Environmental cycles regulate development time via circadian clock mediated gating of adult emergence

Background

Previous studies have implicated a role for circadian clocks in regulating pre-adult development of organisms. Among them two approaches are most notable: 1) use of insects whose clocks have different free-running periods and 2) imposition of artificial selection on either rate of development, timing of emergence or circadian period in laboratory populations. Using these two approaches, influence of clock on rate of development has been elucidated. However, the contribution of circadian clocks in determining time taken for pre-adult development has remained unclear. Here we present results of our studies aimed to understand this influence by examining populations of fruit flies carrying three different alleles of the period gene and hence having different free-running periods. We tried to achieve similarity of genetic background among the three strains while also ensuring that they harbored sufficient variation on loci other than period gene.

Results

We find that under constant conditions, flies with long period have slower development whereas in presence of light-dark cycles (LD) of various lengths, the speed of development for each genotype is influenced by whether their eclosion rhythms can entrain to them. Under LD 12:12 (T24), where all three strains entrain, they do not show any difference in time taken for emergence, whereas under LD 10:10 (T20) where long period flies do not entrain and LD 14:14 (T28) where short period flies do not entrain, they have slower and faster pre-adult development, respectively, compared to the controls. We also show that a prior stage in development namely pupation is not rhythmic though time taken for pupation is determined by both the environmental cycle and period allele.

Conclusion

We discuss how in presence of daily time cues, interaction of the cyclic environmental factors with the clock determines the position and width of the gate available for a fly to emerge (duration of time within a cycle when adult emergence can occur) resulting in an altered developmental duration from that observed under constant conditions. We also discuss the relevance of genetic background influencing this regulation.

     

Adaptive significance of egg size plasticity in response to temperature in the migrant skipper, Parnara guttata guttata (Lepidoptera: Hesperiidae)

Females of the migrant skipper, Parnara guttata guttata, that are reared under lower temperatures lay smaller eggs. The adaptive significance of egg size plasticity in response to temperature is unknown in this species. We suggest, based on the following experimental results, that P. g. guttata uses temperature as an indirect cue to predict the host condition (leaf toughness) of the next generation. First, larvae were reared under the typical conditions of temperature and photoperiod experienced during the immature stages in the first, second, and overwintering (third) generations (LD 16:8 at 25°C, LD 14:10 at 25°C and LD 14:10 at 20°C). Females reared under LD14:10 at 20°C produced more, smaller eggs than those reared under LD14:10 and LD16:8 at 25°C. Secondly, survival rates of first instar larvae derived from females reared under the three photoperiod/temperature treatments were measured on young soft rice leaves (soft), or tough, old rice leaves (tough). Survival rates of hatchlings reared on soft and tough leaves did not differ when females were reared under LD16:8 and LD14:10 at 25°C. However, hatchling survival was significantly higher on soft than on tough leaves when females were reared under LD14:10 at 20°C. Thirdly, we found that egg size plasticity in response to temperature in P. g. guttata may be a threshold response. Temperatures below 20°C experienced during the immature stages may be effective for production of smaller and more eggs in the overwintering generation of P. g. guttata.     

Host suitability of the Mediterranean flour moth for rearing <Emphasis Type="Italic">Meteorus pulchricornis</Emphasis> (Hymenoptera: Braconidae), a polyphagous endoparasitoid of pest lepidopteran larvae

We investigated the host suitability of the Mediterranean flour moth Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) for a polyphagous koinobiont endoparasitoid Meteorus pulchricornis (Wesmael) (Hymenoptera: Braconidae), a common natural enemy of various pest lepidopteran larvae. The estimated probability of adult wasp emergence was 80% or higher when eggs were laid in nearly fully grown larvae of E. kuehniella (fresh weight, >?20.0 mg). The body size of emerged adult wasps increased with the initial weight of the host larvae at oviposition. The fresh weight of adult wasps reared on E. kuehniella was approximately 60% of that when reared on a natural host Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae), and the lifetime fecundity of wasps reared on E. kuehniella was approximately half of that when reared on S. litura. Ephestia kuehniella was shown to be a positive host candidate for the mass rearing of M. pulchricornis, but further investigation is needed to increase the body size of wasps for more practical use of this species as a biocontrol agent.     

Variations in the tendency to diapause among successive generations of laboratory strains of <Emphasis Type="Italic">Trichogramma</Emphasis> species (Hymenoptera,Trichogrammatidae): Endogenous or exogenous?

Variations in the tendency to diapause among successive generations of laboratory strains reared under constant conditions were reported for many insects and mites. However, as far as we know, the relative importance of the endogenous and exogenous components of this variability has not been investigated experimentally. We have conducted such a study with 12 successive generations of laboratory strains of Trichogramma telengai Sor. and T. principium Sug. et Sor. In total, 12 strains originated from different females and / or reared under different photothermal conditions (temperature of 20 and 25°C and day length of 12 and 18 h) were investigated. The tendency to diapause was estimated by the proportion of the progeny diapausing at temperatures of 13, 14 and 15°C. Although the variations in the tendency to diapause in sequential generations of different strains of the same species were positively correlated, the correlation between different species was also very strong suggesting that endogenous (species-specific) factors are not the main reason for the observed changes. The data for different photothermal conditions of incubation of the studied strains and of their progeny were also correlated suggesting that occasional changes in temperature or day length are not the reason of the changes in the proportion of diapausing progeny. Evidently, the differences in the tendency to diapause among generations of the studied Trichogramma species were mainly caused by some uncontrolled environmental factors. This in no way excludes the possible role of the endogenous processes, but suggests the necessity for a critical analysis of the available data on the “spontaneous” dynamics in the tendency to diapause among successive generations of the laboratory strains of insects and mites.     

Development rate,number of mature oocytes at emergence and adult size ofEncarsia tricolor at constant and variable temperatures

The duration of the development of the aphelinidEncarsia tricolor Föerster (a parasitoid of the aleyrodidTrialeurodes vaporariorum (Westwood), adult size and number of mature oocytes at emergence were determined under constant and variable temperature regimes. Females developed successfully from 14 to 32°C, but a 100% of pupal mortality was observed at 34°C. Males developed successfully from 16 to 28°C and they developed faster than females. Female and male, egg to adult development at constant temperatures ranged from 51.1 (14°C) to 14.3 (28°C) days and from 32.6 (16°C) to 11.8 (28°C) days, respectively. Predictions of the rate of development at variable temperatures were more accurate when made from 2nd and 3rd degree polynomials than from linear regressions. The number of mature oocytes at emergence ranged from 0.1 (30°C) to 2.2 (20°C). FemaleE. tricolor attained the greatest size at 20–24°C. The comparison with literature data shows thatE. tricolor develops faster thanT. vaporariorum at temperatures above 15°C.     

The influence of temperature,photoperiod, and diet on development and reproduction in the four-spot lady beetle <Emphasis Type="Italic">Harmonia quadripunctata</Emphasis> (Pontoppidan) (Coleoptera,Coccinellidae)

This is the first report on the effects of temperature (from 17 to 30°C), photoperiod (from 10 to 16 h of light), and diet (the wheat aphid Schizaphis graminum and the green peach aphid Myzus persicae) on preimaginal developmental rates, adult body mass, preoviposition period, and female fecundity of the predatory lady beetle Harmonia quadripunctata (Pontoppidan) under the laboratory conditions. A constant temperature of 30°C was lethal to the embryos. Judging from the data obtained at 17, 20, and 25°C, the lower temperature threshold for total egg-to-adult development equaled 12.0°C and the respective sum of effective temperatures was 274 degree-days. Preimaginal development was somewhat faster under short-day conditions, the threshold for this quantitative photoperiodic response being approximately 14 h at the rearing temperatures of 20 and 25°C. At a temperature of 25°C, the mean body mass of newly emerged adults was greater for those individuals that during the larval stage had been fed on the green peach aphid (16.4 mg in males and 18.4 mg in females) than in those fed with the wheat aphid (13.8 mg in males and 15.3 mg in females). The preoviposition period (measured from adult emergence to the first egg laid) also depended on temperature and had a lower threshold of 13.4°C and a sum of effective temperatures of 152 degree-days. Maximum fecundity was observed at 25°C and 16-h day length; under these conditions, the oviposition period lasted over 100 d, and the average lifetime fecundity was about 800 eggs per female. Under short-day conditions, females of H. quadripunctata entered reproductive diapause. The photoperiodic threshold for this qualitative response at the temperatures of 20 and 25°C was about 14 h when fed on the peach aphid and 15 h when fed on the wheat aphid. Relatively small body size, low fecundity, and a strong photoperiodic response that hinders rapid adaptation to novel climates probably explain the fact that H. quadripunctata, in contrast to H. axyridis, has not become an aggressive invader.     

The effect of temperature on the development,reproduction, and longevity of two common cyclopoid copepods — Eucyclops serrulatus (Fischer) and Cyclops strenuus (Fischer)

The embryonic and postembryonic development times, time interval between clutches, and longevity of two common cyclopoid copepods — Eucyclops serrulatus and Cyclops strenuus — were studied at constant temperatures (5°, 10°, 15°, 20° and 25 °C), using algae and protozoans as food. Both, E. serrulatus and C. strenuus showed a considerable temperature tolerance and adaptability. Embryonic development times were similar in both species but postembryonic development times were shorter in E. serrulatus. Time intervals between successive clutches were relatively variable at all temperatures in both species. Longevity was shorter in E. serrulatus than in C. strenuus. The embryonic and postembryonic development of E. serrulatus was short relative to that of littoral copepods; the development of C. strenuus was similar or shorter than that of other species of the genus Cyclops.     

Concentration distribution around a growing gas bubble in tissue

This paper presents the concentration distribution around a growing nitrogen gas bubble in the blood and other tissues of divers who surface too quickly, when the ambient pressure through the decompression process is variable and constant. This effort is a modification of Sirinivasan et al. model (1999) [9]. The mathematical model is solved analytically to find the growth rate of a gas bubble in a tissue after decompression in the ambient pressure. Moreover, the concentration distribution around the growing bubble is introduced. The growth process is affected by ascent rate , tissue diffusivity D_T, initial concentration difference ΔC₀, surface tension σ and void fraction ?₀.     

Effect of temperature on queen oviposition and seasonal colony development in <Emphasis Type="Italic">Lasius japonicus</Emphasis> (Hymenoptera: Formicidae)

Although day length plays a critical role in the seasonal development of many temperate insects, this study showed that this variable is less important than temperature for seasonal regulation of an ant’s life cycle. Here, we examined the effects of temperature on queen oviposition and seasonal colony development of Lasius japonicus Santschi. Queens were collected soon after their nuptial flight and were reared under constant laboratory conditions; colony development was analyzed. The percentage of larvae- and pupae-emerged colonies was reduced in low rearing temperatures, indicating that temperature is the primary environmental factor for the regulation of seasonal development. Larval diapause was induced at higher temperatures than the reproductive diapause of queens. Larvae need to enter diapause before winter; nevertheless, eggs that were still unhatched in late autumn would not necessarily be wasted because they could be eaten by queens. Cyclic fluctuation in the egg number was observed. At 25 °C, the second increase in egg number was synchronized with pupation, suggesting a social effect on queen oviposition. In contrast, at 20 and 17.5 °C, the clear second peak of oviposition was found in colonies wherein larvae did not emerge, suggesting that endogenous rhythmicity is involved in the regulation of queen oviposition.     

Biochemical Properties of α-Amylase from Midgut of <Emphasis Type="Italic">Alphitobius diaperinus</Emphasis> (Panzer) (Coleoptera: Tenebrionidae) Larvae

The lesser mealworm, Alphitobius diaperinus (Panzer), is the main insect pest in the poultry industry, thus causing serious damage to production. In this work, the properties of midgut α-amylase from larvae of A. diaperinus were characterized, and its in vitro activity to proteinaceous preparations from different cultivars of common bean (Phaseolus vulgaris) was determined, as well as the amylolitic activity of insects reared on different types of poultry diet. In order to establish some assay conditions, time course and enzyme concentration upon the reaction rate were determined. Product proceeded linearly with time, and the activity was directly proportional to the enzyme concentration. Banding patterns in mildly denaturing electrophoresis showed a single band with apparent molecular weight of 42 kDa. α-Amylase reached optimal temperature at 45°C and pH 5.0 as the optimal one. It maintained 34.6% of the activity after being kept at 60°C for 5 min, and 23%, after 60 min. However, at 80°C, only 14 and 6% remained after 5 and 60 min, respectively. The presence of Ca²⁺ and Na⁺ ions decreased the enzyme activity at concentrations higher than 2 and 100 mM, respectively. The activity was significantly inhibited by some proteinaceous extracts from common bean cultivars, and it declined with increasing proteinaceous concentration. No significant difference was observed when the amylolytic activity was determined in A. diaperinus reared on different poultry diets, offered to broilers in the starter, grower, finisher, and layer phases.     

Sub-lethal effects of pesticide residues in brood comb on worker honey bee (Apis mellifera) development and longevity

Background

Numerous surveys reveal high levels of pesticide residue contamination in honey bee comb. We conducted studies to examine possible direct and indirect effects of pesticide exposure from contaminated brood comb on developing worker bees and adult worker lifespan.

Methodology/Principal Findings

Worker bees were reared in brood comb containing high levels of known pesticide residues (treatment) or in relatively uncontaminated brood comb (control). Delayed development was observed in bees reared in treatment combs containing high levels of pesticides particularly in the early stages (day 4 and 8) of worker bee development. Adult longevity was reduced by 4 days in bees exposed to pesticide residues in contaminated brood comb during development. Pesticide residue migration from comb containing high pesticide residues caused contamination of control comb after multiple brood cycles and provided insight on how quickly residues move through wax. Higher brood mortality and delayed adult emergence occurred after multiple brood cycles in contaminated control combs. In contrast, survivability increased in bees reared in treatment comb after multiple brood cycles when pesticide residues had been reduced in treatment combs due to residue migration into uncontaminated control combs, supporting comb replacement efforts. Chemical analysis after the experiment confirmed the migration of pesticide residues from treatment combs into previously uncontaminated control comb.

Conclusions/Significance

This study is the first to demonstrate sub-lethal effects on worker honey bees from pesticide residue exposure from contaminated brood comb. Sub-lethal effects, including delayed larval development and adult emergence or shortened adult longevity, can have indirect effects on the colony such as premature shifts in hive roles and foraging activity. In addition, longer development time for bees may provide a reproductive advantage for parasitic Varroa destructor mites. The impact of delayed development in bees on Varroa mite fecundity should be examined further.     

Univoltine seasonal cycle and obligate diapause in the noctuid moth <Emphasis Type="Italic">Charanyca trigrammica</Emphasis> Hufn. (Lepidoptera,Noctuidae)

Charanyca trigrammica is a univoltine species over its whole distribution range. To study the diapause-inducing factors, caterpillars were reared at 24°C and day lengths of 12, 13, 14, 15, 16, 17, and 18 h. In all the studied regimes, pupation was not recorded, suggesting an obligate diapause. However, the development was faster under short days. The threshold of this short-day photoperiodic response was ca 17 h. This type of environmental regulation of development may be considered a special feature of diapause occurring in the last larval instars. Hence, the obligate diapause in the last larval instars determines the univoltine cycle of Ch. trigrammica in Belgorod Province of Russia, ensuring successful hibernation of all individuals.     

Metabolism of <Superscript>14</Superscript>C-glycine as a substrate for photorespiration of the leaf at different developmental stages of ephemeroides

The metabolism of ¹⁴C-glycine (a substrate for photorespiration) was studied in the light and in darkness under natural CO₂ concentration (0.03%) in the leaves of ephemeroides Scilla sibirica Haw. and Ficaria verna Huds. at different developmental stages. Using one and the same sample, potential photosynthesis (at 1% CO₂), true photosynthesis (at 0.03% CO₂), and leaf respiratory capacity were measured by the radiometric and manometric methods, respectively. All measurements were performed at 15°C, an average temperature during ephemer growth. It was found that, in the white zone of the Scilla leaf, the rate of CO₂ evolution resulting from metabolization of exogenous ¹⁴C-glycine was similar in the light and in darkness. In the green zone of the Scilla leaf and in the green leaf of Ficaria, both ¹⁴C-glycine absorption and ¹⁴CO₂ evolution were lower in the light as compared with darkness, which is explained by CO₂ reassimilation. In all treatments of both plant species, a specific inhibitor of glycine decarboxylase complex (GDC), aminoacetonitrile (5 mM) suppressed CO₂ evolution by 20–40%. It was concluded that in ephemeroides mitochondrial GDC, responsible for CO₂ evolution in photorespiration, is formed at the earliest stage of leaf development. This indicates that photorespiration can occur simultaneously with the development of the leaf photosynthetic activity. On the basis of the assumption that carbon losses in the form of CO₂ evolved during photorespiration comprise 25% of true photosynthesis, it was calculated that, in ephemer leaves, the highest rates of photorespiration and photosynthesis were attained during flowering when the leaf area was the largest and the rate of dark respiration was reduced by 1.5–2.0 times. The highest rates of dark respiration were observed in the beginning of growth. In senescing leaves by the end of the plant vegetation, potential photosynthesis and true photosynthesis were reduced, whereas dark respiration remained essentially unchanged. It is concluded that the high rates of potential and true photosynthesis are characteristic of ephemeroides when they complete their short developmental program in early spring (at 15°C); theoretically, photorespiration also occurs at a high rate during this period, when this process provides for a defense against the threat of photoinhibition at low temperature and high insolation.     

Alaboratory evaluation ofAnthocoris nemorum andA. nemoralis [Hem.: Anthocoridae] as predators ofPhorodon humuli [Hom.: Aphididae]

Anthocoris nemorum (L.) andA. nemoralis (F.) were reared on apterous hop aphids,Phorodon humuli (Schrank), at 20±0.5°C.A. nemorum andA. nemoralis killed an average of 255 and 174 aphids respectively during nymphal development, and the adults killed 37 and 33 per day respectively. Small anthocorid nymphs selectively killed small aphids but adult anthocorids and 5th instar nymphs killed aphids of every size. Mean development times from emergence to final moult were 22 and 16 days forA. nemorum andA. nemoralis respectively.     

Biological characteristics and thermal requirements of a Brazilian strain of the parasitoid Trichogramma pretiosum reared on eggs of Pseudoplusia includens and Anticarsia gemmatalis

A new strain of the parasitoid Trichogramma pretiosum, was collected in Rio Verde County, State of Goiás, Central Brazil, and designated as T. pretiosum RV. This strain was then found to be the most effective one among several different strains of T. pretiosum tested in a parasitoid selection assay. Therefore, its biological characteristics and thermal requirements were studied, aiming at allowing its multiplication under controlled environmental conditions in the laboratory. The parasitoid was reared on eggs of Pseudoplusia includens and Anticarsia gemmatalis at different constant temperatures within an 18–32 °C temperature range. The number of annual generations of the parasitoid was also estimated at those temperatures. Results have shown that T. pretiosum RV developmental time, from egg to adult, was influenced by all temperatures tested within the range, varying from 6.8 to 20.3 days and 6.0 to 17.0 days on eggs of P. includens and A. gemmatalis, respectively. The emergence of T. pretiosum RV from eggs of A. gemmatalis was higher than 94% at all temperatures tested. When this variable was evaluated on eggs of P. includens, however, the figures were higher than that within the 18–30 °C range (more than 98%), and were also statistically higher than the emergence observed at 32 °C (90.2%). The sex ratio of the parasitoids emerged from eggs of A. gemmatalis decreased from 0.55 to 0.29 at 18–32 °C, respectively. However, for those emerged from eggs of P. includens, the sex ratio was similar (0.73, 0.72 and 0.71) at 20, 28 and 32 °C, respectively. The lower temperature threshold (Tb) and thermal constant (K) were 10.65 °C and 151.25 degree-days when the parasitoid was reared on eggs of P. includens; and 11.64 °C and 127.60 degree-days when reared on eggs of A. gemmatalis. The number of generations per month increased from 1.45 to 4.23 and from 1.49 to 4.79 when the parasitoid was reared on eggs of P. includens and A. gemmatalis, respectively, following the increases in the temperature.     

A generalized theory of particulate electron conduction enzymes applied to cytochrome oxidase. A theory of coupled electron and/or ion transport applied to pyruvate carboxylase

A previously developed theory of particulate electron conduction enzymes was based on a model of an enzyme particle catalyzing the oxidation-reduction of two different substrates at two different enzymatic sites on the same particle with conduction of electrons between the two sites through the enzyme particle. Using the simplifying assumption that the percent reduction of the second substrate is held constant, there was previously shown to be a hyperbolic relationship between the first order rate constant (k′) and the sum (C _x ) of oxidized plus reduced substrate, of the formk′=α/(C _x +β), where α and β are positive constants. It is shown here that if this simplifying assumption is omitted, a positive constant is added to the right hand side of this equation, which describes exactly the experimental data of Smith and conrad on cytochrome oxidase. If electron transport is assumed to be coupled to ion transport, this equation becomesk′=(α/C _x )?γ (where γ is a positive constant) which describes the experimental data of Eadie and Gale on pyruvic carboxylase of yeast. It seems probable that the same theory is applicable to coupled ion-ion transport and coupled electron-electron transport in both membranous systems, and in particulate preparations consisting of membrane fragments.     

Deep simple epicotyl morphophysiological dormancy in seeds of two Viburnum species, with special reference to shoot growth and development inside the seed

Background and Aims

In seeds with deep simple epicotyl morphophysiological dormancy, warm and cold stratification are required to break dormancy of the radicle and shoot, respectively. Although the shoot remains inside the seed all winter, little is known about its growth and morphological development prior to emergence in spring. The aims of the present study were to determine the temperature requirements for radicle and shoot emergence in seeds of Viburnum betulifolium and V. parvifolium and to monitor growth of the epicotyl, plumule and cotyledons in root-emerged seeds.

Methods

Fresh and pre-treated seeds of V. betulifolium and V. parvifolium were incubated under various temperature regimes and monitored for radicle and shoot emergence. Growth of the epicotyl and cotyledons at different stages was observed with dissecting and scanning electron microscopes.

Key Results

The optimum temperature for radicle emergence of seeds of both species, either kept continuously at a single regime or exposed to a sequence of regimes, was 20/10 °C. GA₃ had no effect on radicle emergence. Cold stratification (5 °C) was required for shoot emergence. The shoot apical meristem in fresh seeds did not form a bulge until the embryo had grown to the critical length for radicle emergence. After radicle emergence, the epicotyl–plumule and cotyledons grew slowly at 5 and 20/10 °C, and the first pair of true leaves was initiated. However, the shoot emerged only from seeds that received cold stratification.

Conclusions

Seeds of V. betulifolium and V. parvifolium have deep simple epicotyl morphophysiological dormancy, C_1bB (root)–C₃ (epicotyl). Warm stratification was required to break the first part of physiological dormancy (PD), thereby allowing embryo growth and subsequently radicle emergence. Although cold stratification was not required for differentiation of the epicotyl–plumule, it was required to break the second part of PD, thereby allowing the shoot to emerge in spring.