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ISOLATION and IDENTIFICATION of Taphrina Caerulescens in Quercus Eduardii in AGUASCALIENTES, MEXICO

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ISOLATION and IDENTIFICATION of Taphrina Caerulescens in Quercus Eduardii in AGUASCALIENTES, MEXICO ISOLATION AND IDENTIFICATION OF Taphrina caerulescens IN Quercus eduardii IN AGUASCALIENTES, MEXICO AISLAMIENTO E IDENTIFICACIÓN DE Taphrina caerulescens EN Quercus eduardii EN AGUASCALIENTES, MÉXICO Gregg Evans1, Onesimo Moreno-Rico2*, José J. Luna-Ruíz3, Joaquín Sosa-Ramírez3, Celeste E. Moreno-Manzano4 1Ciencias Biológicas, Centro de Ciencias Básicas (CCB), Universidad Autónoma de Aguascalientes (UAA), Avenida Universidad # 940, Colonia Ciudad Universitaria, C.P. 20131, Aguascalientes, Aguascalientes, México ([email protected]). 2Departamento de Microbiología, CCB, UAA, Avenida Universidad # 940, Ciudad Universitaria C.P. 20131, Aguascalientes, Aguascalientes, México ([email protected]). 3Departamento de Disciplinas Agrícolas, Centro de Ciencias Agropecuarias, UAA, Jesús María, Aguascalientes. ([email protected]), ([email protected]). 4CBTA 61, Aquiles Elorduy Garcia, Calvillo, Aguascalientes, México ([email protected]). ABSTRACT RESUMEN Taphrina caerulescens exclusively affects plants of the Quercus Taphrina caerulescens afecta exclusivamente a las plantas del gé- genus. The identification and isolation of this fungus is difficult nero Quercus. La identificación y el aislamiento de este hongo due to its dimorphic nature and extremely slow growth habit es difícil debido a su naturaleza dimórfica y su hábito de cre- in artificial growth media. The objective of this research was to cimiento extremadamente lento en los medios de crecimiento isolate and identify the fungal pathogen T. caerulescens. Three artificial. El objetivo de esta investigación fue aislar e identificar methods were used to isolate the fungus, however, only the spore el patógeno fúngico T. caerulescens. Tres métodos se usaron para fall method was successful. In order to identify the fungus, a aislar el hongo; sin embargo, solo el método de caída de esporas visual inspection of the host plants infected leaves was carried fue exitoso. Para identificar el hongo, se efectuó una inspección out. The symptoms and signs of infection were noted. In visual de las hojas infectadas de las plantas huésped y se observa- addition to the typical blister formation on the infected leaves, ron los síntomas y signos de infección. Además de la formación an atypical symptom comprising of chlorotic spots covered with típica de ampollas en las hojas infectadas, también se observó asci was also observed. Three biochemical tests were carried un síntoma atípico que comprende manchas cloróticas cubiertas out to prove that the pathogen was in the genus Taphrina: The con asci. Tres pruebas bioquímicas se realizaron para demostrar first, tested the ability to utilize urea, the second, its capacity to que el patógeno pertenecía al género Taphrina: la primera, probó produce extra cellular DNase, and the third tested its reaction to la capacidad de utilizar urea, la segunda, su capacidad de produ- Diazonium blue B (rapid blue salts). For species identification cir DNasa extracelular, y la tercera probó su reacción al azul de the following analyses were carried out: 1) carbon assimilation diazonio B (sales azules rápidas). Para identificar las especies se tests using the glucose, saccharose, D-xylose, raffinose, trehalose efectuaron los siguientes análisis: 1) pruebas de asimilación de and sorbitol, as carbon sources; 2) molecular analysis tools were carbono usando la glucosa, sacarosa, D-xilosa, rafinosa, treha- utilized to extract and sequence the genomic DNA. The resulting losa y sorbitol, como fuentes de carbono; 2) se utilizaron herra- sequences when analysed with the NCBI Nucleotide blast tool mientas de análisis molecular para extraer y secuenciar el ADN confirmed, with 99% similitude, that the isolate was similar in genómico. Cuando las secuencias resultantes se analizaron con la nature to T. caerulescens strain CBS 351.35 ascension number herramienta de explosión de nucleótidos NCBI se confirmó, con AF492081.1. Pathogenicity test was also carried out and resulted una similitud del 99%, que el aislamiento era de naturaleza simi- in the infection of 25 % of the inoculated plants. lar a la cepa T. caerulescens CBS 351.35 con número de ascensión AF492081.1. También se realizó una prueba de patogenicidad que resultó en la infección del 25% de las plantas inoculadas. * Author for correspondence v Autor responsable. Received: April, 2018. Approved: August, 2018. Published as ARTICLE in Agrociencia 53: 781-795. 2019. 781 AGROCIENCIA, 1 de julio - 15 de agosto, 2019 Key words: Aguascalientes PAs, oak populations, Quercus Palabras clave: PAs de Aguascalientes, poblaciones de roble, eduardii, pathogenicity, leaf blisters, Taphrina caerulescens. Quercus eduardii, patogenicidad, ampollas de hoja, Taphrina caerulescens. INTRODUCTION INTRODUCCIÓN aphrina caerulescens belongs to the order Taphrinales. The first complete monograph aphrina caerulescens pertenece al orden Ta- of the genus was realized by Mix (1949) who phrinales. La primera monografía completa Tdivided the genus into 100 species, and Taphrina del género fue realizada por Mix (1949), que spp. are dimorphic (Mix, 1949; Kramer, 1960). dividióT el género en 100 especies y las especies de Ta- Dimorphic fungi are those fungi that exist either in phrina son dimorfas (Mix, 1949; Kramer, 1960). Los yeast form or as mold (mycelial form) depending hongos dimórficos son aquellos hongos que existen on environmental conditions, physiological en forma de levadura o como moho (forma micelial) conditions of the fungus or the genetic characteristics según las condiciones ambientales, las condiciones (Flannigan et. al., 2001). When the fungus is within fisiológicas del hongo o las características genéticas its host tissue, it is present in its filamentous state; (Flannigan et al., 2001). Cuando el hongo está den- however, under adverse conditions it is present in its tro de su tejido huésped, se presenta en su estado yeast state and in this state the fungus behaves like filamentoso; sin embargo, en condiciones adversas, a saprophyte. Taphrina spp. were identified based también se presenta en su estado de levadura y en upon geographical distribution, site and type of the este estado el hongo se comporta como un saprófito. infection and symptoms, localization of mycelium, Las especies de Taphrina se identificaron con base a la morphology of the fungal structures and host range distribución geográfica, el sitio, el tipo de infección, (Mix, 1949; Taylor and Birdwell, 2000; Rodríguez los síntomas, la localización del micelio, la morfo- and Fonseca, 2003). In order to identify Taphrina logía de las estructuras fúngicas y el rango del hués- spp. molecular methods were used by Sjamsuridzal et ped (Mix, 1949; Taylor y Birdwell, 2000; Rodríguez al. (1997), Rodríguez and Fonseca (2003), Bacigálová y Fonseca, 2003). Métodos moleculares se usaron et al. (2003), Tavares et al. (2004), Petrýdesová et por Sjamsuridzal et al. (1997), Rodríguez y Fon- al. (2013), Isheng et al. (2014) and Selbmann et seca (2003), Bacigálová et al. (2003), Tavares et al. al. (2014). The most important species include: T. (2004), Petrýdesová et al. (2013), Isheng et al. (2014) deformans causes deformation of leaves, fruits and y Selbmann et al. (2014) para identificar especies de flowers and the defoliation of Prunus persica and Taphrina. Las especies más importantes incluyen: T. Prunus dulcis (Mill.) D.A.Webb; T. communis causes deformans, que causa deformación de hojas, frutos deformations in American plum (Prunus americana); y flores y la defoliación de Prunus persica y Prunus T. pruni affects European plum species; T. wiesneri dulcis (Mill.) D.A.Webb; T. communis, causa defor- (Rathay) Mix, syn. T. cerasi (Fuckel) Sadeb (Farr et maciones en la ciruela americana (Prunus americana); al., 1989) produces leaf blisters and witches broom in T. pruni, afecta a las especies de ciruela europeas; T. all cherry species; T. armeniacae (Georgescu y Badea) wiesneri (Rathay) Mix, syn. T. cerasi (Fuckel) Sadeb causes witches broom in apricot (Prunus armeniaca); (Farr et al., 1989) produce ampollas en las hojas y en- T. confusa (Atk.) Giesen in Prunus virginiana; T. fermedad de escoba de bruja en todas las especies de farlowii Sadeb causes blistering of the leaves of cerezas; T. armeniacae (Georgescu y Badea) causa de various shrubs and hedges, including Prunus serotina. escoba de bruja en el albaricoque (Prunus armeniaca); Taphrina caerulescens causes leaf blister and T. confusa (Atk.), causa escoba de bruja en Prunus deformation in approximately 50 species of oak virginiana; T. farlowii Sadeb, causa ampollas en las (Quercus); Q. acutissima (Carruth), Q. agrijolia hojas de varios arbustos y setos, incluyendo Prunus (Nee) Q. alba (L.), Q. borealis (Michx), Q. cerris serotina. (L), Q. cinerea (Michx), Q. coccijera (L), Q. coccinea Taphrina caerulescens causa ampollas en las hojas (Muench), Q. crispula (Blume), Q. dentata (Thunb), y deformación en aproximadamente 50 especies de Q. douglasii (Hook), Q. ellipsoidalis (Hill) , Q. roble (Quercus): Q. acutissima (Carruth), Q. agrijo- jruticosa (Brot), Q. gambelii (Nutt), Q. geminala lia (Nee) Q. alba (L.), Q. borealis (Michx), Q. cerris 782 VOLUMEN 53, NÚMERO 5 ISOLATION AND IDENTIFICATION OF Taphrina caerulescens IN Quercus eduardii IN AGUASCALIENTES, MEXICO (Small), Q. georgiana (Curtis), Q. ilicifolia (Wangh), (L), Q. cinerea (Michx), Q. coccijera (L), Q. coccinea Q. imbricaria (Michx), Q. kelloggii (Newb), Q. (Muench), Q. crispula (Blume), Q. dentata (Thunb), laurifolia (Michx), Q. lobata (Nee), Q. macrocarpa Q. douglasii (Hook),
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