keys (Table 1) envases de plástico que contenían etanol 96%. En and photographed using an optical microscope. el laboratorio los insectos se separaron con base During the first sampling, insects of the en sus semejanzas morfológicas y se mantuvie- Thysanoptera (207 individuals), Coleoptera (76 ron a -20 °C. Se tomaron de 2 a 3 individuos de individuals) and Hemiptera orders were found. Four cada grupo de insectos y se extrajo DNA total con families of the Hemiptera order were identified: CTAB (Sambrook y Rusell, 2001). El resto de los Membracidae (17 individuals), Pyrrhocoridae insectos de cada grupo se mantuvieron en alcohol (62 individuals from the Dysdercus genus) and para su posterior identificación en caso de resultar Aleyrodidae and Cicadellidae. Pérez et al. (2009) positivos por PCR para begomovirus
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