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AIMS: Entomopathogenic Metarhizium fungi are widely recognized for their biological control potential. In Cuba, several fungus-based bio-insecticides have been developed and are produced as part of integrated pest management (IPM) programmes for economically relevant agricultural pests. Screening of fungal isolates from the INISAV strain collection was used for the development of bio-insecticides against important pest insects as, for example the sweet potato weevil, Cylas formicarius. METHODS AND RESULTS: Six fungal isolates from Cuba were microscopically, morphologically and molecular-taxonomically characterized using marker sequences ef1a, rpb1 and rpb2, and the 5TEF region of the ef1a gene. Five isolates were assigned to the species Metarhizium anisopliae sensu stricto and one isolate to Metarhizium robertsii. The pathogenic potential was evaluated against adults of C. formicarius, and growth and conidial production on different nutritional media were determined. Metarhizium anisopliae strain LBM-267 displayed pronounced virulence against the sweet potato weevil and abundant conidia production on several culture media. CONCLUSIONS: Entomopathogenic fungal isolates from Cuba were assigned to the taxonomic species M. anisopliae sensu stricto and M. robertsii. Virulence assessment with respect to C. formicarius led to the identification of two M. anisopliae isolates holding biocontrol potential. Isolate LBM-11 has previously been developed into the bio-insecticide METASAVE-11 that is widely used to control several species of plant pathogenic weevils, Lepidoptera and thrips in Cuba. Isolate LBM-267 has not been employed previously but is as virulent against C. formicarius as LBM-11; its growth and conidial production capacities on different nutritional media will likely facilitate economically feasible bio-insecticide development. SIGNIFICANCE AND IMPACT OF THE STUDY: Metarhizium anisopliae isolate LBM-267 has been selected as a promising candidate for biocontrol of the sweet potato weevil, an economically important agricultural pest in Cuba, and for further R&D activities within the framework of the Biological Control Program of Cuba.

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Sweet potato is a subsistence crop cultivated worldwide. Although it is generally considered tolerant to different diseases, it is quite susceptible to the fungus Plenodomus destruens that causes foot-rot disease. Plant growth-promoting bacteria associated with sweet potato remain poorly studied, but some Bacillus strains may have potential as biological control agents. Here, we evaluate the persistence of two bacterial strains-Bacillus safensis T052-76 and Bacillus velezensis T149-19-in pot experiments and assess their impact on indigenous bacterial and fungal communities associated with sweet potato. Numbers of cells of both strains introduced into pots remained stable in the rhizosphere of sweet potato over the 180-day experiment. Denaturing gradient gel electrophoresis based on the rrs gene encoding bacterial 16S rRNA and the fungal ribosomal internal transcribed spacer region showed that bands corresponding to the introduced strains were not detected in plant endosphere. PERMANOVA and non-metric multidimensional scaling statistical analyses showed that: (1) strain T052-76 altered the structure of the indigenous bacterial community (rhizosphere and soil) more than strain T149-19; (2) T052-76 slightly altered the structure of the indigenous fungal community (rhizosphere and soil) and (3) strain T149-19 did not disturb the fungal community. Our results demonstrate the stability of both Bacillus strains in the sweet potato rhizosphere and, apart from the influence of B. safensis T052-76 on the bacterial community, their limited impact on the microbial community associated with this important crop plant.

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Ceratocystis wilt is among the most important diseases on mango (Mangifera indica) in Brazil, Oman, and Pakistan. The causal agent was originally identified in Brazil as Ceratocystis fimbriata, which is considered by some as a complex of many cryptic species, and four new species on mango trees were distinguished from C. fimbriata based on variation in internal transcribed spacer sequences. In the present study, phylogenetic analyses using DNA sequences of mating type genes, TEF-1α, and ß-tubulin failed to identify lineages corresponding to the four new species names. Further, mating experiments found that the mango isolates representing the new species were interfertile with each other and a tester strain from sweet potato (Ipomoea batatas), on which the name C. fimbriata is based, and there was little morphological variation among the mango isolates. Microsatellite markers found substantial differentiation among mango isolates at the regional and population levels, but certain microsatellite genotypes were commonly found in multiple populations, suggesting that these genotypes had been disseminated in infected nursery stock. The most common microsatellite genotypes corresponded to the four recently named species (C. manginecans, C. acaciivora, C. mangicola, and C. mangivora), which are considered synonyms of C. fimbriata. This study points to the potential problems of naming new species based on introduced genotypes of a pathogen, the value of an understanding of natural variation within and among populations, and the importance of phenotype in delimiting species.

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El cultivo del boniato presenta una gran importancia, ya que se puede emplear en la alimentación humana y animal, así como en la industria; el mismo produce raíces reservantes de gran valor calórico y nutritivo con alto contenido de carbohidratos. Entre las raíces y tubérculos cultivados es el segundo en importancia y representa más del 80% de la producción mundial. El empleo de las técnicas in vitro constituye una poderosa herramienta en la explotación comercial, propiciando el empleo de la micropropagación en diferentes especies. Para desarrollar el presente trabajo se recolectaron raíces tuberosas pertenecientes al clon Inivit B 93-1. Se procedió a la formación de callos potencialmente embriogénicos, para lo cual se emplearon explantes de limbos foliares, desinfectados con hipoclorito de sodio (1%) y sembrados en el medio de cultivo propuesto por Murashige y Skoog (1962), vitaminas MS (10,0 ml/l-1), mioinositol (100 mg/l-1), sacarosa (3%), gelrite (0,2%), 2,4-D (0,25-2,5 mg/l-1) y 6-BAP (0,25-1,0 mg/l-1), el pH fue ajustado a 5,8 ± 0,01 mantenidos en la oscuridad durante treinta días, lográndose los mejores resultados con el uso del 2,4-D (0,50 mg/l-1) y 6-BAP (0,25 mg/l-1), y en los mismos se evaluó la dinámica del crecimiento y se lograron los mejores resultados entre los 28 y 32 días después de la siembra, para lo cual los resultados obtenidos servirán de base a otros estudios y permitirán evaluar, controlar y desarrollar estrategias para la conservación y el uso de los recursos naturales, dando cumplimiento al objetivo referente a estudiar la dinámica del crecimiento en la formación de callos potencialmente embriogénicos en el cultivo del boniato.

The cultivation of the sweet potato presents a great importance, since you can use in the human feeding, animal as well as in the industry, the same one produces roots reservantes of great caloric and nutritious value with high content of carbohydrates. Between the roots and cultivated tubers it is the second in importance and it represents more than 80% of the world production. The employment of the techniques in vitro constitutes a powerful tool in the commercial, propitiated exploitation the employment of the micropropagación in different species. It is for it that you/they were gathered to develop the present work tuberous roots of the clon INIVIT B 93-1. Was realized the formation of callus with embryogenic structures, explantes of leaves were used, disinfected with hipoclorito of sodium (1%) and inoculated in the tissue culture medium proposed by Murashige and Skoog (1962), vitamins MS (10.0 ml/l-1), myoinositol (100 mg/l-1), sucrose (3%), gelryte (0.2%), 2,4-D (0.25-2.5 mg/l-1) and 6-BAP (0.25-1.0 mg/l-1), the pH was adjusted 5.8 ± 0.01 maintained in the darkness during thirty days, being achieved the best results with the use of the 2,4-D (0.50 mg/l-1) and 6-BAP (0,25 mg/l-1) and was evaluated the grow dynamic and obtained the better resulted between 28 and 32 days after culture, for that which the obtained results will serve from base to other studies and they will allow to evaluate, to control and to develop strategies for the conservation and use of the natural resources, giving execution to the objective with respect to studying the dynamics of the growth potentially in the formation of tripes embriogénicos in the cultivation of the sweet potato.

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Un elemento esencial en la investigación con microorganismos fitopatógenos es su conservación y uso seguro.Desde este punto de vista, en este estudio se evaluaron métodos de conservación de hongos que atacanel ñame, empleando como factores de respuesta el potencial de viabilidad, y la estabilidad y presencia o ausenciade cambios en las características macro y microscópicas. Como resultado del trabajo se proponen los métodos más adecuados para los géneros Colletotrichum y Fusarium, hongos causantes de las mayores pérdidasen las plantaciones de ñame en la Costa Caribe colombiana. Las cepas utilizadas en este estudio provienen de las colecciones de la Universidad de Sucre, del Instituto de Biotecnología de la Universidad Nacional de Colombia (IBUN) y de una donación de la doctora Lucía Afanador de la Universidad Nacional, Sede Medellín.Estas fueron sometidas a diferentes métodos de conservación, entre ellos criopreservación, liofilizacióny cultivo periódico con y sin aceite mineral. Los resultados obtenidos permitieron elegir la criopreservacióncomo el método más eficiente para la conservación de la colección, y el cultivo periódico con aceite mineralcomo método alternativo y complementario. Estos minimizan el riesgo de pérdida del material biológico y brindan condiciones de manejo que conservan las características biológicas bajo estudio desde campos comola microbiología, la bioquímica y la biología molecular, entre otros.

An essential element in phytopathogenic microorganism research is their preservation and safe use. The purposeof this study was to evaluate methods for preserving fungi producing diseases in yam, using potential viability, stability and the presence or absence of macro- and microscopic characteristic changes as response factors. More suitable methods for Colletotrichum and Fusarium genera were proposed as a result of the work; these are fungi causing the greatest losses in yam plantations on the Colombian Caribbean Coast. The strains used in this study came from collections kept at the University of Sucre and the Universidad Nacional de Colombia’s Institute of Biotechnology (IBUN) in Bogota and a donation from Dr Lucia Afanador from the Universidad Nacional in Medellín. These strains were subjected to different preservation methods, including cryopreservation, lyophilisation and periodic culture with and without mineral oil. The results led to choosing cryopreservation as the most efficient method for preserving the collection and the periodic culture withmineral oil as an alternative and complementary method. These minimised the risk of biological material loss and provided handling conditions conserving the biological characteristics of the fungi being studied from the fields of microbiology, biochemistry and molecular biology.

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Atypical interstitial pneumonia in cattle associated with the ingestion of damaged sweet potatoes is reported in the State of Paraíba, Northeastern Brazil. The sweet potatoes were severely damaged by Myzus tersicae, and had an obvious fungal infection. Eighteen milking cows, 1 bull and 1 steer were fed approximately 400 kg of the sweet potatoes. Six days after consumption 13 cows were affected with labored abdominal breathing, extended and lowered head, coughing, expiratory grunt, salivation and protruded tongue. Six animals died and the others recovered in 4-7 d. At necropsy lungs were distended and did not collapse when the thorax was opened. Bullous emphysema and gelatinous exudates were observed in the interlobular, peribronchial and subpleural tissues. Microscopically, the lungs had severe edema and emphysema, congestion, and alveolar epithelial cell hyperplasia. The sweet potatoes were cultured but Fusarium spp was not isolated, probably because a zigomycete fungus covered the plates in 48 h of incubation.

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