Mutation in acetylcholinesterase1 associated with triazophos resistance in rice stem borer, Chilo suppressalis (Lepidoptera: Pyralidae)

Two full-length genes encoding different acetylcholinesterases (AChEs), designated as Ch-ace1 and Ch-ace2, were cloned from strains of the rice stem borer (Chilo suppressalis) susceptible and resistant to the organophosphate insecticide triazophos. Sequence analysis found an amino acid mutation A314S in Ch-ace1 (corresponding to A201 in Torpedo californica AChE) that was consistently associated with the occurrence of resistance. This mutation removed an MspA1 I restriction site from the wild type allele. An assay based on restriction fragment length polymorphism (RFLP) analysis was developed to diagnose A314S genotypes in field populations. Results showed a strong correlation between frequencies of the mutation and phenotypic levels of resistance to triazophos. The assay offers a prospect for rapid monitoring of resistance and assisting with the appropriate choice of insecticide for combating damage caused by C. suppressalis.     

Screen of Bacillus thuringiensis toxins for transgenic rice to control Sesamia inferens and Chilo suppressalis

Transgenic rice to control stem borer damage is under development in China. To assess the potential of Bacillus thuringiensis (Bt) transgenes in stem borer control, the toxicity of five Bt protoxins (Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1Ca) against two rice stem borers, Sesamia inferens (pink stem borer) and Chilo suppressalis (striped stem borer), was evaluated in the laboratory by feeding neonate larvae on artificial diets containing Bt protoxins. The results indicated that Cry1Ca exhibited the highest level of toxicity to both stem borers, with an LC₅₀ of 0.24 and 0.30 μg/g for C. suppressalis and S. inferens, respectively. However, S. inferens was 4-fold lower in susceptibility to Cry1Aa, and 6- and 47-fold less susceptible to Cry1Ab and Cry1Ba, respectively, compared to C. suppressalis. To evaluate interactions among Bt protoxins in stem borer larvae, toxicity assays were performed with mixtures of Cry1Aa/Cry1Ab, Cry1Aa/Cry1Ca, Cry1Ac/Cry1Ca, Cry1Ac/Cry1Ba, Cry1Ab/Cry1Ac, Cry1Ab/Cry1Ba, and Cry1Ab/Cry1Ca at 1:1 (w/w) ratios. All protoxin mixtures demonstrated significant synergistic toxicity activity against C. suppressalis, with values of 1.6- to 11-fold higher toxicity than the theoretical additive effect. Surprisingly, all but one of the Bt protoxin mixtures were antagonistic in toxicity to S. inferens. In mortality-time response experiments, S. inferens demonstrated increased tolerance to Cry1Ab and Cry1Ac compared to C. suppressalis when treated with low or high protoxin concentrations. The data indicate the utility of Cry1Ca protoxin and a Cry1Ac/Cry1Ca mixture to control both stem borer populations.     

Antioxidant responses of Chilo suppressalis (Lepidoptera: Pyralidae) larvae exposed to thermal stress

High temperatures cause a variety of physiological stress responses in insects, including increased generation of reactive oxygen species (ROS), which can cause oxidative damage. This study investigated the effects of thermal stress on ROS generation, the expression of heat shock protein 70 (Hsp70) at the mRNA and protein levels, the activity of antioxidant enzymes (SOD, CAT), and apoptosis in hemocytes of Chilo suppressalis larvae. Results indicated that thermal stress significantly elevated the level of ROS and antioxidant enzyme activity in C. suppressalis larvae. Real-time quantitative PCR showed that hsp70 gene expression was induced by heat stress. Flow cytometric results revealed that the expression profile of Hsp70 at the protein level was in agreement with that at the mRNA level. The expression of Hsp70 at both the mRNA and protein levels reached a maximum at 36 °C in larval hemocytes. Exposure to tested temperatures did not cause any significant change in the rate of apoptosis in larval hemocytes. These results suggest that thermal stress leads to oxidative stress and that antioxidant enzymes and the Hsp70 play an important role in reducing oxidative damage in C. suppressalis larvae.     

Effects of diapause and cold-acclimation on the avoidance of freezing injury in fat body tissue of the rice stem borer, Chilo suppressalis Walker

Overwintering freeze-tolerant larvae of Chilo suppressalis can survive at -25 degrees C, but non-diapausing larvae cannot. We reported earlier that to prevent intracellular freezing, which causes death in overwintering larvae of the Saigoku ecotype distributed in southwestern Japan, water leaves and glycerol enters fat body cells through water channels during freezing. However, it is still unclear how diapause and low-temperature exposure are related to the acquisition of freeze tolerance. We compared the extent of tissue damage, accumulation of glycerol, and transport of glycerol and water in fat body tissues between cold-acclimated and non-acclimated non-diapausing and diapausing larvae. The tissue from cold-acclimated diapausing larvae could survive only when frozen in Grace's insect medium with 0.25 M glycerol at -20 degrees C. The protection provided by glycerol was offset by mercuric chloride, which is a water-channel inhibitor. Fat body tissue isolated from non-acclimated diapausing larvae was injured by freezing even though glycerol was added to the medium, but the level of freezing injury was significantly lower than in non-diapausing larvae. Radiotracer assays in cold-acclimated diapausing larvae showed that during freezing, water left the cells into the medium and glycerol entered the cells from the medium at the same time. Therefore, in Saigoku ecotype larvae of the rice stem borer, both diapause and cold-acclimation are essential to accumulate glycerol and activate aquaporin for the avoidance of freezing injury.     

Diapause termination, post-diapause development and reproduction in the beet webworm, Loxostege sticticalis (Lepidoptera: Pyralidae)

Effects of photoperiod and cold exposure on diapause termination, post-diapause development and reproduction in Loxostege sticticalis were examined. Larvae were reared at diapause inducing condition (22 °C, L:D 12:12) consistently or transferred to long day photoperiod (L:D 16:8) and darkness (L:D 0:24) respectively, after entering into diapause. Diapause was terminated in approximately 40% of the larvae after 36 days, and no significant differences were observed between photoperiods, suggesting larval diapause was terminated spontaneously without being induced by photoperiods. Cold exposure significantly hastened diapause termination. The diapause termination incidence increased significantly with peaks of 98% at both 5 °C and 0 °C exposure for 30 days, as compared to 42% in controls not exposed to cold, while the mortality and number of days required for diapause termination decreased dramatically. The optimal low temperature exposure periods under 5 °C or 0 °C were 20 days and 30 days, showing a higher termination incidence and shorter time for diapause termination. This suggests that the low temperatures in winter play an important role in diapause termination under natural conditions. The threshold temperatures for post-diapause development in prepupae and pupae were 9.13 °C and 10.60 °C respectively, with corresponding accumulations of 125 and 200 degree-days. Adults that experienced larval diapause significantly delayed their first oviposition, oviposition period was prolonged, and the lifetime number of eggs laid was decreased, however both males and females have significantly longer longevity. The field validation of diapause termination, the degree-days model, and the relationship between diapause and migration in L. sticticalis were also discussed.     

Seasonal changes in glycerol content and cold hardiness in two ecotypes of the rice stem borer, Chilo suppressalis, exposed to the environment in the Shonai district, Japan

The rice stem borer, Chilo suppressalis, is divided into at least two ecotypes in Japan, the Shonai ecotype (SN) which is distributed in the northern part of Japan, and the Saigoku ecotype (SG) which is distributed in the southwestern region. Cold hardiness is positively correlated with the level of glycerol in both ecotypes. To investigate whether ecological distribution affects glycerol accumulation and cold hardiness development in these two ecotypes, overwintering larvae of the SN and SG ecotypes were concurrently exposed to the Shonai district. Obvious differences in the progress of glycerol accumulation and cold hardiness development in SN and SG larvae were found in early winter in the Shonai district. The levels of glycerol content and cold hardiness were low in October and high in January in both ecotypes, but those levels were different within this period (November and December) between ecotypes; the levels in SN larvae quickly reached their maximum, whereas, in SG larvae levels increased slowly. Under controlled conditions, the effect of the period of acclimation at 10 degrees C and subsequent low-temperature (5 degrees C) exposure on glycerol accumulation was investigated. These results indicated that glycerol accumulation in SN was stimulated by the progression of diapause termination, whereas a higher cumulative effect on glycerol production in SG was found when diapause was in a deep state.     

Diapause induction and termination in the predatory mite Euseius finlandicus in peach orchards in northern Greece

The course of diapause induction as well as of diapause termination infemales of the predatory mite Euseius (Amblyseius)finlandicus Oudemans (Acari: Phytoseiidae) in Northern Greece wasdetermined by transferring females during winter and early spring from peachtrees to a short day (LD 8:16) or a long day (LD 16:8) photoperiod both at 20°C. The first diapause females were found in mid September andby the first week of October all the females were in diapause. The mean numberof days required for diapause termination under the short day photoperiod LD8:16 was gradually reduced from 93.7 days in the beginning of October to lessthan 20 days in mid February and early March. Under the long day photoperiod ofLD 16:8 diapause was terminated in less than 20 days irrespective of the dateof collection. These results indicate that by mid February photoperiodicsensitivity of females was lost and diapause was terminated. However, femaleswere found in their overwintering sites until the second half of March,probablybecause of the prevailing relatively low temperatures and lack of adequatefood.     

水稻二化螟的交配行为

在室内条件下,对水稻二化螟Chilo suppressalis的交配行为及能力进行了研究.结果表明：大多数二化螟雌蛾一生只交配一次,平均0.92次;而雄蛾具有多次交配能力,最多达4次,平均2.72次.二化螟雌蛾的日龄影响其交配率、交配起始时间和持续时间,随二化螟雌蛾日龄的增加,其交配率逐渐下降,交配起始时间逐渐提前,而交配持续时间逐渐上升.相反,二化螟雄蛾日龄对其交配率、交配起始时间和持续时间没有明显影响.交配日龄对二化螟雌蛾的生殖力也存在显著影响,随着二化螟雌蛾交配日龄的增加,雌蛾产卵量下降,卵孵化率降低,产卵期缩短,它们都与雌蛾交配日龄存在显著的负相关;而雌蛾产卵前期和雌蛾寿命随雌蛾交配日龄的增加而延长,与雌蛾交配日龄存在显著的正相关.但二化螟雄蛾交配日龄对雌蛾的生殖力没有明显影响,二化螟雄蛾一生都具有较强的交配繁殖能力.同时,不同交配史的雄蛾与雌蛾交配,对雌蛾的生殖力也没有显著影响.表明二化螟的交配活动是由雌蛾主导控制的.最后,对这些结果在二化螟性信息素防治中应用的可行性进行了探讨.在应用性信息素控制二化螟的实践中,可以在两方面取得实效,一是性信息素可以阻碍雌雄之间正常交配,降低交配率;二是可以推迟二化螟雌虫的交配,使其产卵量和卵孵化率降低.     

Role of membrane transport of water and glycerol in the freeze tolerance of the rice stem borer, Chilo suppressalis Walker (Lepidoptera: Pyralidae)

Overwintering larvae of the rice stem borer, Chilo suppressalis accumulate glycerol and are freezing tolerant to about -25 degrees C. However, non-diapausing larvae cannot accumulate glycerol and are killed by freezing. We compared the extent of tissue damage, the effects of glycerol concentration, and the transport of glycerol and water in fat body tissues from these larvae at selected freezing temperatures. Tissues from overwintering larvae, but not non-diapausing larvae, survive when frozen at -20 degrees C with 0.25 M glycerol, but the protection afforded by glycerol is offset by the water-channel inhibitor mercuric chloride. Glycerol in higher concentration (0.75 M) affords some protection even to the fat body of non-diapausing larvae. Radiotracer assays of overwintering larvae show that water leaves the tissues during freezing while glycerol enters, and that mercuric chloride disrupts this process. Transport is also disrupted after lethal freezing at -35 degrees C. Therefore, membrane transport of water and glycerol is involved in the avoidance of freezing injury to fat body cells of the rice stem borer, apparently by mediating the replacement of water with glycerol in freezing-tolerant tissues.     

Geographic variation in diapause induction and termination of the cotton bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae)

Overwintering diapause in Helicoverpa armigera, a multivoltine species, is controlled by response to photoperiod and temperature. Photoperiodic responses from 5 different geographical populations showed that the variation in critical photoperiod for diapause induction was positively related to the latitudinal origin of the populations at 20, 22 and 25 °C. Diapause response to photoperiod and temperature was quite different between northern and southern populations, being highly sensitive to photoperiod in northern populations and temperature dependence in southern populations. Diapause pupae from southern population showed a significantly shorter diapause duration than from northern-most populations when they were cultured at 20, 22, 25, 28 and 31 °C; by contrast, overwintering pupae from southern populations emerged significantly later than from northern populations when they were maintained in natural conditions, showing a clinal latitudinal variation in diapause termination. Diapause-inducing temperature had a significant effect on diapause duration, but with a significant difference between southern and northern populations. The higher rearing temperature of 22 °C evoked a more intense diapause than did 20 °C in northern populations; but a less intense diapause in southern population. Cold exposure (chilling) is not necessary to break the pupal diapause. The higher the temperature, the quicker the diapause terminated. Response of diapause termination to chilling showed that northern populations were more sensitive to chilling than southern population.     

Photoperiodism of diapause induction in Thyrassia penangae (Lepidoptera: Zygaenidae)

Thyrassia penangae enters winter diapause as a prepupa in a cocoon. Photoperiodism of diapause induction was systematically investigated in this moth. The photoperiodic response curves under 24-h light-dark cycles showed that this insect is a typical long-day species. The critical daylength was 13 h 30 min at 25 °C, 13 h at 30 °C and 12 h 20 min at 28 °C. Transferring experiments from a short day (LD 12:12) to a long day (LD 15:9) or vice versa indicated that photoperiodic sensitivity mainly occurs during the larval period. In experiments using non-24-h light-dark cycles, when the length of photophase exceeded the critical daylength (13.5 h), was diapause inhibited effectively, even when the length of scotophase exceeded the critical nightlength (10.5 h). Only when a long scotophase was combined with a short photophase, diapause was induced effectively. This result suggests that daylength measurement is more important than nightlength measurement in T. penangae. Night interruption experiments under 24-h light-dark cycles exhibited two points of apparent light sensitivity, but the photosensitive position was highly influenced by temperature and the length of scotophase. Nanda-Hamner experiments failed to reveal the involvement of a circadian system in this photoperiodic time measurement. All light-dark cycles from LD 12:12 to LD 12:72 resulted in a short day response, and all cycles from LD 14:4 to LD 14:72 resulted in a long day response, suggesting that photoperiodic time measurement in this moth is performed by a day-interval timer or an hourglass-like clock.     

Phosphatases in the rice stem borer, Chilo suppressalis Walker (Lepidoptera; Pyralidae): Some properties and changes of the activities during hibernation

To elucidate the regulation of glycerol synthesis from α-glycerophosphate (α-GP) in larvae of the rice stem borer, Chilo suppressalis Walker, phosphatase activities were examined during hibernation. The presence of alkaline and acid phosphatase was demonstrated in hibernating larvae. The activity of the alkaline type decreased during diapause entry while the activity of the acid type increased at the initiation of diapause and then decreased during postdiapause stage. This pattern of acid type activity change coincided with that of glycerol accumulation in hemolymph during hibernation. Thus, the acid type phosphatase may largely be involved in glycerol synthesis.     

Diapause in the egg parasitoid Trichogramma cordubensis: role of temperature

The role of temperature in the induction of diapause in Trichogramma cordubensis (Hymenoptera: Trichogrammatidae), under controlled laboratory conditions, was investigated using Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs as hosts. Results indicate that prestorage temperatures and the duration of exposure of the parasitoids to these temperatures affected the induction of diapause. It was possible to induce diapause in prepupae of T. cordubensis by exposing the preimaginal stages (prior to the prepupal stage) to 10°C for at least 30 days, but adults emerged without diapause when the duration of exposure was of only 10 or 20 days. Parasitoids failed to enter diapause when prestorage temperatures were 7 or 12°C, regardless of the duration of exposure. However, at these two temperatures, preimaginal development of T. cordubensis was delayed, allowing short-term storage (40 days at induction temperatures followed by 30 days at 3°C) by keeping parasitoids in quiescence without reducing the percentages of adult emergence. Good percentages of adult emergence after long-term low-temperature storage (30 or 40 days at 10°C followed by six months at 3°C) occurred only when T. cordubensis was in diapause. The long-term storage of parasitoids in diapause allows an enlargement in the mass rearing potentialities of this species for future biological control releases by allowing producers to stockpile the parasitoids for release in the field season.     

Diapause termination in the spotted stem borer, Chilo partellus (Lepidoptera: Pyralidae) in the laboratory

The effect of temperature, photoperiod, artificial diet and water on the termination of diapause by larvae of the stem borer, Chilo partellus (Lepidoptera: Pyralidae), was studied in the laboratory. Termination of diapause as indicated by pupation was affected mainly by a combination of high temperature and a long day photoperiod. Total darkness did not prevent termination of diapause and pupation occurred also in larvae which were never exposed to water. Long days accelerated pupation, but, under 16 h daylength, termination of diapause was faster than under constant illumination. Provision of artificial diet had no effect or slowed down pupation but water decreased the time to pupation. Under 28°C, 16 h daylength and availability of water, C. partellus diapausing larvae terminated diapause and pupated in about 9 days.     

Diapause induction and clock mechanism in the cabbage beetle, Colaphellus bowringi (Coleoptera: Chrysomelidae)

Photoperiodic control of diapause induction was investigated in the short-day species, Colaphellus bowringi, which enters summer and winter diapause as adult in the soil. Photoperiodic responses at 25 and 28 degrees C revealed a critical night length between 10 and 12 h; night lengths > or =12 h prevented diapause, whereas night lengths <12 h induced summer diapause in different degree. Experiments using non-24-h light-dark cycles showed that the duration of scotophase played an essential role in the determination of diapause. Night-interruption experiments with T=24 h showed that diapause was effectively induced by a 2-h light pulse in most scotophases; whereas day-interruption experiments by a 2-h dark break had a little effect on the incidence of diapause. The experiments of alternating short-night cycles (LD 16:8) and long-night cycles (LD 12:12) during the sensitive larval period showed that the information of short nights as well as long nights could be accumulated. Nanda-Hamner experiments showed three declining peaks of diapause at 24 h circadian intervals. Bünsow experiments showed two very weak peaks for diapause induction, one being 8 h after lights-off, and another 8 h before lights-on, but it did not show peaks of diapause at a 24 h interval. These results suggest that the circadian oscillatory system constitutes a part of the photoperiodic clock of this beetle but plays a limited role in its photoperiodic time measurement.     

Characterization of the complete mitochondrial genome of Chilo auricilius and comparison with three other rice stem borers

The mitogenome of Chilo auricilius (Lepidoptera: Pyraloidea: Crambidae) was a circular molecule made up of 15,367 bp. Sesamia inferens, Chilo suppressalis, Tryporyza incertulas, and C. auricilius, are closely related, well known rice stem borers that are widely distributed in the main rice-growing regions of China. The gene order and orientation of all four stem borers were similar to that of other insect mitogenomes. Among the four stem borers, all AT contents were below 83%, while all AT contents of tRNA genes were above 80%. The genomes were compact, with only 121–257 bp of non-coding intergenic spacer. There are 56 or 62-bp overlapping nucleotides in Crambidae moths, but were only 25-bp overlapping nucleotides in the noctuid moth S. inferens. There was a conserved motif ‘ATACTAAA’ between trnS2 (UCN) and nad1 in Crambidae moths, but this same region was ‘ATCATA’ in the noctuid S. inferens. And there was a 6-bp motif ‘ATGATAA’ of overlapping nucleotides, which was conserved in Lepidoptera, and a 14-bp motif ‘TAAGCTATTTAAAT’ conserved in the three Crambidae moths (C. suppressalis, C. auricilius and T. incertulas), but not in the noctuid. Finally, there were no stem-and-loop structures in the two Chilo moths.     

Effect of anoxia on diapause termination in eggs of the false melon beetle, Atrachya menetriesi

The effect of anoxia on diapause development in the leaf beetle Atrachya menetriesi was investigated to elucidate the role of oxygen in regulation of egg diapause. While anoxia alone had no effect on diapause termination, it decreased diapause intensity before chilling. Such an effect reached a maximum level when anoxia lasted for about 10 days. Anoxia applied during the pre-diapause stage also reduced diapause intensity. On the other hand, anoxia terminated diapause when the diapause intensity had been lowered by sufficient duration of chilling (50 days at 7.5 degrees C). The effect of anoxia was temperature dependent; the larger effect was elicited when anoxia was combined with a higher temperature. A 50-day chilling caused more than 20% of eggs to terminate diapause upon transfer to warm conditions. However, when this chilling period was interrupted on the 20th day by a 5-day exposure to a high temperature of 20, 25 or 30 degrees C, the effect of the former chilling was cancelled partially or completely, suggesting that warming reversed diapause development. This reversing effect of a high temperature, however, was not manifested when the warming was combined with anoxia. The results suggest that anoxia inhibits diapause reversal and facilitates a certain process of diapause development. The sequence of exposure to anoxia and chilling is not important.     

二化螟钙黏蛋白CAD1的原核表达及与Cry1Ac、Cry2Aa蛋白的结合

【目的】二化螟是水稻的重要害虫之一,钙黏蛋白(cadherin,CAD)是一类重要的Bt杀虫蛋白受体,在获得二化螟钙黏蛋白基因(Cs CAD1)的基础上,明确Cs CAD1蛋白与Cry1Ac和Cry2Aa蛋白的结合能力。【方法】利用PCR技术克隆Cs CAD1基因片段,将构建的p ET-28a-(+)-Cs CAD1重组质粒转入原核表达菌株BL21(DE3)中,IPTG诱导表达。目的蛋白经Ni柱亲和纯化后SDS-PAGE电泳检测,利用western blot和ligand blot技术分析其与Cry1Ac和Cry2Aa蛋白的结合能力。【结果】重组载体可在表达菌株BL21中表达一个约44 ku的蛋白,原核表达载体构建成功。SDS-PAGE显示该蛋白条带单一,且纯度较好。Ni柱亲和层析纯化该目的蛋白后进行Ligand blot分析,结果显示Cs CAD1重组蛋白可以与Cry1Ac和Cry2Aa蛋白结合。【结论】Cs CAD1蛋白可以与Cry1Ac和Cry2Aa蛋白结合,是潜在的Cry蛋白受体,所得结果有助于阐明Cry1Ac和Cry2Aa蛋白对二化螟的作用机制。     

The subgenus Persicargas (Ixodoidea: Argasidae: Argas), 26. Argas (P.) arboreus: Effect of photoperiod on diapause induction and termination

Prefeeding exposure of female Argas (Persicargas) arboreus to short photoperiod (SP) induces diapause, expressed as oviposition delay for more than 40 days postfeeding. Diapause incidence increases as prefeeding SP exposure periods increase. Postfeeding SP exposure is not essential to maintain diapause. Postfeeding exposure to SP without prefeeding SP exposure does not induce diapause. A 2 week prefeeding exposure to long photoperiod (LP) terminates diapause in two-thirds of the females exposed to LP after feeding. The remaining one-third, which may differ genetically, require a 6 week LP prefeeding exposure to terminate diapause. An 8 week prefeeding exposure to LP is required to terminate diapause in females exposed to SP after feeding.     

Photoperiodic control of diapause in Pseudopidorus fasciata (Lepidoptera: Zygaenidae) based on a qualitative time measurement

In the zygaenid moth, Pseudopidorus fasciata, both larval diapause induction and termination are under photoperiodic control. In this study, we investigated whether photoperiodic time measurement (with a 24-h light-dark cycle) in this moth is qualitative or quantitative. Photoperiodic response curves, at 22, 25, and 28 degrees C indicated that the incidence of diapause depended on whether the scotophases exceeded the critical night length (CNL) or not. All scotophases longer than the CNL-induced diapause; all scotophases shorter than the CNL-inhibited diapause. The CNL was 10.5h at 25 and 28 degrees C, and 10h at 22 degrees C. By transferring from various short photoperiods (LD 8:16, LD 9:15, LD 10:14, LD 11:13, LD 12:12, and LD 13:11) to a long photoperiod (LD 16:8) at different times, the number of light-dark cycles required for 50% diapause induction at 25 degrees C was 7.14 at LD 8:16, 7.2 at LD 9:15, 7.19 at LD 10:14, 7.16 at LD 11:13, and 7.13 at LD 12:12, without showing a significant difference between the treatments. Only at LD 13:11 (near the CNL), the number of light-dark cycles was significantly increased to 7.64. The intensity of diapause induced under different short photoperiods (LD 8:16, LD 9:15, LD 10:14, LD 11:13, and LD 12:12) at 25 degrees C was not significantly different with an average diapause duration of 36 days. The duration of diapause induced under LD 13:11 was significantly reduced to 32 days. All results indicate that the night-lengths are measured as either "long" or "short" compared with some critical value and suggest that photoperiodic time measurement for diapause induction in this moth is based on a qualitative principle.