RNA Interference of Endochitinases in the Sugarcane Endophyte Trichoderma virens 223 Reduces Its Fitness as a Biocontrol Agent of Pineapple Disease

The sugarcane root endophyte Trichoderma virens 223 holds enormous potential as a sustainable alternative to chemical pesticides in the control of sugarcane diseases. Its efficacy as a biocontrol agent is thought to be associated with its production of chitinase enzymes, including N-acetyl-beta-D-glucosaminidases, chitobiosidases and endochitinases. We used targeted gene deletion and RNA-dependent gene silencing strategies to disrupt N-acetyl-beta-D-glucosaminidase and endochitinase activities of the fungus, and to determine their roles in the biocontrol of soil-borne plant pathogens. The loss of N-acetyl-beta-D-glucosaminidase activities was dispensable for biocontrol of the plurivorous damping-off pathogens Rhizoctonia solani and Sclerotinia sclerotiorum, and of the sugarcane pathogen Ceratocystis paradoxa, the causal agent of pineapple disease. Similarly, suppression of endochitinase activities had no effect on R. solani and S. sclerotiorum disease control, but had a pronounced effect on the ability of T. virens 223 to control pineapple disease. Our work demonstrates a critical requirement for T. virens 223 endochitinase activity in the biocontrol of C. paradoxa sugarcane disease, but not for general antagonism of other soil pathogens. This may reflect its lifestyle as a sugarcane root endophyte.

Potential for entomopathogenic fungi to control Triatoma dimidiata (Hemiptera: Reduviidae), a vector of Chagas disease in Mexico

Introduction The use of entomopathogenic fungi to control disease vectors has become relevant because traditional chemical control methods have caused damage to the environment and led to the development of resistance among vectors. Thus, this study assessed the pathogenicity of entomopathogenic fungi in Triatoma dimidiata. Methods Preparations of 108 conidia/ml of Gliocladium virens, Talaromyces flavus, Beauveria bassiana and Metarhizium anisopliae were applied topically on T. dimidiata nymphs and adults. Controls were treated with the 0.0001% Tween-80 vehicle. Mortality was evaluated and recorded daily for 30 days. The concentration required to kill 50% of T. dimidiata (LC50) was then calculated for the most pathogenic isolate. Results Pathogenicity in adults was similar among B. bassiana, G. virens and T. flavus (p>0.05) and differed from that in triatomine nymphs (p=0.009). The most entomopathogenic strains in adult triatomines were B. bassiana and G. virens, which both caused 100% mortality. In nymphs, the most entomopathogenic strain was B. bassiana, followed by G. virens. The native strain with the highest pathogenicity was G. virens, for which the LC50 for T. dimidiata nymphs was 1.98 x108 conidia/ml at 13 days after inoculation. Conclusions Beauveria bassiana and G. virens showed entomopathogenic potential in T. dimidiata nymphs and adults. However, the native G. virens strain presents a higher probability of success in the field, and G. virens should thus be considered a potential candidate for the biological control of triatomine Chagas disease vectors.

Soilborne fungi and bacteria symbiotically associated with Steinernema spp. acting as biological agents against Fusarium wilt of tomato

An isolate of Gliocladium virens from disease affected soil in a commercial tomato greenhouse proved highly antagonistic to Fusarium oxysporum f.sp. lycopersici, used together with an isolate of the nematophagus fungus Verticillium chlamydosporium. Significant disease control was obtained when young mycelial preparation (on a food-base culture) of the G. virens together with V. chlamydosporium was applied in potting medium. Similar results were observed when a Trichoderma harzianum isolate was treated in combination with the V. chlamydosporium isolate. Most promising, in terms of minimizing the Fusarium wilt of tomato incidence, was also the effect of the bacteria associated with entomopathogenic nematodes (Steinernema spp.), Pseudomonas oryzihabitans and Xenorhabdus nematophilus.

STUDIES ON HAEMOLYMPH CONSTITUENTS OF PILA VIRENS (LAMARCK) AND THE EFFECTS OF PESTICIDES ON THE ACTIVITY PATTERN OF SELECTED ENZYMES

The present study is an attempt to understand the physiological responses of a freshwater gastropod, in terms of haematological parameters, in normal conditions as well as in various natural and man made altered conditions of the environment.Pila virens, a freshwater prosobranch,commonly found in paddy fields, ponds, and streams of Kerala is selected as the test animal for the present investigation. Various haemolymph constituents such as total carbohydrate, glycogen, total protein, total lipid, urea,ammonia,sodium,potasium, calcium, and chloride which are directly involved in the control and maintenance of different physiological systems, were analysed in the present study. Selected haematological parameters like total haemocyte number, and packed cell volume were also determined. Besides , the activity pattern of selected haemolymph enzymes such as acid phosphatase (ACP), alkaline phosphatase (ALP),Glutamate-oxaloacetate transaminase (GOT), and glutamate-pyruvate transaminase (GPT), all having diagnostic value in terms of internal defence system and metabolism of the organism, were also studied.

Predatory behavior of Canthon virens (Coleoptera: Scarabaeidae): A predator of leafcutter ants

We present a detailed description of the predatory behavior of the beetle Canthon virens Mannerheim, 1829, on the leafcutter ant Atta sp. We observed 51 acts of predation, which were also recorded on film and subjected to behavioral analysis. Canthon virens exhibited 28 behaviors while predating upon Atta sp. queens. Adult beetles search for queens while flying in a zigzag pattern, 15 to 20cm above the ground. After catching a queen, the predator stands on its back and starts cutting the queen cervix. Once the prey is decapitated, the predator rolls it until an insurmountable obstacle is reached. The distance from the site of predation to the obstacle can vary widely and is unpredictable. The beetle rolling the queen also buries it in a very peculiar way: first, it digs a small hole and pulls the queen inside, while another beetle is attached to the prey. The burial process takes many hours (up to 12) and may depend on the hardness of the soil and the presence of obstacles. In general, one or two beetles are found in a chamber with the queen after it is buried. They make the brood balls, which serve as food for the offspring. This study contributes to the knowledge of the predatory behavior of Canthon virens, a predator poorly studied in Brazil and widespread in the country. Copyright © 2012 Luiz Carlos Forti et al.

Struktur und Funktion des Lipoproteins von Nereis virens (Annelida)

Ein discoidales Lipoprotein aus dem Polychaeten Nereis virens (Annelida) wurde eingehend charakterisiert. Im Vordergrund standen dabei die transportierten Lipide, sowie die Ultrastruktur des Partikels. Das Nereis-Lipoprotein besitzt eine für Invertebraten atypische Lipidzusammensetzung: Außer den Phospholipiden gibt es keine klar dominierende Lipidklasse. Die Charakterisierung der Apolipoproteine zeigt Gemeinsamkeiten mit den Apolipophorinen der Insekten: Wie diese besitzt das Nereis-Lipoprotein zwei Apolipoproteine, die in einer 1:1-Stöchiometrie angeordnet sind. Das größere Protein (ApoNvLp I) ist dabei stärker zum wässrigen Medium exponiert ist als das kleinere (ApoNvLp II). Beide Proteinuntereinheiten sind N-glycosyliert. ApoNvLp II ist zusätzlich noch O-glycosyliert. Bei den Sekundärstrukturen dominieren β-Strukturen (35%) gegenüber α-Helices (14%); 28% waren ungeordnete Strukturen. Die Masse wurde mit verschiedenen Methoden bestimmt: sie liegt zwischen ~800 kDa (Gelfiltration) und ~860 kDa (Analytische Ultrazentrifugation). Der Sedimentationskoeffizient beträgt 9,7 S. Der zelluläre Lipoproteinrezeptor wurde aus einer großen Anzahl von Zellen und Geweben isoliert. Die biochemische Charakterisierung des Rezeptormoleküls zeigte es als ein monomeres, integrales, N- und O-glycosyliertes Membranprotein mit einer Masse von ~114 kDa. Die Bindungscharakteristika (Abhängigkeit von Ca2+, Disulfidbrücken) weisen es als Mitglied der LDLR-Superfamilie aus. In vitro-Inkubationsversuche mit fluoreszenzmarkierten Lipoproteinen zeigten die Aufnahme sowohl in Oocyten als auch in freie Coelomzellen (Elaeocyten) sowie in Spermatogonien- und Tetradenstadien. Auffällig war, dass die Lipide zusammen mit den Apolipoproteinen in die Dottergranula der Eizellen eingelagert wurden und nicht direkt in die Lipidtropfen. Auch bei den Elaeocyten wurden die Lipide nicht direkt in den Lipidtropfen eingelagert. Intakte Lipoproteine konnten per Dichtegradienten-Ultrazentrifugation nur aus Spermatogonien isoliert werden. Die isolierten Lipoproteine hatten die gleiche ‚Morphologie’ wie die aus der Coelomflüssigkeit isolierten, zeigten jedoch sehr viele Peptidfragmente im SDS-Gel, was auf eine beginnende Degradation hinweist. Es wird ein Modell für den Lipidtransport in Nereis virens vorgeschlagen, bei dem den Elaeocyten eine entscheidende Rolle im Lipidstoffwechsel zufällt.

Saprophytic microorganisms with potential for biological control of Botrytis cinerea on Geraldton waxflower flowers

Saprophytic bacteria, yeasts and filamentous fungi were isolated from Geraldton waxflower flowers and screened to identify potential antagonism towards Botrytis cinerea. Isolates from other sources (e.g. avocado) were also tested. Isolates were initially screened in vitro for inhibition of B. cinerea conidial germination, germ tube elongation and mycelial growth. The most antagonistic bacteria, yeasts and fungi were selected for further testing on detached waxflower flowers. Conidia of the pathogen were mixed with conidia or cells of the selected antagonists, co-inoculated onto waxflower flowers, and the flowers were sealed in glass jars and incubated at 20 degreesC. The number of days required for the pathogen to cause flower abscission was determined. The most antagonistic bacterial isolate, Pseudomonas sp. 677, significantly reduced conidial germination and retarded germ tube elongation of B. cinerea. None of the yeast or fungal isolates tested was found to significantly reduce conidial germination or retard germ tube elongation, but several significantly inhibited growth of B. cinerea. Fusarium sp., Epicoccum sp. and Trichoderma spp. were the most antagonistic of these isolates. Of the isolates tested on waxflower, Pseudomonas sp. 677 was highly antagonistic towards B. cinerea and delayed waxflower abscission by about 3 days. Trichoderma harzianum also significantly delayed flower abscission. However, as with most of the fungal antagonists used, inoculation of waxflower flowers with this isolate resulted in unsightly mycelial growth.

La biodiversidad microbiana del suelo y su rol en el manejo de enfermedades causadas por hongos de suelo en soja. Soil microbial diversity and its key role in the management of soybean diseases caused by fungi.

Las poblaciones microbianas juegan un rol fundamental en la estabilidad de los sistemas agrícolas e indican los niveles de salud de un suelo, tanto que pueden ser utilizadas como indicadores de sustentabilidad de un agroecosistema. Los microorganismos reflejan el efecto que tienen las prácticas agrícolas sobre el suelo a través de modificaciones en la abundancia (biodiversidad estructural) y actividades de sus poblaciones (como control biológico de los patógenos, entre otras). Al cuantificarse la biodiversidad microbiana nativa se puede conocer la riqueza de un agroecosistema y utilizarla para el manejo sustentable de hongos patógenos. En este trabajo se evaluará el efecto de la rotación de cultivo (soja-maíz y soja en monocultivo) y los sistemas de labranza (siembra directa y labranzas reducida) sobre la biodiversidad microbiana. Se cuantificarán a partir de suelo: poblaciones de hongos y bacterias totales; agentes potenciales de biocontrol de como Trichoderma spp., Gliocladium spp. y micorrizas vesículo arbusculares (mediante la cuantificación de glomalina), biomasa y respiracion microbiana, y la biodiversidad de comunidades de microorganismos que habitan en el suelo mediante el análisis de perfiles de ácidos grasos (PLFA). Al final del ciclo de cultivo de soja se cuantificará la incidencia de enfermedades causadas por hongos de suelo. Dada la gran abundancia y diversidad de los microorganismos del suelo, las metodologías que se emplearán permitirán obtener información global de la riqueza microbiana de un agro-ecosistema. Se relacionará la biodiversidad microbiana con la incidencia de enfermedades por hongos de suelo, en respuesta a diferentes prácticas de manejo. Esto permitirá aprender a combinar las tecnologías para mejorar los beneficios de la produccion y preservar el agroecosistema en el marco de una agricultura sustentable, y no de una agricultura sostenida por insumos.

A new barbet from French Indo-China.

v.34:no.21(1953)

Review of the genus Chalcolepidius Eschscholtz, 1829 (Coleoptera, Elateridae, Agrypninae)

The genus Chalcolepidius is revised. Type specimens of 65 nominal species, except C. costatus Pjatakowa, 1941, C. fleutiauxi Pjatakowa, 1941 and C. viriditarsus Schwarz, 1906, are examined. Eighty five species are studied, of which 34 are synonymyzed and 12 new species described; three species, C. alicii Pjatakowa, 1941, C. haroldi Candèze, 1878 and C. unicus Fleutiaux, 1910, formely included in this genus, are not congeneric and are removed; C. validus Candèze, 1857 is revalidated. The genus is now formed by 63 species. Redescriptions, illustrations and a key for the examined species, and a cladistic analysis for groups of species are also included. New synonyms established: C. apacheanus Casey, 1891 = C. simulans Casey, 1907 syn. nov. = C. acuminatus Casey, 1907 syn. nov. = C. nobilis Casey, 1907 syn. nov.; C. approximatus Erichson, 1841 = C. aztecus Casey, 1907 syn. nov. = C. niger Pjatakowa, 1941 syn. nov.; C. attenuatus Erichson, 1841 = C. cuneatus Champion, 1894 syn. nov. = C. tenuis Champion, 1894 syn. nov.; C. aurulentus Candèze, 1874 = C. candezei Dohrn, 1881 syn. nov. = C. grossheimi Pjatakowa, 1941 syn. nov.; C. bomplandii Guérin, 1844 = C. humboldti Candèze, 1881 syn. nov.; C. chalcantheus Candèze, 1857 = C. violaceous Pjatakowa, 1941 syn. nov.; C. cyaneus Candèze, 1881 = C. scitus Candèze, 1889 syn. nov. = C. abbreviatovittatus Pjatakowa, 1941 syn. nov.; C. desmarestii Chevrolat, 1835 = C. brevicollis Casey, 1907 syn. nov.; C. gossipiatus Guérin, 1844 = C. erichsonii Guérin-Méneville, 1844 syn. nov. = C. lemoinii Candèze, 1857 syn. nov.; C. inops Candèze, 1886 = C. murinus Champion, 1894 syn. nov.; C. jansoni Candèze, 1874 = C. mucronatus Candèze, 1889 syn. nov.; C. lacordairii Candèze, 1857 = C. exquisitus Candèze, 1886 syn. nov. = C. monachus Candèze, 1893 syn. nov.; C. lenzi Candèze, 1886 = C. behrensi Candèze, 1886 syn. nov.; C. oxydatus Candèze, 1857 = C. jekeli Candèze, 1874 syn. nov.; C. porcatus (Linnaeus, 1767) = C. peruanus Candèze, 1886 syn. nov. = C. flavostriatus Pjatakowa, 1941 syn. nov. = C. herbstii multistriatus Golbach, 1977 syn. nov.; C. rugatus Candèze, 1857 = C. amictus Casey, 1907 syn. nov.; C. smaragdinus LeConte, 1854 = C. ostentus Casey, 1907 syn. nov. = C. rectus Casey, 1907 syn. nov.; C. sulcatus (Fabricius, 1777) = C. herbstii Erichson, 1841 syn. nov; C. virens (Fabricius, 1787) = C. perrisi Candèze, 1857 syn. nov.; C. virginalis Candèze, 1857 = C. championi Casey, 1907 syn. nov.; C. viridipilis (Say, 1825) = C. debilis Casey, 1907 syn. nov.; C. webbi LeConte, 1854 = C. sonoricus Casey, 1907 syn. nov.; C. zonatus Eschscholtz, 1829 = C. longicollis Candèze, 1857 syn. nov. New species described: C. albisetosus sp. nov. (Ecuador), C. albiventris sp. nov. (Mexico: Veracruz), C. copulatuvittatus sp. nov. (Venezuela), C. extenuatuvittatus sp. nov. (Venezuela), C. fasciatus sp. nov. (Mexico: Durango), C. ferratuvittatus sp. nov. (Ecuador), C. proximus sp. nov. (Mexico: Sinaloa), C. serricornis sp. nov. (Mexico: Veracruz), C. spinipennis sp. nov. (Mexico: Veracruz), C. supremus sp. nov. (Venezuela), C. truncuvittatus sp. nov. (Mexico: Tamaulipas) and C. virgatipennis sp. nov. (Mexico: Durango). Redescribed species: C. angustatus Candèze, 1857, C. apacheanus Casey, 1891, C. approximatus Erichson, 1841, C. attenuatus Erichson, 1841, C. aurulentus Candèze, 1874, C. bomplandii Guérin-Méneville, 1844, C. boucardi Candèze, 1874, C. chalcantheus Candèze, 1857, C. corpulentus Candèze, 1874, C. cyaneus Candèze, 1881, C. desmarestii Chevrolat, 1835, C. dugesi Candèze, 1886, C. erythroloma Candèze, 1857, C. eschscholtzi Chevrolat, 1833, C. exulatus Candèze, 1874, C. fabricii Erichson, 1841, C. forreri Candèze, 1886, C. fryi Candèze, 1874, C. gossipiatus Guérin-Méneville, 1844, C. inops Candèze, 1886, C. jansoni Candèze, 1874, C. lacordairii Candèze, 1857, C. lafargi Chevrolat, 1835, C. lenzi Candèze, 1886, C. limbatus (Fabricius, 1777), C. mexicanus Castelnau, 1836, C. mniszechi Candèze, 1881, C. mocquerysii Candèze, 1857, C. morio Candèze, 1857, C. obscurus Castelnau, 1836, C. oxydatus Candèze, 1857, C. porcatus (Linnaeus, 1767), C. pruinosus Erichson, 1841, C. rodriguezi Candèze, 1886, C. rostainei Candèze, 1889, C. rubripennis LeConte, 1861, C. rugatus Candèze, 1857, C. silbermanni Chevrolat, 1835, C. smaragdinus LeConte, 1854, C. sulcatus (Fabricius, 1777), C. tartarus Fall, 1898, C. validus Candèze, 1857, reval., C. villei Candèze, 1878, C. virens (Fabricius, 1787), C. virginalis Candèze, 1857, C. viridipilis (Say, 1825), C. webbi LeConte, 1854, C. zonatus Eschscholtz, 1829.

Lauluja kokoelmasta "Piae Cantiones"

Äänitetty: [1962?, Yleisradio?], Ä-5.

Fungos antagonistas a Sclerotinia sclerotiorum em pepineiro cultivado em estufa

O mofo branco causado por Sclerotinia sclerotiorum pode inviabilizar o cultivo de olerícolas em ambiente protegido. Para elaborar-se um programa de controle biológico desse patógeno, necessita-se de antagonistas adequados. Este trabalho objetivou selecionar antagonistas fúngicos eficazes no controle de S. sclerotiorum em pepineiro (Cucumis sativus) cultivado em estufa, bem como, analisar a interferência dos antagonistas no crescimento vegetal. Foram utilizados um isolado de S. sclerotiorum obtido de pepineiro e 112 isolados fúngicos de quatro gêneros: Trichoderma, Fusarium, Penicillium e Aspergillus. Em experimento in vitro, foi utilizada a técnica do papel celofane e selecionados oito isolados de Trichoderma virens, os quais promoveram maior inibição no crescimento do patógeno (94 a 100%). Dois experimentos in vivo foram desenvolvidos em estufa utilizando-se substrato autoclavado e não autoclavado, em copos plásticos, e substrato não autoclavado, em sacos plásticos; o substrato foi infestado com S. sclerotiorum e foram utilizados oito isolados de T. virens como antagonistas. Todos os isolados testados controlaram o tombamento de mudas, mas o efeito sobre o crescimento vegetal variou de acordo com os isolados e o tratamento do substrato.

Avaliação antagônica in vitro e in vivo de Trichoderma spp. a Rhizopus stolonifer em maracujazeiro amarelo

Para estudar a potencialidade antagônica de espécies de Trichoderma spp. in vitro e in vivo a Rhizopus stolonifer, patógeno causador da podridão floral do maracujazeiro, foram estudadas as espécies de Trichoderma viride, T. virens, T. harzianum e T. stromaticum. O crescimento micelial do fitopatógeno foi realizado pelo teste do pareamento de culturas, para crescimento individual foram utilizadas cinco temperaturas. Avaliou-se também o crescimento micelial em 24h e 48h, avaliando a taxa de crescimento dos isolados. Na produção de metabolitos voláteis e não voláteis foram utilizados papel celofane e sobreposição de placas. Em condição de campo os frutos/planta foram tratados com a suspensão na concentração de 2 x 10(8) Conídios/mL sendo avaliado o número médio de frutos aos 15 e 30. No pareamento de cultura todos os isolados de Trichoderma spp. apresentaram crescimento micelial, impedindo o desenvolvimento do fitopatógeno, para todos os isolados as temperaturas ideais de crescimento foram de 25ºC e 30ºC. Nos períodos de incubação de 24 e 48h, foram constatadas diferenças significativas no crescimento micelial entre os isolados os antagonistas apresentaram velocidade de crescimento maior que o fitopatógeno. Houve uma produção de metabólitos voláteis e não voláteis de ação antifúngica ao R. stolonifer. No ensaio em campo houve diferença significativa entre os tratamentos, verificando-se que o melhor resultado entre os antagonistas em estudo cujos percentuais de pegamento foram 74% para os tratamentos Trichoderma harzianum e T. virens, e os tratamentos T. viride e T. stromaticum obtiveram um porcentual de 75% enquanto a testemunha obteve um percentual de 42%.