Characteristic features of induced mutagenesis in hybrid dysgenesis systems of Drosophila melanogaster

The mutagenic effect of low-dose gamma-irradiation was studied in Drosophila melanogaster systems of hybrid dysgenesis by estimating polytene chromosome rearrangements, recombination frequency, and viability at the embryonic and postembryonic developmental stages. A dose of gamma-irradiation which had no effect detectable by routine line crossing proved to significantly reduce the number of recombinants in the H-E and P-M systems and mortality at postembryonic stages. However, this combined effect was obtained if irradiation followed transposition, i.e., it depended on the application sequence of the mutagenic factors. The reverse order of the mutagenic treatment led to summation of the effects: as compared to either control, the frequencies of the dominant allele mutations as well as the larval and pupal mortality in F2 increased significantly (at the level of 99.9%). This allowed us to estimate the contribution of extremely low-dose gamma-irradiation into the mutagenic effect, which was impossible under routine conditions.     

Developmental changes in Drosophila melanogaster following exposure to alternating electromagnetic fields

This study investigated the biological effects of alternating electromagnetic fields (EMFs) on developmental stages of Drosophila melanogaster eggs and the first, second and third instar larvae stages. D. melanogaster eggs and larval stages were exposed to a 11 mT 50 Hz field produced by a pair of Helmholtz coils. Each stage was exposed to aEMFs for 2, 4, 6 and 8 h. Features of adult flies such as head, thorax, abdomen and other morphological changes were studied and compared. The frequency of abnormal flies was calculated using statistical methods at P <.05. The results obtained from exposing larvae in different stages of development showed a significant increase in the number of abnormal adult flies, whereas no significant increase was observed in the group arising from eggs exposed to aEMFs. Also, it appeared that duration of exposure correlates with the increase in the number of abnormal flies. There was no significant difference in mortality rate and sex distribution of the abnormal flies between field exposed and the control groups.     

Salinity tolerance of stable fly (Diptera: Muscidae)

Effects of salinity on the survival, growth, and development of stable fly, Stomoxys calcitrans (L.), were investigated in the laboratory. Larvae failed to develop to pupation when reared in media containing a salinity of 40 parts per thousand (ppt) sodium chloride (NaCl). Maximum salinity supporting larval development equaled the salinity of seawater (34 ppt); the larval LC90 was 24.2 ppt. Deleterious effects of high salinity decreased as larvae matured. Six-day-old larvae reared at a salinity of 34 ppt weighed 79% less than controls, compared with a 36% difference in 9-d-old larvae; by pupation, the difference was only 24%. Salinity did not influence the duration of larval, pupal, or adult stages. Survival of pupae was unimpaired despite a slight increase in number of pupal deformities, and normal adults emerged. Eggs were highly tolerant to saline. They hatched at salinity concentrations lethal to larvae; greater than 50% hatch occurred even when eggs were maintained at 80 ppt NaCl. Sensitivity of larvae to salinities close to that of seawater might be important for control of stable flies inhabiting marine areas.     

Somatic Effects of P Element Activity in Drosophila melanogaster: Pupal Lethality

Nonautonomous P elements normally excise and transpose only when a source of transposase is supplied, and only in the germline. The germline specificity depends on one of the introns of the transposase gene which is not spliced in somatic cells. To study the effects of somatic P activity, a modified P element (delta 2-3) lacking this intron was used as a source of transposase. Nonautonomous P elements from a strain called Birmingham, when mobilized in somatic cells by delta 2-3, were found to cause lethality, although neither component was lethal by itself. The three major Birmingham chromosomes acted approximately independently in producing the lethal effect. This lethality showed a strong dependence on temperature. Although temperature sensitivity was limited to larval stages, the actual deaths occurred at the pupal stage. Survivors, which could be recovered by decreasing the temperature or by reducing the proportion of the Birmingham genome present, often showed multiple developmental anomalies and reduced longevity reminiscent of the effects of cell death from radiation damage. Although the genetic damage occurred in dividing imaginal disc cells, the phenotypic manifestations--death and abnormalities--are not observed until later. The survivors also showed gonadal dysgenic (GD) sterility, a well-known characteristic of P-M hybrid dysgenesis. To explain these findings, we suggest that pupal lethality and GD sterility are both caused by massive chromosome breakage in larval cells, resulting from excision and transposition of genomic P elements acting as substrate for the transposase.     

Characteristic Features of Induced Mutagenesis in Hybrid Dysgenesis Systems of Drosophila melanogaster

The mutagenic effect of low-dose gamma-irradiation was studied inDrosophila melanogaster systems of hybrid dysgenesis by estimating polytene chromosome rearrangements, recombination frequency, and viability at the embryonic and postembryonic developmental stages. A dose of gamma-irradiation which had no effect detectable by routine interstrain cross proved to significantly reduce the number of recombinants in the H–E and P–M systems and mortality at postembryonic stages. However, this combined effect was obtained if irradiation followed trasposition, i.e., it depended on the application sequence of the mutagenic factors. The reverse order of the mutagenic treatment led to summation of the effects: as compared to either control, the frequencies of the dominant lethal mutations as well as the larval and pupal mortality in F₂ increased significantly (at the level of 99.9%). This allowed us to estimate the contribution of extremely low-dose gamma-irradiation into the mutagenic effect, which was impossible under routine conditions.     

Changes in polyamine levels in various organs of Bombyx mori during its life cycle

Polyamines in various organs of larval, pupal, and moth stages of Bombyx mori, were assayed by high-performance ion-exchange chromatography and paper and thin-layer chromatography. Putrescine and spermidine were especially abundant in the silk gland, gonads, mucous gland, and sucking stomach; spermine was also present in them, but at much lower concentrations. Both norspermidine and norspermine were detected in almost all organs examined, while their precursor 1,3-diaminopropane was found only in a limited number of organs. Low concentrations of sym-homospermidine were observed in the silk gland and ovary. Cadaverine content was particularly high in the mucous gland which contained diapause eggs and the sucking stomach. Diapause eggs contained much higher levels of cadaverine than non-diapause eggs. The concentrations of most polyamines in the silk glands remained rather constant during the larval stage, and decreased markedly at the pupal stage. Polyamines in gonads, in contrast, did not decrease at the pupal stage, but putrescine, diaminopropane, and norspermidine rather increased during the pupal and moth stages.     

Sublethal effects of Bacillus thuringiensis on the spruce budworm, Choristoneura fumiferana

Exposing larvae of the spruce budworm, Choristoneura fumiferana (Clemens), to sublethal ( 50% lethal dose) levels of Bacillus thuringiensis subsp. kurstaki at various stages of their development significantly increased development time to the pupal stage and reduced pupal size and number of eggs laid per female, but did not affect the proportion of embryonated eggs. The changes in larval development time, pupal weight and fecundity depended on the larval stage that was treated. Exposure of fourth instars delayed larval development and reduced only male pupal weights with no effects on fecundity. Exposure of sixth instars delayed larval development to a lesser extent than exposure of fourth instars but had a pronounced effect on weight of both male and female pupae. The effect on pupal weight was sex dependent, as males tended to be more affected than females. The reduction in male pupal weight did not appear to influence fecundity, because the effect of exposure was explained by the change in female pupal weight. Effects on larval growth and pupal weight were proportional to the dose that was ingested during exposure, and were observed at doses as low as one-tenth of the LD₅₀. Ingestion of an LD₅₀ caused a 29 or 45% delay in development of, respectively, female or male larvae when exposed as fourth instars and a 30% reduction in female pupal weight when larvae were exposed as sixth instars.     

Body size and cell size in Drosophila: the developmental response to temperature

In Drosophila, like most ectotherms, development at low temperature reduces growth rate but increases final adult size. Cultures were shifted from 25 degrees C to low (16.5 degrees C) or to high (29 degrees C) temperature at regular intervals through larval and pupal stages, and the flies of both sexes showed an increase or decrease, respectively, in the size of thorax, wing and abdominal tergite. Size changes in the wing blade resulted from changes in the size of the epidermal cells (with only a small increase in cell number in males reared at low temperature). The temperature-shifts became less effective as they were made at successively later developmental stages, demonstrating a cumulative effect of temperature on adult size. The thorax and wing develop from the same imaginal disc, with most cell division occurring in larval stages, but they differ in timing of temperature sensitivity, which extends only to pupariation or into the late pupal stage, respectively. Growth of the adult abdomen occurs largely after pupariation but its size is temperature-sensitive through both larval and pupal stages. We discuss growth control in Drosophila and the likely effects of temperature on food assimilation, growth efficiency and allocation of nutrients to the production of different tissues.     

Genome‐wide microarray screening for Bombyx mori genes related to transmitting the determination outcome of whether to produce diapause or nondiapause eggs

The bivoltine silkworm Bombyx mori (Lepidoptera: Bombycidae) exhibits a maternally controlled embryonic diapause. Maternal silkworms decide whether to lay diapause or nondiapause eggs depending on environmental factors such as the temperature and photoperiod during the egg and larval stages, and then induce diapause eggs during the pupal stage. However, little is known about the molecular mechanism that conveys the outcome of whether to produce diapause or nondiapause eggs from the egg or larval stages to the pupal stage. This study used microarray analysis to investigate differentially expressed genes in the larval brains of diapause‐ and nondiapause‐egg producers, to which bivoltine silkworms were destined by thermal or photic stimulation during the egg stage. The cytochrome P450 18a1 and Krüppel homolog 1 genes were upregulated in producers of diapause eggs compared with those of nondiapause eggs under both experimental conditions. Cytochrome P450 18a1 encodes a key enzyme for steroid hormone inactivation and Krüppel homolog 1 is an early juvenile hormone‐inducible gene that mediates the repression of metamorphosis. The upregulation of these genes during the larval stage might be involved in the signaling pathway that transmits information about the diapause program from the egg stage to the pupal stage in the silkworm.     

Factors affecting size, longevity and fecundity in the reindeer oestrid flies Hypoderma tarandi (L.) and Cephenemyia trompe (Modeer)

1. Laboratory reared reindeer oestrid flies Hypoderma tarandi and Cephenemyia trompe (Diptera: Oestridae) were weighed to determine progressive weight loss and death weights at treatments with various temperature and humidity conditions.
2. Four individual measurements of size were taken: larval weight, wet weight of newly eclosed flies, wing length, and weight of flies after dehydration and fat extraction. In H. tarandi, males were bigger than females (except for wing length), whereas the reverse was true for C. trompe .
3. Size variation was not significantly related to conditions (temperature, humidity, duration) during the pupal stage, but individual reindeer produced flies (both species) of different mean sizes. These size differences were not correlated with larval burden (= number of larvae per individual host), but are hypothesized to be connected to unknown host quality factors.
4. Longevity of flies kept in vials and subjected to various temperature and humidity conditions revealed that C. trompe lived significantly longer than H. tarandi (range: 4–44 and 1.2–27 days, respectively) at 5–33 °C. Male H. tarandi survived longer than females; female C. trompe survived longer than males. Longevity was not significantly correlated to any of the size measures.
5. Most flies had a large portion of their fat reserves left at death.
6. In H. tarandi , mean number of eggs was 609 ± SD 73 (range 354–772, n = 119). Egg number was slightly dependent on larval size, but not on wet weight of newly eclosed flies or wing length. In C. trompe , mean number of eggs was 960 ± SD 208 (range 493–1349, n = 31).
7. The possible adaptive value of large size in oestrids is questioned. Benefits of flexibility in size in oestrids are hypothesized.     

双委夜蛾气味结合蛋白AdisOBP6的原核表达抗体制备及表达谱分析

【目的】本研究旨在对双委夜蛾Athetis dissimilis气味结合蛋白OBP6进行原核表达、抗体制备以及表达谱分析,便于今后对AdisOBP6功能展开研究。【方法】利用生物信息学软件分析AdisOBP6蛋白结构特征;利用通过原核表达获得的重组蛋白4次免疫新西兰大白兔,制备AdisOBP6抗体;采用荧光定量PCR和Western blot技术检测AdisOBP6在双委夜蛾雌雄成虫触角、不同时期精巢以及受精卵和未受精卵中的表达情况。【结果】同源建模预测显示,AdisOBP6具有6个保守半胱氨酸残基和7个α-螺旋结构,并 折叠成一个结合口袋。原核表达结果显示,在20℃下0.5 mmol/L IPTG诱导重组蛋白表达量最多、最稳定。4次免疫重复中,抗体效价分别超过1∶512 000, 1∶512 000, 1∶512 000和1∶64 000。Westernblot检测获得的抗体与AdisOBP6蛋白能够特异性结合。荧光定量PCR结果显示,AdisOBP6在双委夜蛾精巢中的表达量远高于在触角中的,在幼虫期精巢中AdisOBP6就已经开始表达,在蛹期精巢中表达量低于在幼虫期的, 成虫羽化后精巢中的表达量逐渐进入高峰,在未受精卵和受精卵中几乎不表达或表达量极低。Western blot分析发现,AdisOBP6蛋白也主要在精巢中表达,在雌雄成虫触角、受精卵和未受精卵中表达量很低。【结论】本研究实现了AdisOBP6的原核表达,成功地制备了抗体;并证实AdisOBP6在双委夜蛾精巢中大量表达,成虫羽化后表达量达到高峰,推测AdisOBP6可能参与了授精过程。本研究结果为今后进一步研究AdisOBP6的功能奠定了基础。     

Correlated responses to selection for developmental rate inTribolium

Selection for fast and slow development in flour beetlesTribolium castaneum (CS) andT. confusum (CF) has resulted in a number of correlated responses in other characters. In general, the fast selected lines of both species have responded similarly with respect to these other traits. The fast lines are characterized by decreased length of time spent in all juvenile stages, decreased size, and essentially the same productivity and hatchability of eggs as the synthetic stocks. In CS-fast there was a substantial decrease in larval survival and an increase in the frequency of abnormal adults. The latter observations did not hold for the CF-fast line.The CS-slow line, on the other hand, exhibited an increased duration of egg, larval and pupal stages, increased size (at least as determined by pupal weight), and decreased hatchability and larval survival. Productivity also decreased somewhat when measured as the number of eggs per female per day. The frequency of abnormal beetles was also higher in this line than in the foundation stock.Fitness, as measured by competitive ability, decreased in all selected lines, supporting the notion of an intermediate optimum for developmental rate in the foundation populations.No evidence was found to support the existence of a compensation mechanism for developmental rate such as that found inDrosophila, whereby faster larval development is compensated for by an increase in the length of the pupal stage.     

Larvicidal activity of Bacillus thuringiensis subsp. israelensis in the dipteran Haematobia irritans

A strain of Bacillus thuringiensis subsp. israelensis was found to be larvicidal to horn flies, Haematobia irritans (L. [Diptera:Muscidae]). The toxic activity was particulate, appeared during sporulation, and could be prevented by the addition of streptomycin before sporulation. Density gradient centrifugation in Renografin was used to separate endospores, crystals, and low-density particulate matter (fraction 3) from sporulated preparations. Larvicidal activity was restricted to purified crystals and fraction 3, indicating that delta-endotoxin of B. thuringiensis subsp. israelensis was active against horn fly larvae. Purified crystals produced mortality during larval feeding stages, but not pupal stages. Fraction 3 produced significant mortality during both larval and pupal stages. The mortality data indicated the presence of at least two dipteran-active toxins.     

CHANGES IN ACTIVITIES OF CREATINE KINASE, ARGININE KINASE AND THEIR MULTIENZYME FORMS DURING EMBRYONIC AND LARVAL DEVELOPMENT OF SEA URCHINS

The presence and the changes of CPK and APK have been studied during larval development through metamorphosing of Hemicentrotus pulcherrimus, Anthocidaris crassispina and Pseudo-centrotus depressus . While no CPK activity was found in the unfertilized eggs and the embryos of early developmental stages, APK was quite active throughout these stages. At the late 8-armed pluteus stage just prior to metamorphosis, CPK first became active. Electrophoretically this CPK was identical with one of three CPK forms of sperm, tube feet and esophagus but not with two CPK forms of lantern muscle. APK in the unfertilized eggs and early embryos was electrophoretically separated into two distinct molecular forms, one of which disappeared during the late larval stages. The persisting form of larval APK was identical with a single APK form present in the adult muscular tissues.     

Larvicidal activity of Bacillus thuringiensis subsp. israelensis in the dipteran Haematobia irritans.

A strain of Bacillus thuringiensis subsp. israelensis was found to be larvicidal to horn flies, Haematobia irritans (L. [Diptera:Muscidae]). The toxic activity was particulate, appeared during sporulation, and could be prevented by the addition of streptomycin before sporulation. Density gradient centrifugation in Renografin was used to separate endospores, crystals, and low-density particulate matter (fraction 3) from sporulated preparations. Larvicidal activity was restricted to purified crystals and fraction 3, indicating that delta-endotoxin of B. thuringiensis subsp. israelensis was active against horn fly larvae. Purified crystals produced mortality during larval feeding stages, but not pupal stages. Fraction 3 produced significant mortality during both larval and pupal stages. The mortality data indicated the presence of at least two dipteran-active toxins.     

The effect of NeemAzal-T/S®, a commercial neem product, on different developmental stages of the common greenhouse whitefly Trialeurodes vaporariorum Westwood (Hom., Aleyrodidae)

The effects of NeemAzal-T/S^®, a commercial neem product, on different life development stages of the common greenhouse whitefly Trialeurodes vaporariorum Westwood (Hom., Aleyrodidae) were tested in laboratory and greenhouse experiments. Treatment of eggs of T. vaporariorum did not affect either larval emergence or the time until larval emergence. However, the proportion of pupal formation (based on the numbers of emerged larvae) was significantly reduced after a treatment of the eggs 3, 5 and 7 days after oviposition, respectively. The proportion of emerged adults (based on the number of formed pupae) was only significantly reduced when eggs had been treated 5 and 7 days after oviposition. In all neem treatments the time until adult emergence was significantly delayed. A neem treatment of early larval instars of T. vaporariorum resulted in a significantly reduced proportion of pupal formation, but subsequent adult emergence (based on the number of formed pupae) was not affected. Treating early larval instars of T. vaporariorum significantly prolonged the time until adult emergence. A neem treatment of T. vaporariorum pupae significantly reduced the proportion of emerged adults but had no effect on the time until adult emergence. Exposing female whiteflies to fresh, 24-h-old and 72-h-old residues of NeemAzal-T/S^® had no effect on the mortality of the insects, though the number of eggs laid per female was significantly reduced in T. vaporariorum exposed to fresh neem residues. These findings are discussed within the context of integrated control of whiteflies in the greenhouse environment.     

The effects of temperature on development of the large narcissus fly (Merodon equestris)

Eggs, larvae, pupae and adults of the large narcissus fly (Merodon equestris) were reared at a series of constant temperatures between 9–24°C. Egg development required from 37 days at 9°C to 7 days at 21.5°C. The low-temperature threshold for development was 6.7°C. Larvae reared at 1424°C were fully-grown after 18 weeks, but it took much longer for such insects to pupate, and adult flies emerged only after about 45 weeks of development. Large narcissus flies enter diapause during the larval stage and overwinter as fully-fed larvae, forming pupae in the following spring. Post-winter pupation and pupal development took from 169 days at 10°C to 36 days at 21.5°C. Of this, pupal development required from 91 days at 10°C to 19 days at 21.5°C. The low-temperature threshold for post-winter pupation and pupal development was 7.1°C, and for pupal development alone, 7.2°C. Females maintained at or below 19°C laid few eggs, whereas some females kept at or above 21.5°C laid more than 100 eggs (mean 69 ± 36). Approximately 50% of females maintained at or above 21.5°C laid less than 10 eggs during their lifetime. The mean egg-laying time was 6 to 9 days. Although temperatures at or below 19°C inhibited mating, once a female had mated, such temperatures did not prevent oviposition.     

The ribosomal proteins of Drosophila melanogaster. II. Comparison of protein patterns of ribosomes from larvae, pupae and adult flies by two-dimensional polyacrylamide gel electrophoresis

Summary The ribosomal proteins from two larval and two pupal stages, within 24 hours before and after pupation, respectively, and adult flies were extracted and compared by two-dimensional polyacrylamide gel electrophoresis. This technique resolved 53 larval, 50 pupal and 52 adult ribosomal proteins, forming a complex pattern. Some proteins were found only in one stage or the other. At present it is not possible, however, to classify these proteins as stage-specific. Some spots showed considerable increase in their staining intensities from one stage to the other, whereas, at the same time, other spots faded. In the ribosomal protein pattern of adult flies 3 proteins showed altered electrophoretic mobilities as compared to earlier developmental stages.Hormones involved in insect development, epigenetic control and non-ribosomal proteins are discussed as possible causes of the variations in the ribosomal protein composition.