昆虫的耐寒性及其影响因素

昆虫是变温动物,种群的繁衍面临如何安全度过漫长而寒冷的冬季的挑战.为了安全越冬,昆虫必须适应冬季的低温,增强耐寒能力是昆虫的一种重要的季节适应机制.近年来,关于昆虫耐寒性的研究不断开展,研究内容涉及影响耐寒性的因素、耐寒性的机制等方面.影响昆虫耐寒性的因素包括环境因素,如气候的季节变化、纬度或海拔的差异等,以及昆虫自身的发育阶段、滞育发生等.     

Freeze tolerance in larvae of the winter-active Diamesa mendotae Muttkowski (Diptera: Chironomidae): a contrast to adult strategy for survival at low temperatures

The winter-active Diamesa mendotae Muttkowski (Diptera: Chironomidae) is freeze intolerant in the adult stage with a low mean supercooling point (SCP) of ~−20 °C. However, cold-hardiness strategies for immatures of this species are unknown. In this study, we measured SCP values for D. mendotae larvae, pupae and adults using surface-contact thermometry. In addition, the lower lethal temperature (LLT) was determined for the larval stage. The mean SCPs for larvae (−7.4 °C) and pupae (−9.1 °C) were relatively high compared to adults (−19.7 °C). Our results indicate that the larvae of D. mendotae are freeze tolerant with a LLT₉₉ (−25.4 °C), ~−10 °C lower than their minimum SCP (−15.6 °C). Freeze tolerance in these larvae may be a strategy to provide protection from short-term exposures to ice crystals or to permit diapause within frozen substrates. The change in cold-hardiness strategy from freeze tolerant to freeze intolerant between the larval and adult stages of this species is likely a result of the different habitats occupied by these two life stages.     

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。     

Gut Colonization by an Ice Nucleation Active Bacterium,Erwinia(Pantoea)ananasReduces the Cold Hardiness of Mulberry Pyralid Larvae

To evaluate the suitability of using ice nucleation active (INA) bacteria for the biological control of insect pests, the supercooling point (SCP) of larvae of mulberry pyralid,Glyphodes duplicalis,and silkworm,Bombyx mori,ingesting INA strains ofErwinia(Pantoea)ananasandPseudomonas syringaewas determined. Mean SCP of the guts of silkworm larvae ingesting INA strains ofE. ananasranged from −2.5 to −2.8°C, being 5°C higher than that in control treatments. Similarly, mean SCP of mulberry pyralid larvae ingesting INA strain ofE. ananas,which can grow well in the gut, was −4.7°C at 3 days after treatment, being 6.5°C higher than that in control treatments. On the other hand, mean SCP of the larvae-ingesting INA strain ofP. syringae,which cannot grow in the gut, was −9.0°C at 3 days after treatment, rising by only 2.5°C higher than that in the control treatments. In addition, more than 80% of the larvae of mulberry pyralid ingesting the INA strain ofE. ananasfroze and eventually died when exposed to −6°C for 18 h, while only 36% of the larvae ingesting the INA strain ofP. syringae,or approximately 20% of the control larvae, froze and died. Thus, the gut colonization by INA strains ofE. ananasreduced remarkably the cold hardiness of the insects. These findings suggest that INA strains ofE. ananascould be effective as a potential biological control agent of insect pests.     

Inheritance of a vestigial wing mutation in the alfalfa weevil, Hypera postica

Alfalfa weevils (Hypera postica (Gyllenhal)) with vestigial hind wings were discovered in a population from Wageningen, the Netherlands, and two populations from the United States—an eastern weevil strain from Beltsville, Maryland and an Egyptian weevil strain from Atascadero, California. Such a mutant was absent from 23 other populations surveyed in the United States—three from eastern, seven from western, and 13 from Egyptian weevil strains. This mutation is due to a dominant autosomal gene with normal-wing individuals as recessive. The mutant gene can be transferred from eastern weevil to the western weevil strain. The short-wing trait may be useful for genetic manipulation to control the alfalfa weevil.

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Résumé Des H. postica aux ailes postérieures vestigiales ont été découverts dans une population de Wageningen (Pays Bas) et deux des USA—une lignée orientale de Beltsville (Maryland) et une lignée de H. brunneipennis d'Atascadero (Californie). Ce mutant était absent de 23 autres populations examinées aux USA: 3 de l'est, 7 de l'ouest et 13 de H. brunneipennis. Cette mutation est due à un gène dominant antosomal avec aile normale comme récessif. Le gêne mutant peut être transféré des lignées orientales aux lignées occidentales. Le caractère aile courte peut être pratique pour les manipulations génétiques destinées à maîtriser les populations d'H. postica.

     

Control of diapause and supercooling by the retrocerebral complex inPyrrhocoris apterus

Effects of external (photoperiod, temperature) and internal (retrocerebral complex, diapause condition) factors on supercooling were studied inPyrrhocoris apterus (L.) (Heteroptera: Pyrrhocoridae) adults. An increase in supercooling capacity is associated with the induction of diapause by short-day photoperiod or cardiacallatectomy of long-day non-diapause insects in spite of a high temperature of 26°C. The induction of diapause is a prerequisite for a further increase in supercooling capacity by cold acclimation. Post-diapause adults show low values of the supercooling point also in late January or early February, although their developmental potential is fully restored. Evidently the ability to supercool associated with diapause induction is maintained by a low ambient temperature in spite of diapause termination. The supercooling point increases rapidly when these adults are transferred to a high temperature of 26°C and cold re-acclimation is then no longer possible. The inhibition of morphogenesis (maturation of gonads) by cardiacallatectomy does not prevent the supercooling point from increasing in post-diapause adults at the high temperature.     

Low temperature tolerance and starvation ability of the oak processionary moth: implications in a context of increasing epidemics

• 1 We investigated how modifications in winter and spring temperature conditions may affect the survival of a spring‐hatching Lepidoptera, the oak processionary moth Thaumetopoea processionea.
• 2 Supercooling and chilling injury experiments indicate that eggs are especially cold hardy at the start of the winter period, although this ability is reduced later in the season. In the spring, young larvae are sufficiently cold hardy to ensure no direct mortality as a result of late frosts.
• 3 A comparison of phenological models shows that neonate larvae may await the unfolding of new oak leaves for relatively long periods (e.g. 1–30 days). Under both low (4°C after 5 days at 16°C) and high temperature experimental scenarios (constant 16°C), the majority of neonate larvae can survive starvation for more than 2 weeks.
• 4 Larvae may also suffer from food depletion once their development has been initiated (e.g. during cold springs) if the threshold temperature for feeding is not reached for several consecutive days, or in the case of late frosts affecting foliage availability. When temperature is reduced to 4°C, developing larvae become inactive and do not feed anymore; their starvation survival capability is reduced to approximately 2 weeks (cold spring hypothesis). At 16°C, developing larvae that are deprived of food can only survive for 10 days (late frost hypothesis).
• 5 We conclude that, in the oak processionary moth, neonate larvae are relatively well adapted to early hatching relative to budburst, ensuring them the highest foliage quality for development. In some years, however, phenological asynchrony or cold spring conditions may affect the persistence of populations at the limits of the species' range.

     

Cold hardiness and overwintering of the grain aphid Sitobion avenae

ABSTRACT.

• 1 Cold hardiness as measured by supercooling ability in the active stages of the grain aphid Sitobion avenae (F.) decreased progressively with maturation.
• 2 Aphids showed no acclimation response when maintained at low temperatures.
• 3 Starvation did not improve supercooling ability.
• 4 In a single exposure, surface moisture caused inoculation above the inherent supercooling point in a small proportion of a population.
• 5 Field populations show a seasonal change in supercooling ability, which is at a maximum in summer and a minimum in late winter.
• 6 It is concluded that the act of feeding on healthy plant tissue may confer extensive supercooling ability.

     

Cold tolerance and cold hardening strategy of the Japanese pine sawyer Monochamus alternatus (Coleoptera: Cerambycidae)

The Japanese pine sawyer, Monochamus alternatus , is an important pine forest pest and vector transmitting the pine wilt nematode that causes pine wilt disease. Low temperatures in autumn, winter and spring often differentially affect mortality of M. alternatus larvae. In this paper, we mainly compared the differences of mortality and cold hardening of larvae from different seasons, based on supercooling point (SCP) and cumulative probability of individuals freezing (CPIF). The cold hardening of the larvae from autumn, winter and spring seasons were largely different. Correlations between mortality and CPIF of autumn and spring larvae were highest on day 1/4, and gradually decreased with prolonged exposure duration. This beetle's death mainly resulted from freezing in short exposure duration. However, the correlation between mortality and CPIF of winter larvae increased gradually with the prolonged exposure duration. Death did not mainly result from freezing in long exposure duration. Autumn larvae are more susceptible and adaptable than winter and spring larvae. Winter larvae have a slight freeze-tolerance trend. Our research showed that M. alternatus came into complex cold-hardening strategies under natural selection. Freeze avoidance is the primary strategy; with prolonged exposure duration to above SCP or < 0 °C, chill tolerance is more important; this is followed by freeze tolerance during harsh winters.     

蒲氏钩蝠蛾幼虫血淋巴物质与其耐寒性

作为冬虫夏草寄主,蒲氏钩蝠蛾具重要的经济研究价值,主要分布于青藏高原色季拉山,具有较强的耐寒性,其耐寒性机理尚不明了。通过调查蒲氏钩蝠蛾生境土壤温度与血淋巴物质组成周年变化,以期探讨其耐寒性机理。实验结果表明:土壤温度呈规律性升降趋势,1、2月最低,8月逐步上升至最高,而后逐渐下降;血淋巴中主要物质按含量从高到低排列依次为:蛋白质(8.37—74.2 mg/m L)、总糖(12.65—36.12 mg/m L)、脂肪(7.7—12.32 mg/m L)以及甘油(1.66—3.91 mg/m L);在温度较高的夏季,蛋白质、总糖以及脂肪等物质含量处于较低水平,相反,在寒冷的冬季,此三者的含量均居较高水平;此3类能源物质含量与野外土壤温度均呈极显著的负相关关系,因而血淋巴总热量值在夏季较低而冬季较高,与野外土壤温度亦呈极显著负相关关系;与之相反的是,血淋巴中甘油含量的周年变化规律与野外土壤温度无明显的相关关系。因此,血淋巴中蛋白质、总糖以及脂肪等物质对于蒲氏钩蝠蛾耐寒性有着非常重要的作用,幼虫通过积累3类物质,增加血淋巴能量值,抵御青藏高原的高寒气候;可能由于生境的不同,甘油在蒲氏钩蝠蛾中的作用与其它昆虫有差异,在耐寒性提高中的功能尚需进一步研究。实验结果为了解蒲氏钩蝠蛾的强耐寒性以及适应高寒环境的生理生化机制提供理论依据。     

Cold hardiness of larvae of the southwestern corn borer, Diatraea grandiosella

The cold-hardening capacity of field-collected larvae from southeast Missouri and laboratory-reared larvae of the southwestern corn borer, Diatraea grandiosella Dyar, was examined. Supercooling points of non-diapause and diapause larvae collected from maize plants grown in Missouri (36°30 N lat.) were ca.-7.0°C. The hemolymph melting points of diapause field larvae (-0.8°C) were significantly lower than those of non-diapause larvae collected in July (-0.5°C). The supercooling points of hemolymph from non-diapause and diapause field larvae ranged randomly from-10° to-18°C. Supercooling points of non-diapause laboratory larvae increased from-13° to-10°C prior to pupation, whereas those of diapause larvae increased similarly before the onset of diapause, but then decreased during diapause to ca.-17°C. No change in supercooling points or capacity to survive in the presence of ice was observed in diapause laboratory larvae acclimated at 4°C for 63 days. Laboratory and field larvae began to freeze at ca.-1.5°C in the presence of ice, but survived to several degrees below their melting points. The high supercooling points of field larvae appeared to be due to the presence of an environmental ice-nucleator. Although data for laboratory larvae indicate sufficiently low supercooling points to permit winter survival in southeastern Missouri, considerable larval mortality occurs in the field due to inoculative freezing and the presence of an ice-nucleator.     

五种金花茶组植物的耐寒性比较研究

金花茶组植物是世界珍稀、濒危的观赏植物,具有极高的观赏价值和药用价值.为了比较金花茶组植物的耐寒性,以五种金花茶组植物为材料,采用人工模拟低温环境的方法对其2年生叶片进行低温胁迫处理,应用电导法研究五种金花茶种质在20℃(常温对照)、8℃、-2℃、-7℃、-12℃、-17℃、-22℃和-27℃低温下相对电导率的变化,配合Logistic方程,测定其低温半致死温度(LT50),以及叶片中游离脯氨酸、可溶性糖和丙二醛的含量.结果表明:五种金花茶的低温半致死温度(LT50)范围为-14.58~-12.74℃,其中金花茶为-14.58℃、龙州金花茶为-14.27℃、柠檬黄金花茶为-13.44℃、直脉金花茶为-13.09℃、东兴金花茶-12.74℃.低温半致死温度能反映金花茶种质的耐寒性,金花茶和龙州金花茶耐寒性强,其次为柠檬黄金花茶和直脉金花茶,东兴金花茶耐寒性较弱.在降温过程中,五种金花茶叶片相对电导率随温度降低呈S型上升,与温度呈负相关;脯氨酸、可溶性糖和丙二醛含量均呈现先上升后下降的趋势.同一低温条件下,半致死温度低的金花茶脯氨酸和可溶性糖的含量更高,而丙二醛含量更低.该研究结果为金花茶组植物耐寒种质选育提供了科学依据,为人工种植金花茶提供了技术支持.     

RsICE1基因表达和转基因水稻抗冷通路研究

以萝卜幼苗和水稻T1代转RsICE1基因(从抗寒萝卜植株中分离的一个编码碱性螺旋-环-螺旋低温胁迫转录因子,HQ891287)株系为材料,通过半定量RT-PCR及实时定量PCR等方法,分析RsICE1基因的表达、水稻转基因株系中RsICE1基因的遗传及冷诱导基因的表达情况.结果显示:(1)半定量RT PCR分析表明,RsICE1基因在萝卜的根、茎、叶中为组成型表达,在幼苗根和茎中表达量较强;RsICE1基因的表达水平能够被冷处理和NaCl处理诱导上调表达,但ABA和脱水处理无上调作用.(2)卡方测验表明,水稻转基因T1代潮霉素抗性发生了3∶1分离模式;Southern和Northern杂交结果表明,4个抗冷转基因株系中RsICE1基因均以单拷贝、单位点整合到水稻基因组并正常表达.(3)实时定量PCR分析表明,冷胁迫下,RsICE1基因超表达但对水稻OsDREB1A(AF300970)、OsDREB1B(AF300972)、OsDREB1F(AY785897)基因表达没有影响,表明RsICE1基因对转基因水稻抗寒性的影响不依赖OsDERB1冷反应通路.     

濒危植物缙云黄芩茎叶发育过程的解剖学研究

应用石蜡切片法对濒危植物缙云黄芩茎和叶的发育过程进行解剖学观察,同时获取不同发育阶段的叶表皮,观察其微形态特征,为地方特有珍贵物种资源的保护和利用提供理论参考。结果表明:(1)缙云黄芩的茎尖生长锥呈半球状,由原套和原体构成,是发育的起点。(2)茎的发育经历原分生组织、初生生长、次生生长,木质化程度加深,表面腺毛消失,非腺毛数量增加,最终形成成熟茎的结构。(3)叶源于叶原基,之后分化出原表皮、基本分生组织、原形成层,最终形成成熟的叶肉和叶脉;叶片成熟时栅栏组织占比大大缩减,叶片内腔隙极大;叶柄中央维管束与叶片主脉结构相似,突起处的维管束简化。(4)缙云黄芩的叶表皮气孔指数变化幅度较小,整体略有降低,气孔密度则先骤降后缓慢降低,腺毛逐渐消失,非腺毛逐渐增加。     

Cold hardiness in the Alpine beetles Patrobus septentrionis and Calathus melanocephalus

Alpine Patrobus septentrionis and Calathus melanocephalus (Col., Carabidae) were found to be susceptible to freezing. In the summer, the supercooling points were about ?5 to ?6°C. They were lowered during acclimation in the field and in the laboratory at 0 or ?3°C. Cold hardiness was correlated to, but not determined by, haemolymph osmolality. Thermal hysteresis was not detected. Increase in cold hardiness was concluded to be mainly a result of the influence of acclimation conditions on ice-nucleating compounds. In P. septentrionis, the results indicate that different compounds with ice-nucleation activity at different temperatures determine the limit of supercooling at different times. The haemolymph of both species supercooled well below the intact beetles at all seasons. Changes in haemolymph supercooling points could be ascribed to inactivation of ice nucleators in early autumn and to the effect of changes in solute concentration. In P. septentrionis, myo-inositol increased during cold-acclimation to 80–120 mMol concentrations, whereas C. melanocephalus produced 40–60 mMol trehalose.     

Response of the Carrot Weevil,Listronotus oregonensis(Coleoptera: Curculionidae), to Strains ofBacillus thuringiensis

Mortality and frass production bioassays were used to investigate the toxicity of seven strains ofBacillus thuringiensisagainst the adult carrot weevil,Listronotus oregonensis(Le Conte). A semi-artificial diet of carrot foliage with 4% agar was selected to maximize feeding by the insects.Bacillus thuringiensissubsp.tenebrionis(Krieg, Huger, Langenbruch, and Schnetter) (BTT) and two unidentifiedB. thuringiensisstrains, A30 and A429, gave the lowest LC₅₀values. The frass bioassay supported the conclusions of the mortality assay. Mortality of adults continued after their removal from the insecticidal medium, with the highest mortality being caused by strains A429 and BTT. Survivors from the frass bioassay, initially exposed to strains A30, A429, and BTT, did not resume normal levels of feeding after their removal from the insecticidal medium.     

上海地区红棕象甲的耐寒性研究

红棕象甲Rhynchophorus ferrugineus (Olivier)是我国危害棕榈科植物的重要入侵害虫, 为探明其越冬抗寒性, 利用过冷却点测定仪测定了红棕象甲上海种群不同虫态的过冷却点, 在低温箱内测定了该虫不同虫态的耐寒性, 然后结合田间越冬模拟试验、气象资料和寄主分布情况, 初步确定了红棕象甲在我国的越冬北界。结果表明： 红棕象甲过冷却点随虫态的发育程度的升高而下降, 卵、1龄幼虫、5龄幼虫、9龄幼虫和成虫的平均过冷却点分别为-5.92, -6.42, -7.19, -7.43和-11.84℃, 过冷却点由高到低的顺序依次为: 卵>幼虫>成虫。在6, 24, 48和72 h 4个时间处理下, 各虫态在低温与存活率之间呈显著或极显著的logistic回归, 半致死温度（Ltemp₅₀）均随处理时间的延长而上升, 不同虫态在处理72 h 后, 卵、1龄幼虫、5龄幼虫、9龄幼虫和成虫之间的Ltemp₅₀分别为1.61, -1.67, -2.39, -2.40和-0.40℃, 各虫态耐寒性由弱到强的顺序依次为: 卵<成虫<幼虫。红棕象甲不同发育阶段的过冷却点与其耐寒性并不完全相关, 幼虫和成虫均可能是该虫的越冬虫态。连续两年的田间模拟越冬试验表明, 在上海地区, 红棕象甲的幼虫和成虫的越冬存活率均在60％以上, 说明红棕象甲在上海地区是可以越冬的。根据这些结果, 结合寄主分布情况, 初步将红棕象甲在我国的越冬北界定于北纬35°附近, 即1月份0℃等温线左右。     

沙葱萤叶甲的过冷却能力与抗寒性

【目的】沙葱萤叶甲Galeruca daurica Joannis于2009年开始在内蒙古草原暴发成灾, 发生地区不断扩大, 危害日趋严重, 严重影响内蒙古草原畜牧业的可持续发展和生态安全。低温是影响昆虫生长发育和存活的关键因子, 而昆虫对低温的耐受性决定了其越冬存活率。了解沙葱萤叶甲的过冷却点及抗寒能力有助于预测其分布范围及种群数量动态。【方法】采用热电偶法, 在室内测定了沙葱萤叶甲各发育阶段的过冷却点; 比较了幼虫在不同低温条件下（-6~-14℃）暴露2 h及在-5℃低温条件下暴露不同时间（0.5~8 d）的存活率。【结果】沙葱萤叶甲不同发育阶段的过冷却点存在显著差异, 从低到高依次为卵（-29.8℃）、 1龄幼虫（-14.6℃）、 2龄幼虫（-13.3℃）、 蛹（-12.1℃）、 3龄幼虫（-10.2℃）和成虫（-9.0℃）; 越冬卵12月和1月的过冷却点最低, 2月的过冷却点最高。随着处理温度的降低和处理时间的延长, 幼虫的存活率降低。1, 2和3龄幼虫在-5℃下的半致死时间（Ltime₅₀）分别为3.84, 3.80和2.28 d, 低温处理2 h后半致死温度（Ltemp₅₀）分别为-10.1, -9.1和-8.5℃, 高于其过冷却点。【结论】说明沙葱萤叶甲幼虫为不耐寒冷型(chill-intolerant)。     

Cold tolerance of first-instar nymphs of the Australian plague locust, Chortoicetes terminifera

The cold tolerance of first-instar nymphs of the Australian plague locust, Chortoicetes terminifera, was examined using measures of total body water content, supercooling point and mortality for a range of sub-zero temperature exposure regimes. The supercooling points for starved and fed nymphs were −13.1 ± 0.9 and −12.6 ± 1.6 °C, and freezing caused complete mortality. Above these temperatures, nymphs were cold tolerant to different degrees based on whether they were starved or given access to food and water for 24 h prior to exposure. The rate of cooling also had a significant effect on mortality. Very rapid cooling to −7 °C caused 84 and 87% mortality for starved and fed nymphs respectively, but this significantly decreased for starved nymphs if temperature declined by more ecologically realistic rates of 0.5 and 0.1 °C min⁻¹. These results are indicative of a rapid cold hardening response and are discussed in terms of the likely effects of cold nights and frost on first-instar nymphal survival in the field.