Determinación de la calidad fitosanitaria de la semilla de frijol común en Nicaragua y alternativas de manejo del grano

En Nicaragua el género Phaseolus, representa una fuente importante de nutrientes que la población incluye en su dieta diaria, además provee ingresos a los productores con los que subsanan una parte de sus necesidades, El 95% de la producción de frijol en el país descansa en pequeños y medianos productores que enfrentan problemas como: Falta de asistencia técnica, poca o ninguna disponibilidad de créditos, poseen terrenos no adecuados para este cultivo y el uso constante de semillas remanentes de un ciclo a otro; Esto ha incidido de forma directa en la disminución de los rendimientos y en el aumento de los niveles de inoculo en la semilla. Son muchas las enfermedades que atacan a esta leguminosa y algunas de estas llegan a infestar y/o infectar la semilla logrando así un eficiente mecanismo de dispersión. Con el objetivo de conocer sobre la calidad fitosanitaria de la semilla utilizada por los productores, evaluar algunas técnicas para la preservación de la misma, determinar el nivel de conocimiento de los productores con respecto a las enfermedades y discernir sobre la efectividad de las técnicas de almacenamiento de la semilla utilizadas por los productores, se realizó este trabajo involucrando 75 productores del país, logrando recolectar de sus manos un total de 15 variedades las que corresponden a los nombres de: DOR-364, RAB-310, Honduras-46, Estelí-150, Estelí-90A, Estelí-B, Negro, Blanco, Chiricano, Rojo criollo, DICTA-114, Balin tíco, Revolución-84 y dos variedades del Centro Nacional de Investigación Agropecuaria (CNIA), DOR-805 y DOR-576. El trabajo se dividió en dos fases: una de campo que consistió en la colecta de datos y muestras de semillas en las zonas de estudio en el mes de Marzo y una de laboratorio en la que se determinaron los diferentes tipos de microorganismos presente en las semillas a través de Observación de síntomas, pruebas de laboratorio y identificación de los microorganismos. Se hizo una prueba de almacenaje mediante el uso de botes plásticos con tapadera, los tratamientos utilizados para la preservación fueron• Cal, Ceniza y Ceniza+Cal en dosis de 80gr de producto por libra de semilla, más o menos 25 libras por quintal de grano y un testigo. Los resultados de las encuestas demuestran que los productores reconocen las enfem1edades como tales, pero no pueden diferenciarlas en su totalidad como causadas por hongos, bacterias o virus. Se identificaron los siguientes patógenos: Rhizoctonia sotaní, Thanatephoms cucumeris, Collectotrichum lindemuthianum, Fusarium solani, Fusarium poae, Fusarium tricinctum, l-i1sarium oxyspomm, Penicillium .;pp, Aspergillus ochraceus, Aspergillus ustus, Aspergillus glaucus (Emericella nidulans)(Eim>tium link), Aspergiilus niger, Aspergillus parasitim Aspergillus candidus, Aspergillus terreus, Rhizopus oryzae, Rhizopus stolonifer, Xanthomona;• campestris pv phaseoli, Pseudomonas spp, el virus del mosaico común no se detectó en este trabajo. Los mejores resultados se observaron en los tratamientos de cal y cal+ceniza, siendo cal+ceniza quién presentó alta significancia en la disminución de la infección por hongos, gorgojos (Bruchidae) y bacterias, pudiendo disminuir hasta un 57% las infecciones de hongos, un 68% de las infecciones por bacterias a nivel superficial y un l 00% la población de gorgojos, con respecto a las presentadas por el testigo de laboratorio. El tratamiento de cal+ceniza es más barato, eficaz y menos peligroso que el uso de cualquier químico.

Diagnóstico de los agentes causantes del mal seco en el cultivo del quequisque (Xanthosoma spp) en el municipio de Nueva Guinea, Nicaragua

El objetivo del estudio fue contribuir al esclarecimiento de la etiología de los agentes causales del mal seco. La colecta de muestras de suelo y material vegetal enfermo, fue realizada en 17 fincas, en el municipio de Nueva Guinea. Se realizó un análisis patológico y f��sico - químico de suelo, así como una encuesta sobre el historial del manejo del cultivo. Para el aislamiento de agentes causales se utilizaron los medios: papa dextrosa agar (PDA), agar nutritivo (AN) y agar harina de maíz más antibióticos Pimaricina, Ampicilina y Rifampicina(PARC). Para la prueba de patogenicidad, se sembraron plantas en suelo infectado y en suelo estéril donde se inoculó con los patógenos: Pythium myriotylum, Fusarium solani, Ralstonia solanacearum y Pythium myriotylum + Fusarium solani + Ralstonia solanacearum; los que fueron previamente aislados de raíces. Las variables evaluadas fueron frecuencia de aparición de patógenos y severidad en raíces. Los datos de crecimiento de los patógenos en los diferentes medios decultivos fueron analizados por el método de Chi cuadrado. La frecuencia de aparición de P. myriotylun, en medio PARC, Fusariums olani PDA, y Ralstonia solanacearum en AN, fueron significativos con P< 0.0001 en los aislados de raíces y suelo. Las plantas sembradas en suelo inoculado con Pythium myriotylun presentaron clorosis generalizada en las partes aéreas mientras en F. solani y R. solanacearum, presentaron coloración verde pálido y flacidez en hojas. En raíces P. myriotylum presentó pudrición y descortezamiento, F. solani pudrición seca y R. solanacearum puntos necróticos acuosos. Se observó un 80 % de severidad en raíces en los tratamientos de P. myriotylun y P. myriotylun + F. solani + R. solanacearum con tasa de incremento de la enfermedad de 0.005 y 0.006 respectivamente. El tratamiento P. myriotylun presentó los síntomas característicos del mal seco: clorosis generalizada, caída del pecíolo en forma de arco, pudrición y descortezamiento en raíces.

牛蒡根际土壤致害菌Fusariumsolani分离鉴定

目的:探索牛蒡根际镰刀菌与牛蒡之间的相互关系。方法:从全国30个地区采集牛蒡根际土壤样品,进行了根际土壤真菌数量和群落的生态学研究,测定镰刀菌发酵液对牛蒡幼苗和牛蒡子萌发的影响,并对其中毒性最强的两株镰刀菌F130和F131进行了形态学和分子生物学的鉴定。结果:镰刀菌为牛蒡根际的最优势类群,贡献率为34.297%,其次为木霉,贡献率为22.519%;绝大多数镰刀菌对牛蒡有明显的毒性作用,其中F130和F131被鉴定为Fusarium solani。结论:Fusarium solani是牛蒡根际土壤中的致害菌。

Influência de Bacillus subtilis na promoção de crescimento de plantas e nodulação de raízes de feijoeiro.

Linhagens de B. subtilis têm sido bem estudadas como agentes de controle biológico, e também como produtoras de antibióticos. Uma linhagem de B. subtilis 0G isolada de solos supressivos a Fusarium solani foi avaliada quanto à utilização conjunta com Rhizobium phaseoli em feijoeiro, visando aumento de biomassa e nodulação de raízes. Em condições de casa de vegetação, a aplicação de B. subtilis, via sementes, promoveu um aumento na nodulação, em mais de 100%, em solo natural quando aplicado isoladamente ou em conjunto com R. phaseoli e em mais de 160% em solo esterilizado, quando comparado com o tratamento onde o Rhizobium foi aplicado isoladamente. Em solos esterilizados, a aplicação de B. subtilis, em conjunto com Rhizobium, promoveu um aumento significativo no peso da matéria seca das raízes (89%) e da parte aérea (83%). O tratamento de sementes com B. subtilis não afetou a emergência de plântulas em nenhum dos solos estudados. Pelos resultados obtidos pode-se concluir que a utilização de B. subtilis é bastante promissora para aumentar a nodulação de raízes e promover o crescimento de plantas de feijoeiro.

First report of a novel plant lysozyme with both antifungal and antibacterial activities.

A novel lysozyme exhibiting antifungal activity and with a molecular mass of 14.4 kDa in SDS–polyacrylamide gel electrophoresis was isolated from mung bean (Phaseolus mungo) seeds using a procedure that involved aqueous extraction, ammonium sulfate precipitation, ion exchange chromatography on CM-Sephadex, and high-performance liquid chromatography on POROS HS-20. Its N-terminal sequence was very different from that of hen egg white lysozyme. Its pI was estimated to be above 9.7. The specific activity of the lysozyme was 355 U/mg at pH 5.5 and 30 °C. The lysozyme exhibited a pH optimum at pH 5.5 and a temperature optimum at 55 °C. It is reported herein, for the first time, that a novel plant lysozyme exerted an antifungal action toward Fusarium oxysporum, Fusarium solani, Pythium aphanidermatum, Sclerotium rolfsii, and Botrytis cinerea, in addition to an antibacterial action against Staphylococcus aureus.

Survey of Agricultural Practices and Alternatives to Pesticide Use to Conserve Water Resources in the Mojanda Watershed, Ecuador

Agriculture in the Mojanda Watershed is facing rainfall reductions caused by climate change. Reductions of water availability in the Watershed are also due to constant extension of the agricultural activities into the páramo ecosystem above 3000m a.s.l., with this ecosystem having immanently important functions in the local water balance. The application of pesticides threatens the quality of water and with less precipitation contaminations will further concentrate in the outflow. To analyze problems associated with agricultural practices in the area a questionnaire about agricultural practices (28) was conducted and fields (20) were surveyed for pests and diseases with a focus on potatoes (Solanum tuberosum L.), tree tomatoes (Solanum betaceum Cav.) and peas (Pisum sativum L.). Potatoes were infected to a low degree with Phytophthora infestans and according to the farmers the Andean potato weevil (Premnotrypes spec.) caused biggest losses. To combat the weevil the soils are disinfected with toxic Carbofuran (WHO Class 1B). Tree tomatoes showed symptoms of various fungal diseases. Most important was Fusarium solani causing the branches to rot and Anthracnosis (Colletotrichum gloeosporioides) causing the fruits to rot. Fungicide applications were correspondingly high. Peas were only minorly affected by Ascochyta blight (Mycosphaerella pinodes) and a root rot. Overall 19 active ingredients were applied of which fungicide Mancozeb (WHO class table 5) and insecticide Carbofuran (WHO Class 1B) were applied the most. Approved IPM methods are advised to reduce pesticide use. For tree tomatoes regular cutting of branches infected with F. solani and regular collection and disposal of infected fruits with Anthracnosis are advised. For potatoes plastic barriers around the fields prevent the Andean potato weevil from laying eggs thus reducing infestation with the larvae in the tubers. Local bioinsecticide “Biol” seems effective and without harm to the environment, although not used by many farmers. Organic fertilization promises to restore decreasing soil fertility, water holding capacity and reduce erosion. The here presented alternatives and strategies to reduce pesticide use pose an opportunity to preserve the water resources of the region.

Fragrance release from the surface of branched poly(amide)s

Enzymes are powerful tools in organic synthesis that are able to catalyse a wide variety of selective chemical transformations under mild and environmentally friendly conditions. Enzymes such as the lipases have also found applications in the synthesis and degradation of polymeric materials. However, the use of these natural catalysts in the synthesis and the post-synthetic modification of dendrimers and hyperbranched molecules is an application of chemistry yet to be explored extensively. In this study the use of two hydrolytic enzymes, a lipase from Candida cylindracea and a cutinase from Fusarium solani pisii, were investigated in the selective cleavage of ester groups situated on the peripheral layer of two families of branched polyamides. These branched polyamides were conjugated to simple fragrances citronellol and L-menthol via ester linkages. Hydrolysis of the ester linkage between the fragrances and the branched polyamide support was carried out in aqueous buffered systems at slightly basic pH values under the optimum operative conditions for the enzymes used. These preliminary qualitative investigations revealed that partial cleavage of the ester functionalities from the branched polyamide support had occurred. However, the ability of the enzymes to interact with the substrates decreased considerably as the branching density, the rigidity of the structure and the bulkiness of the polyamide-fragrance conjugates increased.

Osmotin from Calotropis procera latex: New insights into structure and antifungal properties

This study aimed at investigating the structural properties and mechanisms of the antifungal action of CpOsm, a purified osmotin from Calotropis procera latex. Fluorescence and CD assays revealed that the CpOsm structure is highly stable, regardless of pH levels. Accordingly, CpOsm inhibited the spore germination of Fusarium solani in all pH ranges tested. The content of the secondary structure of CpOsm was estimated as follows: alpha-helix (20%), beta-sheet (33%), turned (19%) and unordered (28%). RMSD 1%. CpOsm was stable at up to 75 degrees C, and thermal denaturation (T(m)) was calculated to be 77.8 degrees C. This osmotin interacted with the negatively charged large unilamellar vesicles (LUVs) of 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-rac-1-glycerol (POPG), inducing vesicle permeabilization by the leakage of calcein. CpOsm induced the membrane permeabilization of spores and hyphae from Fusarium solani, allowing for propidium iodide uptake. These results show that CpOsm is a stable protein, and its antifungal activity involves membrane permeabilization, as property reported earlier for other osmotins and thaumatin-like proteins. (C) 2011 Elsevier B.V. All rights reserved.

Vicilinas de sementes de leguminosas selvagens:purificação, caracterização, efeito de deletério e mecanismo de ação para bruquídeos e para fungos fitopatogêncios

Globulins fractions of legume seeds of Crotalaria pallida, Erytrina veluntina and Enterolobium contortisiliquum were isolated and submitted to assays against serine, cysteine and aspartic proteinases, as also amylase present in midgut of C. maculatus and Z. subfasciatus. Hemagglutination assays indicated presence of a lectin in E. veluntina globulin fractions. This lectin had affinity to human erythrocytes type A, B and O. Vicilins were purified by chromatography on Sephacryl S-300 followed of a chromatography on Sephacryl S-200, which was calibrated using protein markers. Vicilins from C. pallida (CpV) and E. veluntina (EvV) seeds had a molecular mass of 124.6 kDa and E. contortisiliquum a molecular mass of 151kDa. Eletrophoresis in presence of SDS showed that CpV was constituted by four subunities with apparent molecular mass of 66, 63, 57 and 45 kDa, EvV with three subunities with apparent molecular mass of 45kDa and EcV four subunities, two with 37.1 kDa and two with 25.8 kDa. Non denaturantig eletrophoresis displayed single bands with high homogeneity, where CpV had lower acidic behavior. All vicilins are glycoproteins with carbohydrate contents at 1 to1.5%. Bioassays were done to detect deleterious effects of vicilins against C. maculatus and Z. subfasciatus larvae. CpV, EvV and EcV exhibited a WD50 of 0.28, 0.19 and 1.03%; LD50 0.2, 0.26, and 1.11% respectively to C. maculatus. The dose responses of CpV, EvV and EcV to Z. subfasciatus were: WD50 of 0.12, 0.14, 0.65% and LD50 of 0.09, 0.1, and 0.43% respectively. The mechanism of action of these proteins to bruchids should be based on their properties of bind to chitin present in mid gut of larvae associated with the low digestibility of vicilin. In assays against phytopatogenous fungus, only EcV was capable of inhibit F. solani growth at concentrations of 10 and 20 µg and its action mechanism should be also based in the affinity of EcV to chitin present in the fungi wall

Growth and enzymatic responses of phytopathogenic fungi to glucose in culture media and soil

The effect of inoculation of Aspergillus flavus, Fusarium verticillioides, and Penicillium sp. in Dystrophic Red Latosol (DRL) and Eutroferric Red Latosol (ERL) soils with or without glucose on the total carbohydrate content and the dehydrogenase and amylase activities was studied. The fungal growth and spore production in culture medium with and without glucose were also evaluated. A completely randomized design with factorial arrangement was used. The addition of glucose in the culture medium increased the growth rate of A. flavus and Penicillium sp. but not of F. verticillioides. The number of spores increased 1.2 for F. verticillioides and 8.2 times for A. flavus in the medium with glucose, but was reduced 3.5 times for Penicillium sp. The total carbohydrates contents reduced significantly according to first and second degree equations. The consumption of total carbohydrates by A. flavus and Penicillium sp. was higher than the control or soil inoculated with F. verticillioides. The addition of glucose to soils benefited the use of carbohydrates, probably due to the stimulation of fungal growth. Dehydrogenase activity increased between 1.5 to 1.8 times (p <0.05) in soils with glucose and inoculated with the fungi (except F. verticillioides), in relation to soil without glucose. Amylase activity increased 1.3 to 1.5 times due to the addition of glucose in the soil. Increased amylase activity was observed in the DRL soil with glucose and inoculated with A. flavus and Penicillium sp. when compared to control.

Variability of non-mutualistic filamentous fungi associated with Atta sexdens rubropilosa nests

A survey of the filamentous fungi other than the symbiotic one found in association with Atta sexdens rubropilosa colonies was carried out. Different fungal species (27 taxa) were isolated a few days after treating the workers with toxic baits (sulfluramid; Mirex-SO), from 40 laboratory and 20 field nests. Syncephalastrum racemosum (54 %) and Escovopsis weberi (21 %), Trichoderma harzianum (38 %) and Fusarium oxysporum (23 %) were the prevalent species in laboratory and field nests, respectively. Acremonium kiliense, Acremonium strictum, E. weberi, F oxisporum, Fusarium solani, Moniliella silaveolens and T harzianum were found in both nests' groups. We revealed that many filamentous fungi can co-exist in a dormant state inside the nests of these insects and some of them appear to be tightly associated with this environment.

Disseminated Amphotericin-Resistant Fusariosis in Acute Leukemia Patients: Report of Two Cases

Disseminated fusariosis has emerged as a significant, usually fatal infection in immunocompromised hosts despite antifungal treatment. We describe here two patients with acute leukemia who developed disseminated amphotericin-resistant fusariosis, and review of six studies of cases series in the literature. Two Fusarium solani strains were isolated from blood and skin cultures of one patient, and one strain from the blood culture of the second patient. Both patients died despite antifungal treatment. Strains were identified by sequencing of ITS1 and ITS4 regions. Random amplified polymorphic DNA analysis of the three F. solani isolates showed a low degree of similarity. Screening for Fusarium spp. contaminants within our facility was negative. Using the CLSI M-38-A2 broth dilution method and E tests®, we found that the MICs were low for voriconazole (0. 12 and 0. 5 mg/L, respectively), unexpectedly high for amphotericin B (≥8 and ≥32 μg/mL, respectively) and itraconazole (≥16 mg/ml). Patients with leukemia or persistent neutropenia should be assessed for disseminated fungal infections, including biopsy and skin cultures. Antifungal susceptibility tests are important due to the possibility of the strains being amphotericin resistant. Treatments must be aggressive, with high doses of antifungals or combined therapy. © 2012 Springer Science+Business Media Dordrecht.

Efeito da inoculação de fungos na atividade enzimática de solos e germinação de sementes de milho

Pós-graduação em Agronomia (Ciência do Solo) - FCAV

Uso e conservação de plantas medicinais nativas por comunidades quilombolas no município de Alcântara, Maranhão

Pós-graduação em Agronomia (Horticultura) - FCA

Reação de araçazeiros a Meloidogyne enterolobii e enxertia da goiabeira ‘Paluma’ em portaenxertos resistentes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)