Impact of mirid (Creontiades spp.) (Hemiptera: Miridae) pest management on Helicoverpa spp. (Lepidoptera: Noctuidae) outbreaks: The case for conserving natural enemies

Creontiades spp. (Hemiptera: Miridae) are sucking pests that attack buds, flowers and young pods in mungbeans, Vigna radiata (L.), causing these structures subsequently to abort. If left uncontrolled, mirids can cause 25-50% yield loss. Traditional industry practice has involved prophylactic applications of dimethoate to control mirids at budding and again a week later. The present trial was initiated to highlight the dangers of such a practice, in particular the risk of a subsequent Helicoverpa spp. lepidopteran pest outbreak. A single application of dimethoate halved the population of important natural enemies of Helicoverpa spp., and caused an above-threshold outbreak of Helicoverpa spp. within 11 days. This shows that even a moderate (e.g. 50%) reduction in natural enemies may be sufficient to increase Helicoverpa spp. populations in mungbeans. As a result, prophylactic sprays should not be used for the control of mirids in mungbeans, and dimethoate should be applied only when mirids are above the economic threshold. Indoxacarb was also tested to establish its effect on Helicoverpa spp., mirids and natural enemies. Indoxacarb showed potential for Helicoverpa spp. control and suppression of mirids and had little impact on natural enemies.

Utilizing the assassin bug, Pristhesancus plagipennis (Hemiptera: Reduviidae), as a biological control agent within an integrated pest management programme for Helicoverpa spp. (Lepidoptera: Noctuidae) and Creontiades spp. (Hemiptera: Miridae) in cotton

Helicoverpa spp. and mirids, Creontiades spp., have been difficult to control biologically in cotton due to their unpredictable temporal abundance combined with a cropping environment often made hostile by frequent usage of broad spectrum insecticides. To address this problem, a range of new generation insecticides registered for use in cotton were tested for compatibility with the assassin bug, Pristhesancus plagipennis (Walker), a potential biological control agent for Helicoverpa spp. and Creontiades spp. Indoxacarb, pyriproxifen, buprofezin, spinosad and fipronil were found to be of low to moderate toxicity on P. plagipennis whilst emamectin benzoate, abamectin, diafenthiuron, imidacloprid and omethaote were moderate to highly toxic. Inundative releases of P. plagipennis integrated with insecticides identified as being of low toxicity were then tested and compared with treatments of P. plagipennis and the compatible insecticides used alone, conventionally sprayed usage practice and an untreated control during two field experiments in cotton. The biological control provided by P. plagipennis nymphs when combined with compatible insecticides provided significant (P<0.001) reductions in Helicoverpa and Creontiades spp. on cotton and provided equivalent yields to conventionally sprayed cotton with half of the synthetic insecticide input. Despite this, the utilization of P. plagipennis in cotton as part of an integrated pest management programme remains unlikely due to high inundative release costs relative to other control technologies such as insecticides and transgenic (Bt) cotton varieties.

The complete mitochondrial genome of the tarnished plant bug, Lygus lineolaris (Heteroptera: Miridae)

The complete mitochondrial genome of the tarnished plant bug, Lygus lineolaris, comprised 17,027 bp. The genome contained 13 protein coding regions, 22 tRNA genes and 2 ribosomal RNA genes. The gene arrangement corresponded to the common order found among insect mtDNAs which was considered to be the ancestral arrangement. The protein coding genes started with ATN and stopped with TAA or TAG. The nucleotide distribution was 76.0% A + T. The control region contained two repeat regions, one was 24 bp and the other was 161 bp. The Genbank accession for the complete L. lineolaris mt genome is EU401991.

Estudios biológicos preliminares sobre Halticus pygmaeus (Miridae : Hemiptera) : I. ciclo biológico y descendencia sobre Medicago sativa

p.111-115

Efecto Insecticida del Extracto de Semillas de Neem (Azadirachta indica A. Juss) sobre Collaria scenica Stal (Hemiptera: Miridae)

This work was developed with the objetive to evaluate the insecticidal effect of the Neem (Azadirachta indica A. Juss) seeds against grass bug nymphs Collaria scenica Stal. For that, an extract was prepared from green fruits of Neem by rotaevaporacion. The extract was diluted in three ppm concentrations corresponding to treatments. Using thin-layer chromatography we identified the presence of Azaridactina. DCA essay was carried out with 4 treatments and 5 repetitions that included the three extract concentrations and a control. In each repetition were used 15 nymphs, randomly placed in plastic boxes with food and the corresponding concentration. Every day an extract application was made and recorded mortality percentage, number of exuviae and number of individuals who came to adulthood in each treatment. The results were analyzed using Kruskal-Wallis and Games-Howell for each variable. The three concentrations of Neem seed extract had a negative effect on the development of the bugs. The more concentrated treatment (250 ppm) was the most effective, showing a 97% mortality, fewer exuvias and and fewer adults in the end of the experiment.

First record of infection by entomopathogenic nematodes of the grass bug Collaria scenica Stal (Hemiptera: Miridae).

NARANJO N, MONTERO DAV, SAENZ APONTE A. 2011. First record of infection by entomopathogenic nematodes of the grass bug Collaria scenica Stal (Hemiptera: Miridae). ENTOMOTROPICA 26(3): 117-125. The study was aimed to test the pathogenicity of Steinernema sp. and Heterorhabditis sp. in Collaria scenica. The effect of different concentrations of infective juveniles (IJ) were tested on nymphs and adults of C. scenica. For this purpose, the bugs were inoculated with 5 000 JI of each nematode species in a factorial design (3x2), and seven concentrations were tested in a JI factorial design (7x2x2). The bugs showed 100% mortality and symptoms of pathogenicity. Infection was found with both species of nematodes and penetration was assumed to be through the spiracles and anus. A higher capacity of pathogenicity was observed with Steinernema sp. Based on the results Heterorhabditis sp. and Steinernema sp. could constitute an efficient tool to control populations of C. scenica in pastures.

Control de la chinche de los pastos Collaria scenica (Hemiptera: Miridae) con nematodos entomopatógenos en invernadero

We studied the efficacy of infective juveniles (IJ) of Steinernema sp. strain JCL024 and Heterorhabditis sp. strain SL0708 on adults and nymphs of Collaria scenica under greenhouse conditions. Five doses were evaluated (2, 16, 78, 160 IJ) cm-2 leaf) for each nematode. The experiment was evaluated for 21 days every 24 hours, making mortality counts and assessment of damage on Kikuyo grass. The bugs showed 100% mortality after 15 days. Each bug produced an average of 13,000 IJ/6 days. As for the damage did not reach level 2 within 21 days prior to treatment without nematodes that damage introduced 3 with necrosis and apical leaf curl. Thus, Steinernema sp. strain JCL024 and Heterorhabditis sp. strain SL0708 exercised control bug on the grass and kept under the level of damage to the foliage.

Revision der paläarktischen Arten der Gattung Lygus Hahn (Heteroptera, Miridae)

Als Ergebnis der Revision der Gattung Lygus Hahn (Heteroptera, Miridae, Insecta) wurden die fünf neuen Arten beschrieben: L. sibiricus Aglyamzyanov, 1990, L. orientis Aglyamzyanov, 1994, L. izyaslavi Aglyamzyanov, 1994, L. monticola Aglyamzyanov, 1994 und L. martensi Aglyamzyanov, 2003. Die vier Speziessnamen wurden synonymisiert: L. dracunculi Josifov, 1992, L. alashanensis Qi, 1993, L. renati Schwartz, 1998 (L. elegans Aglyamzyanov, 1994) = L. poluensis (Wagner, 1967) und L. kerzhneri Qi, 1993 = L. punctatus (Zetterstedt, 1838). Artstatus von L. israelensis Linnavuori, 1962 wurde wiederhergestellt. Nach aktuellen Angaben wurden in der Paläarktis 19 Lygus-Arten festgestellt: L. discrepans Reuter, 1906; L. gemellatus (Herrich-Schaeffer, 1835); L. hsiaoi Zheng & Yu, 1992; L. israelensis Linnavuori, 1962; L. italicus Wagner, 1950; L. izyaslavi Aglyamzyanov, 1994; L. maritimus Wagner, 1949; L. martensi Aglyamzyanov, 2003; L. monticola Aglyamzyanov, 1994; L. orientis Aglyamzyanov, 1994; L. pachycnemis Reuter, 1879; L. paradiscrepans Zheng & Yu, 1992; L. poluensis (Wagner, 1967); L. pratensis (Linnaeus, 1758); L. punctatus (Zetterstedt, 1838); L. rugulipennis Poppius, 1911; L. sibiricus Aglyamzyanov, 1990; L. tibetanus Zheng & Yu, 1992 und L. wagneri Remane, 1955. Es wurden die diagnostischen Merkmale analysiert, eine Bestimmungstabelle erstellt und die Areale der Verbreitung der einigen Arten präzisiert.

Compatibilidad de Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) y Nesidiocoris tenuis (Reuter)(Hemiptera: Miridae), depredadores importantes en cultivos hortícolas protegidos, con nuevas barreras físicas selectivas y modernos plaguicidas

La nueva legislación en materia fitosanitaria se dirige hacia una Gestión Integrada de Plagas (GIP). Estos programas dan preferencia a aquellos métodos más respetuosos y sostenibles con el medio ambiente, siendo piezas claves en ellos el control biológico, el físico y otros de carácter no químico. Sin embargo, el uso de insecticidas selectivos es a veces necesario para el adecuado manejo de plagas en cultivos hortícolas. Por ello, el objetivo general de este estudio es aportar conocimientos para mejorar el control de plagas en cultivos hortícolas, mediante la integración de tres estrategias de lucha: biológica, física y química. Una parte de este trabajo ha consistido en el estudio de los posibles efectos que mallas tratadas con insecticida (bifentrin) pudieran provocar mediante diferentes ensayos de laboratorio, invernadero y campo, en los enemigos naturales Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) (depredador de trips), Nesidiocoris tenuis (Reuter) (Hemiptera: Miridae) (depredador de mosca blanca y Tuta absoluta (Meirick) (Lepidoptera: Gelechiidae)), y otros agentes de biocontrol comúnmente usados en cultivos hortícolas protegidos. Este tipo de mallas se han empleado con éxito en entomología médica para controlar mosquitos vectores de la malaria, y actualmente se está trabajando en su desarrollo para uso agrícola como método de exclusión, y método directo de control de plagas. En los ensayos realizados en laboratorio, O. laevigatus y N. tenuis no fueron capaces de detectar la presencia de bifentrin en el ensayo de preferencia. Además, no se produjo mortalidad a corto plazo (72 horas) en ambos chinches depredadores. Por el contrario, se registró una elevada mortalidad cuando se expusieron por contacto a la malla tratada durante 72 horas en cajas de dimensiones reducidas (10 cm de diámetro X 3 cm de altura). En ensayos llevados a cabo bajo condiciones más reales de exposición, en un invernadero experimental con jaulas de 25 X 25 X 60 cm de altura, no se produjo ningún efecto en la mortalidad a corto plazo (72 horas) o en los parámetros reproductivos de O. laevigatus y N. tenuis. Finalmente, en ensayos de campo realizados en túneles semi-comerciales (8 m de largo X 6,5 m de ancho X 2,6 m de altura), ni las condiciones ambientales [temperatura, humedad relativa, radiación ultravioleta (UV) y fotosintéticamente activa (PAR)], ni los enemigos naturales, se vieron afectados por la presencia de la malla tratada con bifentrin en el cultivo. Sin embargo, los resultados no fueron concluyentes, debido al bajo establecimiento de los agentes de biocontrol liberados. Por lo tanto, más estudios son necesarios en invernaderos comerciales para confirmar los resultados preliminares de compatibilidad. Además, en este trabajo se han evaluado los efectos letales (mortalidad) y subletales (parámetros reproductivos) de seis modernos insecticidas sobre los chinches depredadores O. laevigatus y N. tenuis, mediante ensayos de laboratorio y persistencia. Los ensayos se realizaron por contacto residual, aplicando los insecticidas a la dosis máxima de campo sobre placas de cristal (laboratorio) o plantas (persistencia). Los productos fitosanitarios se seleccionaron por representar a un grupo de modernos plaguicidas con modos de acción en principio más selectivos para los enemigos naturales que antiguos plaguicidas como organoclorados, oroganofosforados o carbamatos, y por su uso frecuente en cultivos hortícolas donde O. laevigatus y N. tenuis están presentes. Todos ellos están incluidos o en proceso de inclusión en la lista comunitaria de sustancias activas para uso agrícola, Anexo I de la Directiva 91/414/CEE: abamectina y emamectina (avermectinas neurotóxicas, activadoras del canal del cloro), deltametrina (piretroide neurotóxico, modulador del canal del sodio, control positivo), flubendiamida (neurotóxico, modulador del receptor de rianodina), spinosad (naturalito neurotóxico, agonistas/antagonistas del receptor de nicotínico acetilcolina) y spiromesifen (inhibidor de la acetil CoA carboxilasa). El estudio mostró que O. laevigatus fue más susceptible a los insecticidas que N. tenuis. Además, los resultados revelaron que flubendiamida y spiromesifen fueron compatibles con los dos enemigos naturales estudiados, y por tanto se podrían usar en programas de GIP. Por el contrario, los insecticidas abamectina, deltametrina, emamectina y spinosad no fueron selectivos para ninguno de los chinches depredadores. Sin embargo, los estudios de persistencia demostraron que a pesar de que estos insecticidas no proporcionaron selectividad fisiológica, pueden proporcionar selectividad ecológica en algunos casos. Abamectina, deltametrina, emamectina y spinosad podrían ser compatibles con N. tenuis si el enemigo natural es introducido en el cultivo 4 días después de su aplicación. En el caso de O. laevigatus, abamectina, deltametrina y spinosad se clasificaron como persistentes, por lo tanto es necesario completar el estudio con experimentos de semi-campo y campo que determinen si es posible su uso conjunto en programas de GIP. Por otro lado, emamectina podría ser compatible con O. laevigatus si el enemigo natural es introducido en el cultivo 7 días después de su aplicación. Por último, se ha comprobado la selectividad de tres insecticidas aceleradores de la muda (MACs) (metoxifenocida, tebufenocida y RH-5849) sobre O. laevigatus y N. tenuis. Además de realizar estudios para evaluar la toxicidad en laboratorio de los insecticidas por contacto residual e ingestión (principal modo de acción de los MAC´s), se extrajo RNA de los insectos y con el cDNA obtenido se secuenció y clonó el dominio de unión al ligando (LBD) del receptor de ecdisona correspondiente a O. laevigatus (OlEcR-LBD) y N. tenuis (NtEcR-LBD). Posteriormente, se obtuvo la configuración en tres dimensiones del LBD y se estudió el acoplamiento de las moléculas de los tres insecticidas en la cavidad que forman las 12 α-hélices que constituyen el EcR-LBD. En el caso de N. tenuis se debe mencionar que no fue posible la obtención de la secuencia completa del LBD. Sin embargo, se obtuvo una secuencia parcial (hélice 6-hélice 11), que mostró una alta conservación de aminoácidos con respecto a la obtenida en O. laevigatus. Los ensayos de toxicidad mostraron que metoxifenocida, tebufenocida y RH-5849 no produjeron ningún efecto nocivo en ambos depredadores. Además, los estudios de modelado por homología y acoplamiento molecular llevados a cabo con O. laevigatus, también indicaron que los MACs no produjeron ningún efecto deletéreo en este enemigo natural. Por lo tanto, estos compuestos pueden ser aplicados de manera segura en programas de GIP en los cuales O. laevigatus y N. tenuis estén presentes. ABSTRACT The new pesticide legislation on pest control is aimed at integrated pest management (IPM). These programs are based on the most environmentally sustainable approaches, where biological, physical control and other non-chemical methods are the cornerstone. However, selective pesticides are often required for pest management on horticultural crops. Therefore, the main goal of this study is to provide knowledge to improve pest control on horticultural crops through the integration of three strategies: biological, physical and chemical. Firstly, the effects of insecticide treated nets (bifenthrin) were evaluated in different laboratory, greenhouse and field experiments on the natural enemies Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) (predator of thrips), Nesidiocoris tenuis (Reuter) (Hemiptera: Miridae) (predator of whiteflies and Tuta absoluta (Meirick) (Lepidoptera: Gelechiidae)), and other biocontrol agents commonly used on protected horticultural crops. These types of nets have been successfully used in medical entomology to control mosquito malaria vectors, and work is currently being done on their use as exclusion barriers and as a direct method of pest control in agriculture. In experiments made under laboratory conditions, O. laevigatus and N. tenuis were not able to detect the presence of bifenthrin in a dual-choice test. Furthermore, no shortterm mortality (72 hours) was recorded on both predatory bugs. In contrast, a high mortality rate was found when they were exposed by contact to the bifenthrin-treated net for 72 hours in small cages (10 cm diameter X 3 cm high). In assays carried out under more realistic conditions of exposure, in an experimental greenhouse with cages of 25 X 25 X 60 cm high, short-term mortality (72 hours) and reproductive parameters were not affected. Lastly, in field experiments carried out in semi-commercial tunnels (8 m long X 6.5 m width X 2.6 m high), neither environmental conditions [temperature, relative humidity, ultraviolet (UV) and photosynthetically active radiation (PAR)] nor natural enemies were affected by the presence of the bifenthrin-treated net on the crop. However, results were not conclusive, mainly due to a low settlement of the released biocontrol agents, and further studies are needed in commercial greenhouses to confirm our preliminary results of compatibility. Secondly, the lethal (mortality) and sublethal effects (reproductive parameters) of six modern pesticides on the predatory bugs O. laevigatus and N. tenuis has been evaluated through laboratory and persistence experiments. Trials were carried out by residual contact, applying the insecticides to the maximum field recommended concentration on glass plates (laboratory) or plants (persistence). Insecticides were chosen as representatives of modern pesticides with a more selective mode of action on natural enemies than organochlorine, organophosphorus and carbamate insecticides. Moreover, they were also chosen because of their frequent use on horticultural crops where O. laevigatus and N. tenuis are present. All of them have been included or have been requested for inclusion in the community list of active substances on the agricultural market, Annex I of the European Directive 91/414/EEC: abamectin and emamectin (neurotoxic avermectins, chloride channel activators), deltamethrin (neutotoxic pyrethroid, sodium channel modulator, positive commercial standard), flubendiamide (neurotoxic, rianodine receptor modulator), spinosad (neurotoxic naturalyte, nicotinic acetylcholine receptor allosteric activator) and spiromesifen (inhibitors of acetyl CoA carboxylase). The study showed that O. laevigatus was more susceptible to all the studied pesticides than N. tenuis. In addition, the research results indicated no impact of flubendiamide and spiromesifen on the two natural enemies studied under laboratory conditions. Consequently, both pesticides are candidates to be included in IPM programmes where these biocontrol agents are present. On the other hand, abamectin, deltamethrin, emamectin and spinosad were not selective for both predatory bugs in laboratory experiments. However, persistence test demonstrated that in spite of the lack of physiological selectivity, these pesticides can provide ecological selectivity in some cases. Abamectin, deltamethrin, emamectin and spinosad could be compatible with N. tenuis if the mirid bug is released 4 days after the insecticide treatment on the crop. With regard to O. laevigatus, abamectin, deltamethrin and spinosad were classified as persistent in our assays, thus the study should be completed with semi-field and field experiments in order to ascertain their possible joint use in IPM programs. In contrast, emamectin could be compatible with O. laevigatus if the pirate bug is released 7 days after the insecticide treatment on the crop. Finally, the selectivity of three moulting accelerating compounds (MACs) (methoxyfenozide, tebufenozide and RH-5849) has also been evaluated on O. laevigatus and N. tenuis. In addition to laboratory experiments to evaluate the toxicity of the insecticides by residual contact and ingestion, molecular approaches were used as well. RNA of both insects was isolated, cDNA was subsequently synthesized and the complete sequence of the ligand binding domain (LBD) of the ecdysone receptor of O. laevigatus (OlEcR-LBD) and N. tenuis (NtEcR-LBD) were determined. Afterwards, the three dimensional structure of LBD was constructed. Finally, the docking of the insecticide molecules in the cavity delineated by the 12 α-helix that composed the EcRLBD was performed. In the case of N. tenuis, it should be noted that in spite of intensive efforts, we did not manage to complete the sequence for the LBD.However, a partial sequence of the LBD was obtained (helix 6-helix 11), and a strong conservation between the amino acids of N. tenuis and O. laevigatus was observed. Results showed no biological activity of methoxyfenozide, tebufenozide and RH-5849, on both predatory bugs. Moreover, modeling of the OlEcR-LBD and docking experiments also suggested that MACs were devoid of any deleterious effect on O. laevigatus. Therefore, our results indicate that these compounds could be safely applied in IPM programs in which O. laevigatus and N. tenuis are present.

Alternativas botánica, biológica y química para el manejo de piojo del tomate (Halticus sp) (Hemíptera: Miridae) y mosca blanca (Bemisia tabaci, Gen) (Hemiptera: Aleyroridae) en el cultivo de tomate (Solanum lycopersicum L.), bajo condiciones de casa malla.

El piojo del tomate (Halticus sp.) y la mosca blanca (Bemisia tabaci) son los principales agentes de daño, causantes de problemas fitosanitarios severos para los productores de tomate (Solanum lycopersicum, L), bajo condiciones de casa malla. Esta plaga provoca importantes pérdidas económicas, disminuyendo así los rendimientos al afectar la calidad de los frutos lo cual incide en mayores costos de producción. En base a esta problemática en el CEVAT (Centro de Experimentación y Validación de Tecnologías), UNA, se realizó un estudio en el período comprendido entre los meses de Septiembre 2015 a Enero 2016, con el objetivo de evaluar insecticidas biológico, botánico y químico contra el piojo del tomate y mosca blanca. Los productos evaluados fueron: Engeo, Madero Negro y Metarhizium anisopliae. Las variables evaluadas fueron: número de Halticus sp por planta, incidencia del daño por Halticus sp por planta, numero de mosca blanca por planta, incidencia del daño de virosis por planta, severidad del daño de virosis por planta. De los tratamientos evaluados, el menor promedio de piojo del tomate por planta lo presentó el tratamiento Engeo y de igual manera en mosca blanca, seguido de madero negro. El menor porcentaje de incidencia y severidad lo presentó el tratamiento Engeo en comparación con los demás tratamientos evaluados. El análisis económico realizado determinó que los tratamientos que presentaron los mejores rendimientos fueron Engeo con 18,156.25 kg/ha, seguido por Madero Negro con 16,891 kg/ha y Metarhizium anisopliae que obtuvo 14,797 kg/ha. Engeo presentó el mayor beneficio neto con 16,642.92 US$/ha, seguido por Madero Negro con 15,240.86 US$/ha. En el análisis de la tasa de retorno marginal resultó que el tratamiento Madero Negro es el que obtuvo la mejor tasa de retorno marginal con 6,307.36 % es decir 63.07 US$ por cada dólar invertido.

Evaluación del efecto insecticida de los extractos vegetales de Sambucus nigra (Caprifoliacea) en el control de Collaria scenica (Hemiptera: Miridae) en condiciones de laboratorio

Bogotá (Colombia): Universidad de La Salle. Facultad de Ciencias Agropecuarias. Programa de Zootecnia

Expression patterns of vascular-specific promoters RolC and Sh in transgenic potatoes and their use in engineering PLRV-resistant plants

The expression patterns of GUS fusion constructs driven by the Agrobacterium rhizogenes RolC and the maize Sh (Shrunken: sucrose synthase-1) promoters were examined in transgenic potatoes (cv. Atlantic). RolC drove high-level gene expression in phloem tissue, bundle sheath cells and vascular parenchyma, but not in xylem or non-vascular tissues. Sh expression was exclusively confined to phloem tissue. Potato leafroll luteovirus (PLRV) replicates only in phloem tissues, and we show that when RolC is used to drive expression of the PLRV coat protein gene, virus-resistant lines can be obtained. In contrast, no significant resistance was observed when the Sh promoter was used.

The M(r) 43K major capsid protein of rice ragged stunt oryzavirus is a post-translationally processed product of a M(r) 67 348 polypeptide encoded by genome segment 8

The nucleotide sequence of DNA complementary to rice ragged stunt oryzavirus (RRSV) genome segment 8 (S8) of an isolate from Thailand was determined. RRSV S8 is 1 914 bp in size and contains a single large open reading frame (ORF) spanning nucleotides 23 to 1 810 which is capable of encoding a protein of M(r) 67 348. The N-terminal amino acid sequence of a ~43K virion polypeptide matched to that inferred for an internal region of the S8 coding sequence. These data suggest that the 43K protein is encoded by S8 and is derived by a proteolytic cleavage. Predicted polypeptide sizes from this possible cleavage of S8 protein are 26K and 42K. Polyclonal antibodies raised against a maltose binding protein (MBP)-S8 fusion polypeptide (expressed in Escherichia coli) recognised four RRSV particle associated polypeptides of M(r) 67K, 46K, 43K and 26K and all except the 26K polypeptide were also highly immunoreactive to polyclonal antibodies raised against purified RRSV particles. Cleavage of the MBP-S8 fusion polypeptide with protease Factor X produced the expected 40K MBP and two polypeptides of apparent M(r) 46K and 26K. Antibodies to purified RRSV particles reacted strongly with the intact fusion protein and the 46K cleavage product but weakly to the 26K product. Furthermore, in vitro transcription and translation of the S8 coding region revealed a post-translational self cleavage of the 67K polypeptide to 46K and 26K products. These data indicate that S8 encodes a structural polypeptide, the majority of which is auto- catalytically cleaved to 26K and 46K proteins. The data also suggest that the 26K protein is the self cleaving protease and that the 46K product is further processed or undergoes stable conformational changes to a ~43K major capsid protein.

Putative full-length clones of the genomic DNA segments of subterranean clover stunt virus and identification of the segment coding for the viral coat protein

Subterranean clover stunt disease is an economically important aphid-borne virus disease affecting certain pasture and grain legumes in Australia. The virus associated with the disease, subterranean clover stunt virus (SCSV), was previously found to be representative of a new type of single-stranded DNA virus. Analysis of the virion DNA and restriction mapping of double-stranded cDNA synthesized from virion DNA suggested that SCSV has a segmented genome composed of 3 or 4 different species of circular ssDNA each of about 850-880 nucleotides. To further investigate the complexity of the SCSV genome, we have isolated the replicative form DNA from infected pea and from it prepared putative full-length clones representing the SCSV genome segments. Analysis of these clones by restriction mapping indicated that clones representing at least 4 distinct genomic segments were obtained. This method is thus suitable for generating an extensive genomic library of novel ssDNA viruses containing multiple genome segments such as SCSV and banana bunchy top virus. The N-terminal amino acid sequence and amino acid composition of the coat protein of SCSV were determined. Comparison of the amino acid sequence with partial DNA sequence data, and the distinctly different restriction maps obtained for the full-length clones suggested that only one of these clones contained the coat protein gene. The results confirmed that SCSV has a functionally divided genome composed of several distinct ssDNA circles each of about 1 kb.

Descripción del comportamiento de insectos y enfermedades asociadas al cultivo de fresa (Fragaria spp, L) en el municipio de la Sabana, departamento de Madriz

La fresa ( Fragaria spp , L), es una planta herbácea, pertenece a la familia de las Rosácea s y al genero Fragariae, esta es hospedera de un sin número de insectos plagas y enfermedades. Actualmente en Nicaragua se cultiva en zonas altas, arriba de los 1200 msnm como una alternativa de diversificación de fincas cafetaleras y a la demanda que existe del producto. Estas alturas se ubican en los departamentos de Madríz, en el municipio de La Sabana, no existiendo hasta la fecha un reporte formal sobre los principales insectos plagas e insectos benéficos y enfermedades presentes en el cultivo. Ante tal situación y debido a la importancia que esta tomando este cultivo, se realizó una investigación con el objetivo de identificar y describir la fluctuación poblacional de todos los insectos e incidencia de enfermedades presentes en el cultivo de fresa. El estudio se realizó en la finca La Patasta, Municipio de la Sabana, Departamento de Madríz, en el periodo comprendido entre Octubre 2004 a Abril 2005. El monitoreo se realizó semanalmente en una parcela la cual constaba de cinco sitios específicos, realizando capturas manuales de especimenes con ayudas de vasos cristalinos y cámaras húmedas en el caso de enfermedades. Los resultados obtenidos fueron los siguientes: se identificó y se describió la fluctuación poblacional de insectos de las principales familias del orden Coleóptera (Chrysomelidae, Scarabaeidae, Curculionidae), así como las principales familias del orden Hemiptera (Pentatomidae, Miridae, Ligaeidae,) también las familias del orden Orthoptera (Gryllidae) y el Orden Homóptera (Cicadellidae), de igual forma se identificò y describió la fluctuación poblacional de depredadores naturales de la familia Coccinellidae y Aracnidae, así como también se identificaron y se describieron las principales enfermedades causadas por (Mycosphaerella fragariae, Xanthomonas fragariae y Botrytis spp) presentes en este cultivo