Mechanisms of premating isolation between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) (Lepidoptera: Noctuidae)

Helicoverpa armigera and Helicoverpa assulta are sympatric sibling species, and in the laboratory they can interbreed and produce viable offspring. To assess the contributions of temporal barriers and sexual barriers to premating isolation, we investigated both the temporal rhythms of calling behavior and pheromone titers of H. armigera and H. assulta females and the behavioral responses of males to conspecific and heterospecific calling females in a wind tunnel. Both H. armigera and H. assulta females called throughout the scotophase, and there was more calling during the second half of the scotophase than during the first half. Maximal pheromone titer and maximal calling activity in H. armigera synchronously occurred at the sixth hour into the scotophase, whereas, in H. assulta, the maximal pheromone titer occurred 2 h before the peak of calling. Pheromone blend ratios of the two species were opposite and, within each species, changes in the ratio within the scotophase and at different ages were relatively small. Males of both H. armigera and H. assulta responded strongly to their conspecific calling females in the wind tunnel and completed the whole courtship sequence. In contrast, they did not land and had no copulation attempts in response to heterospecific calling females. These results show that the two species do not have obvious temporal differences in calling behavior and pheromone production, and the specificity of sex pheromone blend emitted by females plays a key role in their premating isolation. In addition, we summarized the potential isolation mechanisms of H. armigera and H. assulta.     

Tarsal taste neurons of Helicoverpa assulta (Guenée) respond to sugars and amino acids, suggesting a role in feeding and oviposition

Helicoverpa assulta and Helicoverpa armigera are sibling species with different host-plant ranges. We have previously reported electrophysiological and behavioral responses of H.armigera to sugars and amino acids. Here we describe a parallel study performed on H. assulta and compare the results obtained with the two species. In females, fourteen gustatory chemosensilla, identified on one ventrolateral side of the fifth tarsomere were stimulated with sucrose, glucose, fructose, maltose, myo-inositol, and the twenty common amino acids, using the tip-recording technique. The taste receptor neurons in eight chemosensilla were identified sensitive to the sugars, myo-inositol, Lys, Glu, Arg, Trp, and Ser which all induced proboscis extension reflex (PER) when tarsi were stimulated. There was a positive correlation between electrophysiological activities and PER responses triggered by sucrose. No stimulatory effect on oviposition was observed with sugar or amino acid mixtures. In males, three chemosensilla showed responses to the four sugars, but generally weaker than in females. The major difference of the two species was the variety of amino acids triggering electrophysiological responses. The stimulatory effect of sugars and amino acids on H.assulta was also generally weaker than that on H. armigera.     

Molecular characterization of two acetylcholinesterase genes from the oriental tobacco budworm, Helicoverpa assulta (Guenée)

Acetylcholinesterase (AChE) has been known to be the target of organophosphorous and carbamate insecticides. Only a single AChE, however, existed in insects and was involved in insecticide resistance, recently another AChE is reported in mosquitoes and aphids. We have cloned cDNAs encoding two ace genes, designated as Ha-ace1 and Ha-ace2 by a combined degenerate PCR and RACE strategy from adult heads of the oriental tobacco budworm, Helicoverpa assulta. The Ha-ace1 and Ha-ace2 genes encode 664 and 647 amino acids, respectively and have conserved motifs including a catalytic triad, a choline-binding site and an acyl pocket. Both Ha-AChEs were determined to be secretory proteins based on the existence of a signal peptide. The Ha-ace1 gene, the first reported ace1 in lepidopterans, belongs to the ace1 subfamily whereas the Ha-ace2 gene showed high similarity to those in the ace2 subfamily. Phylogenetic analysis showed that the Ha-ace1 gene was completely diverged from the Ha-ace2, suggesting that the Ha-ace genes are duplicated. Quantitative real time-PCR revealed that expression level of the Ha-ace1 gene was much higher than that of the Ha-ace2 in all body parts examined. The biochemical properties of purified proteins by affinity chromatography showed substrate specificity for acetylthiocholine iodide, and inhibitor specificity for BW284C51 and eserine and their peptide sequences partially identified by a MALDI-TOF mass spectrometer demonstrated that two Ha-AChEs were expressed in vivo.     

烟青虫(Helicoverpa assulta Guenée)信息素的结构鉴定及田间试验(英语)

北京地区的烟青虫(Helicover Pa asslta Cucn(?)e)的性信息素腺体提取物经毛细管柱气相色谱分析及GC MS分析,鉴定了6种组分.这6种组分为:十六醛(16:Ald)、顺 9—十六烯醛(Z9—16:Ald)、顺11—十六烯醛(Zll 16:Ald)、顺9—十六烯醇(Z9-16:OH)、顺11—十六烯醇(Zll-16:OH)、顺9—十六烯基乙酸酯(Z9 16:OAc),比例为10.9:58.7:3.9:14.7:1.1:10.7.田间试验表明,只有16:Ald、Z9 16:Ald和Zll 16:Ald(比例为15.3:79.2:5.5)组成的三组分诱芯和Z9—16:Ald和Zll—16:Ald(比例为93.4:6.6)组成的两组分诱芯对于雄蛾有强烈的引诱活性.在3种醛为组分的诱芯中加入Z9 16:OH明显地降低引诱活性.     

Genetic basis of sex pheromone blend difference between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) (Lepidoptera: Noctuidae)

The two closely related moth species, Helicoverpa armigera and H. assulta, are sympatric in China. Both species use a mixture of (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-9-hexadecenal (Z9-16:Ald) as their sex pheromones but in widely different ratios. Hybridization and backcrossing experiments between H. armigera and H. assulta were conducted and sex pheromone compositions of the parent species, their F(1) hybrids and backcrosses were compared to study the genetic basis of the production of their sex pheromone blend composition. Results show that the difference in sex pheromone blend ratios of these Helicoverpa species is mainly controlled by an autosomal locus with two alleles, with the allele from H. armigera being almost completely dominant over that derived from H. assulta.     

Types of Antennal Sensilla of the Asian Corn Borer,Ostrinia furnacalis (Guenéee)

This study was carried out to investigate the types of sensilla and their distribution on the antennae of the Asian corn borer, Ostrinia furnacalis (Guenéee). O. furnacalis had six types of antennal sensilla: sensilla trichodea, sensilla chaetica, sensilla coeloconica, sensilla auricillica, sensilla styloconica and Böohms bristle. S. auricillica, single-walled multiporous sensilla, were presumed to be involved in chemical reception. S. coeloconica were double-walled, multiporous sensilla and could detect chemicals also. S. chaetica appeared to be mechano-receptors. S. styloconica were poreless and were thought to have thermo- and hygro-receptors. S. trichodea were single-walled chemosensilla and two subtypes were identified by transmission electron microscopy. The scape and the pedicel had Böohms bristles, which seem to function as mechanical stimulus receptors.     

Seasonal abundance of two armyworm species, Spodoptera exitgua (Hübner) and Spodoptera litura (F.) in the Philippines

The two armyworm species Spodoptera exigua (Hübner) and S. litura (F.) are Important pests on several agricultural crops in the tropics. Knowledge about species abundance is important for the development of integrated pest management strategies. During 1998 and 1999 population densities were recorded weekly using pheromone traps at San Leonardo and Munoz (Province Nueva Ecija, 150 km north of Metro Manila) on the Philippine island Luzon. Furthermore, meteorological conditions and natural enemies were observed. Both armyworm species were present year-round at both observation sites. No differences in numbers of male insects trapped in pheromone traps between seasons were found for S. litura. Contrary, numbers of male S. exigua trapped during rainy season was significant higher than during dry season. Rainfall explained between 30 and 40% of trapped moths and has to be considered when monitoring population density with pheromone traps. Parasitism rates were below 5% in both years and for both species and can not explain the high variation observed in population densities.     

The effect of amino acids on the C₁₄-sphingosine-triggered germination of Nomuraea rileyi, and surface amino acids on Spodoptera litura fabricius

D-erythro-C??-Sphingosine (C??-Sph) accelerated the germination of Nomuraea rileyi in a solution containing peptone, but activity declined to a large degree in water. This suggests the presence of a co-factor in C??-Sph-triggered germination. Since the main role of peptone is to supply nitrogen constituents, we examined the effects of various nitrogen constituents. It was found that Ala and His were highly effective for C??-Sph-triggered germination.     

Bacillus thuringiensisδ-Endotoxin Proteins Show a Correlation in Toxicity and Short Circuit Current Inhibition Against Helicoverpa zea

Pesticidal activity of Bacillus thuringiensisδ-endotoxins, Cry1Aa, Cry1Ab, Cry1Ac, and Cry2A, was determined by using the force-feeding bioassay method to 4^th instar larvae of Helicoverpa zea. H. zea was susceptible to Bt toxins in the order Cry1Ac > Cry1Ab > Cry1Aa > Cry2A with 63.60, 89.04, 159.65, and 375.78 ng/larvae respectively. The abilities of selected Bacillus thuringiensis toxins to inhibit short circuit current (I_SC) in midgut epithelia of H. zea were also investigated by voltage clamp assay. The voltage-clamp studies were conducted on isolated midguts, measuring the inhibition of short circuit current (I_SC) by activated toxin. A Cry1Aa toxin dilution of 33.3 and 500 ng/ml resulted in inhibition of I_SC of −2.29 μA/min (lag time 15 min) and −4.48 μA/min (lag time, 2 min) respectively. The Cry1Ab dilution of 25 ng/ml inhibited I_SC to −1.39 μA/min, a lag time of 14 min, and 333.3 ng/ml dilution resulted in decay of I_SC−2.49 μA/min, lag time 1 min respectively. The Cry1Ac lower dilution 16.7 ng/ml inhibited I_SC to −1.39 μA/min, lag time 4 min, and a high dilution 333.3 ng/ml decay I_SC to −2.44 μA/min, lag time 1 min. The inhibition of I_SC (−1.10 μA/min, lag time 25) at lower dilution (33.3 ng/ml) and high dilution (500 ng/ml), decay (−2.38 μA/min, lag time 5 min), showed a correlation between toxin concentration and inhibitory response with Cry2A toxin. The lag time decreased with increasing concentration of toxin applied, which is additional evidence of dose response besides direct correlation of toxicity assays and I_SC. Received: 7 April 2000 / Accepted: 2 May 2000     

Temperature-dependent development of Cnaphalocrocis medinalis Guenée (Lepidoptera: Pyralidae) and their validation in semi-field condition

The developmental time and survival of the immature stages of Cnaphalocrocis medinalis Guenée were studied at nine constant temperatures (15, 17.5, 20, 22.5, 25, 27.5, 30, 32.5, and 35 °C), 40 ± 10% relative humidity, and a 16:8 h light:dark cycle. The total developmental time decreased with increasing temperature between 15 (115.6 days) and 32.5 °C (20.9 days), but increased above 32.5 °C. The relationship between the developmental rate and temperature was fitted by a linear model and three nonlinear developmental rate models (Logan 6, Briere 1, and Shi et al.). The nonlinear shape of temperature-dependent development was best described by the Briere 1 model (r² = 0.99), and this was supported by statistical information criteria. The total mortality of immature C. medinalis was lowest at 25 °C (67.2%) and highest at 35 °C (98.1%). The distribution of the developmental times of each stage was described by the two-parameter Weibull distribution equation (r² = 0.84–0.96). The predicted date for the cumulative 50% moth emergence was within a variation of one day using the Briere 1 model. The temperature-dependent developmental models for C. medinalis could be applied to determine an optimal management strategy for C. medinalis in paddy fields, and will be helpful in developing a full-cycle phenology model for C. medinalis.     

Phylogenetic Molecular Species Delimitations Unravel Potential New Species in the Pest Genus Spodoptera Guenée, 1852 (Lepidoptera,Noctuidae)

Nowadays molecular species delimitation methods promote the identification of species boundaries within complex taxonomic groups by adopting innovative species concepts and theories (e.g. branching patterns, coalescence). As some of them can efficiently deal with large single-locus datasets, they could speed up the process of species discovery compared to more time consuming molecular methods, and benefit from the existence of large public datasets; these methods can also particularly favour scientific research and actions dealing with threatened or economically important taxa. In this study we aim to investigate and clarify the status of economically important moths species belonging to the genus Spodoptera (Lepidoptera, Noctuidae), a complex group in which previous phylogenetic analyses and integrative approaches already suggested the possible occurrence of cryptic species and taxonomic ambiguities. In this work, the effectiveness of innovative (and faster) species delimitation approaches to infer putative species boundaries has been successfully tested in Spodoptera, by processing the most comprehensive dataset (in terms of number of species and specimens) ever achieved; results are congruent and reliable, irrespective of the set of parameters and phylogenetic models applied. Our analyses confirm the existence of three potential new species clusters (for S. exigua (Hübner, 1808), S. frugiperda (J.E. Smith, 1797) and S. mauritia (Boisduval, 1833)) and support the synonymy of S. marima (Schaus, 1904) with S. ornithogalli (Guenée, 1852). They also highlight the ambiguity of the status of S. cosmiodes (Walker, 1858) and S. descoinsi Lalanne-Cassou & Silvain, 1994. This case study highlights the interest of molecular species delimitation methods as valuable tools for species discovery and to emphasize taxonomic ambiguities.     

Tissue Distribution and Purification of Prophenoloxidase in Larvae of Asian Corn Borer, Ostrinia furnacalis Guenée (Lepidoptera: Pyralidae)

Phenoloxidase (PO) plays an important role in the de-fense response to the foreign invaders, e.g. pathogens andparasites, in the formation of melanin as well as in cuticlesclerotization in insects [1]. PO is synthesized as azymogen, prophenoloxidase (PPO)…     

γ-Aminobutyric Acid Metabolism in Detached Leaves of Wheat Seedling and Leaves and Cotyledons of Sunflower

With a prerequisite of 5 hours in the light the detached leaves of wheat seedling and detached leaves and cotyledons of sunflower were able to utilize exogenous γ-aminobutyric acid, the amount of alanine and glutamine formed were increased. It was found that deamination of γ-aminobutyric acid was associated with γ-aminobutyric-pyruvate transaminase in the leaves of wheat seedling. The specific activity of this transaminase was enhanced about 5.6 fold by adsorption with calcium phosphate gel-Some properties of this transaminas were as follows: the optimum pH was 8.9. The produced amount of alanine was linear with time up to a period of one hour. The primary rate of reaction was proportional to enzyme concentration in the first hour. The relation of substrate concentration to the primary rate of alanine formation was determinated and Michaelis constant was evaluated about 2.6 × 10-3 M. The action of this transaminase in metabolism of γ-aminobutyrie acid was discussed.     

Binding specificity of OBPs in the yellow peach moth Conogethes punctiferalis (Guenée)

Odorant-binding proteins (OBPs) are translators of the external chemical signals, which are critical for maintaining insect life. However, few OBPs were reported in the yellow peach moth (YPM), Conogethes punctiferalis (Guenée). In the current study, five OBPs (CpunOBP1, CpunOBP2, CpunOBP7, CpunPBP2 and CpunPBP4) were expressed and purified from the antennae of the YPM. The results showed that the proteins encoded by five CpunOBPs had six conserved cysteine residues, which were typical structural features of classic OBPs. Moreover, the fluorescence competitive binding assays indicated that the binding affinity of five CpunOBPs to the selected YPM female sex pheromones, host plant volatiles and Penicillium-inoculated apple volatiles was obviously different. The binding affinities of CpunOBP1 and CpunOBP2 with β-ionone were the strongest. CpunOBP7 could bind with 12 host plant volatiles but was unable to interact with any one of the three tested female sex pheromones. CpunPBP2 and CpunPBP4 exhibited the highest binding affinity to female sex pheromone trans-10-hexadecenal among 30 tested compounds. In conclusion, these results suggest the functional differentiation of the CpunOBPs in recognizing sex pheromones, host plant volatiles and fungus-infected host plant volatiles, which will provide new insights into selecting target proteins for YPM biocontrol.     

Metabolism of l-Threonic Acid in Rumex x acutus L. and Pelargonium crispum (L.) L'Hér

l-Threonic acid is a natural constituent in leaves of Pelargonium crispum (L.) L'Hér (lemon geranium) and Rumex x acutus L. (sorrel). In both species, l-[(14)C]threonate is formed after feeding l-[U-(14)C]ascorbic acid to detached leaves. R. acutus leaves labeled with l-[4-(3)H]- or l-[6-(3)H]ascorbic acid produce l-[(3)H]threonate, in the first case internally labeled and in the second case confined to the hydroxymethyl group. These results are consistent with the formation of l-threonate from carbons three through six of l-ascorbic acid. Detached leaves of P. crispum oxidize l-[U-(14)C] threonate to l-[(14)C]tartrate whereas leaves of R. acutus produce negligible tartrate and the bulk of the (14)C appears in (14)CO(2), [(14)C]sucrose, and other products of carbohydrate metabolism. R. acutus leaves that are labeled with l-[U-(14)C]threonate release (14)CO(2) at linear rate until a limiting value of 25% of the total [U-(14)C]threonate is metabolized. A small quantity of [(14)C]glycerate is also produced which suggests a process involving decarboxylation of l-[U-(14)C]threonate.     

Metabolism of (−)-deprenyl and PF-(−)-deprenyl in brain after central and peripheral administration

We examined the cerebral metabolism of L-deprenyl and its fluoro-derivative pF-deprenyl, assaying the parent compounds, their metabolites desmethyl deprenyl, L-amphetamine, and L-methamphetamine, and the fluoro analogs of these metabolites. We compared the levels of the metabolites after subcutaneous injection with those after intracerebral administration (via microdialysis) of the parent compounds. The assay of the parent compounds and their metabolites was by GC-MS measurement of the components of brain microdialysate samples. After their subcutaneous administration, deprenyl and F-deprenyl rapidly entered the brain and then their concentration decreased, with an approximate half-life of 4.5 h. After the intracerebral administration the diffusion from the site of administration was minor. A small fraction (a few percent) of the intracerebrally administered deprenyl was metabolized in situ in the brain possibly by a nonenzymatic process. Metabolism of pF-deprenyl was somewhat more rapid. The higher cerebral levels of metabolites after the subcutaneous administration indicate their exogenous origin—metabolism of parent compounds in the periphery and penetration of the brain by the metabolites.     

Nucleases of the Metallo-β-lactamase Family and Their Role in DNA and RNA Metabolism

ABSTRACT

Proteins of the metallo-β-lactamase family with either demonstrated or predicted nuclease activity have been identified in a number of organisms ranging from bacteria to humans and has been shown to be important constituents of cellular metabolism. Nucleases of this family are believed to utilize a zinc-dependent mechanism in catalysis and function as 5′ to 3′ exonucleases and or endonucleases in such processes as 3′ end processing of RNA precursors, DNA repair, V(D)J recombination, and telomere maintenance. Examples of metallo-β-lactamase nucleases include CPSF-73, a known component of the cleavage/polyadenylation machinery, which functions as the endonuclease in 3′ end formation of both polyadenylated and histone mRNAs, and Artemis that opens DNA hairpins during V(D)J recombination. Mutations in two metallo-β-lactamase nucleases have been implicated in human diseases: tRNase Z required for 3′ processing of tRNA precursors has been linked to the familial form of prostate cancer, whereas inactivation of Artemis causes severe combined immunodeficiency (SCID). There is also a group of as yet uncharacterized proteins of this family in bacteria and archaea that based on sequence similarity to CPSF-73 are predicted to function as nucleases in RNA metabolism. This article reviews the cellular roles of nucleases of the metallo-β-lactamase family and the recent advances in studying these proteins.