RESUMO
Brazil and Argentina have a combined soybean area of 53.6 million hectares, which accounts for over half of the total global production. The soybean crop in South America extends from latitude 8-10° S to 32-36° S. Such a vast, almost contiguous area imposes a serious sanitary risk to the crop. Currently, the prevalence of anthracnose is increasing, with recurring reports of severe epidemics and expressive yield losses. Soybean anthracnose is mainly associated with Colletotrichum truncatum, although other Colletotrichum species have also been reported as causal agents of this disease. Knowledge about the morphological, cultural, and molecular variability of C. truncatum in South America is crucial for disease management. Here, we present data on the molecular, morphological, biological, cultural, and pathogenicity of C. truncatum isolates collected in Brazil and Argentina. Light microscopy and randomly-amplified polymorphic DNA (RAPD) analysis were used for estimating the variability of isolates. Colletotrichum truncatum displayed three types of conidiogenesis, viz. conidial formation from conidiogenous cells on hyphal extremities, in conidiomas in acervuli, and directly from fertile setae (a mechanism yet-unreported for C. truncatum). RAPD profiling was effective in revealing the genetic diversity among C. truncatum isolates. The intra-group similarity was greater among the Argentinian isolates when compared to the Brazilian group. Furthermore, the results indicated a strong correlation between geographical origin and molecular grouping, with the exclusive or semi-exclusive assembling of Brazilian and Argentinian isolates in distinct clades. Finally, a preliminary account of the reaction of soybean accessions to C. truncatum is also included.
RESUMO
Diaporthe (teleomorpho)-Phomopsis - (anamorph) (DP) is a fungal group of great genetic diversity with over 900 species associated to a wide host range that includes cultivated and uncultivated species, forest, fruit trees and weeds. DP isolates are hemibiotrophs and have different sources of primary inoculum as stubble and seeds to restart cycles of parasitism - saprophytism. They colonize host tissues from early plant stages and establish different nutritional relationships, acting as endophytic and necrotrophic fungi. The plasticity of the Phomopsis genus has favored its expansion to different agro-ecosystems and various hosts constituting an epidemiological risk. The objective was to validate the identity and evaluate the biological relationships among 12 isolates of P longicolla and D. phaseolorum var. sojae (anamorph P phaseoli var. sojae) obtained in different tempered and subtropical agro-environments of Argentina, in order to analyze the variability and strategies for preserving fungal biodiversity. Macromorphological attributes (such as texture and color of colonies, stroma shape and distribution, pycnidia and perythecia shape and distribution) and micro-morphological characteristics (such as size and shape of conidia, asci and ascospores) allowed identifying three new isolates as P longicolla. A complementary molecular analysis was also made to overcome the limitations derived from the morphological analysis, thus the AFP.8413 isolate was finally identified as P longicolla. The molecular characterization was useful to identify the evaluated isolates and to group them in four taxa of the Diaporthe-Phomopsis complex: ten isolates were included in P. longicolla, one isolate was included in D. phaseolorum var. sojae (anamorph P. phaseoli var. sojae), one isolate was identified as D. phaseolorum var. caulivora and two isolates were included in D. phaseolorum var. meridionalis. The use of phenotipic and molecular tools have contributed to an accurate identification of P longicolla, and comprehension about the biological relationships (homo or heterothallic hibridizations) among D. phaseolorum varieties (P phaseoli) and species of Diaporthe-Phomopsis. This allowed also a better understanding of the mechanisms of fungic plasticity, to colonize and expand their host range and genetic variability, promoting thus their biodiversity conservation. Rev. Biol. Trop. 63 (3): 871-884. Epub 2015 September 01.
Assuntos
Ascomicetos , Biodiversidade , Variação Genética , Argentina , Ascomicetos/classificação , Ascomicetos/citologia , Ascomicetos/genética , FilogeniaRESUMO
Diaporthe(telomorfo) - Phomopsis(anamorfo) (DP) constituye un grupo fúngico de amplia diversidad genética con más de 900 especies distribuidas en un amplio rango de hospedantes que incluye especies cultivadas y no cultivadas, forestales, frutales y malezas. Los aislamientos de DP son hemi-biótrofos y disponen de diferentes fuentes de inóculo primario, como el rastrojo y las semillas, para reiniciar sus ciclos de parasitismo-saprofitismo. Ellos colonizan los tejidos del hospedante desde los estadios tempranos del desarrollo y establecen relaciones nutricionales de endofitia y necrotrofia fúngica. La plasticidad del género Phomopsisha favorecido su expansión a diferentes agro-ecosistemas y hospedantes constituyendo un importante riesgo epidemiológico. El objetivo fue validar la identidad y evaluar las relaciones biológicas de 12 aislamientos de P. longicollay D. phaseolorumvar. sojaeobtenidos en distintos agro-ambientes templados y subtropicales de Argentina, para analizar la variabilidad y estrategias de conservación de la bio-diversidad fúngica. Las cualidades macro-morfo-lógicas(textura y color de colonias, forma y distribución de estromas, desarrollo, forma y distribución de cuerpos fructíferos), y los caracteres micro-morfológicos(tamaño y forma de conidios, ascos y ascosporas) permitieron identificar a tres nuevos aislamientos de P. longicollaincluidos en el complejo D/P. El análisis molecular complementario corrigió las limitaciones derivadas de la caracterización basada sólo en marcadores morfológicos y logró reubicar al aislamiento AFP.8413 de identidad dudosa, en el nodo correspondiente a P. longicolla.De esta manera, la caracterización molecular definió la identidad de los aislamientos y los ubicó en los 4 taxones del complejo DP: diez aislamientos fueron incluidos en Plo(AFP.Gpo 4.4, AFP.Gpo 3.5, AFP.Gpo 4.3, AFP.Gpo 3.4, AFP.CaA, AFP. CaB, AFP.B5L16, AFP.B4L17, AFP.227B2, AFP.8413), un aislamiento incluido en Dps(AFP.Qcol7), un aislamiento en Dpc(AFP.Dpc16) y dos aislamientos incluidos en Dpm(AFP.Dpm109 y AFP.Dpm112). La adecuada identificación de P. longicollay el avance en el conocimiento de las relaciones biológicas (hibridaciones homo o heterotálicas) entre variedades de D. phaseolorum (P. phaseoli)y especies de Diaporthe- Phomopsispermiten comprender la plasticidad para colonizar un amplio rango de hospedantes, los mecanismos de variabilidad genética y la preservación de la diversidad fúngica.
Diaporthe(teleomorpho)-Phomopsis (anamorph) (DP) is a fungal group of great genetic diversity with over 900 species associated to a wide host range that includes cultivated and uncultivated species, forest, fruit trees and weeds. DP isolates are hemi-biotrophs and have different sources of primary inoculum as stubble and seeds to restart cycles of parasitism -saprophytism. They colonize host tissues from early plant stages and establish different nutritional relationships, acting as endophytic and necrotrophic fungi. The plasticity of the Phomopsisgenus has favored its expansion to different agro-ecosystems and various hosts constituting an epidemiological risk. The objective was to validate the identity and evaluate the biological relationships among 12 isolates of P. longicollaand D. phaseolorumvar. sojae(anamorph P. phaseolivar. sojae)obtained in different tempered and subtropical agro-environments of Argentina, in order to analyze the variability and strategies for preserving fungal biodiversity. Macro-morphological attributes (such as texture and color of colonies, stroma shape and distribution, pycnidia and perythecia shape and distribution) and micro-morphological characteristics (such as size and shape of conidia, asci and ascospores) allowed identifying three new isolates as P. longicolla.A complementary molecular analysis was also made to overcome the limitations derived from the morphological analysis, thus the AFP.8413 isolate was finally identified as P. longicolla.The molecular characterization was useful to identify the evaluated isolates and to group them in four taxa of the Diaporthe-Phomopsiscomplex: ten isolates were included in P. longicolla,one isolate was included in D. phaseolorumvar. sojae(anamorph P. phaseolivar. sojae),one isolate was identified as D. phaseolorumvar. caulivoraand two isolates were included in D. phaseolorumvar. meridi-onalis.The use of phenotipic and molecular tools have contributed to an accurate identification of P. longicolla,and comprehension about the biological relationships (homo or heterothallic hibridizations) among D. phaseolo-rumvarieties (P. phaseoli)and species of Diaporthe-Phomopsis.This allowed also a better understanding of the mechanisms of fungic plasticity, to colonize and expand their host range and genetic variability, promoting thus their biodiversity conservation.
Assuntos
Ascomicetos , Biodiversidade , Variação Genética , Argentina , Ascomicetos/classificação , Ascomicetos/citologia , Ascomicetos/genética , FilogeniaRESUMO
The aims of this study were to select microbial isolates from phyllosphere of maize and to examine their antagonistic activity against Exserohilum turcicum. Selection was performed through the ability of isolates to compete with the pathogen using an index of dominance and to affect growth parameters of E. turcicum. Most of the epiphytic populations obtained for the screening were bacteria. These isolates were found in the order of 6 log CFU/g of leaf fresh weight. According to similar morphological characteristics and staining, 44 out of 111 isolates obtained were selected for testing antagonistic effects. At water potential, ψ, -1.38MPa and -4.19MPa, three Bacillus isolates showed dominance at a distance (5/0) and a significant reduction of growth rate of the pathogen. Three Bacillus isolates only decreased the growth rate of E. turcicum at -1.38MPa. At -4.19MPa the growth rate decreased with three isolates of Pantoea and three Bacillus. In this study a negative and significant correlation was observed between the growth rate of E. turcicum and the dominance index in the interaction of the pathogen with some bacteria. These results show that with decreasing growth rate of the pathogen the dominance index of the interaction increases. Eleven potential biocontrol agents against E. turcicum were selected.
Assuntos
Ascomicetos , Bactérias , Agentes de Controle Biológico , Doenças das Plantas/prevenção & controle , Zea mays/microbiologiaRESUMO
The aims of this study were to select microbial isolates from phyllosphere of maize and to examine their antagonistic activity against Exserohilum turcicum. Selection was performed through the ability of isolates to compete with the pathogen using an index of dominance and to affect growth parameters of E. turcicum. Most of the epiphytic populations obtained for the screening were bacteria. These isolates were found in the order of 6 log CFU/g of leaf fresh weight. According to similar morphological characteristics and staining, 44 out of 111 isolates obtained were selected for testing antagonistic effects. At water potential, ψ, −1.38 MPa and −4.19 MPa, three Bacillus isolates showed dominance at a distance (5/0) and a significant reduction of growth rate of the pathogen. Three Bacillus isolates only decreased the growth rate of E. turcicum at −1.38 MPa. At −4.19 MPa the growth rate decreased with three isolates of Pantoea and three Bacillus. In this study a negative and significant correlation was observed between the growth rate of E. turcicum and the dominance index in the interaction of the pathogen with some bacteria. These results show that with decreasing growth rate of the pathogen the dominance index of the interaction increases. Eleven potential biocontrol agents against E. turcicum were selected.(AU)
El objetivo de este estudio fue seleccionar aislamientos microbianos de la filósfera de maíz y examinar su actividad antagonista contra Exserohilum turcicum. La selección se realizó a través de la capacidad de los aislamientos de competir con el patógeno usando un índice de dominancia y también la capacidad de afectar los parámetros de crecimiento de E. turcicum. La mayoría de las poblaciones epifíticas aisladas para la selección fueron bacterias. Estos aislamientos se encontraron en el orden de 6 log de UFC por gramo de peso fresco de hoja de maíz. En base a características morfológicas y tintóreas similares, se seleccionaron 44 de 111 aislamientos obtenidos para evaluar su capacidad antagónica. A los potenciales agua, ψ, −1,38 MPa y −4,19 MPa, tres aislados del género Bacillus mostraron dominancia a distancia (5/0) y una reducción significativa de la velocidad de crecimiento del patógeno. Tres aislamientos de Bacillus disminuyeron la velocidad de crecimiento de E. turcicum a −1,38 MPa. A −4,19 MPa la velocidad de crecimiento disminuyó con tres aislamientos de Pantoea y tres de Bacillus. En este estudio se observó una correlación negativa y significante entre la velocidad de crecimiento de E. turcicum y el índice de dominancia cuando el patógeno interactuó con algunas bacterias. Esto estaría indicando que cuando disminuye la velocidad de crecimiento del patógeno se incrementa el índice de dominancia de la interacción. Se seleccionaron once posibles agentes de biocontrol contra E. turcicum.(AU)
RESUMO
The aims of this study were to select microbial isolates from phyllosphere of maize and to examine their antagonistic activity against Exserohilum turcicum. Selection was performed through the ability of isolates to compete with the pathogen using an index of dominance and to affect growth parameters of E. turcicum. Most of the epiphytic populations obtained for the screening were bacteria. These isolates were found in the order of 6 log CFU/g of leaf fresh weight. According to similar morphological characteristics and staining, 44 out of 111 isolates obtained were selected for testing antagonistic effects. At water potential, ψ, −1.38 MPa and −4.19 MPa, three Bacillus isolates showed dominance at a distance (5/0) and a significant reduction of growth rate of the pathogen. Three Bacillus isolates only decreased the growth rate of E. turcicum at −1.38 MPa. At −4.19 MPa the growth rate decreased with three isolates of Pantoea and three Bacillus. In this study a negative and significant correlation was observed between the growth rate of E. turcicum and the dominance index in the interaction of the pathogen with some bacteria. These results show that with decreasing growth rate of the pathogen the dominance index of the interaction increases. Eleven potential biocontrol agents against E. turcicum were selected.
El objetivo de este estudio fue seleccionar aislamientos microbianos de la filósfera de maíz y examinar su actividad antagonista contra Exserohilum turcicum. La selección se realizó a través de la capacidad de los aislamientos de competir con el patógeno usando un índice de dominancia y también la capacidad de afectar los parámetros de crecimiento de E. turcicum. La mayoría de las poblaciones epifíticas aisladas para la selección fueron bacterias. Estos aislamientos se encontraron en el orden de 6 log de UFC por gramo de peso fresco de hoja de maíz. En base a características morfológicas y tintóreas similares, se seleccionaron 44 de 111 aislamientos obtenidos para evaluar su capacidad antagónica. A los potenciales agua, ψ, −1,38 MPa y −4,19 MPa, tres aislados del género Bacillus mostraron dominancia a distancia (5/0) y una reducción significativa de la velocidad de crecimiento del patógeno. Tres aislamientos de Bacillus disminuyeron la velocidad de crecimiento de E. turcicum a −1,38 MPa. A −4,19 MPa la velocidad de crecimiento disminuyó con tres aislamientos de Pantoea y tres de Bacillus. En este estudio se observó una correlación negativa y significante entre la velocidad de crecimiento de E. turcicum y el índice de dominancia cuando el patógeno interactuó con algunas bacterias. Esto estaría indicando que cuando disminuye la velocidad de crecimiento del patógeno se incrementa el índice de dominancia de la interacción. Se seleccionaron once posibles agentes de biocontrol contra E. turcicum.
Assuntos
Ascomicetos , Bactérias , Agentes de Controle Biológico , Doenças das Plantas/prevenção & controle , Zea mays/microbiologiaRESUMO
The aims of this study were to select microbial isolates from phyllosphere of maize and to examine their antagonistic activity against Exserohilum turcicum. Selection was performed through the ability of isolates to compete with the pathogen using an index of dominance and to affect growth parameters of E. turcicum. Most of the epiphytic populations obtained for the screening were bacteria. These isolates were found in the order of 6log CFU/g of leaf fresh weight. According to similar morphological characteristics and staining, 44 out of 111 isolates obtained were selected for testing antagonistic effects. At water potential, ¤ê, -1.38MPa and -4.19MPa, three Bacillus isolates showed dominance at a distance (5/0) and a significant reduction of growth rate of the pathogen. Three Bacillus isolates only decreased the growth rate of E. turcicum at -1.38MPa. At -4.19MPa the growth rate decreased with three isolates of Pantoea and three Bacillus. In this study a negative and significant correlation was observed between the growth rate of E. turcicum and the dominance index in the interaction of the pathogen with some bacteria. These results show that with decreasing growth rate of the pathogen the dominance index of the interaction increases. Eleven potential biocontrol agents against E. turcicum were selected.
