杂拟谷盗的热适应特性

为了解杂拟谷盗Tribolium confusum Jacquelin du Val的热适应特性,将杂拟谷盗分别于15,25和35℃下驯化2周后,用温度梯度仪测量在不同温度驯化下杂拟谷盗的最适温度、临界低温和临界高温。结果表明,驯化温度对杂拟谷盗最适温度、临界低温和临界高温的影响极显著(P<0.01),最适温度、临界低温、临界高温均随着驯化温度的升高而升高。最适温区的范围随着驯化温度的升高而扩大。驯化温度对杂拟谷盗最适温度的影响最大(0.317),对临界低温的影响(0.310)大于临界高温(0.255)。     

Digestive proteolysis organization in two closely related Tenebrionid beetles: red flour beetle (Tribolium castaneum) and confused flour beetle (Tribolium confusum)

The spectra of Tribolium castaneum and T. confusum larval digestive peptidases were characterized with respect to the spatial organization of protein digestion in the midgut. The pH of midgut contents in both species increased from 5.6–6.0 in the anterior to 7.0–7.5 in the posterior midgut. However, the pH optimum of the total proteolytic activity of the gut extract from either insect was pH 4.1. Approximately 80% of the total proteolytic activity was in the anterior and 20% in the posterior midgut of either insect when evaluated in buffers simulating the pH and reducing conditions characteristic for each midgut section. The general peptidase activity of gut extracts from either insect in pH 5.6 buffer was mostly due to cysteine peptidases. In the weakly alkaline conditions of the posterior midgut, the serine peptidase contribution was 31 and 41% in T. castaneum and T. confusum, respectively. A postelectrophoretic peptidase activity assay with gelatin also revealed the important contribution of cysteine peptidases in protein digestion in both Tribolium species. The use of a postelectrophoretic activity assay with p‐nitroanilide substrates and specific inhibitors revealed a set of cysteine and serine endopeptidases, 8 and 10 for T. castaneum, and 7 and 9 for T. confusum, respectively. Serine peptidases included trypsin‐, chymotrypsin‐, and elastase‐like enzymes, the latter being for the first time reported in Tenebrionid insects. These data support a complex system of protein digestion in the Tribolium midgut with the fundamental role of cysteine peptidases. © 2009 Wiley Periodicals, Inc.     

杂拟谷盗体内 Wolbachia 感染密度的时间和空间分布状况

为搞清杂拟谷盗体内沃尔巴克氏体(Wolbachia)感染密度的时间和空间分布状况,本研究采用实时荧光定量PCR方法测定了杂拟谷盗不同发育阶段、不同日龄、不同性别和不同身体部位的Wolbachia感染密度。结果表明,杂拟谷盗在卵和成虫期Wolbachia感染密度高于其幼虫和蛹期,成虫腹部的Wolbachia感染密度高于其头部和胸部,而成虫不同日龄和雌雄之间的Wolbachia感染密度均没有显著性差异。本研究明确了Wolbachia在杂拟谷盗体内的时空分布和动态变化规律,这对于揭示寄主与共生菌之间的互作关系有着重要意义。     

Effects of terpinen-4-ol fumigation on protein levels of detoxification enzymes in Tribolium confusum

Terpinen-4-ol has high fumigating activity to stored-grain pests including Tribolium confusum. To understand the detoxification of terpinen-4-ol in insects, proteomic analysis was performed to identify related proteins and pathways in response to terpinen-4-ol fumigation in T. confusum. By using isobaric tags for relative and absolute quantitation (iTRAQ)-based strategy, 4,618 proteins were obtained from T. confusum adults in the present study. Comparative proteomic analysis showed that 148 proteins were upregulated and 137 proteins were downregulated in beetles under the LC₅₀ of terpinen-4-ol treatment for 24 hr. According to functional classifications, differentially expressed proteins (DEPs) were enriched in xenobiotic metabolism pathways. In the detoxification pathway, the levels of 25 cytochrome P450s, 5 glutathione S-transferases, and 2 uridine diphosphate (UDP)-glucuronosyltransferases were changed, most of which were upregulated in T. confusum exposed to terpinen-4-ol. The results indicated that terpinen-4-ol was potentially metabolized and detoxified by enzymes like P450s in T. confusum.     

Avidin expressed in transgenic rice confers resistance to the stored-product insect pests Tribolium confusum and Sitotroga cerealella

Rice (Oryza sativa var. Nipponbare) was transformed with an artificial avidin gene. The features of this construct are as follows: (1) a signal peptide sequence derived from barley alpha amylase was added at the N-terminal region, (2) codon usage of the gene was optimized for rice, and (3) the gene was driven by rice glutelin GluB-1, an endosperm-specific promoter. Avidin was produced in the grain of the transgenic rice but not in the leaves. The concentration of avidin in the kernels was about 1,800 ppm. All larvae of the confused flour beetle (Tribolium confusum) and Angoumois grain moth (Sitotroga cerealella) died when fed transgenic avidin rice powder or kernels, respectively, whereas most of the test insects developed into adults when they were fed a nontransgenic rice control diet. Avidin extracted from the transgenic rice kernel lost most biotin-binding activity after 5 min heating at 95 degrees C.     

The Influence of Oxidizing Agents on Macromolecular Synthesis for Chitinous Material

Summary

The 5th instar larvae of the common flour beetle, Tribolium confusum, was utilized as an invertebrate model to test the larvicidal potential of oxidizing agents used as flour preservatives. Larvae were exposed to 10 ppm of either diflubenzuron or benzoyl peroxide. Results showed decreases in both total protein and glucose concentrations. Histological examination also indicated smaller deposition of chitinous material in the endocuticle. Both treatments had similar effects. Thus, these data indicate that oxidizing agents mixed in flour may be used for the control of development of flour beetle larvae.     

Pace‐of‐life: Relationships among locomotor activity,life history,and circadian rhythm in the assassin bug,Amphibolus venator

The pace‐of‐life syndrome (POLS) hypothesis means that animal behavior is correlated with life history strategies. Studies have reported that the free‐running period of the circadian rhythm (length of the period) is correlated with life history strategies in some animals. Although the length of the circadian rhythm may be associated with the POLS hypothesis, few studies have investigated the relationships among animal behavior, life history traits, and circadian rhythm. We tested the POLS hypothesis in the assassin bug, Amphibolus venator, which shows individual variation in locomotor activity. We found higher repeatability of differences in locomotor activity between individuals. Moreover, we found a trade‐off between locomotor activity and developmental period such that active individuals developed faster. However, locomotor activity was not correlated with the length of the circadian rhythm in A. venator. Therefore, this study suggests that the length of the circadian rhythm in A. venator does not support the POLS hypothesis.     

Suppression of the stored-product pest Sitophilus zeamais (Coleoptera: Curculionidae) by simultaneous release of a predatory bug and a parasitoid wasp

The control of the maize weevil, Sitophilus zeamais in brown rice was estimated by using a reduviid bug, Amphibolus venator, and a pteromalid wasp, Theocolax elegans. Results showed that the simultaneous release of both species of natural enemies could enhance biological control compared with the release of either species alone.     

Mitochondrial DNA phylogeography of the Cotesia flavipes complex of parasitic wasps (Hymenoptera: Braconidae)

The Cotesia flavipes species complex of parasitic wasps are economically important worldwide for the biological control of lepidopteran stem borers. The complex currently comprises three species: Cotesia flavipes Cameron, C. sesamiae (Cameron) and C. chilonis (Matsumura) (Hymenoptera: Braconidae), which appear morphologically similar. Despite their economic importance, little is known about the genetic diversity and phylogeography of these parasitoids. Differences in the biology of geographic populations have generally been interpreted as genetic divergence among strains, but direct genetic evidence is lacking. In Australia, several stem borer pests in neighbouring countries have been identified as significant threats to the sugar industry. However, the status of C. flavipes in Australia is unknown. To examine the genetic variation among worldwide populations of the C. flavipes complex and investigate the status of the Australian C. flavipes-like species, partial sequence data were generated for mitochondrial gene regions, 16S rRNA and COI. Parsimony, minimum evolution and Bayesian analyses based on 21 geographic populations of the complex and four outgroups supported the monophyly of the complex and the existence of genetically divergent populations of C. flavipes and C. sesamiae. The geographically isolated Australian haplotypes formed a distinct lineage within the complex and were ~3.0% divergent from the other species. The results indicated that historical biogeographic barriers and recent biological control introductions play an important role in structuring lineages within these species. This study provides a phylogeographical context for examining adaptive evolution and host range within biologically divergent strains of the C. flavipes complex.     

Tribolium confusum responses to blends of cereal kernels and plant volatiles

In this study, we report on the behavioural responses of the Tribolium confusum to 6 blends of cereal volatiles. There were tested four doses (1 ng/min, 10 ng/min, 100 ng/min and 1000 ng/min in 50 μl of hexane applied on filter paper). A Y‐tube experiment revealed that T. confusum females and males were attracted to the blend 1, 4 and 5 at the concentrations between 1 and 10 ng/min. Confused T. confusum females and males were not attracted to any concentration of blend 2 and 3. Yet the beetle females and males were repelled by the highest concentrations (1000 ng/min) for all the blends tested and also by the concentration 100 ng/min of all the blends tested, except for blend 1 and 4. Females and males were also repelled by the three blend 6 concentrations tested (10, 100 and 1000 ng/min).     

茶翅蝽沟卵蜂和黄足沟卵蜂对茶翅蝽的寄生潜能研究

茶翅蝽Halyomorpha halys(Stl)是水果和蔬菜生产中的重要害虫。为了寻找优势卵寄生蜂用于其生物防治,本文进行了相关的野外调查和室内试验。通过定期野外采集茶翅蝽卵块,培育、收集和鉴定寄生蜂,发现在自然环境中茶翅蝽卵粒的寄生率为56.29%,其中茶翅蝽沟卵蜂比例最高,占77.66%;其次是黄足沟卵蜂占20.44%。Y型嗅觉仪测定结果显示,茶翅蝽沟卵蜂和黄足沟卵蜂对茶翅蝽卵均具有明显的选择趋性。室内寄生能力试验结果表明,茶翅蝽沟卵蜂与黄足沟卵蜂单独作用的寄生率分别为94.06%和84.21%,与两种寄生蜂混合寄生茶翅蝽卵的寄生率91.65%均无显著性差异。两种寄生蜂混合寄生时,茶翅蝽沟卵蜂与黄足沟卵蜂的寄生比例分别为58.95%和41.05%,二者之间无显著差异。     

Introduction of Beauveria bassiana as a biological control agent for Tribolium castaneum in Kerman province

Barley, Hordeum vulgare, one of the important crops in the word, is used in malting, feed and food industries. The red flour beetle, Tribolium castaneum, was found wherever grains or other dried foods are stored. Disinfestations of barley using chemical methods to kill insects, in this research, for the first time we isolated the pathogenic KB512 of entomopathogenic fungi Beauveria bassiana from soil and insects, which produced aerial and submerged conidia and blastospores in laboratory conditions. We investigated the best conditions for the production and utilisation of spore suspension to spray the larvae of T. castaneum, which is one of the important pests in Kerman province (Iran). One hundred and eighty isolates that naturally infected by T. castaneum were reared during spring and summer seasons 2010–2011. The pathogenicity test was carried out with direct spray. To bioassay the isolates, three concentrations of the spore suspension were prepared as follows: 1?×?10⁶ and 1?×?10⁸ conidia/ml. The pests were sprayed by aerial conidial suspension, which was prepared by 0.01% Tween 80 in distilled water, and the controls were sprayed by 0.01% Tween 80 in distilled water. After spraying the pests, the plates were incubated at 25?±?1?°C and 80% of relative humidity. Then, the treated pests were monitored every day for the fungal growth and mortality.     

Use of ablated stalks to assess field rates of parasitism of Diatraea saccharalis (Lepidoptera: Crambidae) by Cotesia flavipes (Hymenoptera: Braconidae)

We evaluated a technique that used ablated sections of corn stalks infested with larvae of Diatraea saccharalis F. to assess parasitism by Cotesia flavipes (Cameron) in a corn field. Stalkborer larvae were retrieved successfully from the artificially infested, ablated stalks for at least 96 h after deployment in the field. Levels of parasitism in ablated stalks attached to corn plants were comparable to levels of parasitism measured using whole plants that were artificially infested. Olfactometer comparisons showed preference by female C. flavipes for both whole plants and ablated stalks containing larval D. saccharalis, over uninfested stalks and plants. This technique provides a means to estimate parasitism of stalkborer larvae by C. flavipes without destructive sampling of agronomic plants.     

Spatial and temporal population interactions between the parasitoids Cotesia flavipes and Tachinidae flies: considerations on the adverse effects of biological control practice

Biological control of Diatraea saccharalis is regarded as one of the best examples of successful classical biological control in Brazil. Since the introduction of the exotic parasitoid Cotesia flavipes, the decrease of D. saccharalis infestation in sugarcane fields has been attributed to the effectiveness of this agent. Recently, the native tachinid fly parasitoids (Lydella minense and Paratheresia claripalpis) have also been implicated in the success. Here, we investigated the spatial and temporal population interactions between C. flavipes and the tachinid flies, and provide a critical analysis of the biological control practice, focusing on the undesirable effects of introductions of exotic natural enemies. To investigate these questions, a large data set comprising information from two sugarcane mills located in the state of São Paulo, Brazil (Barra and São João Mills), was analysed. Analysis of the correlation between C. flavipes and tachinid fly population densities through time revealed that such populations were inversely correlated in the São João Mill and not correlated in the Barra Mill. Logistic regressions were computed to investigate the proportion of sites occupied by the parasitoid species at both mills as a function of time. An increasing trend in the proportion of sites occupied by C. flavipes was observed, with a concomitant decrease of the sites occupied by tachinid flies. This effect was more intense in the São João Mill. Thus, there is a convincing possibility that constant releases of C. flavipes decreased the tachinid fly populations, resulting in an undesirable effect of biological control practice.     

常规棉花粉和转Cry1Ac+CpTI棉花粉对拟澳洲赤眼蜂繁殖和存活的影响

在室内评价了常规棉花粉和转Cry1Ac CpTI棉(Bt棉)花粉通过不同处理方式作为食物源对拟澳洲赤眼蜂Trichogrammaconfusum寿命、寄生卵数、子代羽化数和性比等繁殖和存活特征的影响。共设计7个饲喂处理,即不饲喂(UNFED)、水(W)、水 常规棉花粉(W P)、水 Bt棉花粉(W BtP)、10%蜂蜜水(H)、10%蜂蜜水 常规棉花粉(H P)和10%蜂蜜水 转Bt棉花粉(H BtP)。在提供或不提供米蛾Corcyracephalonica卵条件下,经W P或W BtP饲喂的雌蜂寿命与W或UNFED间无显著差异,但均显著短于H和H P或H BtP饲喂的雌蜂,而经H P或H BtP饲喂的雌蜂寿命又均显著长于经H饲喂的雌蜂(p<0.05)。比较每雌寄生卵数和子代羽化数来看,各处理明显分为3类,即UNFED、W、W P和W BtP为最低,H为居中,H P和H BtP为最高。比较每雌子代性比来看,各处理也明显分为3类,即H、H P和H BtP为最低,W P和W BtP为居中,UNFED和W为最高。饲喂W P和W BtP的雌蜂寿命、寄生卵粒数、子代羽化数和性比分别与W BtP和H BtP饲喂间无显著差异(p<0.05)。因而,棉花花粉需与蜂蜜组合才能成为拟澳洲赤眼蜂实现其最大存活和繁殖力的食物;Bt棉花粉对拟澳洲赤眼蜂无影响。     

Insecticidal effect of the combined application of spinosad,Beauveria bassiana and diatomaceous earth for the control of Tribolium confusum

Laboratory bioassays were carried out to evaluate the insecticidal effect of Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) germinated conidia, a commercial formulation of diatomaceous earth and a liquid formulation of spinosad, applied alone or in all possible combinations on wheat, against adults of the confused flour beetle, Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae). Diatomaceous earth was applied at one (200 ppm), the fungus at two (100 and 300 ppm) and spinosad at three (0.01, 0.1 and 0.5 ppm) dose rates, whereas all combinations of two or three of the insecticide concentrations were also tested. Insect mortality was determined after 2, 7 and 14 d of exposure. The simultaneous use of B. bassiana germinated conidia with diatomaceous earth notably increased mortality of T. confusum adults by sixfold 7 d after exposure, as compared with either B. bassiana or diatomaceous earth alone. When diatomaceous earth and spinosad were combined, diatomaceous earth did not increase the efficacy of spinosad. In general, the combination of all three agents did not differ from either the application of the agents tested alone, or their binary combinations, in most treatments examined.     

Influence of processing and substrate variety on survival and development of Tribolium confusum (Coleoptera: Tenebrionidae)

Tribolium confusum is a major pest of grains and grain products. Some legumes hitherto not used as major food are gaining interest due to increase in human population and the need of food and feeds. Susceptibility of fresh and processed Glycine max and Cajanus cajan flours, and white wheat flour (control) to T. confusum was studied towards effective storage techniques and cheap insect diet. Ten copulating pairs of the beetle were introduced into twenty grams of each flour. Number of eggs laid, emerged larvae, pupae, eclosed adults, and duration of development of each stage was used as indices of survival of T. confusum. Proximate composition, saponin; and α-amylase and protease inhibitors determined suitability of flours as insect diet. This work discusses the significance of proximate composition and anti-nutrient properties in fresh and roasted processed flours on the development and survival of T. confusum.     

The stimulation of immune defence accelerates development in the red flour beetle (Tribolium castaneum)

The timing of the transition between life stages is of key importance for an organism. Depending on the environmental conditions, maturing earlier at a smaller size or maturing later at a larger size can be advantageous for fitness. Exposure to parasites and subsequent immune activation may lead to alterations in development. Immune defence often comes at a cost, such as energy drain towards immune function, which is likely to delay development. On the other hand, animals may react to an anticipated risk of infection with a phenotypically plastic shift in life history, which may more likely lead to accelerated development and earlier maturation. We tested these alternatives in the red flour beetle, Tribolium castaneum. Young larvae were exposed to a non-infectious immune challenge with heat-killed bacteria (either Escherichia coli or Bacillus thuringiensis) and they were followed up for their development, survival, adult size and reproduction. We found that animals that had experienced a bacterial challenge developed into adults earlier than sham-treated beetles, while they did not differ significantly in survival or adult size. Beetles exposed to E. coli produced fewer offspring, while exposure to B. thuringiensis did not affect offspring number. Taken together, our results indicate that T. castaneum is able to speed up its development when facing a risk of infection.     

The effects of Farinocystis tribolii on the growth and development of the flour beetle Tribolium castaneum

Farinocystis tribolii multiplied only in the fatbody of Tribolium castaneum larvae. In the advanced stages of infection, the fatbody was destroyed and the albuminoid granules were completely depleted. Larvae in late stages of infection were “C” shaped. Diseased pupae were flat and straight with the head and mouth parts not completely flexed with the pupal body. Infected pupae failed to develop into adults. Heavily infected adults were distended. Larvae of third, fourth, and fifth instars, although succumbing to the infection ultimately, continued to live even after the healthy larvae of the same age had entered pupation, indicating a prolongation of larval period. Molting was either affected or delayed in infected insects. F. tribolii infection induced a juvenile hormonal effect producing larval-pupal and pupal-adult intermediate forms. Ether extract of spores of F. tribolii when applied on pupae of T. castaneum and final instar nymphs of Dysdercus cingulatus produced adultoids.