Identification and up-regulation of three small heat shock proteins in summer and winter diapause in response to temperature stress in Pieris melete.

Small heat shock proteins (sHSPs) are conserved proteins that play key roles in organismal adaptation to adversity stressors. However, little is known about sHSPs during summer diapause. Three sHSP genes: PmHSP19.5, PmHSP19.9, and PmHSP20.0 were identified and cloned from Pieris melete. Sequence alignment and phylogenetic analysis revealed that the three sHSPs have a typical, conserved α-crystallin domain. PmHSP19.5 and PmHSP20.0 were both upregulated in summer diapause (SD) and winter diapause (WD), compared to non-diapause (ND) pupae. All three sHSPs were upregulated and showed similar trends in response to thermal stress. The 0 °C chilling treatment slightly affected sHSP transcripts in ND pupae, whereas both PmHSP19.5 and PmHSP19.9 were upregulated and PmHSP20.0 was downregulated after chilling at 0 °C for 24-96 h in both SD and WD pupae. The transcripts of PmHSP19.5 and PmHSP19.9 were significantly induced at 31 °C for 30 d in SD and WD pupae. The PmHSP20.0 transcript gradually decreased during the SD and WD programs. This is the first time that sHSPs have been linked to both overwintering and summer diapause processes. These findings suggest that sHSPs are involved in both summer and winter diapause maintenance and play a possible key role in temperature stress.

Overexpression of klotho suppresses growth and pulmonary metastasis of osteosarcoma in vivo.

Klotho is originally discovered as an anti-aging gene and knock-out of klotho accelerates aging in mice. Subsequent studies support the anti-carcinogenesis role of klotho in a variety of human malignancies. The present study investigated the role of klotho on growth and metastasis of osteosarcoma cells. The osteosarcoma cells were transduced with lentivirus particles encoding klotho or scramble control. The reconstructed osteosarcoma cells were injected into the femoral medullary cavity of nude mice to establish a xenograft animal model. The anti-tumor properties of klotho were evaluated in terms of tumor growth, apoptosis, glycogen production, and pulmonary metastasis. Lentivirus-mediated overexpression of klotho significantly decreased tumor volume and weight in osteosarcoma mice. Determination of PCNA and Ki67 expression revealed that overexpression of klotho inhibited cell proliferation in tumor tissues obtained from osteosarcoma xenografts. PAS staining also showed that overexpression of klotho significantly decreased the production of glycogen in osteosarcoma. Moreover, TUNEL positive cells were significantly increased after lentivirus-mediated overexpression of klotho. Furthermore, lentivirus-mediated upregulation of klotho reduced the number of pulmonary metastatic lesions in mice compared to control mice. These findings demonstrated that elevated klotho could inhibit osteosarcoma cell growth and pulmonary metastasis in vivo, suggesting that klotho may be a valuable therapeutic target for osteosarcoma.

Effects of starvation on respiratory metabolism and energy metabolism in the cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

Intermittent food shortages are commonly encountered in the wild. To cope with the threat of starvation, insects initiate a suite of behavioral activities and physiological countermeasures. The cotton bollworm, Helicoverpa armigera, is a major agricultural pest worldwide, but how H. armigera modulates its metabolism under starvation remains ambiguous. In the present study, the respiratory rates (V̇O2; ml g-1 h-1) from the third-larval instar to the pupal stage were first determined. Our results highlighted a transient rise during the larval-larval molt and larval-pupal transition, followed by a sharp decline in the pupal stage and, finally, an upward trend before eclosion. When subjected to food deprivation, the starved larvae experienced a significant decline in the rates of O2 consumed and CO2 produced, as well as in respiratory quotient (RQ) values, indicative of severe metabolic depression during starvation and a shift of metabolic substrates with prolonged starvation. For metabolic substrate analysis, an apparent decline in triglyceride and glycogen contents was observed in the starved larvae, and the hemolymph trehalose content was significantly reduced throughout starvation. In addition, comparative transcriptome analysis showed that 48 h of larval starvation caused substantial transcriptional regulations in several energetically costly processes, wherein the marked up-regulations were detected in the pathways of glycolysis and fatty acid metabolism. Overall, our work makes a comprehensive study on the respiratory rate and energy metabolism in the starved H. armigera larvae, and provides a deep insight into the physiological adaptive strategies to alleviate nutritional stress.

[Regulating effect of agricultural landscape pattern on ecological pest control by natural enemies.] / å†œç”°æ™¯è§‚æ ¼å±€å¯¹å®³è™«å¤©æ•Œç”Ÿæ€æŽ§å®³åŠŸèƒ½çš„è°ƒæŽ§ä½œç”¨.

It's well known that regional agricultural landscape pattern can significantly regulate the relationship between plants, herbivorous insects and natural enemies. The improvement of the ecosystem services of regional agricultural landscape pattern has profound impacts on pest control, which has received much interests in current landscape ecology and pest control studies. This review systematically summarized the impacts of agricultural landscape pattern, as well as its succession on the diversity and ecological pests control of important natural enemies. From the perspective of different natural enemies, the impacts of agricultural landscape pattern on natural predators, parasi-toids, beneficial microorganisms and its ecological pests control functions were all recapped. We outlined crop spatial distribution and vegetation habitat management strategies. This review could provide useful insights in improving the ecosystem services of natural enemies in agricultural landscape based on the vegetation habitat management, and offer theoretical support for regional ecological agriculture in response to the strategy of 'chemical pesticide reduction application'.

Molecular characterization of three Hsp90 from Pieris and expression patterns in response to cold and thermal stress in summer and winter diapause of Pieris melete.

Heat shock proteins (Hsps) have been linked to stresses and winter diapause in insects, but whether they are components of summer diapause is still unknown. In this study, complementary DNAs of Hsp90 from Pieris melete, Pieris rapae and Pieris canidia named PmHsp90, PrHsp90 and PcHsp90, respectively, were cloned and sequenced. The deduced amino acid sequence consisted of 718 amino acid residues with a putative molecular mass of 82.6, 82.6 and 82.7 kDa, respectively. The amino acid sequences contained all of the five conserved signature motifs in the Hsp90 family and a bHLH protein folding activity region. The differential expression pattern of PmHsp90 in response to summer diapause and winter diapause, which are related to heat/cold stress, was investigated. Cold stress induced Hsp90 up-regulation in summer and winter diapause pupae, but not in non-diapause individuals. Heat shock up-regulated PmHsp90 gradually with an increase in temperature in summer diapause, and PmHsp90 was rapidly up-regulated in winter diapause. After 30 min heat shock at 39°C, substantial up-regulation of PmHsp90 transcript levels were observed both in summer and winter diapause. However, in non-diapause a relatively stable expression was found under different durations of 39°C heat shock. Compared to the optimal treatment of 18°C for diapause development, a high temperature acclimation of 31°C induced PmHsp90 up-regulation in summer diapause, whereas a low temperature acclimation of 4°C induced up-regulation in winter diapause. The current results indicate that Hsp90 may play an important role in response to heat/cold stress both in summer and winter diapause.

Dynamics of MMP­9, MMP­2 and TIMP­1 in a rat model of brain injury combined with traumatic heterotopic ossification.

The present study aimed to detect early changes in the concentration of matrix metalloproteinase-9 (MMP-9), matrix metalloproteinase­2 (MMP­2) and tissue inhibitor of metalloproteinase­1 (TIMP­1) in a rat model of brain injury combined with traumatic heterotopic ossification (HO). A total of 132 male Sprague­Dawley rats were used to establish the experimental and control groups. Anatomy and sample collection were conducted on postoperative days 1, 2, 3, 4, 5, 6 and 7. Hematoxylin and eosin and immunohistochemical staining were performed for local tissues. MMP­9, MMP­2 and TIMP­1 levels and gene expression level were measured by ELISA and reverse transcription­quantitative polymerase chain reaction. Radiological investigation of the rat lower limbs was conducted at weeks 5 and 10 following modeling to observe the occurrence of HO. The incidence of HO for rats in the experimental group was higher compared with the control group. The serum MMP­9 levels of the experimental group were notably higher on postoperative days 5­7 compared with the control group. The MMP­9 gene expression of the experimental group was higher on postoperative days 3­7 compared with the control group. The TIMP­1 gene expression levels were markedly higher compared with the control group at each time point. Thus, an increase in inflammatory response is closely associated with brain injury, in addition to an increase in the number of inflammatory cells with the incidence of HO. The pathological elevation of MMP­9 and the altered dynamic equilibrium between MMP­9 and TIMP­1 contributed to the degradation, remodeling and calcification of the extracellular matrix, resulting in the induction of osteoblast precursor cells in HO. MMP­9 is a predictive marker of HO.

Life-history responses of the rice stem borer Chilo suppressalis to temperature change: Breaking the temperature-size rule.

Temperature is a key environmental factor for ectotherms and affects a large number of life history traits. In the present study, development time from hatching to pupation and adult eclosion, pupal and adult weights of the rice stem borer, Chilo suppressalis were examined at 22, 25, 28 and 31°C under L18:D 6. Larval and pupal times were significantly decreased with increasing rearing temperature and growth rate was positively correlated with temperature. Larval and pupal developmental times were not significantly different between females and males. The relationship between body weight and rearing temperature in C. suppressalis did not follow the temperature-size rule (TSR), both males and females gained the highest body weight at 31°C. Females were significantly larger than males at all temperatures, showing a female biased sex size dimorphism (SSD). Contrary to Rensch's rule, SSD and body weight in C. suppressalis tended to increase with rising temperature. Male pupae lost significantly more weight at metamorphosis compared to females. We discuss the adaptive significance of the reverse-TSR in the moth's life history.

Synchronous vitellogenin expression and sexual maturation during migration are negatively correlated with juvenile hormone levels in Mythimna separata.

Annual migration of pests between different seasonal habitats can lead to serious crop damage. Reproductive immaturity is generally associated with the migratory process (oogenesis-flight syndrome), but the mechanism of reproductive development during migration varies unpredictably. Here, the vitellogenin gene (MsVg) and three key regulatory enzyme genes (MsJhamt, MsJheh and MsJhe) related to juvenile hormone (JH) synthesis and degradation were identified and characterized in Mythimna separata. The relative expression of MsVg varied significantly in response to seasonal changes and was significantly correlated with stages of ovarian development. The relatively low levels of JH titer did not differ significantly in male moths but slightly increased in female adults during the migratory season, which was consistent with changes in mRNA levels for MsJhamt, MsJheh and MsJhe. JH titer was negatively associated with relative seasonal levels of vitellogenin mRNA transcripts and with ovarian development in migrating M. separata. The synchrony of MsVg expression with sexual maturation highlighted the potential of MsVg transcript levels to serve as an index to monitor the adult reproductive status. In addition, the level of JH and sexual maturity were correlated with the extent of JH in regulating the MsVg expression and reproduction during seasonal northern and southern migration.

Dynamic transcriptome analysis and volatile profiling of Gossypium hirsutum in response to the cotton bollworm Helicoverpa armigera.

In response to insect herbivory, plants emit elevated levels of volatile organic compounds for direct and indirect resistance. However, little is known about the molecular and genomic basis of defense response that insect herbivory trigger in cotton plants and how defense mechanisms are orchestrated in the context of other biological processes. Here we monitored the transcriptome changes and volatile characteristics of cotton plants in response to cotton bollworm (CBW; Helicoverpa armigera) larvae infestation. Analysis of samples revealed that 1,969 transcripts were differentially expressed (log2|Ratio| ≥ 2; q ≤ 0.05) after CBW infestation. Cluster analysis identified several distinct temporal patterns of transcriptome changes. Among CBW-induced genes, those associated with indirect defense and jasmonic acid pathway were clearly over-represented, indicating that these genes play important roles in CBW-induced defenses. The gas chromatography-mass spectrometry (GC-MS) analyses revealed that CBW infestation could induce cotton plants to release volatile compounds comprised lipoxygenase-derived green leaf volatiles and a number of terpenoid volatiles. Responding to CBW larvae infestation, cotton plants undergo drastic reprogramming of the transcriptome and the volatile profile. The present results increase our knowledge about insect herbivory-induced metabolic and biochemical processes in plants, which may help improve future studies on genes governing processes.

Chemosensillum immunolocalization and ligand specificity of chemosensory proteins in the alfalfa plant bug Adelphocoris lineolatus (Goeze).

Insect chemosensory proteins (CSPs) are a family of small soluble proteins. To date, their physiological functions in insect olfaction remain largely controversial in comparison to odorant binding proteins (OBPs). In present study, we reported the antenna specific expression of three CSPs (AlinCSP4-6) from Adelphocoris lineolatus, their distinct chemosensillum distribution as well as ligand binding capability thus providing the evidence for the possible roles that they could play in semiochemical detection of the plant bug A. lineolatus. The results of qRT-PCR and western blot assay clearly showed that all of these three CSPs are highly expressed in the adult antennae, the olfactory organ of insects. Further cellular investigation of their immunolocalization revealed their dynamic protein expression profiles among different types of antennal sensilla. In a fluorescence competitive binding assay, the selective ligand binding was observed for AlinCSP4-6. In ad`dition, a cooperative interaction was observed between two co-expressed CSPs resulting in an increase of the binding affinities by a mixture of AlinCSP5 and AlinCSP6 to terpenoids which do not bind to individual CSPs. These findings in combination with our previous data for AlinCSP1-3 indicate a possible functional differentiation of CSPs in the A. lineolatus olfactory system.

A comparison of the life-history traits between diapause and direct development individuals in the cotton bollworm, Helicoverpa armigera.

In order to understand the differences of life-history traits between diapause and direct development individuals in the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), the development time, body size, growth rate, and adult longevity were investigated between the two populations, which were induced under 12:12 L:D and 16:8 L:D photoperiods, respectively, at 20, 22, and 25°C. The results indicated that the larval development time, pupal weight, adult weight, and growth rate were significantly different between diapause and direct developing individuals. The diapause developing individuals had a significantly higher pupal and adult weight and a longer larval time compared with direct developing individuals. However, the growth rate in diapause developing individuals was lower than that in the direct developing individuals. Analysis by GLM showed that larval time, pupal and adult weight, and growth rate were significantly influenced by both temperature and developmental pathway. The pupal and adult weights were greater in males than females in both developmental pathways, exhibiting sexual size dimorphism. The dimorphism in adult weight was more pronounced than in pupal weight because female pupae lost more weight at metamorphosis compared to male pupae. Protogyny was observed in both developmental pathways. However, the protogyny phenomenon was more pronounced at lower temperatures in direct developing individuals, whereas it was more pronounced in diapause developing individuals when they experienced higher temperatures in their larval stage and partial pupal period. The adult longevity of diapause developing individuals was significantly longer than that of direct developing individuals. The results reveal that the life-history strategy was different between diapause and direct developing individuals.

Anti-inflammatory and anti-apoptotic effect of combined treatment with methylprednisolone and amniotic membrane mesenchymal stem cells after spinal cord injury in rats.

This study was undertaken to investigate the synergistic effects of methylprednisolone (MP) administration and transplantation of amniotic membrane mesenchymal stem cells (AM-MSCs) following T11 spinal cord clip compressive injury in rats. The combination treatment with MP (50 mg/kg) and delayed transplantation of AM-MSCs after rat spinal cord injury, significantly reduced (1) myeloperoxidase activity, (2) the proinflammatory cytokines: tumor necrosis factor-α, interleukin (IL)-1ß, IL-6, IL-17, interferon-γ and (3) the cell apoptosis [terminal deoxynucleotidyl transferase, dUTP nick end labeling (TUNEL) staining, and caspase-3, Bax and Bcl-2 expressions]; increased: (1) the levels of the anti-inflammatory cytokines (IL-10 and transforming growth factor-ß1) and (2) the survival rate of AM-MSCs in the injury site. The combination therapy significantly ameliorated the recovery of limb function (evaluated by Basso, Beattie and Bresnahan score). Taken together, our results demonstrate that MP in combination with AM-MSCs transplantation is a potential strategy for reducing secondary damage and promoting functional recovery following spinal cord injury.

The preferential binding of a sensory organ specific odorant binding protein of the alfalfa plant bug Adelphocoris lineolatus AlinOBP10 to biologically active host plant volatiles.

Semiochemicals such as sex pheromones and plant volatiles are crucial components of insect mating systems and host plant localization. In the olfactory signal transduction pathway, odorant-binding proteins (OBPs) are important elements that function in the first step of the pathway by carrying hydrophobic semiochemicals across the sensillum lymph to the olfactory receptors (ORs). In this study, we examined the binding affinities of semiochemicals to AlinOBP10, a putative OBP from the alfalfa plant bug, Adelphocoris lineolatus, that we demonstrate is expressed mainly in sensory organs. We then characterized the biological activities of the high affinity semiochemicals by measuring their electrophysiological activities in antennae and behavioral responses in the plant bug. AlinOBP10 displayed weak binding affinities to two major putative pheromone components, hexyl butyrate and (E)-2-hexenyl butyrate. In contrast, AlinOBP10 exhibited higher binding affinities to six host plant volatiles, namely myrcene, ß-pinene, ß-ionone, 3-hexanone, (E)-2-hexenal, and 1-hexanol. The biological activities of these six putative ligands were further studied in electroantennogram recordings and Y-tube olfactometer trials. The three compounds, (E)-2-hexenal, 1-hexanol, and 3-hexanone elicited strong electrophysiological responses, but elicited distinct behaviors. While 3-hexanone was attractive to female adults, (E)-2-hexenal and 1-hexanol were significant repellents. Although a weak electrophysiological response was elicited with ß-pinene, it was a strong repellent. These results demonstrate that AlinOBP10 can interact with attractants, as well as repellents, with some specificity toward plant volatiles over sex pheromones.

[Treatment of intra-articular calcaneal fractures with unilateral external fixator combined with limited internal fixation].

OBJECTIVE: To study the clinical effects of the unilateral external fixator combined with limited internal fixation such as cannulated lag screws for the treatment of intra-articular calcaneal fractures. METHODS: From May 2007 to February 2012,52 patients (58 feet) with intraarticular calcaneal fractures were treated with unilateral external fixator and limited internal fixation such as lag screws. Forty-five patients (51 feet) were male,7 patients (7 feet) were female,ranging in age from 18 to 64 years,averaged 36.5 years. Time from injury to surgery was 4 to 13 (6.2 +/- 0.8) days. According to Sanders classification, there were 36 cases (40 feet) with type II fractures, 13 cases (15 feet) with type III fractures, 3 cases (3 feet) with type IV fractures. All patients were examed with X-ray and CT. Böhler angle, Gissane angle, length, width, height were measured on the radiographs, respectively. The function of the feet was assessed by the Maryland grading system. RESULTS: Fifty-one patients(57 feet) were followed up for 5 to 16 months,with an average of 7.2 months. Calcaneal Böhler angle increased from (12.65 +/- 5.32) degrees preoperatively to (30.63 +/- 4.28) degrees postoperatively. The calcaneal Gissane angle decreased from (129.87 +/- 6.25) degrees preoperatively to (122.11 +/- 4.89) degrees postoperatively. The calcaneal length increased from (64.5 +/- 7.1) mm preoperatively to (71.3 +/- 5.7) mm postoperatively. The calcaneal width decreased from (34.6 +/- 5.7) mm preoperatively to (28.2 +/- 6.1) mm postoperatively. The calcaneal height increased from (30.2 +/- 5.2) mm preoperatively to (39.3 +/- 6.4) mm postoperatively. All of the comparison between preoperative and postoperative measurements revealed statistically significant (P < 0.05). The length,width, height, Böhler angle and Gissane angle were improved significantly (P < 0.05). According to the Maryland grading system, the result was excellent in 20 feet, good in 31 feet, fair in 4 feet, pour in 2 feet,excellent and good rate was 89.5%. Three feet with wound infection or flap necrosis were healed after wound debridement and dressing exchange. CONCLUSION: The unilateral external fixation combined with limited internal fixation is a satisfactory treatment for intra-articular calcaneal fractures in short term with less damage, few complications and good functional recovery.

Inheritance of photoperiodic control of pupal diapause in the cotton bollworm, Helicoverpa armigera (Hübner).

Pupae of the cotton bollworm, Helicoverpa armigera display a diapause in response to the exposure of their larvae to short photoperiods and relatively low temperatures. Due to geographic variation in photoperiodic response, moths from a northern population, Langfang (39°32'N, 116°41'E), enter diapause in response to short daylengths (D strain) while moths from a southern population, Ledong (18°28'N, 108°53'E), exhibit no diapause under the same conditions (N strain). In the present study, crosses between the two strains are utilized to evaluate the inheritance of diapause under different photoperiods at temperatures 20, 22 and 25°C. The moths in both reciprocal crosses and backcrosses to D strain showed a clear long-day response, similar to that of the D strain, suggesting that the photoperiodic response controlling diapause in this moth is heritable. The incidences of diapause for all F(1) hybrids were intermediate between those of their parents. However, the incidences of diapause at 20°C in F(1) (N×D) strain were significantly higher than those in F(1) (D×N) strain, indicating that the male parent plays a more important role in the determination of diapause. The N strain also showed a short-day photoperiodic response at the lower temperature of 20°C, indicating that the N strain still has the capability to enter a photoperiodically induced diapause, depending on the rearing temperature. Results from all crosses under photoperiods LD 12:12 or LD 13:11 at 22°C showed that inheritance of diapause in H. armigera did not fit an additive hypothesis and that the capacity for diapause was transmitted genetically in the manner of incomplete dominance with non-diapause characteristic partially dominant over the diapausing. Diapause duration in hybrid pupae was also influenced by their inheritance from both parents. Diapause duration in hybrid pupae was intermediate between those of their parents. These results reveal that both diapause induction and duration are under the control of polygene.

Diapause induction, maintenance and termination in the rice stem borer Chilo suppressalis (Walker).

The rice stem borer, Chilo suppressalis, enters facultative diapause as fully grown larvae in response to short-day conditions during the autumn. Our results showed that the critical night length for diapause induction in C. suppressalis was between 10 h 22 min and 10h 45 min at 22, 25 and 28 degrees C, 11 h 18 min at 31 degrees C, and between 10 h 5 min and 10 h 20 min under field conditions (average temperature ranged from 27.2 to 30.7 degrees C). The diapause incidence declined in ultra-long nights (18-22 h scotophases) and DD, and increased in ultra-short nights (2-6 h scotophases) and LL. Moreover, we found that the third instar was the stage most sensitive to the photoperiod, and night length played an essential role in the initiation of diapause. Night-interruption experiments with a 1-h light pulse at LD 12:12 (light 12:dark 12) exhibited two troughs of diapause inhibition, with one occurring in early scotophase and the other in late scotophase. Field observations for six years showed that most larvae entered winter diapause in August in response to declining day lengths, despite the high temperatures prevailing during August. By periodically transferring the field-collected overwintering larvae to different photoperiods and temperatures, the results showed that photoperiod had a significant influence on diapause development during the early phase of diapause, while high temperature significantly accelerated the termination of larval diapause.

Diapause induction and clock mechanism in the cabbage butterfly Pieris melete Ménétriés.

Photoperiodic control of diapause induction was systematically investigated in the cabbage butterfly, Pieris melete, which enters summer and winter diapause as a pupa. Summer and winter diapause are induced principally by short and long scotophases, respectively; the intermediate scotophases (11-12 h) permit pupae to develop without diapause. Photoperiodic responses under 24-h light-dark cycles at 16.9, 18, 20 and 22 degrees C showed that the hibernation response was temperature compensated, whereas aestivation response was strongly temperature-dependent. The incidence of diapause for both aestivation and hibernation showed a decline at the ultra-short and ultra-long scotophases. Experiments using non-24-h light-dark cycles showed that the length of the scotophase played an essential role in the determination of diapause. The highest photosensitivity differed under hibernation and aestivation conditions. With a 3 x LD 12:12 interruption, a maximal inhibition of aestivation occurred in the L3/2 stage, and of hibernation it occurred in the L4/0 stage. A long-night of LD 10:14 induced hibernation diapause but inhibited aestivation diapause and, conversely, a short-night of LD 14:10 inhibited hibernation diapause but induced aestivation diapause. With a 1-h light pulse at LD 11:13, a maximal inhibition of hibernation occurred 3 h before lights-on (late scotophase), whereas, with a 1-h light pulse at LD 12.5:11.5, a maximal induction of aestivation occurred 2-3 h after the onset of darkness (early scotophase). Nanda-Hamner and Bünsow experiments failed to reveal the involvement of a circadian system, suggesting that the photoperiodic time measurement for diapause induction in this butterfly resembles an hourglass-like timer or a damped circadian oscillator.

A comparison of photoperiodic control of diapause between aestivation and hibernation in the cabbage butterfly Pieris melete.

In the cabbage butterfly, Pieris melete, summer and winter diapause are induced principally by long and short daylengths, respectively; the intermediate daylengths (12-13 h) permit pupae to develop without diapause. In this study, photoperiodic control of summer and winter diapause was systematically investigated in this butterfly by examining the photoperiodic response, the number of days required to induce 50% summer and winter diapause and the duration of diapausing pupae induced under different photoperiods. Photoperiodic response curves at 18 and 20 degrees C showed that all pupae entered winter diapause at short daylengths (8-11 h), the incidence of diapause dropped to 82.3-85.5% at 22 degrees C without showing a significant difference between short daylengths, whereas the incidence of summer diapause induced by different long daylengths (14-18 h) was varied and was obviously affected by temperature. By transferring from various short daylengths (LD 8:16, LD 9:15, LD 10:14 and LD 11:13) to an intermediate daylength (LD 12.5:11.5) at different times after hatching, the number of cycles required to induce 50% winter diapause (7.28 at LD 8:16, 7.16 at LD 9:15, 7.60 at LD 10:14 and 6.94 at LD 11:13) showed no significant difference, whereas by transferring from various long daylengths (LD 14:10, LD 15:9, LD 16:8 and LD 17:7) to an intermediate daylength (LD 12.5:11.5) at different times, the number of cycles required to induce 50% summer diapause (5.95 at LD 14:10, 8.02 at LD 15:9, 6.80 at LD 16:8, 7.64 at LD 17:7) were significantly different. The intensity of winter diapause induced under different short daylengths (LD 8:16, LD 9:15, LD 10:14 and LD 11:13) was not significantly different with an average diapause duration of 87 days at a constant temperature of 20 degrees C and 92 days at a mean daily temperature of 19.0 degrees C, whereas the intensity of summer diapause induced under different long daylengths (LD 14:10, LD 15:9, LD 16:8 and LD 17:7) was significantly different (the diapause duration ranged from 75 to 86 days at a constant temperature of 20 degrees C and from 76 to 88 days at a mean daily temperature of 19.0 degrees C). All results suggested that photoperiodic control of diapause induction and termination is significantly different between aestivation and hibernation.

Photoperiodic counter of diapause induction in Pseudopidorus fasciata (Lepidoptera: Zygaenidae).

Induction of larval diapause is a photoperiodically controlled event in the life history of the moth Pseudopidorus fasciata. In the present study, the photoperiodic counter of diapause induction has been systematically investigated. The required day number (RDN) for a 50% response was determined by transferring from a short night (LD 16:8) to a long night (LD 12:12) or vice versa at different times after hatching, The RND differed significantly between short- and long-night cycles at different temperatures. The RDN for long-night cycles at 20, 22, 25 and 28 degrees C was 11.5, 9.5, 7.5 and 8.5 days, respectively. The RDN for short-night cycles was 3 days at 22 degrees C and 5 days at 20 degrees C indicating that the effect of one short night was equivalent to the effect of 2-3 long nights effect. Night-interruption experiments of 24h photoperiods by a 1 h light pulse showed that the most crucial event for the photoperiodic time measurement in this moth was whether the length of pre-interruption (D(1)) or the post-interruption (D(2)) scotophases exceeded the critical night length (10.5 h). If D(1) or D(2) exceeded 10.5 h diapause was induced. The diapause-averting effect of a single short-night cycle (LD 16:8) against a background of long nights (LD 12:12) showed that the photoperiodic sensitivity was greatest during the first 7 days of the larval period and the highest sensitivity was on the fourth day. Both non-24 and 24 h light-dark cycle experiments revealed that the photoperiodic counter in P. fasciata is able to accumulate both long and short nights during the photosensitive period, but in different ways. The information from short-night cycles seems to be accumulated one by one in contrast to long-night cycles where six successive cycles were necessary for about 50% diapause induction and eight cycles for about 90% diapause. These results suggest the accumulation of long-night and short-night cycles may be based on different mechanisms.