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Confirmation of the Identity of Exserohilum Turcicum, Causal Agent of Maize Leaf Blight in Sinaloa

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Confirmation of the Identity of Exserohilum Turcicum, Causal Agent of Maize Leaf Blight in Sinaloa Confirmation of the identity of Exserohilum turcicum, causal agent of maize leaf blight in Sinaloa Confirmación de la identidad de Exserohilum turcicum, agente causal del tizón foliar del maíz en Sinaloa Rubén Félix-Gastélum*, Glenda Judith Lizárraga-Sánchez, Universidad de Occidente, Unidad Los Mochis, Departamento de Ciencias Biológicas, Bulevard Macario Gaxiola y Carretera Internacional s/n Los Mochis, Sinaloa, CP. 81223, México; Ignacio Eduardo Maldonado-Mendoza, Karla Yeriana Leyva-Madrigal, Ins- tituto Politécnico Nacional. Centro Interdisciplinario de Investigación para el Desarrollo Integral Regional- IPN Unidad Sinaloa. Departamento de Biotecnología Agrícola. Bulevard Juan de Dios Bátiz Paredes No. 250. Guasave, Sinaloa, CP. 81101, México; Gabriel Herrera- Rodríguez, Junta Local de Sanidad Vegetal del Valle del Fuerte. Lázaro Cárdenas, PTE. 315, Colonia Centro, Los Mochis Sinaloa, CP. 81200, México; Silvia Espinoza-Matías, Laboratorio de Microscopía Electrónica de Barrido, Facultad de Ciencias, Universidad Na- cional Autónoma de México. Delegación Coyoacán. CDMX, CP. 04510. *Autor para correspondencia: ruben. [email protected]. Recibido: 09 de Marzo, 2018. Aceptado: 03 de Julio, 2018. Félix-Gastélum R, Lizárraga-Sánchez GJ, Maldonado- Abstract. Fungal diseases constitute an important Mendoza IE, Leyva-Madrigal KY, Herrera-Rodríguez maize production constraint in Sinaloa, Mexico. In G, Espinoza-Matías S. 2018. Confirmation of the iden- recent growing seasons, a severe foliar epiphytotic tity of Exserohilum turcicum, causal agent of maize leaf disease has occurred in the northern coastal region blight in Sinaloa. Revista Mexicana de Fitopatología of this state. The symptoms of the disease resembled 36(3): 468-478. those caused by Exserohilum turcicum, reported in DOI: 10.18781/R.MEX.FIT.1803-1 the high humid valleys in the states of the central plateau of Mexico and other parts of the world with Primera publicación DOI: 07 de Agosto, 2018. temperate to subtropical climate. The objective First DOI publication: August 07, 2018 of this research was to confirm the identity of E. turcicum associated to maize leaf blight and to assess the disease severity in 17 maize hybrids in Resumen. Las enfermedades de origen fungoso commercial fields. Variation in color and colony constituyen una limitante en la producción de maíz morphology of the fungal isolates was observed in en Sinaloa, México. En ciclos agrícolas recientes, various media. The maize leaf extract agar was the una enfermedad foliar ha ocurrido en niveles epi- most favorable medium for fast mycelial growth démicos en la región costera del Norte de Sinaloa. of the fungal isolates. The isolates collected were Los síntomas de la enfermedad se asemejan a los used in pathogenicity tests, where they exhibited a causados por Exserohilum turcicum previamente variation in pathogenicity on the commercial maize Publicación en línea, septiembre 2018 468 Revista Mexicana de FITOPATOLOGÍA Fully Bilingual Mexican Journal of Phytopathology reportados en los valles altos en estados de la Mesa hybrid DK-3000. Koch´s postulates were fulfilled Central de México y otras partes del mundo con by re-isolation of the pathogen from inoculated clima subtropical o templado. El objetivo de ésta plants. Although the teleomorph (Setosphaeria investigación fue confirmar la identidad de E. tur- turcica) was not observed, morphometric and cicum asociado al tizón foliar del maíz y evaluar molecular studies indicated that E. turcicum is la severidad de la enfermedad en 17 híbridos de the causal agent of maize leaf blight. Differential maíz en lotes comerciales. Variación en color y disease severity occurred in maize hybrids and was morfología de los aislados se observó en diferen- highly dependent on prolonged periods of high tes medios. El medio extracto de maíz-agar fue el relative humidity. Future research should focus on más favorable para el desarrollo micelial de los determining the formae specialis crucial for maize aislados. En pruebas de patogenicidad, los aislados breeding programs to achieve genetic resistance exhibieron variación en patogenicidad en el híbri- against the disease. do comercial de maíz DK-3000. Los postulados de Koch se cumplieron mediante el re-aislamiento del Key words: fungus, pathogenicity, severity, patógeno de las plantas inoculadas. Aun cuando el anamorph, identification, morphology. teleomorfo (Setosphaeria turcica) no se observó, estudios morfométricos y moleculares confirmaron la identidad de E. turcicum como agente causal del In recent growing seasons, a severe foliar tizón del maíz. Los híbridos de maíz mostraron una epiphytotic disease occurred in several maize severidad diferencial de la enfermedad, lo cual es- hybrids in the northern coastal regions of Sinaloa tuvo relacionado con periodos prolongados de alta during the months of November through January, humedad relativa. Estudios futuros deberían enfo- including periods of relative humidity ≥ 90% carse a la determinación de las formae specialis for 13-17 h, and an average daily temperature importantes para la implementación de un progra- between 18 and 26 °C. The symptoms of the ma de mejoramiento genético para el control de la disease developed initially in the lower leaves enfermedad. causing oval, long, elliptical, grayish-green or tan necrotic lesions of different sizes. Later, symptoms Palabras clave: hongo, patogenicidad, severidad, spread to the middle and upper leaves of the plants. anamorfo, identificación, morfología. Although yield losses are yet to be estimated, personal observations by the author indicate the disease has destroyed up to 65% of the foliage En ciclos agrícolas recientes, una severa enfer- of some commercial maize hybrids grown in the medad foliar atacó a varios híbridos de maíz en ni- northern coastal region of the state. Although maize veles epifíticos en las regiones costeras del norte de leaf blight caused by Exserohilum turcicum (Et) is Sinaloa en los meses entre noviembre y enero, que one of the most important foliar diseases in maize incluyeron periodos diarios con humedad relativa ≥ grown in Mexico, at the present time, there is not 90% por 13-17 h, y una temperatura diaria prome- enough scientific evidence on the etiology of the dio de entre 18 y 26 °C. Los síntomas de la enferme- disease, although maize growers and field advisors dad se desarrollaron inicialmente en las hojas infe- have shown interest in determining its etiology. riores, causando lesiones ovaladas, largas, elípticas, Consequently, the main goal of the present study Publicación en línea, septiembre 2018 469 Revista Mexicana de FITOPATOLOGÍA Mexican Journal of Phytopathology Fully Bilingual verde grisáceo o marrones de diferentes tamaños. was to confirm the identity of the causal agent of Más tarde, los síntomas alcanzaron a las hojas me- maize leaf blight at the species level, based on dias y superiores de las plantas. Si bien todavía es- morphometric and molecular data of the anamorph. tán por determinarse las pérdidas de rendimiento, Fifteen symptomatic samples were collected observaciones personales del autor indican que la from the same number of maize fields in the enfermedad ha destruido hasta 65% del follaje de municipalities of Ahome and El Fuerte, Sinaloa, algunos híbridos comerciales de maíz sembrados from December 16, 2013 through January 15, en la región costera del norte del estado. Aunque 2014. One sample from each field consisted in five el tizón foliar del maíz causado por Exserohilum symptomatic leaves collected from the corners turcicum (Et) es una de las enfermedades foliares and center of each field (n=25). Fragments of más importantes en el maíz sembrado en México, symptomatic leaves (5-6 cm long) were disinfested hoy en día no existe suficiente evidencia científica in a 0.5% sodium hypochlorite solution for 2 min, acerca de la etiología de la enfermedad, aunque los washed for 3 min in sterile distilled water and dried productores de maíz y asesores de campo demues- on Whatman No. 1 filter paper. Fragments were tran interés en determinar su etiología. En conse- placed in a moist chamber, for 48 h at 25 °C. From cuencia, el objetivo principal del presente estudio conidia formed on the leaf tissue, monosporic fue confirmar la identidad del agente causal del ti- cultures were obtained through direct transference zón foliar del maíz a nivel de especie, con base en of them with a dissecting needle to Petri plates with datos morfométricos y moleculares del anamorfo. potato dextrose agar (PDA), under a dissecting Quince muestras sintomáticas fueron colectadas microscope. del mismo número de campos de maíz en los mu- Mycelial growth of eight fungal monoconidial nicipios de Ahome y El Fuerte, Sinaloa, entre el 16 isolates were evaluated on the following culture de diciembre de 2013 y el 15 de enero de 2014. media: potato dextrose agar (PDA), Czapek- Una muestra de cada campo consistió en cinco ho- Dox agar (CDA), glucose peptone agar (GPA) jas sintomáticas recolectadas de las esquinas y el and Richard´s synthetic agar (RSA), and maize centro de cada campo (n=25). Fragmentos de hojas leaf extract agar (CLEA) (Dhingra and Sinclair, sintomáticas (5-6 cm de longitud) fueron desinfes- 1985). After eight days of incubation, under 14-h tados en una solución de hipoclorito de sodio al light/10-h dark regime at 25 °C, the surface color 0.5% por 2 min, lavados por 3 min en agua desti- of the fungal colonies in the different culture media lada estéril y secados con papel de filtro Whatman were in the olivaceous and dark grayish-olive Núm. 1. Los fragmentos fueron colocados en una colors. cámara húmeda por 48 h a 25 °C. De los conidios All isolates presented light olive pigmentation formados en el tejido de las hojas se obtuvieron on the back of the colony in CDA and RSA, but cultivos monospórico a través de su transferencia no pigmentation was observed in other media. con una aguja de disección a placas de Petri con The radial mycelial growth rate of the isolates on papa dextrosa y agar (PDA), bajo un microscopio CLEA ranged from 3.7 to 4.9 mm/day and was de disección.
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