Adaptations to cold in Canadian arctic insects

The survival of insects that inhabit Canadian arctic regions depends on a number of factors which have important ecological, behavioral, physiological, and biochemical components. The ability to withstand low winter temperatures is one of the most conspicuous adaptations of northern insects and the one most closely studied in the laboratory. Most species studied so far conform to one or other of the two major overwintering strategies, namely, frost susceptibility, the ability to avoid freezing by supercooling to a considerable degree, or frost tolerance, the survival of actual ice formation within the body. The Arctic beetle, Pytho americanus Kirby, is frost tolerant in both larval and adult stages, a situation which would be congruous with its northern distribution and allow it to spread its life cycle over a number of growing seasons. The main biochemical correlates during the cold-hardening process in this species are increasing glycerol and decreasing glycogen concentrations. In addition to its normally assumed roles in cryoprotection there is evidence to suggest that glycerol may further serve to minimize dehydration in the overwintering insect by increasing the level of bound water. P. americanus larvae and adults have narrow supercooling ranges and maintain their supercooling points in the region of ?4 to ?8 °C. It is hypothesized that these elevated supercooling points are a result of the presence in the hemolymph of nucleating agents which ensure ice formation at high subzero temperatures.Low temperature tolerance strategies of some other arctic and alpine species have been examined and compared with those of relatives from more southerly latitudes. P. americanus has been collected in the Canadian Rockies at elevations of over 6000′, and its frost-tolerant attributes are identical to those of the population collected in the Arctic. A closely related species, P. deplanatus, from the Rockies, however, although it too exhibits frost tolerance in the larval stage, differs markedly from P. americanus in its ability to depress its supercooling range to ?54 °C. It appears that P. deplanatus does not have the ability to synthesize ice-nucleating agents and, therefore, can overwinter in a supercooled condition. Two congeneric species of willow leaf gall sawflies (Pontania spp.), one from Tuktoyaktuk, N.W.T., and the other from southern Vancouver Island have also been compared and contrasted. Pontania sp. on Salix glauca (Tuk., ca. 70 °N) is frost tolerant in its larval stage, has relatively high supercooling points (ca. ?9.0 °C), but does not accumulate glycerol. Pontania sp. from Salix lasiandra (Victoria, ca. 48 °N) has almost identical overwintering properties, indicating the close phylogenetic affinities of cold tolerance in this genus rather than independent adaptation to widely different climatic conditions. Some of the lowest supercooling points ever recorded are from willow stem gall forming insects. Rhabdophaga sp. (Cecidomyiidae) forms potato galls on the stems of Salix lanata in the Inuvik area, N.W.T. After low temperature acclimation, supercooling points down to ?66 °C have been recorded from individual larvae. This is a record, and it indicates that we may be dealing with a system in which most water is in a metabolically bound state. Glycerol levels reach 20% of the fresh body weight during this period. Diastrophus kincaidii Cynipidae) forms stem galls on Thimble Berry (Rubus parviflorus) on southern Vancouver Island. Both of the forementioned species overwinter as larvae in their galls and are, therefore, exposed to ambient air temperatures. A more benign winter climate on Vancouver Island is reflected in the fact that D. kincaidii has supercooling points only in the ?30 to ?33 °C range at the peak of low temperature acclimation, and glycerol levels just below 4% of fresh body weight. Both species are frost susceptible and depend on their supercooling abilities to survive low winter temperatures.     

Change in overwintering mechanism of the Cucujid beetle, Cucujus clavipes

Overwintering larvae of the Cucujid beetle, Cucujus clavipes, were freeze tolerant, able to survive the freezing of their extracellular body fluids, during the winter of 1978–1979. These larvae had high levels of polyols (glycerol and sorbitol), thermal hysteresis proteins and haemolymph ice nucleators that prevented extensive supercooling (the supercooling points of the larvae were ? 10°C), thus preventing lethal intracellular ice formation. In contrast, C. clavipes larvae were freeze suspectible, died if frozen, during the winter of 1982–1983, but supercooled to ～ ? 30°C. The absence of the ice nucleators in the 1982–1983 larvae, obviously essential in the now freeze-susceptible insects, was the major detected difference in the larvae from the 2 years. However, experiments in which the larvae were artifically seeded at ? 10°C (the temperature at which the natural haemolymph ice nucleators produced spontaneous nucleation in the 1978–1979 freeze tolerant larvae) demonstrated that the absence of the ice nucleators was not the critical factor, or at least not the only critical factor, responsible for the loss of freeze tolerance in the 1982–1983 larvae. The lower lethal temperatures for the larvae were approximately the same during the 2 winters in spite of the change in overwintering strategy.     

Seasonal changes in supercooling capacity and major cryoprotectants of overwintering Asian longhorned beetle (Anoplophora glabripennis) larvae

相似文献   

寄主植物对甜菜夜蛾三龄幼虫抗寒力的影响

寄主植物是影响昆虫抗寒性的主要因子之一。研究了不同温度下甜菜夜蛾Spodoptera exigua(Hübner)3龄幼虫取食小白菜、甘蓝、葱和菠菜后,对过冷却能力和体内冷冻保护剂的影响。结果表明,寄主植物对甜菜夜蛾3龄幼虫的过冷却能力有显著性影响,其中以取食甘蓝的幼虫过冷却点最低。温度和寄主植物对其过冷却点、结冰点和虫体含水量有明显的交互作用。寄主植物对其体内的海藻糖含量有显著性影响,而对甘油和糖原含量没有显著性影响。温度和寄主植物仅对海藻糖含量有显著的交互作用。随着温度的升高,取食不同寄主的幼虫体内海藻糖和糖原含量的变化趋势完全相反,认为海藻糖是由糖原转化而来。研究结果提示冬季合适的寄主植物有利于甜菜夜蛾低龄幼虫越冬。     

Global and comparative proteomic profiling of overwintering and developing mountain pine beetle,Dendroctonus ponderosae (Coleoptera: Curculionidae), larvae

BackgroundMountain pine beetles, Dendroctonus ponderosae Hopkins (Coleoptera: Curculionidae), are native to western North America, but have recently begun to expand their range across the Canadian Rocky Mountains. The requirement for larvae to withstand extremely cold winter temperatures and potentially toxic host secondary metabolites in the midst of their ongoing development makes this a critical period of their lives.ResultsWe have uncovered global protein profiles for overwintering mountain pine beetle larvae. We have also quantitatively compared the proteomes for overwintering larvae sampled during autumn cooling and spring warming using iTRAQ methods. We identified 1507 unique proteins across all samples. In total, 33 proteins exhibited differential expression (FDR < 0.05) when compared between larvae before and after a cold snap in the autumn; and 473 proteins exhibited differential expression in the spring when measured before and after a steady incline in mean daily temperature. Eighteen proteins showed significant changes in both autumn and spring samples.ConclusionsThese first proteomic data for mountain pine beetle larvae show evidence of the involvement of trehalose, 2-deoxyglucose, and antioxidant enzymes in overwintering physiology; confirm and expand upon previous work implicating glycerol in cold tolerance in this insect; and provide new, detailed information on developmental processes in beetles. These results and associated data will be an invaluable resource for future targeted research on cold tolerance mechanisms in the mountain pine beetle and developmental biology in coleopterans.     

Cold tolerance of the montane Sierra leaf beetle,Chrysomela aeneicollis

Small ectothermic animals living at high altitude in temperate latitudes are vulnerable to lethal cold throughout the year. Here we investigated the cold tolerance of the leaf beetle Chrysomela aeneicollis living at high elevation in California’s Sierra Nevada mountains. These insects spend over half their life cycle overwintering, and may therefore be vulnerable to winter cold, and prior studies have demonstrated that survival is reduced by exposure to summertime cold. We identify overwintering microhabitat of this insect, describe cold tolerance strategies in all life stages, and use microclimate data to determine the importance of snow cover and microhabitat buffering for overwinter survival. Cold tolerance varies among life history stages and is typically correlated with microhabitat temperature: cold hardiness is lowest in chill-susceptible larvae, and highest in freeze-tolerant adults. Hemolymph osmolality is higher in quiescent (overwintering) than summer adults, primarily, but not exclusively, due to elevated hemolymph glycerol. In nature, adult beetles overwinter primarily in leaf litter and suffer high mortality if early, unseasonable cold prevents them from entering this refuge. These data suggest that cold tolerance is tightly linked to life stage. Thus, population persistence of montane insects may become problematic as climate becomes more unpredictable and climate change uncouples the phenology of cold tolerance and development from the timing of extreme cold events.     

The role of macromolecular antifreeze in the darkling beetle,Meracantha contracta

Summary Macromolecular antifreeze solutes are present in the hemolymph of the overwintering larvae of the darkling beetle,Meracantha contracta. These antifreeze solutes produce a thermal hysteresis in the hemolymph of overwinteringMeracantha larvae whereby the freezing point of the hemolymph may be 3–4 °C below the melting point. This thermal hysteresis is very similar to that produced by proteinaceous and glycoproteinaceous antifreezes which are used by many cold water, marine teleost fishes to prevent freezing. One function of the macromolecular antifreeze inMeracantha may be to hinder inoculative freezing which might otherwise occur because of the dampness of the hibernaculae. A probably more important function is to depress the supercooling point of the frost susceptibleMeracantha larvae, thereby preventing lethal ice formation in the larvae's body fluids down to temperatures of approximately –11 °C.     

The ontogeny of cold tolerance in the gall fly, Eurosta solidagensis

The lower lethal temperature of many insects indicates an overwintering flexibility as a result of either extensive supercooling or production of cryoprotectants. Ontogenetically, the gall fly (Eurosta solidagensis) utilizes both means of seasonal cryoprotection. All stages except third instar larvae demonstrate supercooling points well below the lowest temperature normally experienced by that particular stage. The third instar larvae exhibit a high supercooling point but are well protected by a cryoprotectant system consisting of glycerol, sorbitol, and trehalose. Glycerol is accumulated, possibly from triglyceride sources, during early autumn and reaches plateau levels (0·6 M) by early winter. Sorbitol synthesis is delayed until freezing exposures and reaches a plateau with glycerol at 0·3 M. It is not until mid-winter that peak trehalose levels are reached (300 mg %). All cryoprotectant levels are a reflection of haemolymph concentrations.Laboratory acclimation experiments further quantify these results. Trehalose synthesis is time and temperature dependent and appears to be affected by developmental processes.     

Seasonal changes in the composition of storage and membrane lipids in overwintering larvae of the codling moth,Cydia pomonella

The codling moth (Cydia pomonella) is a major insect pest of apples worldwide. It overwinters as a diapausing fifth instar larva. The overwintering is often a critical part of the insect life-cycle in temperate zone. This study brings detailed analysis of seasonal changes in lipid composition and fluidity in overwintering larvae sampled in the field. Fatty acid composition of triacylglycerol (TG) depots in the fat body and relative proportions of phospholipid (PL) molecular species in biological membranes were analyzed. In addition, temperature of melting (T_m) in TG depots was assessed by using differential scanning calorimetry and the conformational order (fluidity) of PL membranes was analyzed by measuring the anisotropy of fluorescence polarization of diphenylhexatriene probe in membrane vesicles. We observed a significant increase of relative proportion of linoleic acid (C18:2n6) at the expense of palmitic acid (C16:0) in TG depots during the larval transition to diapause accompanied with decreasing melting temperature of total lipids, which might increase the accessibility of depot fats for enzymatic breakdown during overwintering. The fluidity of membranes was maintained very high irrespective of developmental mode or seasonally changing acclimation status of larvae. The seasonal changes in PL composition were relatively small. We discuss these results in light of alternative survival strategies of codling moth larvae (supercooling vs. freezing), variability and low predictability of environmental conditions, and other cold tolerance mechanisms such as extending the supercooling capacity and massive accumulation of cryoprotective metabolites.     

泰安地区松阿扁叶蜂越冬幼虫抗寒性

松阿扁叶蜂越是松树的重要食叶害虫之一。其幼虫在松林土壤中越冬,越冬幼虫的抗寒能力影响翌年的种群密度和危害程度。用NTC-TD热敏电阻 数字电表法研究了越冬幼虫的过冷却能力并调查了越冬场所、龄期、死亡率等。结果表明:越冬幼虫的过冷却点随外界温度的高低而变化。4、5、6龄越冬幼虫平均过冷却点依次由越冬初期的-19.3℃-、20.3℃-、21.6℃降至越冬期的-22.5℃-、24.8℃-、23.1℃,冰点依次由越冬初期的-12.8℃-、12.9℃-、12.1℃降至越冬期的-14.1℃、-16.8℃-、15.3℃。从越冬期进入早春出蛰期后,随着温度的回升,过冷却点也随之升高。调查林间越冬幼虫龄期表明,4龄、5龄、6龄皆有,以5龄为最多,占53%;越冬幼虫以山坡中部和顶部土壤瘠薄处虫口密度较小,分别占28.6%和29.3%,山坡基部土层较厚处密度较大,占42.1%;越冬幼虫在树冠内的分布,以南部方位最多(27.0%),东部方位(26.3%)和西部方位(25.6%)略少,北部方位最少(21.1%);调查幼虫死亡因子,以真菌感染致死最多,占总死亡数的42.4%,细菌和其它因子分别为28%和29.7%。     

Reprint of: Seasonal changes in the composition of storage and membrane lipids in overwintering larvae of the codling moth,Cydia pomonella

The codling moth (Cydia pomonella) is a major insect pest of apples worldwide. It overwinters as a diapausing fifth instar larva. The overwintering is often a critical part of the insect life-cycle in temperate zone. This study brings detailed analysis of seasonal changes in lipid composition and fluidity in overwintering larvae sampled in the field. Fatty acid composition of triacylglycerol (TG) depots in the fat body and relative proportions of phospholipid (PL) molecular species in biological membranes were analyzed. In addition, temperature of melting (T_m) in TG depots was assessed by using differential scanning calorimetry and the conformational order (fluidity) of PL membranes was analyzed by measuring the anisotropy of fluorescence polarization of diphenylhexatriene probe in membrane vesicles. We observed a significant increase of relative proportion of linoleic acid (C18:2n6) at the expense of palmitic acid (C16:0) in TG depots during the larval transition to diapause accompanied with decreasing melting temperature of total lipids, which might increase the accessibility of depot fats for enzymatic breakdown during overwintering. The fluidity of membranes was maintained very high irrespective of developmental mode or seasonally changing acclimation status of larvae. The seasonal changes in PL composition were relatively small. We discuss these results in light of alternative survival strategies of codling moth larvae (supercooling vs. freezing), variability and low predictability of environmental conditions, and other cold tolerance mechanisms such as extending the supercooling capacity and massive accumulation of cryoprotective metabolites.     

光周期对中华通草蛉自然越冬成虫及实验种群幼虫耐寒能力的影响

【目的】滞育诱导期进行短光照处理可影响昆虫耐寒性。为明确光周期对中华通草蛉Chrysoperla sinica (Tjeder)耐寒性的影响, 针对中华通草蛉滞育解除过程及非滞育虫态的耐寒性进行了一系列研究。【方法】测定了中华通草蛉自然越冬成虫的过冷却点（supercooling point, SCP）以及长光周期（15L∶9D）和短光周期（9L∶15D）条件下自然越冬成虫在滞育解除过程中在-12℃下的死亡率, 并测定了室内长、 短两种光周期下实验种群2龄和3龄幼虫的过冷却点（SCP）、 结冰点（freezing point, FP）以及-7℃下的死亡率。【结果】中华通草蛉12月份的自然越冬成虫SCP集中在-10~-14℃之间。SCP低于-12℃的个体占43.70%, 且-12℃处理1 d死亡率为62.00%。-12℃处理1 d条件下的长、 短光周期处理自然越冬成虫, 除处理0 d外, 长光周期处理死亡率均高于短光周期处理的, 且在处理15 （P=0.012）, 20 （P=0.01）和25 d （P=0.001）差异显著。中华通草蛉试验种群相同龄期幼虫在短光周期下的SCP和FP均高于长光周期下, 但差异不显著(P>0.05); 但在-7℃下, 2龄幼虫短光周期下的低温死亡率为67.00%±4.04%, 显著低于长光周期下的低温死亡率（78.00%±1.33%）（P=0.011）, 3龄幼虫短光周期条件下低温死亡率为24.33%±1.33%, 显著低于长光周期下的低温死亡率（53.00%±3.46%）（P=0.002）。【结论】中华通草蛉为结冰敏感型, 诱导滞育的短光照处理可提高其幼虫期及滞育解除过程中成虫的耐寒能力。     

The inhibition of ice nucleators by insect antifreeze proteins is enhanced by glycerol and citrate

Antifreeze proteins depress the freezing point of water while not affecting the melting point, producing a characteristic difference in freezing and melting points termed thermal hysteresis. Larvae of the beetle Dendroides canadensis accumulate potent antifreeze proteins (DAFPs) in their hemolymph and gut, but to achieve high levels of thermal hysteresis requires enhancers, such as glycerol. DAFPs have previously been shown to inhibit the activity of bacterial and hemolymph protein ice nucleators, however, the effect was not large and therefore the effectiveness of the DAFPs in promoting supercooling of the larvae in winter was doubtful. However, this study demonstrates that DAFPs, in combination with the thermal hysteresis enhancers glycerol (1 M) or citrate (0.5 M), eliminated the activity of hemolymph protein ice nucleators and Pseudomonas syringae ice-nucleating active bacteria, and lowered the supercooling points (nucleation temperatures) of aqueous solutions containing these ice nucleators to those of water or buffer alone. This shows that the DAFPs, along with glycerol, play a critical role in promoting hemolymph supercooling in overwintering D. canadensis. Also, DAFPs in combination with enhancers may be useful in applications which require inhibition of ice nucleators.     

Effects of transgenic Bt cotton on overwintering characteristics and survival of Helicoverpa armigera

The effects of transgenic Bt cotton on the overwintering generation of the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), are unknown. We hypothesized that a Bt cotton diet may adversely affect fitness of this generation and examined fresh weight, lipids, glycogens, low-molecular-weight sugars and SCPs (supercooling points) of pupae, as well as survival of larvae, diapausing pupae and adult emergence in comparison with controls. Field and laboratory experiments showed that larvae fed on Bt cotton had a decreased pupation rate, and fewer entered diapause and emerged as adults compared with larvae fed non-Bt cotton. Furthermore, larvae fed Bt cotton had reduced pupal weight, glycogen content and trehalose levels both in diapausing and in non-diapausing pupae, and only diapausing pupae had an increased SCP compared to controls. The SCPs of diapausing pupae reared on Bt cotton were significantly higher than those reared on non-Bt cotton. The trehalose levels of diapausing pupae reared on Bt cotton were significantly lower than those of larvae reared on non-Bt cotton. Thus, these results suggest that a Bt cotton diet weakens the preparedness of cotton bollworm for overwintering and reduces survival of the overwintering generation, which will in turn reduce the density of the first generation in the following year. Effects of transgenic Bt cotton on the overwintering generation of cotton bollworm appear to have significantly contributed to the suppression of cotton bollworm observed throughout northern China in the past decade.     

越冬期不同阶段二点委夜蛾越冬幼虫耐寒性变化

【目的】二点委夜蛾Athetis lepigone (Moschler)是我国夏玉米苗期的新害虫,随着耕作制度的改变,二点委夜蛾的发生危害区域和面积逐渐扩大。本研究探讨二点委夜蛾的抗寒能力,为揭示抗寒机理提供理论基础。【方法】分别在越冬期的3个不同阶段,即越冬初期（2012年11月7日）、越冬期（2012年1月20日）和越冬末期（2013年3月5日）,对二点委夜蛾老熟幼虫的体重、过冷却点（supercooling point, SCP）、结冰点（freezing point, FP）、含水量、脂肪和糖原含量进行测定。【结果】过冷却点在这3个时期有显著差异,最低值（-23.16±0.38℃）出现在1月份,最高值（-16.24±1.24℃）出现在越冬初期,结冰点变化趋势与过冷却点一致。通过定量检测发现,虫体鲜重与过冷却点无相关性（r=0.17, P=0.12）;脂肪含量在越冬初期含量最高,而在越冬末期最低;糖原含量在越冬期含量最低;自由水的含量随着过冷却能力升高而降低,随其降低而升高,而结合水含量恰好相反。【结论】二点委夜蛾的抗寒性在越冬期不同阶段出现明显的变化,即随着冬季低温的到来,其抗寒能力逐渐增强,冬季过后又随气温的回升,其抗寒力逐渐减弱。     

Diapause development and acclimation regulating enzymes associated with glycerol synthesis in the Shonai ecotype of the rice stem borer larva,Chilo suppressalis walker

Overwintering larvae of the Shonai ecotype of the rice stem borer, Chilo suppressalis, enter diapause in early September and terminate diapause at the end of October. Cold acclimation at 0°C did not influence glycerol, trehalose or glycogen content in larvae collected on 22 September. Acclimation at 0°C increased the glycerol content and reduced the glycogen content significantly in larvae collected on 2 October and 22 November compared with acclimation at 15°C. These results indicate that overwintering larvae at different phases of diapause development respond differently to the low temperature stimulus for glycerol synthesis. Thus, we evaluated the metabolic rearrangements associated with glycerol synthesis during diapause development and after temperature acclimation. Larvae collected on 2 October were acclimated at 15°C for 15 and 60 days. Some of those acclimated at 15°C were then moved to 0°C for 15 days. The larvae acclimated at 15°C for 15 days were in deep diapause and accumulated little glycerol, while larvae acclimated at 15°C for 60 days were nearly ready to emerge from diapause and accumulated glycerol at 155.5 μmol/g. When larvae acclimated to 15°C for 15 days were transferred to 0°C, glycerol accumulation was stimulated to the same extent (ca 140 μmol/g) as it was in larvae that were acclimated to 15°C for 60 days and then transferred to 0°C. These results indicate that low temperature has a cumulative effect on glycerol production in larvae at different phases of diapause development. Glycerol accumulation was accomplished by activation of glycogen phosphorylase and inhibition of fructose-1,6-bisphosphatase, and activation of enzymes associated with glycerol synthesis, mainly glyceraldehyde-3-phosphatase and polyol dehydrogenase with glyceraldehyde activity.     

Overwintering strategy of endoparasitoids in Chilo suppressalis: a perspective from the cold hardiness of a host

Although parasitoids ultimately kill their host, koinobiont parasitoids must protect not only themselves but also their hosts against extreme environments. In this study, the parasitism rate of Chilo suppressalis Walker (Lepidoptera: Pyralidae) was investigated, and the average body weights, supercooling points, and concentrations of glycerol (acting as a cryoprotectant) in the hemolymph were compared between parasitized and non‐parasitized larvae. Five species of koinobiont endoparasitoids parasitized the overwintering C. suppressalis larvae and the total parasitism rate was 47.6% (n = 1 537). Average body weight of parasitized larvae was significantly lower than that of non‐parasitized larvae, and the parasitism rate of the lighter group (20–30 mg) was highest. The supercooling point of parasitized C. suppressalis larvae (?15.7 ± 0.3 °C) was significantly lower than that of the non‐parasitized larvae (?14.3 ± 0.2 °C). In addition, supercooling points were not correlated with body weights between parasitized and non‐parasitized larvae, indicating that cold hardiness of parasitized larvae was enhanced by endoparasitoids. Furthermore, the concentration of glycerol in the hemolymph was significantly higher in parasitized larvae (205.0 ± 7.1 μmol ml^?1) than in non‐parasitized larvae (169.8 ± 14.4 μmol ml^?1), which suggests that the mechanism that decreases the supercooling point of parasitized larvae was associated with glycerol. All these results indicated that the cold hardiness of parasitized C. suppressalis larvae was enhanced by their endoparasitoids, which benefitted overwintering endoparasitoids.     

Seasonal patterns of the thermal response in relation to sugar and polyol accumulation in overwintering adults of elm leaf beetle, Xanthogaleruca luteola (Coleoptera: Chrysomelidae)

Elm leaf beetle, Xanthogaleruca luteola (Muller), is one of the key pests of elm trees that survive winter in reproductive diapause in sheltered locations. Overwintering adults of the elm leaf beetle showed a complex sugar/polyol cryoprotectant system. The major components of the multiple systems were glucose, myo-inositol and trehalose. In this study, we investigated the seasonal profile of low molecular weight compounds and glycogen in natural population and also in response to thermal constant regimes (5 and 15 °C). Among these components, a remarkable seasonal pattern of accumulation/depletion was observed in myo-inositol over the course of hibernation with the development of diapause progress. Incubating at 5 °C only elicited a strong response in myo-inositol synthesis during diapause. It suggests that the elm leaf beetle accumulates myo-inositol not only in relation to entering diapause but also in response to low temperatures and their interactions. The laboratory acclimation experiments showed that adults exposed to 15 °C had no chance for accumulation of low molecular weight carbohydrate even during diapause. The results of this study illustrated that overwintering adults of elm leaf beetle produce myo-inositol as the primary substance which plays a specific role in some biochemical adjustments in overwintering adults of X. luteola.     

Influence of soil moisture on supercooling capacity and associated physiological parameters of overwintering larvae of rice stem borer

Influence of soil moisture on the supercooling capacity and associated physiological parameters of overwintering larvae of the rice stem borer Chilo suppressalis was examined by exposing larvae to soil moistures of 25, 50, 75 and 100% of saturated soil water content (SSWC) at ambient temperature for 30 d from December 2007 to January 2008 in Beijing, China. At the end of the exposure, supercooling points (SCPs) varied significantly among the treatments, the lowest being in the larvae exposed to soil moisture of 25% SSWC. Fresh weight was significantly higher in the larvae exposed to soil moisture of 100% SSWC than in those kept at 25 and 50% SSWC. Dry weight and body water content (% fresh weight) were not different among the treatments. Glucose and trehalose contents were markedly lower, and glycerol content was significantly higher in the larvae confined to soil moisture of 25% SSWC than in those exposed to the other soil moisture treatments. It is suggested that variation in body water content (% fresh weight) contributes to the differences in SCPs of the overwintering C. suppressalis larvae in all treatments, but the influence of soil moisture treatments on supercooling capacity are caused through changes in glycerol content.     

Reduction of insect cold-hardiness using ice-nucleating active fungi and surfactants

The supercooling point (SCP) of an insect model, the lady beetle Hippodamia convergens Guérin-Menéville (Coleoptera, Coccinellidae) was markedly elevated by treatment with aqueous suspensions of the filamentous, ice nucleation active (INA) fungi Fusarium avenaceum and slightly elevated by Fusarium acuminatum. Addition of the surfactant Tween 80 to the fungal suspensions further reduced the supercooling capacity of adult beetles. When used alone the surfactant Triton X-100 produced a greater SCP elevation than Tween 20 or Tween 80. The emulsifier gum arabic was ineffective in elevating beetle SCPs when applied alone and when added to INA fungal preparations it decreased their efficacy. Aqueous suspensions of both viable sporulating and viable pleomorphic (a permanent, degenerative, nonsporulating cultural state) forms of both fungal species were more effective in elevating the SCP than killed preparations except for the pleomorphic F. acuminatum suspension in which the killed form was slightly more active. Application of INA fungi applied in combination with surfactants may be useful in the development of methods for the biological control of overwintering freeze-susceptible insect pests by decreasing their capacity to avoid lethal freezing by supercooling.