Adaptive latitudinal cline of photoperiodic diapause induction in the parasitoid Nasonia vitripennis in Europe

Living in seasonally changing environments requires adaptation to seasonal cycles. Many insects use the change in day length as a reliable cue for upcoming winter and respond to shortened photoperiod through diapause. In this study, we report the clinal variation in photoperiodic diapause induction in populations of the parasitoid wasp Nasonia vitripennis collected along a latitudinal gradient in Europe. In this species, diapause occurs in the larval stage and is maternally induced. Adult Nasonia females were exposed to different photoperiodic cycles and lifetime production of diapausing offspring was scored. Females switched to the production of diapausing offspring after exposure to a threshold number of photoperiodic cycles. A latitudinal cline was found in the proportion of diapausing offspring, the switch point for diapause induction measured as the maternal age at which the female starts to produce diapausing larvae, and the critical photoperiod for diapause induction. Populations at northern latitudes show an earlier switch point, higher proportions of diapausing individuals and longer critical photoperiods. Since the photoperiodic response was measured under the same laboratory conditions, the observed differences between populations most likely reflect genetic differences in sensitivity to photoperiodic cues, resulting from local adaptation to environmental cycles. The observed variability in diapause response combined with the availability of genomic tools for N. vitripennis represent a good opportunity to further investigate the genetic basis of this adaptive trait.     

烟夜蛾滞育蛹和非滞育蛹的耐寒性

对烟夜蛾Helicoverpa assultaGuen?e不同日龄滞育蛹和非滞育蛹的过冷却点和结冰点测定结果表明,在预蛹至5日龄蛹期,过冷却点和结冰点均逐渐下降。在预蛹期和3日龄蛹期,滞育蛹的过冷却点与非滞育蛹差异不显著;5日龄蛹时,滞育蛹和非滞育蛹的过冷却点分别下降至(-15.7±1.8)℃和(-13.5±1.2)℃,结冰点分别降至(-12.7±2.7)℃和(-9.5±1.3)℃,两类蛹间的差异均达极显著水平。自然越冬后,滞育蛹的存活率显著高于非滞育蛹,在土壤不同深处越冬的蛹的存活率存在差异,距土表15cm深的蛹存活率最高。     

丽蝇蛹集金小蜂的生物学及其滞育调控机制研究进展

丽蝇蛹集金小蜂可以蛹外寄生多种双翅目蝇科害虫,在防治卫生害虫及牲畜害虫方面效果显著。该蜂是实验室内研究寄生蜂的理想模式昆虫,其发育学、行为学、生态学以及遗传学等的研究已有70多年的历史,近年来,随着金小蜂基因组测序的完成,系统的RNAi方法和CRISPR-Cas9技术在丽蝇蛹集金小蜂中的成功应用,该蜂现已成为优良的新型遗传学研究模式昆虫。丽蝇蛹集金小蜂以末龄幼虫进行兼性滞育,其滞育由母代经历的短光照和低温决定,研究该蜂的滞育,不仅有助于揭示昆虫滞育的分子机制,也可帮助解决有益昆虫实际应用中的贮存、运输和适时防治等问题,为提高天敌昆虫的应用效果提供技术支撑。本文总结了近年来丽蝇蛹集金小蜂的研究文献,主要介绍了该蜂的基本生物学特征,概括了现代分子生物学前沿技术在金小蜂研究中的应用情况,并重点讨论了丽蝇蛹集金小蜂滞育的最新研究进展,以期为深入金小蜂的滞育研究和促进该蜂其他研究领域的发展提供参考。     

Maternal control of cold and desiccation tolerance in eggs of the band-legged ground cricket Dianemobius nigrofasciatus in relation to embryonic diapause

Cold and desiccation tolerance was investigated in the eggs of the band‐legged ground cricket Dianemobius nigrofasciatus in relation to embryonic diapause. Diapause eggs were more tolerant to both desiccation and cold than non‐diapause eggs. In addition, diapause‐destined eggs on day zero (0–12 h after being laid) already showed high tolerance to these stresses before entering diapause. This clearly indicates that stress tolerance, like diapause, is controlled by photoperiod, but is not directly associated with diapause itself. Because the acquisition of stress tolerance predates the onset of diapause, it is plausible that diapause programming during some period before the onset of diapause is involved in the acquisition of stress tolerance. Weights and sizes were nearly identical in short‐day and long‐day eggs until day five. Sorbitol, a major sugar alcohol in eggs of D. nigrofasciatus, was accumulated at the same level in short‐day and long‐day eggs on days zero and five. These results indicate that the surface‐to‐volume ratio as well as the accumulation of sugar alcohol is not involved in the acquisition of stress tolerance. Maternal factors are clearly involved in the acquisition of stress tolerance in D. nigrofasciatus eggs, but the physiological mechanisms underlying the tolerance are still unclear.     

Influence of ecdysterone, precocene and compounds with juvenile hormone activity on induction, termination and maintenance of diapause in the parasitoid wasp, Nasonia vitripennis

ABSTRACT. Using the Galleria bioassay, no difference could be found between the JH titre of diapausing and developing Nasonia larvae. Compared to the values found in some Lepidoptera, the JH titres in Nasonia larvae are low, c. 14 Galleria units/g live weight. Induction of diapause could not be brought about by topical application of JH I, JH analogues, precocene, or ecdysterone to the maternal generation, nor by treating eggs or larvae with compounds with JH activity. Diapause was easily terminated by topical application of ecdysterone, however. If ecdysterone treatment was preceded by JH-treatment, the percentage of larvae terminating diapause was reduced: JH II seems to be more potent than JH I or JH III in this effect if the interval between JH and ecdysterone treatment is 72 h.     

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.     

Proteomic and metabolomic profiles of larval hemolymph associated with diapause in the cotton bollworm,Helicoverpa armigera

Background

Diapause is programmed developmental arrest coupled with the depression of metabolic activity and the enhancement of stress resistance. Pupal diapause is induced by environmental signals and is prepared during the prediapause phase. In the cotton bollworm, Helicoverpa armigera, the prediapause phase, which contains two sub-phases, diapause induction and preparation, occurs in the larval stage. Here, we performed parallel proteomic and metabolomic analyses on H. armigera larval hemolymph during the prediapause phase.

Results

By two-dimensional electrophoresis, 37 proteins were shown to be differentially expressed in diapause-destined larvae. Of these proteins, 28 were successfully identified by MALDI-TOF/TOF mass spectrometry. Moreover, a total of 22 altered metabolites were found in diapause-destined larval hemolymph by GC-MS analysis, and the levels of 17 metabolites were elevated and 5 were decreased.

Conclusions

The proteins and metabolites with significantly altered levels play different roles in diapause-destined larvae, including diapause induction, metabolic storage, immune response, stress tolerance, and others. Because hemolymph circulates through the whole body of an insect, these differences found in diapause-destined larvae most likely correspond to upstream endocrine signals and would further influence other organ/tissue activities to determine the insect’s fact: diapause or development.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-14-751) contains supplementary material, which is available to authorized users.     

大猿叶虫滞育进程中成虫内生殖系统的形态特征

大猿叶虫Colaphellus bowringi Baly是十字花科蔬菜的一种重要害虫,以成虫在土壤中滞育越冬和越夏。本研究通过解剖大猿叶虫非滞育成虫,观察描述了雌雄成虫内生殖系统的结构特点,并绘制了雌雄成虫内生殖系统模式图。对滞育初期、滞育期间、滞育解除后未取食和滞育解除后取食成虫解剖显示,滞育初期,雌雄成虫内生殖系统几乎与羽化初期成虫一致。滞育期间雄成虫的附腺和射精管亦不发达,滞育解除后未取食雄成虫的附腺膨大且粗于输精管,射精管略膨大,取食雄成虫的射精管呈不透明淡桔黄色。滞育期间雌成虫的卵巢小,略大于滞育初期卵巢,少数卵巢小管的基部可见具卵黄原沉淀的卵粒。滞育解除后未供食雌成虫卵巢明显膨大可见大量成形卵粒,部分卵粒可见卵黄原沉淀;取食雌成虫的卵巢膨大,可见大量成熟卵粒,侧输卵管和总输卵管中可见待产的卵。作者认为,大猿叶虫成虫在滞育期间能够缓慢发育,部分滞育前积累的代谢物质被用于滞育后发育,但只有经过取食,成虫才能正常交配和产卵。     

The spatio-temporal partitioning of sperm by males of the prospermatogenic parasitoid Nasonia vitripennis is in line with its gregarious lifestyle

Male fitness depends on the number of lifetime progeny of their mates and could be constrained by the chance of finding a mate, lifespan and temporal patterns of sperm production and allocation. Here, we used the parasitic wasp Nasonia vitripennis with a two-week lifespan and a gregarious lifestyle, to analyze how the reproductive system is organized to allocate spermatozoa over consecutive matings. Results show that spermatogenesis is synchronized and completed one day before emergence so that males emerge with a full sperm complement. We also found a regulation of spermatozoa transfer between testis and seminal vesicles that allows males to partition small ejaculates over multiple matings. Overall, this study shows that for N. vitripennis, male fertilization potential is determined (1) at the pupal stage, when spermatogenesis takes place to generate a complete life-long stock, (2) on emergence, when transport of spermatozoa from testes to seminal vesicles is initiated and (3) in adulthood, during which spermatozoa are partitioned over successive copulations. Such life history-traits are consistent with the gregarious lifestyle of N. vitripennis.     

The influence of temperature and photoperiod on the reproductive diapause and cold tolerance of spotted‐wing drosophila,Drosophila suzukii

Knowledge regarding the reproductive status of spotted‐wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), is of critical importance in predicting potential infestations of this invasive pest, as eggs are laid in ripe or ripening fruit of several commercially important small‐fruit crops. Token environmental stimuli for the induction of reproductive diapause and improved cold tolerance were identified for D. suzukii. Diapause induction was evaluated by assessing, via dissection, the number of mature eggs carried by field‐captured D. suzukii and laboratory‐reared D. suzukii held under various temperature and photoperiod regimes. Egg load decreased over time in females trapped from July to December at sites in Benton County, OR, and Ontario County, NY, both USA, and reached zero eggs by December at all sites. Photoperiods below 14 h of day length led to reduced egg maturation in laboratory‐reared flies held at moderate temperatures (15 or 20 °C). Whereas very few mature eggs were found in females held at 10 °C under short‐ or long‐day photoperiods for several weeks after eclosion, a spontaneous return to ovarian maturity was observed in short‐day‐entrained females after 7 weeks. Diapause termination was investigated by evaluating fecundity in diapausing females returned to optimal environmental conditions. Whereas long‐day‐entrained flies began producing offspring immediately upon return to optimal conditions, short‐day‐entrained flies returned after 1 and 6 weeks at 10 °C were slower to produce offspring than colony flies or short‐day‐entrained flies returned after 7 weeks. Cold tolerance was evaluated by observing chill coma recovery rates after 24 h exposure to ?1 °C. Cold‐acclimated (diapausing) females recovered from chill coma faster than cold‐hardened or unacclimated females.     

Improved salt tolerance of melon (Cucumis melo L.) by the addition of proline and potassium nitrate

A pot experiment was carried out under glasshouse conditions with melon (Cucumis melo) cv. “Tempo F1” in a mixture of peat, perlite and sand (1:1:1) to investigate the effects of external proline and potassium nitrate applications to salinity-treated (150 mM) plants with respect to fruit yield, plant growth, some physiological parameters and ion uptake. Treatments were—(i) control (C): plants receiving nutrient solution, (ii) salinity treatment, as for control plus 150 mM NaCl. Salinity treatment was combined with or without either 5 mM supplementary KNO₃ or 10 mM proline. The salt treatment (150 mM NaCl) led to significant decreases in plant growth, fruit yield, relative water content (RWC), stomatal density, uptake of Ca²⁺, K⁺ and N, and chlorophyll a and b contents, accompanied by significant increases in Na⁺ uptake, proline concentration and membrane permeability. Supplementary KNO₃ and proline treatments significantly ameliorated the adverse effects of salinity on plant growth, fruit yield and the physiological parameters examined. This could be attributed to the effects of all the external supplements in maintaining membrane permeability, and increasing concentrations of Ca²⁺, N and K⁺ in the leaves of plants subjected to salt stress.     

两种圆柏属植物不同季节显微和超微结构变化与耐寒性的关系

以祁连圆柏和圆柏为材料,研究了圆柏属植物的抗寒性与解剖结构变化,以及淀粉与可溶性糖粉转化的关系.结果表明,祁连圆柏和圆柏叶片的上下表皮细胞外覆盖一层角质膜,叶肉细胞间隙大,形成发达的通气组织.在生长季节,二者叶绿体内均积累了很多淀粉粒,在冷冻休眠季节淀粉粒消失或变小,同时叶片可溶性糖分含量增加,祁连圆柏的可溶性糖分增幅高于圆柏;而休眠季节圆柏叶绿体受到低温伤害,脂质球数目增多,出现脂质小滴,部分叶绿体变形,祁连圆柏类囊体的结构受到影响较小,脂质球数量少,叶绿体形状没有发生明显的变化.因此,祁连圆柏和圆柏抗寒性与发达的通气组织和淀粉粒生长季节的积累有关,在低温胁迫下祁连圆柏叶绿体表现出高的稳定性,通过可溶性糖分的调节力强,耐寒性增强.     

Physiological and biochemical analysis of overwintering and cold tolerance in two Central European populations of the spruce bark beetle, Ips typographus

Overwintering success is one of the key aspects affecting the development and outbreaks of the spruce bark beetle, Ips typographus (L.) populations. This paper brings detailed analysis of cold tolerance, and its influence on overwintering success, in two Central European populations of I. typographus during two cold seasons. Evidence for a supercooling strategy in overwintering adults is provided. The lower lethal temperature corresponds well to the supercooling point that ranges between −20 and −22 °C during winter months. The supercooled state is stabilized by the absence of internal ice nucleators and by seasonal accumulation of a mixture of sugars and polyols up to the sum concentration of 900 mM. The cryoprotective function of accumulated metabolites is probably based on increasing the osmolality and viscosity of supercooled body fluids and decreasing the relative proportion of water molecules available for lethal formation of ice nuclei. No activity of thermal hysteresis factors (stabilizers of supercooled state) was detected in hemolymph. Lethal times for 50% mortality (Lts50) in the supercooled state at −5, −10 or −15 °C are weeks (autumn, spring) or even months (winter), suggesting relatively little mortality caused by chill injury. Lts50 at −15 °C are significantly shorter in moist (6.9 days) than in dry (>42 days) microenvironment because there is higher probability of external ice nucleation and occurrence of lethal freezing in the moist situation.     

Brown adipose tissue specific lack of uncoupling protein 3 is associated with impaired cold tolerance and reduced transcript levels of metabolic genes

Uncoupling protein 3 (Ucp3) is located within the mitochondrial inner membrane of brown adipose tissue and skeletal muscle. It is thought to be implicated in lipid metabolism and defense against reactive oxygen species. We previously reported on a mutation in our breeding colony of Djungarian hamsters (Phodopus sungorus) that leads to brown adipose tissue specific lack of Ucp3 expression. In this study we compared wildtype with mutant hamsters on a broad genetic background. Hamsters lacking Ucp3 in brown adipose tissue displayed a reduced cold tolerance due to impaired nonshivering thermogenesis. This phenotype is associated with a global decrease in expression of metabolic genes but not of uncoupling protein 1. These data implicate that Ucp3 is necessary to sustain high metabolic rates in brown adipose tissue.     

Cloning of heat shock protein genes (hsp70, hsc70 and hsp90) and their expression in response to larval diapause and thermal stress in the wheat blossom midge,Sitodiplosis mosellana

Sitodiplosis mosellana Géhin, one of the most important pests of wheat, undergoes obligatory diapause as a larva to survive unfavorable temperature extremes during hot summers and cold winters. To explore the potential roles of heat shock proteins (hsp) in this process, we cloned full-length cDNAs of hsp70, hsc70 and hsp90 from S. mosellana larvae, and examined their expression in response to diapause and short-term temperature stresses. Three hsps included all signature sequences of corresponding protein family and EEVD motifs. They showed high homology to their counterparts in other species, and the phylogenetic analysis of hsp90 was consistent with the known classification of insects. Expression of hsp70 and hsp90 were highly induced by diapause, particularly pronounced during summer and winter. Interestingly, hsp70 was more strongly expressed in summer than in winter whereas hsp90 displayed the opposite pattern. Abundance of hsc70 mRNA was comparable prior to and during diapauses and was highly up-regulated when insects began to enter the stage of post-diapause quiescence. Heat-stressed over-summering larvae (⩾30 °C) or cold-stressed over-wintering larvae (⩽0 °C) could further elevate expression of these three genes, but temperature extremes i.e. as high as 45 °C or as low as −15 °C failed to trigger such expression patterns. Notably, hsp70 was most sensitive to heat stress and hsp90 was most sensitive to cold stress. These results suggested that hsp70 and hsp90 play key roles in diapause maintenance and thermal stress; the former may be more prominent contributor to heat tolerance and the latter for cold tolerance. In contrast, hsc70 most likely is involved in developmental transition from diapause to post-diapause quiescence, and thus may serve as a molecular marker to predict diapause termination.     

Effects of photoperiod and temperature on the rate of larval development, food conversion efficiency, and imaginal diapause in Leptinotarsa decemlineata

Larvae of Leptinotarsa decemlineata developed faster and consumed less food under short-day (Sd, 12:12 h light:darkness) than under long-day (Ld, 18:6 h L:D) conditions. The average index of food conversion efficiency was 5.4 in the Ld (25 degrees C), and 7.2 and 11.9 (at 20 and 25 degrees C, respectively) in the Sd insects. Pupae were smaller under the Ld conditions due to a greater loss of biomass during the prepupal period that was nearly twice longer than in the Sd insects. Virgin Ld females laid eggs for 6 months and survived 13 months. The lack of oviposition, reduced food intake, and behavioural changes characterised diapause in the Sd adults. Application of 100 microg JH III to newly ecdysed adults was used to probe diapause intensity. At 25 degrees C, the treatment elicited oviposition most effectively in females that were just transferred from the Ld to the Sd conditions. A distinctly lower response occurred in insects that had been kept under Sd conditions since hatching; their transfer to Ld conditions at the time of treatment had little effect on JH sensitivity. JH application to Sd females reared at 20 degrees C caused enlargement of the germaria but no eggs were formed.     

Seasonal and geographical variation in diapause and cold hardiness of the Asian corn borer,Ostrinia furnacalis

Asian corn borer, Ostrinia furnacalis (Guenée), is a key corn pest in the Asian‐Western Pacific countries. It overwinters as full‐grown larvae in plant stalks or in a spun‐silk covering located in the plant debris in the temperate regions of China. Supercooling point (SCP) and survival rate after low sub‐zero temperature treatment were assessed for field‐collected populations in the laboratory using a cool bath with a 1°C/min cooling rate until ?40°C. Mean SCPs were varied among geographical populations, with a significant decline from ?22.7°C of Haikou, the multivoltine tropical population in the south, to ?28.5°C of Gongzhuling, the univoltine temperate population in the northeast of China. In addition, there was more than 1°C difference in SCP between Gongzhuling univoltine and bivoltine populations that were from the same geographic origin. Mean SCPs of the Guangzhou population fluctuated over the year, with significantly lower SCPs in winter than in other seasons, which correlated with a significantly higher proportion of diapausing larvae in winter than in other seasons. Over 41% of overwintering larvae from the northeast population could withstand to be supercooled for a few minutes to the low sub‐zero temperature of ?40°C, but only 6.7% of their southern counterparts did so. The findings from this study suggest that O. furnacalis mostly takes advantage of freeze avoidance as diapausing larvae for overwintering in the southern region, whereas it exhibits freeze tolerance in diapause in the northeastern region.     

Endocrine changes associated with metamorphosis and diapause induction in the yellow-spotted longicorn beetle, Psacothea hilaris

At 25 degrees C and under a long-day photoperiod, all 5th instar Psacothea hilaris larvae pupate at the next molt. Under a short-day photoperiod, in contrast, they undergo one or two additional larval molts and enter diapause; the 7th instar larvae enter diapause without further molt. The changes in hemolymph juvenile hormone (JH III) titers, JH esterase activity, and ecdysteroid titers in pupation-destined, pre-diapause, and diapause-destined larvae were examined. JH titers of the 5th instar pupation-destined larvae decreased continuously from 1.3 ng/ml and became virtually undetectable on day 13, when JH esterase activity peaked. Ecdysteroids exhibited a small peak on day 8, 1 day before gut purge, and a large peak on day 11, 2 days before the larvae became pre-pupae. The two ecdysteroid peaks are suggested to be associated with pupal commitment and pupation, respectively. JH titers of the 5th instar pre-diapause larvae were maintained at approximately 1.5 ng/ml for 5 days and then increased to form a peak (3.3 ng/ml) on day 11. JH esterase activity remained at a low level throughout. Ecdysteroid levels exhibited a large peak of 40 ng/ml on day 18, coincident with the larval molt to the 6th instar. JH titers of the 7th instar diapause-destined larvae peaked at 1.9 ng/ml on day 3, and a level of approximately 1.1 ng/ml was maintained even 30-60 days into the instar, when they were in diapause. Ecdysteroid titers remained approximately 0.02 ng/ml. Diapause induction in this species was suggested to be a consequence of high JH and low ecdysteroid titers.