Upregulation of two actin genes and redistribution of actin during diapause and cold stress in the northern house mosquito, Culex pipiens

Two actin genes cloned from Culex pipiens L. are upregulated during adult diapause. Though actins 1 and 2 were expressed throughout diapause, both genes were most highly expressed early in diapause. These changes in gene expression were accompanied by a conspicuous redistribution of polymerized actin that was most pronounced in the midguts of diapausing mosquitoes that were exposed to low temperature. In nondiapausing mosquitoes reared at 25 degrees C and in diapausing mosquitoes reared at 18 degrees C, polymerized actin was clustered at high concentrations at the intersections of the muscle fibers that form the midgut musculature. When adults 7-10 days post-eclosion were exposed to low temperature (-5 degrees C for 12 h), the polymerized actin was evenly distributed along the muscle fibers in both nondiapausing and diapausing mosquitoes. Exposure of older adults (1 month post-eclosion) to low temperature (-5 degrees C for 12 h) elicited an even greater distribution of polymerized actin, an effect that was especially pronounced in diapausing mosquitoes. These changes in gene expression and actin distribution suggest a role for actins in enhancing survival of diapausing adults during the low temperatures of winter by fortification of the cytoskeleton.     

Oxygen consumption in relation to sorbitol utilization at the termination of diapause in eggs of the silkworm,Bombyx mori

Rates of oxygen consumption were followed throughout the entire period of diapause in eggs of Bombyx mori. In non-diapause eggs at 25 degrees C, O(2) uptake was divisible into three phases, corresponding to morphogenetic processes. In diapause eggs at 25 degrees C, O(2) uptake showed a peak (100 &mgr;l/g eggs/h) at 1 day and then suddenly dropped to reach a level of 8-10 &mgr;l/g eggs/h at 10 days and thereafter. To break diapause, eggs were exposed to 5 degrees C for varying periods. When O(2) uptake was measured at 5 degrees C, it remained at 6 &mgr;l/g eggs/h. When eggs were chilled for increasing periods and O(2) uptake was measured immediately after warming to 25 degrees C, the rates increased after a lag phase. In HCl-treated eggs, O(2) uptake increased immediately after acid-treatment. In all cases, highly increasing O(2) uptake at 25 degrees C coincided with termination of diapause. These results were discussed in relation to sorbitol utilization at the termination of diapause.     

Population growth and development of the psocid Liposcelis rufa (Psocoptera: Liposcelididae) at constant temperatures and relative humidities

We investigated the effects of eight temperatures (22.5, 25.0, 27.5, 30.0, 32.5, 35.0, 37.5, and 40.0 degrees C) and four relative humidities (43, 55, 63, and 75%) on population growth and development of the psocid Liposcelis rufa Broadhead (Psocoptera: Liposcelididae). L. rufa did not survive at 43% RH, at all temperatures tested; at 55% RH, at the highest four temperatures; and at 63% RH and 40.0 degrees C. The greatest population growth was recorded at 35.0 degrees C and 75% RH (73-fold growth). At 40.0 degrees C, L. rufa populations declined or barely grew. L. rufa males have two to four nymphal instars, and the percentages of males with two, three, and four instars were 31, 54, and 15%, respectively. Female L. rufa have two to five instars, and the percentages of females with two, three, four, and five instars were 2, 44, 42, and 12%, respectively. The life cycle was shorter for males than females. We developed temperature-dependent developmental equations for male and female eggs, individual nymphal, combined nymphal, and combined immature stages. The ability of L. rufa to reproduce at a relative humidity of 55% and temperatures of 22.5-30.0 degrees C and at relative humidities of 63-75% and temperatures of 22.5-37.5 degrees C, in addition to being able to survive at 40.0 degrees C, suggests that this species would be expected to have a broader distribution than other Liposcelis species. These data provide a better understanding of L. rufa population dynamics and can be used to help develop effective management strategies for this psocid.     

Effects of photoperiod and temperature on seasonal morph development and diapause egg oviposition in a bivoltine race (Daizo) of the silkmoth,Bombyx mori L

A bivoltine race (Daizo) of the silkmoth, Bombyx mori, has two types of adults which are classified on the basis of the number of dark-brown and brown scales distributed along the outer lines of the anterior wings. Development of these two types of adults is determined by photoperiod and temperature experienced during the embryonic and larval stages. The two types of adults are thought to be seasonal (summer and autumn) morphs. All adults developed under short days at low temperature (15 degrees C and 23 degrees ) were classified as summer morphs and oviposited only nondiapause eggs (ND-eggs), and those that developed under long days at high temperature (28 degrees C and 25 degrees C) were autumn morphs ovipositing only diapause eggs (D-eggs), only ND-eggs or a mixture of D- and ND-eggs. Larval exposure to long days at 25 degrees C shifted the wing pattern towards the autumn morphs and decreased the incidence of female adults ovipositing only D-eggs. Larval exposure to long days elicits an opposite effect on D-egg oviposition to that observed when long days are received only during the embryonic stage.     

The Race for Females: The Mating System of the Red Mason Bee, Osmia rufa (L.) (Hymenoptera: Megachilidae)

The ecology of a species strongly influences the strategies with which males and females maximize their lifetime reproductive success. When males and females do not invest equally in offspring, the sex with the higher parental investment becomes a rare resource for the other. The spatial and temporal distribution of the limiting sex forms the basis of the mating system. In nest-constructing Aculeata such as the red mason bee, Osmia rufa, females perform intensive brood care, whereas males do not invest in their offspring but instead compete for access to mates. Receptive females of this species are widely distributed and do not assemble at certain places. Therefore, territorial behavior is not an advantageous mating tactic for males, which instead search for females within individual home ranges usually centered around food plants. The unmarked or defended home ranges of different males may completely overlap. Competitive searching leads to a random distribution of matings among males that is largely independent of body size. The mating system of O. rufa can be described as scramble competition polygyny.     

Larval diapause duration and fat metabolism in three geographical strains of the blow fly,Calliphora vicina

Diapausing larvae of the blow fly, Calliphora vicina, from three geographical strains exposed, as adults, to short days, were maintained under identical conditions (darkness, 11-12 degrees C) and examined for changes in wet weight, dry weight, water and fat content during diapause development to the emergence of post-diapause adults. Larvae produced by flies originating from northern Finland (Nallikari, 65 degrees N) showed a longer, more intense, diapause than those from localities further south (Edinburgh, Scotland, 55 degrees N and Barga, Italy, 44 degrees N), but all three strains showed similar rates of loss of the parameters measured. This was also the case for post-diapause adults, flies of the Barga strain with its relatively short diapause emerging with greater residual fat reserves than flies from the Edinburgh or Nallikari strains with their more protracted diapause. It was concluded that the rates of water and fat loss were functions of the conditions used for diapause larval maintenance (probably temperature) rather than the maternally programmed degree of diapause incidence, or of its 'depth' or 'intensity'.     

The role of frass and cocoon volatiles in host location by Monodontomerus aeneus, a parasitoid of Megachilid solitary bees

Monodontomerus aeneus (Fonscolombe) is a parasitic wasp that oviposits on the prepupae and pupae of Osmia cornuta (Latreille) and other solitary bee species. A two-armed olfactometer was used to test the olfactory attractiveness of O. cornuta prepupae, cocoon, and larval frass to female M. aeneus. Both cocoon and frass attracted the female parasitoids, but frass alone was more attractive than the cocoon and the cocoon with frass was more attractive than frass alone. Female parasitoids were not attracted by the host prepupa. M33 (methanol) was the organic volatile most emitted by cocoons and m61 (acetic acid) was the compound most emitted by frass. However, cocoons showed higher emission for almost all compounds, including m61 (acetic acid). Although acetic acid alone attracted M. aeneus, a complex volatile signal is probably involved in the attraction process because the ratio of acetic acid and acetaldehyde characteristic of the frass was more attractive than other ratios.     

Seasonal change of the thermal response in relation to myo-inositol metabolism in adults of Aulacophora nigripennis (Coleoptera Chrysomelidae)

The thermal response of Aulacophora nigripennis adults in relation to myo-inositol metabolism changed seasonally. Myo-inositol accumulation was stimulated at 15 and 5 degrees C in October (autumn), but only 5 degrees C was effective for the accumulation in December (early winter). In February (mid winter), myo-inositol degradation was prominent at both temperatures. Myo-inositol metabolism of this beetle thus gradually shifts from synthesis to degradation as the season progresses. This may be related to the progress of diapause development and may contribute to the seasonal profile of the myo-inositol pool in a natural population. Post-diapause adults in March (spring) and June (early summer) regained the ability for myo-inositol accumulation at 5 degrees C. Thus, there are at least two types of myo-inositol accumulation in this beetle; one is under the control of diapause and the other responds directly to environmental conditions independently of diapause.     

Duration of prepupal summer dormancy regulates synchronization of adult diapause with winter temperatures in bees of the genus Osmia

Osmia (Osmia) bees are strictly univoltine and winter as diapausing adults. In these species, the timing of adult eclosion with the onset of wintering conditions is critical, because adults exposed to long pre-wintering periods show increased lipid loss and winter mortality. Populations from warm areas fly in February-March and are exposed to longer growth seasons than populations from colder areas, which fly in April-May. Given their inability to produce an extra generation, early-flying populations should develop more slowly than late-flying populations and thus avoid the negative consequences of long pre-wintering periods. In this study we compare the development under natural and laboratory conditions of phenologically-distinct populations in two Osmia species. Early-flying populations took ～2 months longer to develop than late-flying populations. Differences between populations in larval and pupal period duration were very small, whereas the prepupal period was much longer in early-flying populations. In contrast to the larval and pupal stages, the prepupal stage showed a non-linear response to temperature, was strongly affected by thermoperiod, and exhibited minimum respiration rates. Coupled with other lines of evidence, these results suggest that the prepupal period in Osmia corresponds to a summer diapause, and its duration may be under local selection to synchronize adult eclosion with the onset of winter temperatures. We discuss the implications of our results relative to current expectations of global warming.     

The antioxidant system in diapausing and active red mason bee Osmia bicornis

The red mason bee Osmia bicornis L. (Hymenoptera: Megachilidae) belongs to a group of insects that undergo an obligatory diapause in the imago stage. Red mason bees (O. bicornis) consume oxygen during overwintering diapause, and reactive oxygen species can be produced despite substantial inhibition of metabolism. These are first studies to investigate the antioxidant system in diapausing red mason bees and to compare the antioxidant systems of overwintering and active imagines. The present study analyzes total antioxidant status, glutathione and ascorbic acid levels, and the activity of superoxide dismutase, catalase and peroxidase in overwintering (October to March) and active (April) female and male O. bicornis. Diapause phases (prediapause, diapause and post‐diapause) cannot be distinguished based on the parameters of the antioxidant system of the bees. During overwintering, a significant decrease is noted only in ascorbic acid content. The remaining antioxidants remain fairly stable, which indicates the absence of oxidative stress in diapausing specimens. The analyzed parameters distinguish diapausing bees from active insects. Excluding total antioxidant status, the evaluated parameters are significantly higher in active individuals than in overwintering specimens. Sex‐related differences are found only for catalase, the level of which is consistently higher in males than in females.     

Low temperature storage of larvae and synchronization of adult emergence in the predatory midge Aphidoletes aphidimyza.

Diapause larvae of Aphidoletes aphidimyza were stored at a temperature of 3 degrees C under continuous darkness for up to 7 months with survival rates above 50%; after storage for 1 year the survival rate dropped to 12%. Diapause was terminated in the majority of individuals within 120 days of chilling under storage conditions. Brief exposure (10-60 s) to the vapor of n-hexane appeared to be a useful alternative to chilling for the termination of diapause. The larvae with terminated diapause required, on average, an additional 31 days at 22 degrees C and long-day conditions in order to reach the adult stage. The 10-90% adult emergence spanned a period of 21.1 days. When the larvae with terminated diapause were exposed to 30 degrees C for 1 week after the end of low temperature storage, the survival rate was not affected, the average "time-to-adult" shortened moderately to 28 days, and the synchrony of adult emergence improved considerably to 10 days. Low temperature storage of nondiapause larvae resulted in a decrease in survival from 98 to 31% during the first 60 days of storage. Nondiapause larvae did not enter diapause during low temperature storage and, as a consequence, the adults emerged relatively rapidly (after 14-15 days) and synchronously (within 2-3 days) after the end of storage. Directions for future research, which might bring further improvement in low temperature storability and synchrony of adult emergence in A. aphidimyza, are proposed.     

Physiology of diapause and cold hardiness in the overwintering pupae of the fall webworm Hyphantria cunea (Lepidoptera: Arctiidae) in Japan

The fall webworm Hyphantria cunea Drury, which was accidentally introduced to Japan in 1945, overwinters on the ground in pupal diapause. Diapause termination, as indicated by the respiration rate and the period required for adult emergence, began in March and ended in April. Cold hardiness (the ability to survive exposure to -15 degrees C) decreased linearly with diapause development from November to the following April under field conditions. Cold hardiness of diapause pupae (DP) decreased as the acclimation temperature decreased from 15 to -10 degrees C, whereas cold hardiness of non-diapause pupae (NDP) remained high as the acclimation temperature decreased from 5 to -5 degrees C. However, H. cunea in Japan can survive exposure to -5 degrees C for two weeks, whether it is in a diapause or non-diapause state. Trehalose was the main sugar detected in the body, but its level was less than 0.8%. Trehalose levels increased in field-collected pupae from January to March. DP accumulated less trehalose than NDP, as the acclimation temperature was decreased from 5 to -5 degrees C. The alanine content in field-collected pupae increased from November to February. Both diapause and low temperature caused an accumulation of alanine. These results suggest that under field conditions, overwintering pupae of H. cunea in Japan do not accumulate high levels of sugars and polyols and do not develop a high level of cold hardiness. Furthermore, DP do not accumulate high levels of sugars and polyols and their ability to survive exposure to -15 degrees C is not greater than that of NDP. The physiological and biochemical bases of diapause in H. cunea from Japan are discussed.     

Temperature-dependent activation of ERK/MAPK in yolk cells and its role in embryonic diapause termination in the silkworm Bombyx mori

The silkworm Bombyx mori requires 2-3 months of low temperature (5 degrees C) to terminate embryonic diapause. The molecular mechanisms, however, are unknown. Extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK) is temperature-dependently activated in the yolk cells of diapausing eggs after 45 days at 5 degrees C, coincident with the acquisition of developmental competence of the embryos at 25 degrees C. Yolk cell granulation and dissociation also begin in diapause eggs incubated at 5 degrees C for 45 days. We used dechorionated egg culture as a model system of diapause termination and observed that both yolk cell dissociation and embryonic development are inhibited by MAPK-ERK kinase (MEK) inhibitor U0126. Therefore, we suggest that ERK in yolk cells has a role in regulating changes in yolk morphology and termination of embryonic diapause in B. mori.     

Cold-Hardiness and Supercooling Capacity in Diapausing and Nondiapausing Stages of the Cabbage Root Fly Delia radicum

Supercooling point (SCP) values and cold-hardiness were measured in individual ontogenetic stages of Delia radicum (Diptera:Anthomyiidae) in various physiological states (winter diapause, summer quiescence, and normal development). Winter diapause-destined mature third-instar larvae had a lower SCP (-9.9 degrees C) than their nondiapause counterparts (-5.2 degrees C), and more of them survived exposure to -10 degrees C for 5 h to pupariation and adult emergence. Values of SCPs were equal in both diapause and nondiapause states of prepupal and pupal stages. The lowest SCP (ca. -20 degrees C) was found in the stage of phanerocephalic pupa (PCP) regardless of the physiological state. The cold-hardiness of PCP corresponded with a low SCP value only in diapausing pupae stored for 80 days at 3 degrees C and in pupae which had terminated their diapause and whose further development was inhibited by storage at low temperatures (3 degrees C). Such pupae survived exposure to temperatures close to their SCP (14 days at -17 degrees C). However, this high cold-hardiness was only acquired after some time and/or exposure to 3 degrees C, as the PCP at the beginning of diapause showed significantly impaired cold-hardiness despite the fact that their SCP was low. The cold-hardiness of nondiapausing PCP did not correspond at all to that of low SCP, as no pupa survived the exposure to -17 degrees C for 1 day; survival rates at temperatures of -13.5 and -10 degrees C were also remarkably lower than those in diapausing pupae. Cold-hardiness in D. radicum was closely connected with the diapause syndrome but the changes in SCP value corresponded rather with the ontogeny of this insect. Copyright 1993, 1999 Academic Press.     

Variations of hydrogen peroxide and catalase expression in Bombyx eggs during diapause initiation and termination

For diapause eggs of the silkworm, Bombyx mori, diapause initiation is prevented with hydrochloric acid (HCl) at around 20 h post-oviposition while diapause status is terminated with chilling around 5°C. To investigate whether hydrogen peroxide (H(2)O(2)) and catalase expression are involved in diapause initiation and termination, the concentration of H(2)O(2), relatively higher levels of catalase mRNA and activity of catalase were compared between (1) 20-h-old diapause eggs and the HCl-treated diapause eggs, and (2) 10-day-old diapause eggs and the 5°C-chilled diapause eggs. Compared to diapause eggs, the HCl-treated eggs had significantly higher H(2)O(2) concentrations (up from approximately 1-3 μmol/g fresh mass to 5-8 μmol/g fresh mass), higher relative level of catalase mRNA (up from 0 to 35.2%) and higher catalase activity (up from 2.51 units/mg protein to 4.97 units/mg protein) at 96 h post-treatment. On the other hand, the 5°C chilling resulted in significant increases of H(2)O(2) concentration (up from 0.79 μmol/g fresh mass to 5.57 μmol/g fresh mass), relative level of catalase mRNA (up from 0 to 71.4%) and catalase activity (up from 0.88 units/mg protein to 3.42 units/mg protein) within 120 days. The results obtained in this work suggest that variations of H(2)O(2) and catalase expression in Bombyx eggs are involved in diapause initiation and termination.     

Role of the Corpus Allatum in the Control of Adult Diapause in the Blow Fly,Protophormia terraenovae

Adults of the blow fly, Protophormia terraenovae developed their ovaries promptly after emergence under long-day conditions at 25 degrees C, although they entered diapause under short-day conditions at 20 degrees C. Application of a juvenile hormone analog (JHA), implantation of the corpus allatum (CA) from reproductive adults, or transection of the nervus corporis allati induced ovarian development under diapause-inducing conditions. Removal of the CA suppressed ovarian development under diapause-averting conditions, and JHA application induced ovarian development in allatectomized adults. Therefore adult diapause in P. terraenovae is caused by a reduction of juvenile hormone secretion from the CA, and the endocrine activity of the CA is inhibited through a nervous pathway by the brain in diapause adults. Copyright 1997 Elsevier Science Ltd. All rights reserved     

Effect of thermoperiod on diapause intensity in Pyrrhocoris apterus (Heteroptera Pyrrhocoridae)

The intensity of adult diapause in Pyrrhocoris apterus was measured in two series of experiments as the duration of pre-oviposition period at a constant temperature of 25 degrees C after transfer from short (12L:12D) to long day conditions (18L:6D). Higher diapause intensity was induced with a thermoperiod than at constant temperatures. After the induction throughout larval instars 3-5 and during 4 weeks of adult life at short days and a thermoperiod of 25/15 degrees C the pre-oviposition period was 30+/-4 and 26+/-3 days. After induction at constant 25 degrees C the pre-oviposition period was 22+/-3 and 23+/-4 days, while after induction at constant 20 degrees C it was 17+/-4 and 19+/-4 days. Induction at a lower constant temperature of 20 degrees C was thus followed by a less intense diapause than the induction at a higher constant temperature of 25 degrees C. These counterintuitive results are discussed. The oxygen consumption rate measured at experimental temperatures prior to transfer from short to long days was higher at thermoperiodic conditions than at constant temperatures and it was similar at constant 20 and 25 degrees C. Thus, the oxygen consumption rate measured prior to the transfer was highest (indication of the least intense diapause) in the insects that showed later, after the transfer to long days, the longest pre-oviposition period (indication of the most intense diapause). Within the first two days after transfer to constant 25 degrees C, oxygen consumption rate measured at 25 degrees C decreased in the thermoperiodic insects, while it transiently increased in insects from constant 20 degrees C. Two days and later after the transfer, oxygen consumption rate was similar in all groups. Cold hardiness was not correlated with diapause intensity. The low lethal temperature in diapausing insects was correlated with the night temperature during diapause induction.     

A true summer diapause induced by high temperatures in the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)

Summer diapause in the cotton bollworm, Helicoverpa armigera (Hübner), which prolongs the pupal stage, particularly in males, is induced by high temperatures. In the laboratory, summer-diapausing pupae of H. armigera were induced at high temperatures (33-39 degrees C) with a photoperiod of LD8:16; winter-diapausing and non-diapausing pupae, cultured at 20 degrees C with a photoperiod of LD8:16 and at 27 degrees C, LD16:8, respectively, acted as a control. Retention time of eye spots, weight, and lipid and glycogen levels were compared. At high temperatures, both body weight and energy storage capacity were much higher in summer-diapausing pupae than in non-diapausing pupae reared at 33-39 degrees C. At temperatures (>33 degrees C) high enough to maintain summer diapause, the eye spots of summer-diapausing pupae did not move during the 30-day experiment. However, eye spots of summer-diapausing pupae placed at 30 degrees C began to move about 10 days after they were transferred, significantly later than in non-diapausing pupae reared at 33-39 degrees C or non-diapausing pupae reared at 27 degrees C, which initiated eye spot movement 2 days after pupation. The differences in retention time of eye spots between summer- and winter-diapausing pupae shows that winter diapause is more intense than summer diapause in this insect. The weight loss, and lipid and glycogen metabolism curves indicate that the summer-diapausing pupae's metabolism is very low. We conclude that summer diapause in the cotton bollworm is a true diapause and that the summer diapause enables the cotton bollworm to withstand the high temperatures of summer.     

Osmia cornifrons vitellogenin: cDNA cloning,structural analysis and developmental expression

Osmia cornifrons plays a major role in the pollination of orchards, but basic information on vitellogenin and oocyte development is limited. To better understand vitellogenin in hymenopteran insects, we cloned a cDNA encoding vitellogenin from the hornfaced bee O. cornifrons. Osmia cornifrons vitellogenin cDNA contains 5477 bp with an open reading frame of 1783 amino acid residues, and has a predicted molecular mass of approximately 200.21 kDa and a pI of 6.55. Osmia cornifrons vitellogenin possesses four consensus (RXXR/S) cleavage sites and has conserved DGXR and GL/ICG motifs in the C‐terminus. The deduced amino acid sequence of the O. cornifrons vitellogenin cDNA showed a 66% identity with Megachile rotundata, 53% to Apis mellifera, 51% to Bombus ignitus and 42%–30% with other hymenopteran insect vitellogenins. Phylogenetic analysis showed that O. cornifrons vitellogenin clustered with vitellogenins from Megachilidae, Apidae, Vespidae and Formicidae species but not with those from Pteromalidae, Aphelinidae or Ichneumonidae species. The expression profile of O. cornifrons vitellogenin mRNA during development revealed that O. cornifrons vitellogenin was first detected in the pupal stage and was continuously detected during the adult stage. Interestingly, O. cornifrons vitellogenin mRNA expression was low in mid‐diapause, then gradually increased beginning on day 3 of the newly emerged adult stage, and subsequently declined. These results suggest that the expression level of O. cornifrons vitellogenin mRNA is stage‐specific.