Analysis of orientation behaviour of Tribolium castaneum and T. confusum to synthetic aggregation pheromone

Adult male T. castaneum (Herbst) and T. confusum (du Val) secretes an aggregation pheromone that is attractive to both sexes. Orientation behaviour of the two Tribolium species responding to different concentrations of synthetic aggregation pheromone in still and moving air was studied in an 2.5 m×0.4 m olfactometer. Analysis of Tribolium tracks indicated that the aggregation pheromone stimulated the beetles to walk faster at higher concentrations to increase the frequency and magnitude of turning and to decrease track reversal distances and distances between turns. The mean walking speed of both species was lowest at the highest air speed. The behavioural responses of the beetles to the pheromone in still and moving air were similar, indicating chemotaxis as the major orientation mechanism used by both species to locate an odour source. The beetles showed greater orientation efficiency within a discrete pheromone plume than a diffuse plume.     

How varying pest and trap densities affect Tribolium castaneum capture in pheromone traps

The red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), is an important insect pest in food processing facilities. Pheromone trapping is frequently used to monitor red flour beetle populations in structures; however, the optimal trap density and the relationship between trap captures and beetle density is not known. Two experiments were performed concurrently in environmentally controlled 30‐m² walk‐in chambers to determine the relationship between aggregation pheromone trap captures of red flour beetles and beetle and trap number. In one experiment, beetle density was kept constant at 200 individuals per chamber while trap number was varied from 1 to 8, and in the other experiment trap number remained constant at one per chamber while beetle density varied from 20 to 800 individuals. Results indicated that approximately one out of 23 red flour beetles were captured in a trap. Number of beetles captured in traps increased significantly as beetle density increased; however, the proportion of beetles captured remained consistent across beetle densities with a mean of 4.7 ± 0.6% of individuals captured. Trap captures varied significantly with trap placement within experimental chambers, indicating that subtle differences in the trapping environment can influence trap captures. Data suggested that trap densities of 0.07–0.10 m^?2 (2–3 traps per chamber) would maximize trap capture, whereas a trap density of 0.13 m^?2 (four traps per chamber) would maximize the predictive ability of a trapping equation estimating beetle density from trap captures. Results provide information needed to more thoroughly explore how environmental factors might influence red flour beetle trap capture in the absence of changes in beetle density. Further understanding of these relationships will allow for more accurate assessments of absolute beetle density from pheromone trap capture data.     

Response of German cockroaches to aggregation pheromone emitted by adult females

The response of early, middle and late instar (mixed sexes), adult male, and gravid and non-gravid female German cockroaches to aggregation pheromone on filter papers conditioned by gravid and non-gravid females was studied. Cockroaches were attracted to and settled on each of the conditioned papers. Classes varied in the strength of their response to conditioned papers, relative response to gravid versus non-gravid female conditioned paper, and in clustering behavior. A significantly greater attraction of all classes except middle instars and non-gravid females to paper conditioned by non-gravid females suggests that non-gravid females emit more aggregation pheromone than gravid females. Possible explanations of differences in response with age/sex class, including an enhanced attraction of males from female sex phermone, are discussed.

---

Résumé L'étude a porté sur les réponses de six groupes/âges/sexes de B. germanica à des papiers filtres imprégnés de phéromones émises par des femelles pleines ou non. Tous les groupes ont été attirés par les papiers traités, mais les attractions des stades larvaires moyens ou âgés ont été inférieures aux autres. 4 groupes sur 6 ont été plus attirés par le papier traité avec des femelles ne contenant pas d'oeufs mûrs que par les femelles pleines, ce qui suggère une plus forte émission par les femelles sans oeufs mûrs. Les stades larvaires moyens ont été les seuls à être plus attirés par le papier traité avec les femelles pleines. Les femelles ne contenant aucun oeuf ne manifestent aucune préférence. La tendance à former des amas inégaux était moins nette pour les stades larvaires moyens et âgés que pour les autres groupes. Les premiers stades ont montré une tendance très nette à se rassembler sur le même papier. Le rassemblement de mâles adultes sur du papier traité avec des femelles ne contenant pas d'oeufs peut avoir été partiellement une réponse à la phéromone sexuelle femelle qui avait imprégné le papier pendant le conditionnement.

     

First evidence of DNA methylation in insect Tribolium castaneum

DNA methylation has been studied in many eukaryotic organisms, in particular vertebrates, and was implicated in developmental and phenotypic variations. Little is known about the role of DNA methylation in invertebrates, although insects are considered as excellent models for studying the evolution of DNA methylation. In the red flour beetle, Tribolium castaneum (Tenebrionidae, Coleoptera), no evidence of DNA methylation has been found till now. In this paper, a cytosine methylation in Tribolium castaneum embryos was detected by methylation sensitive restriction endonucleases and immuno-dot blot assay. DNA methylation in embryos is followed by a global demethylation in larvae, pupae and adults. DNA demethylation seems to proceed actively through 5-hydroxymethylcytosine, most probably by the action of TET enzyme. Bisulfite sequencing of a highly abundant satellite DNA located in pericentromeric heterochromatin revealed similar profile of cytosine methylation in adults and embryos. Cytosine methylation was not only restricted to CpG sites but was found at CpA, CpT and CpC sites. In addition, complete cytosine demethylation of heterochromatic satellite DNA was induced by heat stress. The results reveal existence of DNA methylation cycling in T. castaneum ranging from strong overall cytosine methylation in embryos to a weak DNA methylation in other developmental stages. Nevertheless, DNA methylation is preserved within heterochromatin during development, indicating its role in heterochromatin formation and maintenance. It is, however, strongly affected by heat stress, suggesting a role for DNA methylation in heterochromatin structure modulation during heat stress response.     

Field trapping of Metamasius hemipterus with synthetic aggregation pheromone

The five synthetic pheromone components of the West Indian sugarcane borer (WISB), Metamasius hemipterus (L.) (Coleoptera, Curculionidae) were tested in the field. The combination of sugarcane (SC) and the major pheromone compound, 4-methyl-5-nonanol (1) was attractive. However, the addition of 2-methyl-4-heptanol (2) or 2-methyl-4-octanol (3) was required to reach high catch levels while 5-nonanol (4) or 3-hydroxy-4-methyl-5-nonanone (5) did not enhance WISB attraction. The redundancy phenomenon, here reported for the first time in rhynchophorinous species, was observed between compounds 2 and 3. SC +1 +3 was more attractive than living male baits, however, the sex-ratio of the catches was equivalent between both treatments. The sex-ratio of catches was affected by the qualitative composition of the pheromone formulation. Compound 3 had a sexual role, attracting more females while 5 seemed to play an aggregation role, luring both sexes in the same proportion.     

Biochemical effects of sublethal doses of cypermethrin on the sixth-instar larvae of Tribolium castaneum (Herbst.)

Effects of sublethal doses ie, 2, 4, 10, and 20 ppm of cypermethrin, were studied on the sixth-instar larvae of Tribolium castaneum (Herbst.). Of all the biochemical parameters tested, the free amino acids and cholesterol content and the activity of amylase were found to be the most sensitive components. Glutamate pyruvate transaminase activity was elevated at all doses except 2 ppm. The activities of alkaline phosphatase and glutamate oxaloacetate transaminase and glucose content were raised significantly at doses of 10 and 20 ppm. Acid phosphatase activity and the soluble protein content increased at a dose of 20 ppm. Total lipid and triglycerides, however, decreased significantly at this sublethal dose. Other biochemical parameters, such as cholinesterase and lactate dehydrogenase activities and the total protein, urea, glycogen, DNA, and RNA contents, were not significantly affected by exposure to different doses of cypermethrin.     

赤拟谷盗和杂拟谷盗的生殖隔离研究

赤拟谷盗和杂拟谷盗是两个同域发生的近缘储粮害虫。为明确它们的生殖隔离程度和机制,本研究比较分析了赤拟谷盗和杂拟谷盗雄虫对同种和异种雌虫的交配选择;赤拟谷盗与杂拟谷盗交配后,测定了精子在异种雌体内的存活情况;将赤拟谷盗和杂拟谷盗进行正反杂交,研究其F1代、F2代和回交代杂种是否产生及其雌雄比。结果显示,赤拟谷盗和杂拟谷盗雄虫对同种雌虫的爬跨率和交配率高于其对异种雌虫的爬跨率和交配率,表明赤拟谷盗和杂拟谷盗种间性隔离不完全;赤拟谷盗和杂拟谷盗交配后精子在异种雌体内是存活的,杂交所产的F1代卵为受精卵,说明交配后完成了精子传送和受精过程,表明种间机械隔离和配子隔离不完全;赤拟谷盗和杂拟谷盗的杂种F1代自交和回交产生了F2代和回交代,表明种间不存在杂种不育;一些杂交组合产生的F2代和回交代数量少且存在雌雄性比偏离,表明种间存在部分杂种不活和杂种衰败。本研究明确了赤拟谷盗和杂拟谷盗的生殖隔离机制,这对于揭示它们的物种进化关系有着重要意义。     

An unusual barrier to gene flow: perpetually immature larvae from inter-population crosses in the flour beetle, Tribolium castaneum

We genetically characterize an unusual hybrid incompatibility phenotype manifest in F₁ offspring of crosses between two populations of Tribolium castaneum. Hybrid larvae cease development at the third larval instar, persisting as ‘perpetually immature larvae’ thereafter. Although unable to produce viable adult hybrid offspring with one another, each population produces abundant, fertile hybrids with other populations, indicating a recent origin of the incompatibility and facilitating genetic studies. We mapped the paternal component of the hybrid phenotype to a single region, which exhibits two characteristics common to hybrid incompatibility: marker transmission ratio distortion within crosses and elevated genetic divergence between populations. The incompatible variation and an elevation in between‐population genetic divergence is associated with a region containing the T. castaneum ecdysone receptor homologue, a major regulatory switch, controlling larval moults, pupation and metamorphosis. This contributes to understanding the genetics of speciation in the Coleoptera, one of the most speciose of all arthropod taxa.     

Female mate choice in relation to heterozygosity in Tribolium castaneum

Abstract Female mate choice, both before and after copulation, is pervasive among insect species. It is often hypothesized that females would preferentially mate with males that are genetically dissimilar to promote the genetic variability of the offspring. We used various strains of red flour beetle, Tribolium castaneum, and tested the effect of male and female genetic backgrounds on precopulatory and post‐copulatory female mate choice. Simultaneous mate choice experiments using previously well established pheromone assays did not detect female preference for males of different strains. Post‐copulatory female mate choice was examined through paternity analysis. Two parameters were used to measure post‐copulatory female mate choice, including male defence capacity (P₁, proportion of offspring sired by the first male when a female mated with two males consecutively) and offence capacity (P₂, proportion of offspring sired by the second of two males to mate with a female). When female and male beetle strains were same, defence capacity was significantly higher than when female and male strains were different. However, such a pattern was not observed for offence capacity. The results suggest that female precopulatory mate choice is not affected by genetic background, but the outcome of post‐copulatory processes depends on the genetic background of male and female beetles.     

Maternal effect of a hybrid inviability gene in Tribolium castaneum

The Tribolium castaneum hybrid inviability gene, H, was selectively introgressed into a genetic background lacking H through serial paternal backcrosses. This revealed a poor viability phenotype (partial paralysis and poor control of the limbs, referred to as tremor) not present in the parent strains. Tremor cosegregated with H, but was expressed only when transmitted paternally and only when H was not also present maternally. The inferred maternal, self‐suppressive effect of H may explain nonreciprocal incompatibilty previously observed between H and H‐incompatible strains. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cocoon-spinning larvae of Oriental fruit moth and Indianmeal moth do not produce aggregation pheromone

1 Mature larvae of the Oriental fruit moth (OFM) Grapholita molesta (Lepidoptera: Tortricidae) and the Indianmeal moth (IMM) Plodia interpunctella (Lepidoptera: Pyralidae) leave their food source in search of suitable pupation sites in which to spin cocoons. These sites are typically well-concealed cracks and crevices within the environment. Such cocooning behaviour is also observed in larvae of the codling moth (CM) Cydia pomonella (Lepidoptera: Tortricidae), which aggregate prior to pupation in response to a pheromone blend produced by cocoon-spinning conspecific larvae.
2 In laboratory experiments, we tested whether cocoon-spinning OFM and IMM larvae produce aggregation pheromones and whether CM larvae are cross-attracted to closely-related OFM larvae.
3 Fifth-instar OFM and IMM larvae were not attracted to, or arrested by, cocoon-spinning conspecifics. Moreover, fifth-instar CM larvae were not cross-attracted to either cocoon-spinning OFM or IMM larvae.
4 Analyses of volatiles released from cocoon-spinning OFM and IMM larvae revealed that both OFM and IMM lack components that are present in the aggregation pheromone of CM larvae. This information may help explain why CM larvae are not cross-attracted to cocooning OFM or IMM larvae.     

Developmental trajectories and breakdown in F1 interpopulation hybrids of Tribolium castaneum

When hybrid inviability is an indirect by‐product of local adaptation, we expect its degree of severity between pairs of populations to vary and to be sensitive to the environment. While complete reciprocal hybrid inviability is the outcome of the gradual process of local adaptation, it is not representative of the process of accumulation of incompatibility. In the flour beetle, Tribolium castaneum, some pairs of populations exhibit complete, reciprocal F1 hybrid incompatibility while other pairs are fully or partially compatible. We characterize this naturally occurring variation in the degree and timing of expression of the hybrid incompatible phenotype to better understand the number of genes or developmental processes contributing to speciation. We assessed the morphological and developmental variation in four Tribolium castaneum populations and their 12 possible F1 hybrids at each life‐history stage from egg to adult. We find that the rate of hybrid larval development is affected in all interpopulation crosses, including those eventually producing viable, fertile adults. Hybrid incompatibility manifests early in development as changes in the duration of instars and diminished success in the transition between instars are relative to the parent populations. Parent populations with similar developmental profiles may produce hybrids with disrupted development. The degree and timing of expression of hybrid inviability depends upon populations crossed, direction of the cross, and environment in which hybrids are raised. Our findings suggest that the coordinated expression of genes involved in transitional periods of development is the underlying cause of hybrid incompatibility in this species.     

多种植物挥发物及马铃薯甲虫聚集素对马铃薯甲虫的引诱作用

马铃薯甲虫Leptinotarsa decemlineata (Say)是马铃薯上一种最主要的毁灭性害虫。由于其严重的抗药性, 急需寻找一种替代杀虫剂的防治方法。本实验从开发马铃薯甲虫引诱剂的目的出发,用“Y”型嗅觉仪测定了马铃薯甲虫对来自植物的7种挥发物单体、8个挥发物混合物配方以及马铃薯甲虫聚集素的行为反应,并进一步进行了田间诱集试验。室内生测结果表明,2-苯乙醇对马铃薯甲虫雌雄虫都有明显的引诱作用。芳樟醇+水杨酸甲酯+顺乙酸-3-己烯酯的混合物对马铃薯甲虫雄虫有很强的引诱作用(81.67%), 但是对雌虫引诱作用不明显(63.33%)。在其中加入马铃薯甲虫聚集素不但提高了对雄虫的引诱率（88.33%）,还消除了之前的雌雄性别间反应差异现象,使对雌虫的引诱率达到了83.33%。田间诱集结果显示,芳樟醇+水杨酸甲酯+顺乙酸-3-己烯酯+马铃薯甲虫聚集素在所研究的引诱剂配方中引诱效果最好。从而为马铃薯甲虫发生的预测预报和诱杀技术提供了研究基础。     

Identification of microsatellite markers in the red flour beetle,Tribolium castaneum

We isolated and characterized microsatellite markers for the red flour beetle Tribolium castaneum, an important model species for studies in various areas of evolutionary biology and ecology. A microsatellite‐enriched genomic library was constructed and screened with single stranded oligonucleotide probes [(CCT)₁₇, (AAT)₁₇ and (CAG)₁₇]. Forty‐five primer pairs were designed of which 19 pairs produced successful amplification. Polymorphism screening involved beetles from five beetle strains and revealed 15 polymorphic and four monomorphic markers. The development of polymorphic microsatellite markers will facilitate future ecological and genetic studies involving T. castaneum beetles.     

Crinkled employs wingless pathway for wing development in Tribolium castaneum

Crinkled is associated with embryonic denticle formation and auditory organ development in Drosophila melanogaster. However, the functions of Crinkled have not been fully investigated. Additionally, the genes that participate in the Crinkled pathway are unknown. Phylogenetic analysis indicates that crinkled exhibits a one‐to‐one orthologous relationship in insects. In Tribolium castaneum, the crinkled gene is 6,498 bp in length and consists of six exons. Crinkled expression peaked during two phases in Tribolium: late embryonic and pupal stages. High levels of crinkled mRNA were detected in the fat body, head, epidermis, ovary, and accessory gland of late adults. Knockdown of crinkled using RNA interference (RNAi) severely affected wing morphogenesis in T. castaneum. We further showed that crinkled silencing reduced forked expression through wingless and shaven‐baby, and RNAi of forked phenocopied the effects of crinkled knockdown in T. castaneum. This study investigated the development role of crinkled in postembryonic stages and indicated that forked mediates the functions of crinkled during wing morphogenesis in T. castaneum.     

七种糕点对赤拟谷盗(鞘翅目：拟步甲科)感染的抗性（英文）

Biscuits are flour-based products,which areconsumed by majority of people,both old and young,throughout the world and most especially by children,teenagers and mid-adults(Majumder,1970).It can beserved as a desert or snacks and as a breakfast foodwhentaken with beverages.Its ready-to-serve state andlong shelf life contribute to its acceptability by themasses.Wheat as humanfoodis used principallyintheformof flour for baked products such as bread,bis-cuits,pastries and crackers(Kent,1983).In th…