ABSTRACT
Charcoal rot caused by Macrophomina phaseolina is an important disease of sugarcane in Mexico. This study was carried out to characterize isolates of M. phaseolina obtained from sugarcane by the combination of morphological and molecular analyses. The morphological characterization of 10 isolates was performed using scanning electron microscopy and light microscopy. To confirm the morphological identification, rDNA from two representative isolates was extracted, and the internal transcribed spacer (ITS) region was amplified by polymerase chain reaction and sequenced using specific primers MpKF1 and MpKR1. Based on their morphological characteristics, all isolates were identified as M. phaseolina. Moreover, the analysis of two ITS sequences showed 100% similarity with the M. phaseolina sequences deposited in the GenBank. To our knowledge, this is the first study in the world aimed at characterizing isolates of M. phaseolina obtained from sugarcane.
Subject(s)
Ascomycota/isolation & purification , Saccharum/microbiology , Ascomycota/genetics , Ascomycota/physiology , Ascomycota/ultrastructure , DNA, Fungal/analysis , DNA, Ribosomal Spacer , Mexico , Microscopy, Electron, Scanning , Mycological Typing Techniques , Plant Diseases/microbiology , Ribotyping , Spores, FungalABSTRACT
Mango is one of the most popular and nutritious fruits in the world and Mexico is the world's largest exporter. There are many diseases that directly affect fruit yield and quality. During the period 2016-2017, leaves with grey leaf spots were collected from 28 commercial mango orchards distributed in two main production areas in Sinaloa State of Mexico, and 50 Neopestalotiopsis isolates were obtained. Fungal identification of 20 representative isolates was performed using morphological characterization and phylogenetic analysis based on the internal transcribed spacer (ITS) region of ribosomal DNA, part of the translation elongation factor 1-alpha (TEF) and the ß-tubulin (TUB) genes. Phylogenetic analysis indicated that the 20 isolates from this study formed four consistent groups, however, overall tree topologies do not consistently provide a stable and sufficient resolution. Therefore, even though morphological and phylogenetic separation is evident, these isolates were not assigned to any new taxa and were tentatively placed into four clades (clades A-D). Pathogenicity tests on detached mango leaves of cv. Kent showed that the 20 isolates that belong to the four Neopestalotiopsis clades from this study and induce lesions on mango leaves. This is the first report of species of Neopestalotiopsis causing mango grey leaf spot disease in Mexico.
ABSTRACT
La pudrición carbonosa, causada por Macrophomina phaseolina, es una enfermedad importante de la caña de azúcar en México. Este estudio se realizó con el objetivo de caracterizar aislados de M. phaseolina obtenidos de caña de azúcar mediante análisis morfológicos y moleculares. La caracterización morfológica de 10 aislados se llevó a cabo con el uso de microscopia electrónica de barrido y microscopia de luz. Para confirmar la identificación, se extrajo el ADNr de 2 aislados representativos, y la región del espaciador interno transcrito (ITS) se amplificó mediante la reacción en cadena de la polimerasa y se secuenció usando los iniciadores específicos MpKF1 y MpKR1. Los aislados se identificaron como M. phaseolina con base en la morfología. El análisis de secuencias ITS mostró 100% de similitud con las secuencias de M. phaseolina depositadas en el GenBank. Para nuestro conocimiento, este es el primer estudio del mundo enfocado a caracterizar aislados de M. phaseolina obtenidos de caña de azúcar
Charcoal rot caused by Macrophomina phaseolina is an important disease of sugarcane in Mexico. This study was carried out to characterize isolates of M. phaseolina obtained from sugarcane by the combination of morphological and molecular analyses. The morphological characterization of 10 isolates was performed using scanning electron microscopy and light microscopy. To confirm the morphological identification, rDNA from two representative isolates was extracted, and the internal transcribed spacer (ITS) region was amplified by polymerase chain reaction and sequenced using specific primers MpKF1 and MpKR1. Based on their morphological characteristics, all isolates were identified as M. phaseolina. Moreover, the analysis of two ITS sequences showed 100% similarity with the M. phaseolina sequences deposited in the GenBank. To our knowledge, this is the first study in the world aimed at characterizing isolates of M. phaseolina obtained from sugarcane