Effect of temperature and photoperiod on diapause development in a Douglas fir seed chalcid, Megastigmus spermotrophus

Douglas fir seeds infested by diapausing larvae of a chalcid, Megastigmus spermotrophus, were transferred under different temperature and photoperiod regimes at various dates during autumn and winter. Both winter diapause and prolonged diapause were compared to the diapause patterns observed in control lots kept under natural conditions. The results showed that exposure to low temperatures in autumn and winter is required for the completion of diapause development. Median adult emergence dates and the date of transfer of larvae to 20°C were negatively correlated. The frequency of prolonged diapause of larvae transferred to 20°C decreased significantly after November. When larvae were submitted to different constant temperatures, the longer the daylength, the higher and faster the reactivation. These results suggest that chalcid larvae can perceive daylength through the seed coat. This phenomenon is probably related to the fact that most of the infested seeds falling in autumn are usually covered by a layer of leaf litter during the winter. Received: 14 June 1996 / Accepted: 24 February 1997     

Induction of larval diapause in the blowfly, Calliphora vicina R.-D. (Diptera, Calliphoridae) under field and laboratory conditions

Field experiments conducted in the environs of St. Petersburg (Russia) with a local population of Calliphora vicina showed that induction of larval diapause under natural conditions was significantly dependent on day lengths and temperature. The maternal photoperiodic response had a distinct threshold: the first diapausing larvae hatched from the eggs laid in the middle of August when the day length was 16 h; at shorter photoperiods, the fraction of diapausing larvae depended only on temperature. At the mean temperature of 16°C, larval diapause was rarely recorded; at 12–13°C, about 50% of the larvae entered diapause; at 7–9°C, nearly all the larvae entered diapause. These results of the field experiments agree well with the parameters of photoperiodic and thermal responses studied in the laboratory at constant temperatures and photoperiods.     

Photo-thermal regulation of diapause in Trichogramma piceum Djur. (Hymenoptera, Trichogrammatidae)

Interaction of the photoperiodic conditions of development of maternal females (day lengths of 2 to 22 h at 20°C) with the thermal regime of development of their progeny (temperature of 12 to 15°C at day length of 12 h) in determination of prepupal diapause in Trichogramma piceum was studied under laboratory conditions. At 15°C the diapause was practically absent. At lower temperatures, the proportion of diapausing prepupae was maximal (25% of larvae at 14°C, 70% of larvae at 13°, and 80% of larvae at 12°C) if the maternal females developed under short day conditions (10–12 h). When maternal females developed at day lengths of 18–20 h, diapause was rarely recorded at all temperatures, while ultra-short (less than 8–10 h) days also caused a decrease in the proportion of diapausing progeny. The right (ecologically important) threshold of this maternal long-day photoperiodic response was about 14–15 h independently of the temperature during the progeny development. These results make it possible to clarify the mechanism of the “maternal photoperiodic correction of the progeny thermal response.” Although the impact of the maternal photoperiodic response can be revealed only within a very narrow thermal range, the relative strength of the diapause-inducing effect of different day lengths is independent of the temperature regimen of the progeny development.     

Reproductive and developmental consequences of a diapause maternal effect in the flesh fly, Sarcophaga bullata

Abstract Progeny of Sarcophaga bullata produced from mothers with a history of short day will not enter pupal diapause even if they are reared in a strong diapause-inducing environment (LD 12:12 h at 20^oC). Short-day exposure and diapause commitment are normally inseparable, but this maternal effect provides a tool for examining separately the effect of photoperiod and diapause commitment. Duration of the wandering period of the third instar is longer in diapause-destined larvae than in non-diapause-destined larvae, and fecundity of flies that have experienced pupal diapause is lower than in long-day flies that have not been through diapause. The puparia of diapausing pupae contain more hydrocarbons than puparia of nondiapausing pupae, and this contributes to higher rates of net transpiration for the nondiapausing flies. Flies showing the maternal effect (short-day experience but no diapause) show an intermediate response: length of wandering, fecundity rate and quantities of puparial hydrocarbon are between the extremes observed in the other two groups of flies. Thus, the maternal effect switches the developmental programme to nondiapause, but the progeny retain some characteristics of diapause. Evidence from reciprocal crosses indicates that the photoperiodic history of the female, rather than the male, is responsible for the influence on fecundity.     

The effect of thermal stress on Ostrini00a nubilalis HBN (Lepidoptera, Pyralidae)—I. The orle of the neurosecretory cells in the survival of diapausing larvae

1. 1.|The effect of short-term exposure (up to 12 h) to upper limiting temperature (45°C) on A-type medial neurosecretory cells (MNSC) of the protocerebrum of Ostrinia nubilalis larvae was studied.

2. 2.|For evaluation of the activity of MNSC, cytological parameters were used: quantity of the neurosecretory material, size of the neurosecretory granules, shape of the nuclei and size of the nucleoli.

3. 3.|After 1 h exposure to 45°C the activity of the MNSC was increased, after 3 h it was decreased, while after 6 h exposure to the same temperature some renewed increase in the activity was observed.

4. 4.|The significance of oscillatory changes in the activity of A-type MNSC in survival of the thermal stress of O. nubilalis diapausing larvae is discussed.

Maternal thermal effect on diapause in Trichogramma species (Hymenoptera: Trichogrammatidae)

Both direct thermal and maternal photoperiodic effects on diapause induction have been thoroughly investigated in many insect species, while maternal thermal effects have been infrequently studied. We studied the effect of temperature during development of maternal generation on the proportion of diapausing progeny in four species of the genus Trichogramma Westw., minute egg parasitoids, widely used for biological control of lepidopteran pests. The maternal generations were reared at day lengths of 12 and 18 h and temperatures of 17, 20, 25 and 30°C, and their progeny developed under day length of 12 h and temperatures of 13 and 14°C. In T. evanescens and T. piceum, the proportion of diapausing progeny decreased with increasing temperature under all tested photoperiods and thermal regimes of progeny development; the high temperature of 30°C totally averted diapause of progeny. In T. buesi and T. principium, low temperatures of 17 and 20°C resulted in relatively high proportion of diapausing progeny only when the maternal generation developed under short‐day conditions. The threshold of the maternal thermal response varied from 17–18 to 22–23°C. Under field conditions, Trichogramma females are exposed to such high temperatures only during summer, when diapause in their progeny is in any case prevented by the maternal photoperiodic response and by the thermal response of the larvae. We conclude that the maternal thermal effect on diapause induction, although to a different extent, is inherent to Trichogramma species but, at least as suggested by laboratory experiments, it does not play any role in the regulation of seasonal development under natural conditions. However, during mass rearing of Trichogramma wasps, it should be taken into account that high temperature, even when combined with short photoperiod, can avert diapause in the next generation.     

二化螟滞育幼虫的蛋白和核酸含量以及保护酶活性的变化

为了阐明二化螟Chilo supprressalis滞育幼虫的分子特征及滞育期间保护酶活性的变化规律, 本研究应用Trizol法、 总量DNA提取法和蛋白定量试剂盒, 测定了在长光周期16L∶8D和25℃下发育的非滞育老熟幼虫、 在短光周期12L∶12D 和25℃下诱导滞育51 d的幼虫（称为滞育0个月）、 滞育1, 2和3个月幼虫的核酸含量和总蛋白含量; 同时应用试剂盒测定了老熟幼虫、 滞育0, 1和2个月的二化螟幼虫5种保护酶（POD, CAT, SOD, LDH和ATP酶）的活性。结果表明： 滞育幼虫的总RNA含量显著低于非滞育的老熟幼虫（P<0.05）, 而滞育1, 2和3个月的幼虫之间没有显著差异（P≥0.05）。老熟幼虫的总DNA含量显著高于滞育幼虫（P<0.05）。老熟幼虫的RNA/DNA比值较低, 滞育幼虫的RNA/DNA比值较高, RNA/DNA比值随着滞育时间的推移呈现出先上升后下降的趋势。滞育期大于1个月的幼虫中蛋白含量均显著高于非滞育的老熟幼虫（P<0.05）, 而滞育1, 2和3个月的幼虫之间没有显著差异（P≥0.05）。二化螟幼虫体内5种保护酶活性随发育阶段不同而存在差异。滞育幼虫中POD, CAT和SOD的活性随滞育时间延长而逐渐增强, 滞育2个月幼虫中的活性最高, 而非滞育老熟幼虫中的活性最低; LDH和ATP酶的活性则相反, 非滞育老熟幼虫中的活性最高, 滞育2个月幼虫中的活性最低。这些结果说明, 总RNA和DNA含量降低、 RNA/DNA比值先升后降、 总蛋白含量升高以及保护酶活性的变化是二化螟幼虫滞育过程中的主要生理特征。     

Changes in microRNA abundance may regulate diapause in the flesh fly,Sarcophaga bullata

Diapause, an alternative developmental pathway characterized by changes in developmental timing and metabolism, is coordinated by molecular mechanisms that are not completely understood. MicroRNA (miRNA) mediated gene silencing is emerging as a key component of animal development and may have a significant role in initiating, maintaining, and terminating insect diapause. In the present study, we test this possibility by using high-throughput sequencing and qRT-PCR to discover diapause-related shifts in miRNA abundance in the flesh fly, Sarcophaga bullata. We identified ten evolutionarily conserved miRNAs that were differentially expressed in diapausing pupae compared to their nondiapausing counterparts. miR-289-5p and miR-1-3p were overexpressed in diapausing pupae and may be responsible for silencing expression of candidate genes during diapause. miR-9c-5p, miR-13b-3p, miR-31a-5p, miR-92b-3p, miR-275-3p, miR-276a-3p, miR-277-3p, and miR-305-5p were underexpressed in diapausing pupae and may contribute to increased expression of heat shock proteins and other factors necessary for the enhanced environmental stress-response that is a feature of diapause. In S. bullata, a maternal effect blocks the programming of diapause in progeny of females that have experienced pupal diapause, and in this study we report that several miRNAs, including miR-263a-5p, miR-100-5p, miR-125-5p, and let-7-5p were significantly overexpressed in such nondiapausing flies and may prevent entry into diapause. Together these miRNAs appear to be integral to the molecular processes that mediate entry into diapause.     

大斑芫菁滞育幼虫在滞育不同阶段体内糖类和醇类含量的变化

在以卵滞育的昆虫中昆虫滞育时的生理代谢特点已经得到了大量研究。本文对以末龄幼虫（5龄）滞育的大斑芫菁Mylabris phalerate（Pallas）在不同滞育阶段体内糖类和醇类代谢的特征进行了研究。结果表明： 滞育个体血淋巴中的海藻糖含量高于非滞育个体,且随滞育时间的加大逐渐升高,滞育5个月时达到最大值,为5.61 μmol/mL。糖原的含量随滞育的进程逐渐减少,滞育初期（0.5个月）为0.72 mg/mL,到滞育末期（5个月）时仅为0.1 mg/mL。滞育个体脂肪体中的海藻糖含量都高于非滞育个体,滞育1个月时为非滞育个体的3倍,至滞育末期时达非滞育个体的5倍,为2.5 μmol/g脂肪体。糖原含量总体变化趋势是随滞育时间的加大逐渐减少,滞育早期和中期都高于非滞育个体。在滞育过程中血淋巴积累的小分子多元醇主要为甘油,其次是山梨醇;而在脂肪体中主要为甘油,其次是甘露醇,少量积累山梨醇：表明大斑芫菁滞育幼虫主要积累的是海藻糖和一些小分子多元醇。滞育幼虫在准备滞育时储备了大量糖原,这些糖原可能为滞育期间海藻糖、山梨醇和甘油的代谢提供了原料。     

Maternal thermal and photoperiodic effects on the progeny diapause in Trichogramma telengai Sorokina (Hymenoptera: Trichogrammatidae)

Although maternal photoperiodic and maternal thermal effects on the progeny diapause have been demonstrated in a number of insect species, their interaction has been rarely studied. We investigated this interaction in Trichogramma telengai. In a series of experiments, maternal females were reared at day lengths of 12–18 h and at temperatures of 17, 20, 25 and 30°C. Their progeny developed under day length of 12 h and temperatures of 13, 14 and 15°C. The experiments showed that both short day and low temperature experienced by the maternal generation significantly increased the proportion of diapausing progeny. In particular, the threshold of the maternal photoperiodic response decreased with temperature. Under combinations of photoperiod with daily thermoperiod, the role of the “night” temperature in the induction of diapause in the progeny was much more important than that of the “day” temperature. We conclude that the interaction pattern between the photoperiodic and thermal maternal effects in T. telengai is generally the same as that between the photoperiodic and thermal responses directly influencing diapause induction in other long‐day insects. The threshold temperature of the maternal thermal response of T. telengai was about 25–27°C, while diapause can be induced if larvae develop at temperatures not higher than 15–16°C. This suggests that, at least in the studied Trichogramma species, the maternal thermal effect has no ecological value. In the practice of biocontrol, however, rearing of Trichogramma wasps at high temperature can drastically reduce the proportion of diapausing progeny.     

Maternal influence on the incidence and duration of larval diapause in Calliphora vicina

ABSTRACT. The photoperiodic mechanism governing larval diapause in the blowfly Calliphora vicina R.-D. (Diptera: Calliphoridac) is largely maternal. The critical daylength for adult flies at 20C, and larvae derived from day 12 eggs and maintained in darkness at 11C, is about 14.5 h/24. At short daylength (LD 12:12 h) and a range of temperatures, eggs laid on or before day 9 gave rise to a low incidence of diapause, whereas those laid on day 10 and later gave rise to a high incidence; the photoperiodic 'counter' therefore shows a temperature-compensated mechanism for 'accumulating' successive short days. The rate of switching from non-diapause to diapause progeny is hardly affected by the timing of protein meals, although an early supply of meat accelerates egg development and leads to an increasing proportion of early, non-diapausing progeny. There is no evidence for a maternal effect preventing diapause induction in the first post-diapause ('spring') generation of flies. Diapausing larvae terminate diapause spontaneously in darkness at 11C after a variable period which is positively correlated with the incidence of diapause in the group. Diapause incidence and duration therefore are both determined maternally.     

Observations on the intensity of diapause and cold tolerance in larvae from twelve populations and two reciprocal crosses of the Indian meal moth, Plodia interpunctella

ABSTRACT. The duration of diapause in larvae of Plodia interpunctella (Hübner) (Lepidoptera, Pyralidae) was assessed at 20°C in LD 11:13. Mean times from hatch to pupation for diapausing larvae from different populations ranged from 88 to 236 days. Most non-diapausing larvae pupated within 70 days at this temperature. Transferring diapausing larvae to 25°C and LD 9:15, or to 20°C and LD 15:9, 70 days after hatch reduced the subsequent mean time to pupation by 18–82% and 9–63% respectively. Only two population samples terminated diapause faster under LD 15:9 at 20°C than under LD 9:15 at 25°C. The mortality of diapausing larvae caused by 6- or 10-week exposures at 5, 7.5 or 10°C was generally less than 25%. Hybrids produced from a reciprocal cross between a temperate and a tropical African stock survived well. For other stocks there was some correlation between survival and diapause intensity. The low temperature regime which resulted in the greatest shortening of pupation time after return to the conditions used to induce diapause, did not always coincide with the temperature permitting the best survival. Results, however, indicate that some individuals of all stocks but one from the tropics are likely to survive in the U.K.     

Methods of short-term storage of cultures of the blowfly <Emphasis Type="Italic">Calliphora Vicina</Emphasis> R.-D. (Diptera,Calliphoridae)

Three methods of short-term storage of the blowfly Calliphora vicina strains are considered based on the experimental study of 21 strains originating from different parts of the species range. The colony can be preserved as diapausing adults at 6° and darkness for 2–3 months or more, depending on the geographical origin of the population. During the first five days of adult life the flies should be kept at 12° and short day on a sugar diet, after which they should be transferred into a refrigerator. During artificial hibernation the flies also require periodic sugar feeding every 20 days (3–4 h at 20°C) to maintain their vital functions. The combination of temperatures of 20–23°C and a protein diet terminates reproductive diapause, and oviposition starts in 10–17 days. The fly strain may be preserved as reproductive females at 6°C and darkness with sugar feeding. Flies also require periodic sugar feeding at 20°C (3–4 hours). In this case the flies start laying eggs 2–3 days after being transferred to 20–23°C. The preservation of diapausing larvae is a more reliable method of prolonged strain storage. In this case the flies of maternal generation are maintained at 20–23°C on sugar and protein diet. The egg rafts laid during 5–6 hours are then transferred into 12°C and short day until hatchment. The hatched larvae should be immediately placed into a refrigerator (2–3 or 5–6°C), where they feed during 1–1.5 months and enter diapause. For strain restoration, the diapausing larvae should be transferred into 20–25°C, where they pupariate in 3–5 days and the flies emerge in nearly 10 days.     

Variations in the “spontaneous” dynamics of the tendency to diapause among generations of Trichogramma telengai Sor. (Hymenoptera, Trichogrammatidae)

Variations in the tendency to diapause were investigated in 5 successive generations of 6 parthenogenetic clones of Trichogramma telengai Sor. reared under constant laboratory conditions (the temperature of 20°C and the day length of 18 h). The tendency to diapause was estimated by the proportion of the progeny which diapaused at temperatures of 13, 14 and 15°C. Although the variations in the tendency to diapause in successive generations of different clones were positively correlated, the dynamics of the proportion of diapausing progeny in one of the studied clones was significantly different from that in the other clones reared under the same conditions. Moreover, the differences among subclones originated from the same female were also revealed. Analysis of the results suggests that the interclonal variations in the proportion of diapausing progeny were at least partly caused by endogenous factors. The differences between clones can arise during the development of one generation but can also damp very fast or, at least, they are not accumulated over generations. These variations are probably based on the same epigenetic mechanisms that form the basis of the maternal effect on the progeny diapause in Trichogramma species.     

Water and carbohydrate levels at different developmental stages and dynamics in hibernating pupae of Pieris melete (Lepidoptera: Pieridae)

For insight into the physiological indicators of diapause in Pieris melete, water and carbohydrate (glycogen and trehalose) levels were measured under both natural and laboratory conditions. The highest water content (3.71–3.79 mg/mg dry weight) was found in larvae and developing pupae, which was substantially higher than in diapausing pupae (2.59 mg/mg dry weight). Water content was almost stable during diapause, except for individuals approaching diapause termination (3.43–3.58 mg/mg dry weight). The total carbohydrate level was significantly higher in pre‐pupae (47.41 μg/mg) compared to larvae (22.80 μg/mg) and developing pupae (21.48 μg/mg). The highest level of trehalose was detected in winter diapausing pupae, and no trehalose was found in larvae or developing pupae. Levels of glycogen were highest in pre‐pupae and lowest in diapausing pupae. Levels of total carbohydrate decreased as diapause proceeded, and no significant changes were found in trehalose levels for diapausing pupae under natural conditions or treated for 60–90 days at 5°C. Pupae treated at 20°C for 60–90 days had significantly lower levels of trehalose than those treated for 30 days. Glycogen content was relatively stable at 5°C, but increased after treatment under natural conditions and 20°C for more than 60 days. These results suggest that the dynamics of water and carbohydrate levels are potential physiological diapause indicators, which show metabolic differences between trehalose and glycogen during diapause development.     

高温条件下棉铃虫化蛹率、夏滞育率和蛹重的变化

在预蛹期,高温处理能诱导棉铃虫蛹进入夏滞育。本实验着重就33～39℃的变温下滞育蛹和未滞育蛹的失重动态进行了对比研究,同时以常温（27℃）下蛹作为参照。研究发现： 在33～39℃的变温条件下,棉铃虫化蛹率显著低于其在常温下的化蛹率,且所化蛹中有63.2%的雄性和10.9%的雌性进入高温夏滞育,其中高温滞育蛹和未滞育蛹分别都轻于正常发育蛹。化蛹后第2日至第5日期间,高温滞育蛹失重量显著低于高温未滞育蛹和正常发育蛹的失重量,分别为3.62、13.30和5.49 mg;蛹期总失重量结果与化蛹后第2～5日间蛹失重量趋势一致,高温滞育蛹、未滞育蛹和正常发育蛹失重量分别为15.60、49.35和26.30 mg。蛹失重动态研究发现高温滞育蛹在夏滞育期间其失重曲线平缓,显著低于高温未滞育蛹和正常发育蛹;高温滞育蛹滞育解除后,其失重曲线与正常发育蛹的失重趋势基本一致。结果表明,棉铃虫夏滞育蛹能通过维持低的代谢水平来度过不利环境,具有一定的生态适应意义。     

Effect of soil burial depth and wetting on mortality of diapausing larvae and patterns of post-diapause adult emergence of sorghum midge, Stenodiplosis sorghicola (Coquillett) (Diptera: Cecidomyiidae)

Abstract  In south-eastern Queensland, Australia, sorghum planted in early spring usually escapes sorghum midge, Stenodiplosis sorghicola , attack. Experiments were conducted to better understand the role of winter diapause in the population dynamics of this pest. Emergence patterns of adult midge from diapausing larvae on the soil surface and at various depths were investigated during spring to autumn of 1987/88–1989/90. From 1987/88 to 1989/90, 89%, 65% and 98% of adult emergence, respectively, occurred during November and December. Adult emergence from larvae diapausing on the soil surface was severely reduced due to high mortality attributed to surface soil temperatures in excess of 40°C, with much of this mortality occurring between mid-September and mid-October. Emergence of adults from the soil surface was considerably delayed in the 1988/89 season compared with larvae buried at 5 or 10 cm which had similar emergence patterns for all three seasons. In 1989/90, when a 1-cm-deep treatment was included, there was a 392% increase in adult emergence from this treatment compared with deeper treatments. Some diapausing larvae on the surface did not emerge at the end of summer in only 1 year (1989/90), when 28.0% of the larvae on the surface remained in diapause, whereas only 0.8% of the buried larvae remained in diapause. We conclude that the pattern of emergence explains why spring plantings of sorghum in south-eastern Queensland usually escape sorghum midge attack.     

Effects of photoperiod and low temperature on diapause termination in the yellow-spotted longicorn beetle (Psacothea hilaris)

Abstract. .The effects of photoperiod and low temperature on diapause termination in the yellow-spotted longicorn beetle, Psacothea hilaris (Pascoe) (Coleoptera: Cerambycidae), were examined using a population from Ino, Japan. Diapausing insects obtained by rearing larvae under short daylength (12 or 13 h) at 25^oC were subjected to various treatments. When the photoperiod was changed at the same temperature, diapausing larvae showed a long-day response with a critical daylength between 13.5 and 14h. The diapause was terminated and consequently pupation occurred if the daylength was longer than 13.5 h. Chilling the diapausing larvae at 10^oC for 30 or more days also terminated diapause in most larvae irrespective of the photoperiods during and after chilling treatment. In contrast, the post-chilling photoperiod had a critical effect on development of diapausing larvae chilled for only 15 days.