荒漠昆虫光滑鳖甲的耐寒性季节变化及其生理机制

光滑鳖甲Anatolica polita borealis生活于温差大的新疆荒漠环境, 为探讨其耐寒性及耐寒机制, 本研究测定了其3-9月份成虫在－10℃的耐寒性、过冷却点（SCP）、含水量、甘油含量和血淋巴热滞活性(thermal hysteresis activity, THA)以及冷驯化对增强光滑鳖甲成虫耐寒性的效果, 还测定了光滑鳖甲不同发育阶段幼虫的SCP。结果表明: 光滑鳖甲成虫的耐寒性和SCP具有明显的季节性变化, 3月初SCP为－12.5℃, 7月为－6℃, 9月底为－13.6℃。4℃冷驯化能够提高光滑鳖甲成虫在－10℃的存活率, 未驯化组在40 min的存活率为50%, 而驯化2 h的为70%, 驯化24 h的为90%。虫体含水量在夏季有显著降低, 3月、7月和9月结合水与自由水的比值分别为10.8∶1,2.6∶1和5.4∶1。成虫甘油含量与SCP的回归方程为y=－0.6204x－5.681, R²=0.7714。成虫血淋巴THA与过冷却点的回归方程为y=－5.26x－1.713, R²=0.9049。血淋巴THA比甘油浓度更能影响过冷却点降低的程度。随着幼虫的发育, 其SCP逐渐降低。结果提示, 光滑鳖甲通过提高结合水与自由水的比值、增加抗冻蛋白和甘油的含量使虫体保持较低的SCP, 因而具有较高的耐寒性。     

不同强度快速冷驯化对广聚萤叶甲成虫耐寒性生理指标的影响

【目的】快速冷驯化能在短时间内迅速提高昆虫的耐寒性,是昆虫应对外界温度急剧变化以及短时低温胁迫的重要途径。本研究旨在探究入侵杂草豚草Ambrosia artemisiifolia生防天敌广聚萤叶甲Ophraella communa对不同强度快速冷驯化的生理响应机制。【方法】分别对广聚萤叶甲成虫进行了不同温度（-4, 0, 4和8℃）下4 h及0℃下不同时间（1, 4 , 8和16 h）的快速冷驯化处理,并对其体内的生理物质含量和保护酶活性进行了测定。【结果】除8℃/4 h,0℃/1 h和0℃/8 h外,其余冷驯化处理均使广聚萤叶甲成虫过冷却点显著降低（P<0.05）,其中0℃/4 h处理组最低。而且,随着冷驯化温度降低、持续时间的增长,广聚萤叶甲成虫体甘油含量以及过氧化氢酶（CAT）、过氧化物酶（POD）和超氧化物歧化酶（SOD）活性呈曲线变化,并于0℃/4 h处理时达到极值,但冷驯化处理对虫体自由水和总糖含量的影响并不显著（P≥0.05）。【结论】广聚萤叶甲快速冷驯化的诱导具有其临界强度值和最适条件,过大强度的驯化处理反而不利于其耐寒性的提高。本研究结果对于深入阐明广聚萤叶甲越冬策略以及人工培育耐寒种群的实践具有一定参考价值。     

驯化对始红蝽(Pyrrhocoris apterus)耐寒能力的影响及越冬适应策略

为阐明越冬期间始红蝽应对低温胁迫的耐寒策略及其影响因素,从生理生化水平探讨始红蝽成虫的耐寒能力,逐月测定了12月至翌年3月始红蝽低温驯化前后的过冷却点、低温存活率、LT_(50)以及始红蝽体内耐寒物质含量。结果表明,越冬期间始红蝽自然种群过冷却点最低为(-14.01±0.53)℃,-5、-10℃驯化30min后的始红蝽过冷却点最低降至为(-19.32±0.86)℃、(-25.56±1.09)℃。0℃驯化30min后暴露于-5、-10、-15℃1h的最高存活率依次为100%、39.1%±8.6%、10%;始红蝽自然种群LT_(50)最低为-8.53℃,0℃驯化后降至-9.21℃。越冬期间雌雄始红蝽体内自由水/结合水比值和游离蛋白质含量先下降后上升,12月达到最大值,雌雄分别为144.50±26.22和140.32±21.92,(15.81±0.10)mg/g和(15.47±0.01)mg/g;脂肪、海藻糖和甘油含量先上升后下降,2月达到最大值,雌雄脂肪含量分别为(16.33±0.48)mg/g和(13.15±1.32)mg/g,海藻糖含量分别为(11.98±0.01)mg/g和(10.88±0.02)mg/g,甘油含量分别为(14.74±0.01)mg/g和(15.06±0.03)mg/g。研究证明,低温驯化后始红蝽的过冷却点和LT_(50)明显降低,低温存活率显著提高,越冬期间始红蝽可通过调整体内抗逆物质含量以增强虫体耐寒能力。     

冷驯化对油松毛虫越冬幼虫过冷却点及主要耐寒物质的影响

【目的】冷驯化可增强昆虫的耐寒性,本文研究旨在明确不同冷驯化条件下油松毛虫Dendrolimus tabulaeformis Tsai et Liu越冬幼虫的过冷却点和主要耐寒物质的变化规律。【方法】利用热电偶方法测定越冬幼虫的过冷却点,分别采用差量法、氯仿甲醇法、苯酚硫酸法及毛细管气相测谱法测定其含水率、脂肪、糖原和小分子糖醇的含量。【结果】冷驯化会导致幼虫含水率显著降低;过冷却点和脂肪含量在低于环境气温5℃冷驯化后显著降低,当驯化温度低于环境气温10℃及以上则升高;糖原含量在9月份显著增加,越冬中期(1、3月份)含量略有降低但不显著;小分子糖醇含量的变化均不显著;海藻糖含量略降低;甘油、葡萄糖和半乳糖含量在低于环境气温5℃冷驯化后略降低,低于环境气温10℃冷驯化则升高。【结论】冷驯化使幼虫虫体含水率和脂肪含量降低,糖原含量提高,从而导致其过冷却点降低,耐寒能力提高;冷驯化的温度和时间均会影响其过冷却能力,在最适合的温度和时长可以最大程度提高其耐寒能力。研究结果为揭示油松毛幼虫的耐寒机制及潜在分布区预测提供了科学依据。     

异色瓢虫成虫冷驯化反应及体内几种酶活力的相关变化

为明确冷驯化反应对异色瓢虫Harmonia axyridis (Pallas) 实验种群成虫耐寒性及其生殖能力的影响, 本研究测定了成虫低温存活率、过冷却点(supercooling point, SCP)、体内含水量及雌虫繁殖能力等。结果表明: 冷驯化（在5℃下诱导3 d, 5 d）后, 成虫再在-5℃下暴露3 d的存活率由对照（预先未进行冷驯化）的46%分别提高至60%和67%, 而诱导10 d后的存活率(51%)反而下降。冷驯化效应在其成虫转移至饲养条件下7 d后就消失。随着低温诱导时间的延长过冷却点及体内含水量均呈现下降趋势, 短时间(5, 10 d)的诱导不能使成虫的SCP明显降低, 但可以使含水量极显著下降。冷驯化后异色瓢虫雌虫产卵前期延长; 虽然冷驯化对雌虫首次产卵量没有影响, 但是随着诱导时间的延长连续观察72 h内单头雌虫累计产卵量却降低。冷驯化过程中成虫体内几种酶活力的检测结果表明: 两种细胞保护酶超氧化物歧化酶(SOD)与过氧化氢酶(CAT)活性升高, 与新陈代谢有关的乳酸脱氢酶(LDH)及Na⁺, K⁺-ATP酶活性却降低。结果显示, 低温胁迫前异色瓢虫成虫经过不同时间的诱导后有可能提高其低温抵抗能力, 而且冷驯化诱导成虫耐寒性增加是一种复杂的生理生化过程, 这一过程对其生存和繁殖具有重要的适应意义。     

低温驯化对海滨斯氏线虫生化物质含量及其生存活力的影响

【目的】本文为明确海滨斯氏线虫Steinernema litorale开发应用的潜能和价值,研究了由本实验室从哈尔滨市香坊区原生态榆树根际土壤中分离到的海滨斯氏线虫的适低温特性。【方法】采用25℃(6 d)、10℃(6 d)、4℃(6 d)和25℃(2 d)-10℃(2 d)-4℃(2 d)处理海滨斯氏线虫,分析该线虫冷冻存活率、不同温度处理后的贮存稳定性以及体内生化物质—可溶性糖、糖原、海藻糖、脂肪与蛋白质含量的影响。【结果】海滨斯氏线虫经过25℃(6 d)、10℃(6 d)、4℃(6 d)和25℃(2 d)-10℃(2 d)-4℃(2 d)低温驯化,于﹣20℃冷冻36 h后的存活率分别为0.9%、23.6%、20.0%和49.2%,经25℃(2 d)-10℃(2 d)-4℃(2 d)处理的线虫冷冻存活率明显高于其它3组,阶段性降温显著提高了线虫耐寒力;将上述4组低温驯化后的海滨斯氏线虫置于4℃贮存7个月,其存活率差异不显著(P0.05),均为65%以上;25℃(6 d)驯化的线虫杀虫活性显著低于其他3个处理组线虫的杀虫活性。海滨斯氏线虫经25℃(2 d)-10℃(2 d)-4℃(2 d)阶段降温驯化后体内可溶性糖、脂肪、海藻糖含量最高且显著高于25℃(6 d)驯化的线虫体内可溶性糖、脂肪、海藻糖含量;25℃(6 d)驯化的线虫体内蛋白质、糖原含量与其他3个处理组差异不显著。【结论】阶段降温的低温驯化更有利于提高海滨斯氏线虫耐寒能力,并能用于后期长期贮存。     

低温处理对亚洲玉米螟幼虫抗寒性的诱导效应

室内条件下将亚洲玉米螟Ostrinia furnacalis (Guenée)幼虫放置在5℃(LT1处理组)和0℃(LT2处理组)下低温处理2 h后,分别测定了其低温诱导识别温度、存活率、抗寒性、过冷却点、体内水分和脂质含量百分率,并进行抗冻特异蛋白的诱导; 利用SDS-PAGE方法分析了低温诱导后亚洲玉米螟5 龄幼虫血清中抗冻特异蛋白。结果表明：亚洲玉米螟3、4和5龄幼虫的低温诱导识别温度分别为-13.5℃、-16.5℃和-18.5℃; 3、4和5龄幼虫存活率LT2组>LT1组>对照组(P<0.05);随虫龄增加,幼虫抗寒性逐步增强;对幼虫过冷却点无明显影响(P>0.05); 幼虫水分和脂质含量百分率为LT2组>LT1组>对照组,且随虫龄增加,虫体含水率和脂质含量百分率增高(P<0.05); 低温诱导产生了一种分子量约为29.0 kD的抗冻特异蛋白。研究结果表明低温诱导可以增强亚洲玉米螟幼虫的抗寒性。     

绿盲蝽越冬卵的耐寒能力

近年来,随着转基因棉花的大面积推广,绿盲蝽成为了我国棉花和果树生产的重要害虫。为了阐明绿盲蝽越冬生态适应性,研究了绿盲蝽卵在越冬过程中的耐寒力变化,测定了卵内生化物质的含量,结果表明:绿盲蝽越冬卵低温存活力呈现出明显的月份变化,在-10℃、-20℃处理下,绿盲蝽的越冬卵的半致死时间从大到小的顺序依次为:1月>2月>12月>3月>4月,在-30℃处理下从大到小的顺序依次为:1月>12月>3月>2月>4月。在极端低温(-20℃)下1、2、3、4和5 d后,4月份保护卵死亡率明显低于相同处理下的4月份裸露卵的死亡率,枣枝中的越冬卵死亡率分别为4.32%、5.36%、5.42%、6.79%和7.63%,剥离出来的越冬卵死亡率分别为46.06%、51.84%、54.59%、63.07%和74.41%。人工滞育卵的耐寒性强于正常发育卵的耐寒性,弱于自然越冬过程中滞育卵的耐寒性。冷驯化可以显著提高绿盲蝽越冬卵的低温存活力,在0℃冷驯化20 min后,越冬卵的低温(-20℃、100 h)死亡率显著降低,冷驯化40 min后,低温死亡率趋于平稳,均在37%左右。越冬卵体内物质呈明显的季节性变化,越冬卵越冬期的含水量显著高于越冬后,其中1月份最高(43.98%),4月份达到最低(38.79%)。越冬卵体内总脂肪含量在整个越冬过程中逐渐降低,从12月份的38.24%,降低到4月份的27.08%。蛋白质和糖的含量均是先降低后升高,其中蛋白质从12月份的78.77μg/mg,降低到1月份的59.80μg/mg,然后又逐渐升高至4月份的73.62μg/mg,体内总糖含量由12月份的39.60μg/mg降低至1月份的21.17μg/mg,然后又逐渐升高至4月份的35.10μg/mg。绿盲蝽越冬卵的耐寒力随着月份的变化而变化,在整个越冬期间表现出较强的抗寒性,能够抵御冬季低温,而其越冬场所的保护作用增加了其对于冬季严寒的适应性。     

冷驯化对茶尺蠖抗寒性生理指标的影响

【目的】冷驯化作为一种有效提高耐寒能力的途径,在昆虫对环境温度变化的适应中发挥重要作用,是当前昆虫耐寒性研究的热点。为了解茶尺蠖Ectropis oblique(Prout)对冷驯化的生理响应机制,本文研究了不同冷驯化对茶尺蠖抗寒性生理指标的影响。【方法】分别对茶尺蠖快速冷驯化0℃2 h、5℃2 h,长时间冷驯化5℃24 h、5℃48 h,利用热电偶方法测定了茶尺蠖的过冷却点,采用生理生化测定法研究了其体内的抗寒性生理指标的变化。【结果】长时间冷驯化处理使茶尺蠖过冷却点显著降低;虫体内含水量显著降低;脂质和糖原含量显著增加;蛋白质含量略有增加但无显著差异;过氧化物酶(POD)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)3种保护酶活性显著升高及ATP能量代谢酶活显著降低。短时间冷驯化使茶尺蠖过冷却点显著降低;虫体内含水量降低但差异不显著;脂质、糖原和蛋白质含量显著升高;POD、CAT、SOD 3种保护酶活性及ATP能量代谢酶活分别显著升高和降低。【结论】长时间冷驯化和快速冷驯化均能提高茶尺蠖耐寒性,这是虫体内的水分、脂肪、糖原、蛋白质等含量以及抗逆酶活性变化的综合反映。本研究结果对深入研究茶尺蠖越冬策略以及结合气候条件确定茶尺蠖的分布,为评价田间死亡率及预测翌年发生程度提供重要依据,对制定防治措施具有重要现实意义。     

西花蓟马的快速冷驯化及其生态学代价

西花蓟马是我国蔬菜、果树和观赏植物上的一种重要入侵害虫.该害虫通过取食寄主汁液和传播多种植物病毒造成危害,而后者危害造成的经济损失更大.温度是影响西花蓟马生长发育和繁殖的一个重要非生物因子,而该虫对温度的耐受性决定了它的越冬存活率和地理分布.为探明低温对西花蓟马的不利影响,研究了西花蓟马若虫和成虫的快速冷驯化对其存活、发育和繁殖的影响.结果表明,随着温度降低,西花蓟马若虫和成虫存活率逐渐下降,但若虫对低温更为敏感.当成虫和若虫暴露于-13℃和-13.5℃下2h后,其存活率分别为25％和27％.根据识别温度定义,这两个温度分别被定义为若虫和成虫的识别温度.将西花蓟马成、若虫在0℃或5℃驯化2h后,再置于各自识别温度下,其存活率都得到了明显提高,但雌雄成虫间的存活率并无差异;然而,在0℃下驯化2h后,若虫和雌雄成虫的存活率得到了最大幅度的地提高,分别达46％、54％、49％.西花蓟马若虫经不同低温处理后,其发育历期、羽化后的成虫寿命、产卵时间与对照相比无显著差异,但产卵量显著降低;成虫经过低温处理后,其寿命,产卵量和产卵时间明显降低.研究结果支持昆虫快速冷驯化与其适合度之间存在平衡的假说;同时,也可为该虫的分布和治理研究提供相应的基础信息.     

Rapid cold hardening in the predatory mite Euseius (Amblyseius) finlandicus (Acari: Phytoseiidae)

A rapid cold hardening response was studied in diapause and non-diapause females of the predatory mite Euseius finlandicus. When laboratory reared diapause and non-diapause females were transferred and maintained from the rearing temperature of 20 degrees C for 2 h to -11.5 degrees C and -10 degrees C, 10 to 20% survived respectively. However, conditioning of diapause females for 4 h at a range of temperatures from 0 to 10 degrees C before their exposure for 2 h to -11.5 degrees C, increased survival to approximately 90%. Similarly, conditioning of non-diapause females for 4 h at 5 degrees C before their exposure for 2 h to -10 degrees C increased survival to 90%. A similar rapid cold hardening response in both diapause and non-diapause females was also induced through gradual cooling of the mites, at a rate of approximately 0.4 degrees C per min. The rapid increase in cold tolerance after prior conditioning of the mites to low temperatures, was rapidly lost when they returned to a higher temperature of 20 degrees C. Rapid cold hardening extended the survival time of diapause and non-diapause females at sub-zero temperatures. The cost of rapid cold hardening in reproductive potential after diapause termination was negligible. In non-diapause females, however, the increase in cold tolerance gained through gradual cooling could not prevent cold shock injuries, as both fecundity and survival were reduced.     

Rapid cold hardening in the western flower thrips Frankliniella occidentalis

A rapid cold hardening process is reported in first instar larvae of Frankliniella occidentalis. When larvae are transferred directly from 20 degrees C to -11.5 degrees C for 2h there is 78% mortality, whereas exposure to 0 degrees C for 4h prior to transfer to -11.5 degrees C reduces mortality to 10%. The response can also be induced by exposure to 5 degrees C for 4h or by gradual cooling at rates between 0.1 and 0.5 degrees C min(-1.) The acquired cold tolerance is transient and is rapidly lost (after 1h at 20 degrees C). Rapid cold hardening extends survival times at -11.5 degrees C and depresses lethal temperatures in short (2h) exposures. Rearing at 15 degrees C (12L:12D), (a cold acclimation regime for F. occidentalis), does not protect against the cold shock induced by direct transfer to -11.5 degrees C (which rapid cold hardening does) but does extend survival time at -5 degrees C (i.e. increased chill tolerance) whilst rapid cold hardening does not. The rapid and longer term cold hardening responses in F. occidentalis therefore appear to have different underlying mechanisms.     

Cold shock injury and ecological costs of rapid cold hardening in the grain aphid Sitobion avenae (Hemiptera: Aphididae)

The ability of first instar nymphs and newly moulted pre-reproductive adults of the grain aphid S. avenae to rapidly cold harden was investigated. When nymphs reared at 20 degrees C were transferred directly to -8 degrees C for 3 h, there was 18% survival. This exposure was selected as the discriminating temperature. Maximum increases in survival were achieved by acclimating nymphs for 2 h at 0 degrees C and adults for 3 h at 0 degrees C, resulting in survival of 83% and 68%, respectively. Cooling nymphs from 10 to 0 degrees C at different rates (1, 0.1 and 0.05 degrees C min(-1)) also increased cold hardiness, with the slowest rate of 0.05 degrees C min(-1) conferring the highest survival following exposure to the discriminating temperature. Adult aphids also expressed a rapid cold hardening response but to a lesser extent, with survival increasing from 16% to 68% following 3 h at 0 degrees C. There were no 'ecological costs' associated with rapid cold hardening in terms of development, longevity or fecundity. The data support the hypothesis that rapid cold hardening can be induced during the cooling phase of natural diurnal temperature cycles, allowing insects to track daily changes in environmental temperatures.     

Rapid cold hardening in the olive fruit fly Bactrocera oleae under laboratory and field conditions

A rapid cold hardening response was studied in females and males of the olive fruit fly Bactrocera (Dacus) oleae. When laboratory-reared females and males were transferred and maintained from the rearing temperature of 24 °C for 2 h to –6.5 °C approximately 5% survived. However, conditioning of both females and males for 2 h at various temperatures from 0 to 10 °C before their exposure for 2 h to –6.5 °C increased survival to 80 to 92%. A similar rapid cold hardening response in both females and males was also induced through gradual cooling of the flies at a rate of approximately 0.4 °C per min. The rapid increase in cold tolerance after prior conditioning of the flies to low temperatures, was rapidly lost when they returned to a higher temperature of 24 °C. In the field, in late February and early March, females and males were capable of a rapid cold hardening response. After exposure to the critical temperature they suffered a high mortality when tested in the afternoon and low mortality early in the morning on consecutive days, probably because of differences in the prevailing field temperatures a few hours before testing. This plasticity of cold tolerance gained through rapid cold hardening may allow the flies to survive during periods of the year with great fluctuation in circadian temperatures.     

Anoxia blocks thermotolerance and the induction of rapid cold hardening in the flesh fly, Sarcophaga crassipalpis

Abstract. Anoxia induced by nitrogen or carbon dioxide, or hypoxic/hypobaric conditions generated by a partial vacuum sensitizes red-eye pharate adults of Sarcophaga crassipalpis Macquart to a high temperature exposure that is normally nonlethal (40C for 2–3 h). Thermotolerance induced by a2h exposure to 40C (under aerobic conditions) doubles the pharate adults' tolerance to 45C but provides no protection against a combined exposure to 45C and anoxia, and only modest protection against a combined exposure to 40C and anoxia. Under aerobic conditions, exposing pharate adults to 0C for 2 h increases their tolerance to -10C (rapid cold hardening). Rapid cold hardening at 0C is not induced under anoxia. These results imply that tolerance to high temperatures and rapid cold hardening are dependent on aerobic processes and suggest that certain forms of temperature stress can be further exacerbated with anoxia.     

Effect of long-term and rapid cold hardening on the cold torpor temperature of an aphid

Abstract.  The effect of long-term (seasonal) acclimation and rapid cold hardening is investigated on the cold torpor temperature ( CT _min) of adult grain aphids, Sitobion avenae, reared at 20 or 10 °C for more than 6 months before experimentation. Rapid cold hardening is induced by exposing aphids reared at 20 to 0 °C for 3 h and aphids reared at 10 to 0 °C for 30 min (acclimation regimes previously found to induce maximum rapid cold hardening). The effect of cooling aphids from the same rearing regimes from 10 to −10 °C at 1, 0.5 and 0.1 °C min⁻¹ is also investigated. In the 20 °C acclimated population, rapid cold hardening and cooling at 0.1 °C min⁻¹ both produce a significant decrease in CT _min from 1.5 ± 0.3 to –0.9 ± 0.3 and –1.3 ± 0.3 °C, respectively. Rapid cold hardening also results in a significant reduction in CT _min of the population reared at 10 °C from 0.8 ± 0.1 to –0.9 ± 0.2 °C. However, none of the cooling regimes tested reduces the CT _min of the winter-acclimated (10 °C) population. The present study demonstrates that rapid cold-hardening induced during the cooling phase of natural diurnal temperature cycles could lower the movement threshold of S. avenae , allowing insects to move and continue feeding at lower temperatures than would otherwise be possible.     

Rapid cold hardening increases cold and chilling tolerances more than acclimation in the adults of the sycamore lace bug, Corythucha ciliata (Say) (Hemiptera: Tingidae)

The sycamore lace bug, Corythucha ciliata is a new, invasive pest of Platanus trees in China. Although C. ciliata is often subjected to acute low temperatures in early winter and spring in northern and eastern China, the cold tolerance of C. ciliata has not been well studied. The objectives of this study were to determine whether adults of C. ciliata are capable of rapid cold hardening (RCH), and to compare the benefits of RCH vs. cold acclimation (ACC) in the laboratory. When the adult females incubated at 26 °C were transferred directly to the discriminating temperature (−12 °C) for 2 h, survival was only 22%. However, exposure to 0 °C for 4 h before transfer to −12 °C for 2 h induced RCH, i.e., increased survival to 68%. RCH could also be induced by gradual cooling of the insects at rates between 0.1 and 0.25 °C min⁻¹. The protection against cold shock obtained through RCH at 0 °C for 4 h was lost within 1 h if the adults were returned to 26 °C before exposure to −12 °C. Survival at both −12 and −5 °C was greater for RCH-treated than for ACC-treated adults (for ACC, adults were kept at 15 °C for 5 days), and the lethal temperature (2 h exposure) was lower for RCH-treated than for ACC-treated adults. The results suggest that RCH may help C. ciliata survive the acute low temperatures that often occur in early winter and early spring in northern and eastern China.     

Hardening trumps acclimation in improving cold tolerance of Drosophila melanogaster larvae

Abstract Chill‐susceptible insects are able to improve their survival of acute cold exposure over both the short term (i.e. hardening at a relatively severe temperature) and longer term (i.e. acclimation responses at milder temperatures over a longer time frame). However, the mechanistic overlap of these responses is not clear. Four larval stages of four different strains of Drosophila melanogaster are used to test whether low temperature acclimation (10 °C for 48 h) improves the acute cold tolerance (LT₉₀, ～2 h) of larvae, and whether acclimated larvae still show hardening responses after brief exposures to nonlethal cold or heat, or a combination of the two. Acclimation results in increased cold tolerance in three of four strains, with variation among instars. However, if acclimation is followed by hardening pre‐treatments, there is no improvement in acute cold survival. It is concluded that short‐term thermal responses (e.g. hardening) may be of more ecological relevance to short‐lived life stages such as larvae, and that the mechanisms of low temperature hardening and acclimation in D. melanogaster may be antagonistic, rather than complementary.     

Relationship between rapid cold-hardening and cold acclimation in the eggs of the yellow-spotted longicorn beetle, Psacothea hilaris

Rapid cold-hardening (RCH) and cold acclimation (ACC) were examined in eggs of the yellow-spotted longicorn beetle, Psacothea hilaris (Pascoe) (Coleoptera: Cerambycidae). When eggs incubated at 25 degrees C were transferred directly to conditions of -22 degrees C for 2h, less than 30% survived, whereas exposure to 0 degrees C for 4h prior to transfer to -22 degrees C increased survival to nearly 60%. The rapidly enhanced cold tolerance (RCH) was transient and lost rapidly after 1h at 25 degrees C. Incubation at 15.5 degrees C for 9 days (ACC) also enhanced cold tolerance. Comparison of the cold tolerance of non-treated eggs and eggs pre-treated to give RCH, ACC, or ACC+RCH allowed the relationship between the two hardening processes to be determined. At a mild subzero temperature (-10 degrees C) an RCH effect was not detected, whereas only RCH is effective at the severest subzero temperature just above the SCP (-26 degrees C). At intermediate temperatures (-16, -22 and -25 degrees C), ACC and RCH enhanced survival in combination. Therefore, the two hardening processes have different physiological bases but operate concomitantly over a wide temperature range.     

The influence of developmental stage on cold shock resistance and ability to cold-harden in Drosophila melanogaster

Thermal sensitivity and ability to rapidly cold- and heat-harden may change during ontogeny. This study reports how inherent cold tolerance and ability to rapidly cold-harden change across eight developmental stages in both genders of Drosophila melanogaster using a similar experimental approach for all stages. Inherent cold tolerance was estimated as LT50 by assaying cold shock survival over a wide range of temperatures (-16 to 5 degrees C). Rapid cold-hardening (RCH) was applied by cooling from 25 to 0 degrees C at -0.25 degrees C min(-1) followed by 1 h at 0 degrees C. Individuals were cold shocked either directly or after RCH to estimate the effect of RCH. We found large variation in cold tolerance among developmental stages and minor differences between genders. Eggs were most tolerant followed by adults, pupae and larvae. In the light of this and other studies it is suggested that there is a general pattern of stage specific thermal stress resistance in Drosophila. The capacity to rapidly cold-harden was found in both sexes of larval, pupal and adult stages, though some developmental stages showed negative or neutral effects of RCH which was probably due to the cost associated with the hardening treatment in these cold susceptible stages. The early presence of RCH indicates that the mechanisms behind hardening are not stage specific and that RCH may be an ecologically important trait in early stages of ontogeny.