THE KNICKKOPF DOMON DOMAIN IS ESSENTIAL FOR CUTICLE DIFFERENTIATION IN Drosophila melanogaster

The dopamine monoxygenase N-terminal (DOMON) domain is found in extracellular proteins across several eukaryotic and prokaryotic taxa. It has been proposed that this domain binds to heme or sugar moieties. Here, we have analyzed the role of four highly conserved amino acids in the DOMON domain of the Drosophila melanogaster Knickkopf protein that is inserted into the apical plasma membrane and assists extracellular chitin organization. In principal, we generated Knickkopf versions with exchanged residues tryptophan(299,) methionine(333), arginine(401), or histidine(437), and scored for the ability of the respective engineered protein to normalize the knickkopf mutant phenotype. Our results confirm the absolute necessity of tryptophan(299,) methionine(333), and histidine(437) for Knickkopf function and stability, the latter two being predicted to be critical for heme binding. In contrast, arginine(401) is required for full efficiency of Knickkopf activity. Taken together, our genetic data support the prediction of these residues to mediate the function of Knickkopf during cuticle differentiation in insects. Hence, the DOMON domain is apparently an essential factor contributing to the construction of polysaccharide-based extracellular matrices.

Keystone food resources for honey bees in South Indian west coast during monsoon

The low level, denuded, laterite landscape of coastal Uttara Kannada has a rich diversity of monsoon herbs, including threatened and newly discovered ones. Our study reveals that honey bees congregate on the ephemeral herb community of Utricularias, Eriocaulons and Impatiens during their gregarious monsoon flowering period. Apis dorsata had highest visitations on Utricularias, Impatiens and Flacourtia indica, whereas Trigona preferred Eriocaulons. Laterite herb flora merits conservation efforts as a keystone food resource for the insect community, especially for honey bees.

Biomechanics of substrate boring by fig wasps

Female insects of diverse orders bore into substrates to deposit their eggs. Such insects must overcome several biomechanical challenges to successfully oviposit, which include the selection of suitable substrates through which the ovipositor can penetrate without itself fracturing. In many cases, the insect may also need to steer and manipulate the ovipositor within the substrate to deliver eggs at desired locations before rapidly retracting her ovipositor to avoid predation. In the case of female parasitoid ichneumonid wasps, this process is repeated multiple times during her lifetime, thus testing the ability of the ovipositioning apparatus to endure fracture and fatigue. What specific adaptations does the ovipositioning apparatus of a female ichneumonoid wasp possess to withstand these challenges? We addressed this question using a model system composed of parasitoid and pollinator fig wasps. First, we show that parasitoid ovipositor tips have teeth-like structures, preferentially enriched with zinc, unlike the smooth morphology of pollinator ovipositors. We describe sensillae present on the parasitoid ovipositor tip that are likely to aid in the detection of chemical species and mechanical deformations and sample microenvironments within the substrate. Second, using atomic force microscopy, we show that parasitoid tip regions have a higher modulus compared with regions proximal to the abdomen in parasitoid and pollinator ovipositors. Finally, we use videography to film wasps during substrate boring and analyse buckling of the ovipositor to estimate the forces required for substrate boring. Together, these results allow us to describe the biomechanical principles underlying substrate boring in parasitoid ichneumonid wasps. Such studies may be useful for the biomimetic design of surgical tools and in the use of novel mechanisms to bore through hard substrates.

Parasites exert conflicting selection pressures to affect reproductive asynchrony of their host plant in an obligate pollination mutualism

1. Plant reproductive phenology is generally viewed as an individual's strategy to maximize gamete exchange and propagule dispersal and is often considered largely dependent on patterns of floral initiation. Reproductive phenology, however, can be affected by proximate responses to pollinators, parasites and herbivores which could influence floral longevity or fruit development time. 2. We examined the influence of insect interactants on within-plant reproductive phenology in the fig-fig wasp nursery pollination mutualism in Ficus racemosa (Moraceae). Most figs support a wasp community comprised of a mutualistic pollinator, with several host-plant-specific non-pollinating herbivorous gallers and parasitoids. These wasps reproduce within enclosed inflorescences called syconia, which develop into fruit after pollination. While different wasp species oviposit into syconia at varying times during its ontogeny, all wasp progeny are constrained to exit syconia simultaneously just prior to fruit ripening. Developing larvae of early-ovipositing wasps may hasten syconium ontogeny through formation of earlier and larger nutrient sinks, whereas larvae of late-arriving parasites may lengthen syconium ontogeny to complete their development successfully. Seeds are also important nutrient sinks. The number of seeds and the type and number of developing wasps may therefore be expected to influence syconium development times, thereby affecting the reproductive synchrony of syconia on a plant. 3. Observations on naturally pollinated and parasitized syconia indicated that their seed and wasp content affected syconium development time. Experimental manipulations of syconia to produce only seeds or various combinations of wasps confirmed this finding. Early-ovipositing galler progeny reduced syconium development times, while gallers ovipositing concurrently with pollinators had no effect on syconium development. Late-ovipositing parasitoid progeny, the presence of only seeds within the syconium, or delayed pollination increased syconium development time. The differential development of syconia, which was influenced by mutualistic or parasitic progeny, accordingly contributed to within-tree reproductive asynchrony. 4. Synthesis. Individual reproductive units in fig trees called syconia, which also function as brood sites for pollinating and parasitic fig wasps, have plastic development durations dependent on pollination timing and species of wasps developing within them. Syconium development times are a likely compromise between conflicting demands from developing seeds and different wasp species.

TIME VARYING LINEAR PREDICTION USING SPARSITY CONSTRAINTS

Time-varying linear prediction has been studied in the context of speech signals, in which the auto-regressive (AR) coefficients of the system function are modeled as a linear combination of a set of known bases. Traditionally, least squares minimization is used for the estimation of model parameters of the system. Motivated by the sparse nature of the excitation signal for voiced sounds, we explore the time-varying linear prediction modeling of speech signals using sparsity constraints. Parameter estimation is posed as a 0-norm minimization problem. The re-weighted 1-norm minimization technique is used to estimate the model parameters. We show that for sparsely excited time-varying systems, the formulation models the underlying system function better than the least squares error minimization approach. Evaluation with synthetic and real speech examples show that the estimated model parameters track the formant trajectories closer than the least squares approach.

A Kunitz-type serine protease inhibitor from Butea monosperma seed and its influence on developmental physiology of Helicoverpa armigera

A protease inhibitor from the seeds of Butea monosperma (BmPI) was purified, characterized and studied for its influence on developmental physiology of Helicover-pa armigera. BmPI on two-dimensional separations indicated the presence of a 14 kDa protein with an isoelectric point in the acidic region (pl 5.6). Multiple Sequence Analysis data suggested that the BmPI contains a sequence motif which is conserved in various trypsin and chymotrypsin inhibitors of Kunitz-type. The inhibitor exhibited trypsin inhibitory activity in a broad range of pH (4-10) and temperature (10-80 degrees C). The enzyme kinetic studies revealed BmPI as a competitive inhibitor with a K-i value of 1.2 x 10(-9) M. In vitro studies with BmPI indicated measurable inhibitory activity on total gut proteolytic enzymes of H. armigera (IC(50)2.0 mu g/ml) and bovine trypsin. BmPI supplemented artificial diet caused dose dependent mortality and reduction in growth and weight. The fertility and fecundity of H. armigera, declined whereas the larval-pupal duration of the insect life cycle extended. These detrimental effects on H. armigera suggest the usefulness of BmPl in insect pest management of food crops. (C) 2014 Elsevier Ltd. All rights reserved.

A cis-Regulatory Mutation in Troponin-I of Drosophila Reveals the Importance of Proper Stoichiometry of Structural Proteins During Muscle Assembly

Rapid and high wing-beat frequencies achieved during insect flight are powered by the indirect flight muscles, the largest group of muscles present in the thorax. Any anomaly during the assembly and/or structural impairment of the indirect flight muscles gives rise to a flightless phenotype. Multiple mutagenesis screens in Drosophila melanogaster for defective flight behavior have led to the isolation and characterization of mutations that have been instrumental in the identification of many proteins and residues that are important for muscle assembly, function, and disease. In this article, we present a molecular-genetic characterization of a flightless mutation, flightless-H (fliH), originally designated as heldup-a (hdp-a). We show that fliH is a cis-regulatory mutation of the wings up A (wupA) gene, which codes for the troponin-I protein, one of the troponin complex proteins, involved in regulation of muscle contraction. The mutation leads to reduced levels of troponin-I transcript and protein. In addition to this, there is also coordinated reduction in transcript and protein levels of other structural protein isoforms that are part of the troponin complex. The altered transcript and protein stoichiometry ultimately culminates in unregulated acto-myosin interactions and a hypercontraction muscle phenotype. Our results shed new insights into the importance of maintaining the stoichiometry of structural proteins during muscle assembly for proper function with implications for the identification of mutations and disease phenotypes in other species, including humans.

Effect of mass loading on ionic polymer metal composite actuators and sensors

Ionic polymer metal composites (IPMC) actuator for flapping insect scale wing is advantageous due to its low mass, high deflection and simple actuation mechanism. Some of the factors that affect the actuation of IPMC are the amount of hydration in the polymer membrane and the environmental conditions such as temperature, humidity etc. In structural design, the attachment of wing on the IPMC actuators is an important concern as the attached wing increases the mass of actuators thereby affecting the parameters like displacement, stiffness and resonant frequencies. Such IPMC actuators have to produce sufficient actuation force and frequency to lift and flap the attached wing. Therefore, it is relevant to study the influence of attachment of wing on the actuator parameters (displacement, resonant frequency, block force and stiffness) and performance of the actuators. This paper is divided into two parts; the first part deals with the modeling of the IPMC actuators for its effect on the level of water uptake and temperature using energy based method. The modeling method adapted is validated with the experimental procedure used to actuate the IPMC. The second part deals with the experimental analysis of IPMC actuation at dry, wet and in water conditions. The effect of end mass loading on the performance of 20 Hz, high frequency actuator (HFA) and 8.7 Hz, low frequency IPMC actuators (LFA) and sensors is studied. The IPMC actuators are attached with IPMC flapping wing at its free end and performance analysis on the attached wing is also carried out.

Dynamic response of a piezoelectric flapping wing

Piezo-composite membranes have advantages over motorized flapping where frequencies are high and certain coupling between bending and twisting is useful to generate lift and forward flight. We draw examples of fruit fly and bumble bee. Wings with Piezo ceramic PZT coating are realized. The passive mechanical response of the wing is characterized experimentally and validated using finite element simulation. Piezoelectric actuation with uniform electrode coating is characterized and optimal frequencies for flapping are identified. The experimental data are used in an empirical model and advanced ratio for a flapping insect like condition for various angular orientations is estimated.

A carboxy terminal domain of the L protein of rinderpest virus possesses RNA triphosphatase activity - The first enzyme in the viral mRNA capping pathway

The large protein L of negative-sense RNA viruses is a multifunctional protein involved in transcription and replication of genomic RNA. It also possesses enzymatic activities involved in capping and methylation of viral mRNAs. The pathway for mRNA capping followed by the L protein of the viruses in the Morbillivirus genus has not been established, although it has been speculated that these viruses may follow the unconventional capping pathway as has been shown for some viruses of Rhabdoviridae family. We had earlier shown that the large protein L of Rinderpest virus expressed as recombinant L-P complex in insect cells as well as the ribonucleoprotein complex from purified virus possesses RNA triphosphatase (RTPase) and guanylyltransferase activities, in addition to RNA dependent RNA polymerase activity. In the present work, we demonstrate that RTPase as well as nucleoside triphosphatase (NTPase) activities are exhibited by a subdomain of the L protein in the C terminal region (a.a. 1640 1840). The RTPase activity depends absolutely on a divalent cation, either magnesium or manganese. Both the RTPase and NTPase activities of the protein show dual metal specificity. Two mutant proteins having alanine mutations in the glutamic acid residues in motif-A of the RTPase domain did not show RTPase activity, while exhibiting reduced NTPase activity suggesting overlapping active sites for the two enzymatic functions. The RTPase and NTPase activities of the L subdomain resemble those of the Vaccinia capping enzyme D1 and the baculovirus LEF4 proteins. (C) 2015 Elsevier Inc. All rights reserved.

Evidence for N-7 guanine methyl transferase activity encoded within the modular domain of RNA-dependent RNA polymerase L of a Morbillivirus

Post-transcriptional modification of viral mRNA is essential for the translation of viral proteins by cellular translation machinery. Due to the cytoplasmic replication of Paramyxoviruses, the viral-encoded RNA-dependent RNA polymerase (RdRP) is thought to possess all activities required for mRNA capping and methylation. In the present work, using partially purified recombinant RNA polymerase complex of rinderpest virus expressed in insect cells, we demonstrate the in vitro methylation of capped mRNA. Further, we show that a recombinant C-terminal fragment (1717-2183 aa) of L protein is capable of methylating capped mRNA, suggesting that the various post-transcriptional activities of the L protein are located in independently folding domains.

How to be a fig wasp parasite on the fig-fig wasp mutualism

The nursery pollination mutualism between figs and pollinating fig wasps is based on adaptations that allow wasps to enter the enclosed inflorescences of figs, to facilitate seed set, and to have offspring that develop within the nursery and that leave to enter other inflorescences for pollination. This closed mutualistic system is not immune to parasitic fig wasps. Although the life histories and basic biology of the mutualists have been investigated, the biology of the fig wasp parasites has been severely neglected. This review brings together current knowledge of the many different ways in which parasites can enter the system, and also points to the serious lacunae in our understanding of the intricate interactions between gallers, kleptoparasites, seed eaters and parasitoids within this mutualism.

Perceptual thresholds for detecting modifications applied to the acoustical properties of a violin

This study is the first step in the psychoacoustic exploration of perceptual differences between the sounds of different violins. A method was used which enabled the same performance to be replayed on different "virtual violins," so that the relationships between acoustical characteristics of violins and perceived qualities could be explored. Recordings of real performances were made using a bridge-mounted force transducer, giving an accurate representation of the signal from the violin string. These were then played through filters corresponding to the admittance curves of different violins. Initially, limits of listener performance in detecting changes in acoustical characteristics were characterized. These consisted of shifts in frequency or increases in amplitude of single modes or frequency bands that have been proposed previously to be significant in the perception of violin sound quality. Thresholds were significantly lower for musically trained than for nontrained subjects but were not significantly affected by the violin used as a baseline. Thresholds for the musicians typically ranged from 3 to 6 dB for amplitude changes and 1.5%-20% for frequency changes. interpretation of the results using excitation patterns showed that thresholds for the best subjects were quite well predicted by a multichannel model based on optimal processing. (c) 2007 Acoustical Society of America.

Evaluación preliminar de diecinueve lineas de sorgo (Sorghum bicolor (L) Moench) por su reacción a las principales plagas: Cogollero (Spodoptera frugiperda J.E. Smith), mosquita del sorgo (Stenodiplosis sorghicola coquillet) y enfermedades, CNIA-INTA, 2001

La presente investigación se llevó a efecto en el Km 14 de la carretera Norte, en el Centro Nacional de Investigación Agropecuaria (CNIA – INTA), 2 Km. al sur en el municipio de Managua, departamento de Managua. El ensayo se estableció en la época de postrera del 2001, en suelos de origen volcánico y textura franco arenosa. El propósito del experimento fue evaluar de forma preliminar 19 líneas de sorgo Sorghum bicolor (L) Moench por su reacción a la mosquita del sorgo ( Stenodiplosis sorghicola Coquillet), el cogollero ( Spodoptera frugiperda J:E:Smith) y enfermedades. Los materiales utilizados son parte de un ADIN (All Diseases and Insect Nursery). La parcela experimental tuvo un tamaño de 3.75 m 2 y constó de 1 surco de 5 metros lineales (1 línea por surco). El ensayo se estableció utilizando la metodología de la Universidad de Texas A & M con19 tratamientos (líneas) y 2 repeticiones. Las variables evaluadas fueron: a) daño foliar causado por el gusano cogollero ( Spodoptera frugiperda J. E. Smith ), b) días a floración, c) número de mosquitas d el sorgo ( Stenodiplosis sorghicola Coquillett ) por panoja, d) número de parasitoides de mosquita del sorgo ( Aprostocetus diplosidis Crawford) por panoja, e) severidad de las enfermedades foliares y vasculares. Para la primera y la última variable se utilizó la escala de daño propuesta por la Universidad de Texas A & M. A estos datos se les efectuó un análisis estadístico descriptivo. Para las variables b, c y d se realizó un análisis de varianza y prueba de rangos múltiples de Duncan (P ≤ 0.05), además se les efectuó un análisis de correlación de Pearson. En cuanto al daño foliar causado por el gusano cogollero ( Spodoptera frugiperda J. E. Smith ), no existieron diferencias en el comportamiento de las líneas. Este aspecto no representó una incidencia relevante. Con respecto al período de floración, la línea 96CA5986 presentó el mayor número de días a floración, en tanto que la línea Tx2880 presentó el menor número de días a floración. Los resultados obtenidos indican que el comportamiento de las 19 líneas de sorgo [ Sorghum bicolor ( L.) Moench ] ante el ataque de mosquita del sorgo ( Stenodiplosis sorghicola Coquillett ) y la incidencia de su parasitoide ( Aprostocetus diplosidis Crawford ) es similar para cada línea; La línea 98BRON122 fue la que presentó el mayor valor con respecto a esta variable, mientras que la línea que presentó el menor número de mosquitas por panoja fue B9104; Existe relación entre la variable número de mosquitas ( Stenodiplosis sorghicola Coquillett ) por panoja y días a floración y esta relación es afectada por el parasitoide de la mosquita ( Aprostocetus diplosidis Crawford ). Puede identificarse que la Linea 87EO366*9EO328 fue la que presentó la mayor severidad de enfermedades foliares tales como antracnosis ( Colletotrichum graminicola G. W. Wilson ), mancha zonada ( Gloeocercospora sorghi D.Bain.& Edgerton ex Deighton) , mancha gris de la hoja (Cercospora sorghi Ellis y Everth) tizón de la hoja ( Exserohilum turcicum (pass.). La línea que presentó la menor severidad fue 94B1055. La enfermedad que presentó mayor severidad en el ensayo fue Antracnosis.

Evaluación de veinte lineas de sorgo (Sorghum bicolor (L) Moench) por su reacción a enfermedades en tres localidades de Nicaragua, postrera 2003

Para contribuir a resolver la problemática varietal en el país, se han sumado esfuerzos en evaluar líneas de sorgo por lo que se realizó un experimento en diferentes zonas productoras de sorgo en Nicaragua utilizando un vivero de líneas para evaluar plagas y enfermedades ADIN (All Diseases and Insect Nursery), facilitado por el Dr. Larry E. Claflin, fitopatológo de la Universidad del Estado Kansas (KSU), mediante el programa INTSORMIL (International Sorghum and Millet Project. El estudio se desarrolló de agosto a diciembre del 2003. Se utilizó, parcelas experimentales de 5 m. lineales con un área total de 75 m2 con dos repeticiones en cada localidad; Centro Nacional de Investigaciones Agropecuarias-Instituto Nicaragüense de Tecnología Agropecuaria, (CNIA–INTA), Managua; Tisma, Masaya y Ranchería, Chinandega. Entre las variables estudiadas se evaluó severidad de enfermedades foliares, severidad de mohos en panoja y rendimiento de grano. Para la evaluación de las enfermedades se utilizó una escala de daño que incluye valores de 0 %-100 % de severidad propuesta por (Frederiksen, 2000); modificada por (Pichardo, 2001). Los datos colectados fueron procesados y sometidos aun ANDEVA, y separación de medias utilizando la prueba de Tukey (α=0.05). Se realizó análisis de regresi ón lineal para determinar el rendimiento predictivo por localidad y líneas. Los resultados obtenidos mostraron que existen diferencias estadísticas entre las líneas para las tres localidades. Las líneas evaluadas en Ranchería presentaron los menores niveles o valores de severidad de en fermedades (7-21 %); valores intermedios (15-39%) las líneas evaluadas en Tisma y los más altos valores (39-58 %) las líneas evaluadas en el CNIA–INTA. Las enfermedades presentes en el estudio fueron: mancha gris de la hoja causada por Cercospora sorghi, Ellis & Everth, antracnosis causada por Colletotrichum graminicola G. W. Wilson y mancha zonada de la hoja causada por Gloeocercospora sorghi D. Bain & Edgerton ex Deighton. Los mayores rendimientos fueron obtenidos por la línea Sureño con 9 037.31, 5 296.57 y 3 566.18 kg ha-1 en CNIA-INTA, Tisma y Ranchería respectivamente; y los menores rendimientos por BLD6(w+y) en CNIA-INTA, con 1 184 kg ha-1, B.9955 en Tisma, con 870.53 kg ha-1 y en Ranchería la línea 98BRON122 con 1 352.55 kg ha -1.