昆虫滞育后的生物学特性

从昆虫滞育后性比、寿命、生殖力和重复滞育等方面对昆虫滞育后的生物学特性进行了概括,并分析了影响昆虫滞育后生物学特性数量表达的因子。这些因子包括:(1)滞育前昆虫的发育速率、取食行为和个体大小;(2)滞育期间的环境条件及昆虫取食行为;(3)滞育持续期;(4)滞育后的取食需求;以及(5)滞育后的温度和光周期。     

真菌学的二百五十年

本文对真菌学的发展历史作了简要的回顾并将其分为4个发展阶段:(1)前真菌学阶段(B.C.5000—A.D.1700);(2)古真菌学阶段(1701—1850);(3)近代真菌学阶段(1851—1950);(4)现代真菌学的发展(1951—现在)。     

真菌学的二百五十年*

本文对真菌学的发展历史作了简要的回顾并将其分为4个发展阶段:(1)前真菌学阶段(B.C.5000—A.D.1700);(2)古真菌学阶段(1701—1850);(3)近代真菌学阶段(1851—1950);(4)现代真菌学的发展(1951—现在)。     

意大利蝗卵发育过程保幼激素和蜕皮激素含量的变化及其代谢相关基因的表达谱

【目的】鉴于保幼激素(juvenile hormone, JH)与蜕皮激素(ecdysone, Ecd)在昆虫生长发育中的重要作用,本研究旨在明确这两种激素及其代谢相关基因对意大利蝗Calliptamus italicus卵发育过程的调控机制。【方法】采用ELISA技术检测意大利蝗卵发育过程中JH与Ecd含量的变化;采用qRT-PCR技术检测JH与Ecd代谢通路的重要基因(JHE,JHEH,DIB和EcR)在意大利蝗卵发育过程的表达模式。【结果】意大利蝗卵滞育阶段(阶段Ⅳ-Ⅵ)的JH含量显著高于早期发育阶段(阶段Ⅰ-Ⅲ)的,滞育后发育阶段(阶段Ⅶ-Ⅸ) JH含量显著下降;Ecd含量于滞育早期(阶段Ⅳ)显著上升,而后显著下降,滞育后发育阶段再次上升。JHE的表达量在意大利蝗卵早期发育阶段和滞育后发育阶段均呈先升后降的趋势,滞育阶段JHE的表达量较低;JHEH表达量在意大利蝗卵早期发育阶段也先升后降,滞育阶段变化不显著,滞育后发育阶段显著升高;DIB表达量在意大利蝗卵滞育阶段高于早期发育阶段的,在滞育后发育阶段下降;EcR表达量在意大利蝗卵早期发育阶段及滞育阶段均无显著变化,滞育后发育阶段...     

沙葱萤叶甲成虫不同夏滞育阶段的转录组分析(英文)

【目的】本研究旨在揭示内蒙古草原新害虫沙葱萤叶甲Galeruca daurica专性夏滞育相关的重要基因以及代谢通路。【方法】应用RNA-Seq技术,对沙葱萤叶甲成虫不同夏滞育阶段［滞育前期(PD)、滞育期(D)及滞育后期(TD)］进行转录组测序、分析及基因功能预测,基于RNA-Seq数据筛选夏滞育不同阶段差异表达基因;利用qPCR对基于RNA-Seq数据筛选的10个差异表达基因的表达水平进行验证。【结果】从9个文库中获得202 770 198 clean reads,将12 078 060条转录本组装获得82 292 条unigene,平均长度为783.59 bp,N50为1 545 bp。沙葱萤叶甲D vs PD和TD vs D 比较组分别有2 395(2 119上调和277下调)和62(59上调和3下调)个差异基因。KEGG分析表明,D vs PD和TD vs D比较组差异表达基因分别显著富集于糖孝解/糖异生通路和脂肪酸生物合成通路;此外,许多与钙离子信号转导相关的基因在滞育期间差异表达。10个差异表达基因的qPCR分析表明,RNA-Seq与qPCR结果高度一致。【结论】糖孝解/糖异生、脂肪酸生物合成及钙离子信号通路可能在沙葱萤叶甲滞育调节中起着重要的作用。本研究为进一步研究沙葱萤叶甲成虫专性夏滞育的分子机理奠定了基础。     

苜蓿切叶蜂滞育的诱导因素研究

苜蓿切叶蜂Megachile rotundata(F.)预蛹的滞育主要受光周期和温度的影响。雌蜂感受光周期的变化是决定子代预蛹是否进入滞育的一个主要因素;当代的幼期,特别是高龄幼虫(约3龄)到预蛹这一阶段所感受的温度变化,对预蛹滞育有很大的影响,此期如果环境温度低于(21.59±1.03)℃,可有超过50%的个体进入滞育。此外,放蜂地区的纬度及蜂的代次对预蛹的滞育亦有重要的影响。     

昆虫滞育诱导的分子调控机制

滞育是昆虫长期适应不利环境的重要对策之一,对昆虫的生存、繁衍和进化都具有重要意义。滞育一般分为滞育前期、滞育期和滞育后期3个阶段,其中,滞育前期包括滞育诱导期和滞育准备期,是确保昆虫顺利进入滞育的重要时期。本文首先简要概括了光周期、温度、湿度、食料等外界环境因素在不同昆虫滞育诱导中的作用;然后系统综述了滞育激素、保幼激素、蜕皮激素、促前胸腺激素和胰岛素等内分泌激素,生物钟通路的节律基因,以及DNA甲基化、小RNA和组蛋白修饰等表观遗传修饰在昆虫滞育过程,尤其是滞育诱导调控中的作用,并以黑腹果蝇Drosophila melanogaster的生殖滞育为例,总结了昆虫滞育诱导调控的分子通路;最后就目前昆虫滞育诱导调控研究中亟待解决的问题和后续重点研究方向进行了探讨,展望了将昆虫滞育研究应用于害虫防控实践的主要策略和前景。     

小麦吸浆虫保幼激素酯酶和保幼激素环氧水解酶基因的克隆及在滞育和变态发育过程中的表达动态

【目的】保幼激素(juvenile hormone, JH)在小麦吸浆虫Sitodiplosis mosellana滞育诱导及滞育后静息状态的维持中发挥着重要作用。保幼激素酯酶(hormone esterase, JHE)和保幼激素环氧水解酶(juvenile hormone epoxide hydrolase, JHEH)是调控JH滴度的重要降解酶。本研究旨在探讨JHE和JHEH在小麦吸浆虫滞育和变态发育中潜在功能。【方法】通过RT-PCR和RACE技术从小麦吸浆虫滞育前幼虫克隆JHE和JHEH全长cDNA序列;利用生物信息学软件分析其核苷酸及编码蛋白特性;采用qPCR技术分析其在小麦吸浆虫滞育不同时期(滞育前、滞育期、滞育后静息期和滞育后发育)3龄幼虫及1龄幼虫到成虫不同发育阶段(1-2龄幼虫、预蛹、初蛹、中蛹、后蛹、雌成虫和雄成虫)中的表达水平。【结果】克隆获得了cDNA全长分别为3 102和1 980 bp的小麦吸浆虫SmJHE和SmJHEH基因(GenBank登录号分别为MG876768和MG876769),其开放阅读框分别长1 740和1 371 bp,分别编码579和456个氨基酸,预测蛋白分子量分别为65.67和51.65 kD。SmJHE蛋白含有5个JHE家族特有的保守模块,SmJHEH含有催化三联体Asp228, Asp404和His431及组成阴氧离子洞的两个Tyr(Tyr299和Tyr374)和HGWP花样结构。序列比对和进化分析表明,SmJHE和SmJHEH均与双翅目(Diptera)长角亚目(Nematocera)昆虫同源蛋白氨基酸序列一致性较高,亲缘关系最近。不同滞育时期的表达模式表明,SmJHE和SmJHEH在滞育前期(1龄到滞育前的3龄幼虫早期)表达量变化不明显,进入滞育后表达量基本维持恒定,但均在滞育后静息阶段的当年12月至翌年1月最低。发育表达模式表明,幼虫恢复发育后SmJHE表达量逐渐升高,预蛹期达到最高,在雌成虫中的表达量显著低于雄成虫中的;SmJHEH表达量则在预蛹期最低,在雌成虫中最高。【结论】SmJHE和SmJHEH参与小麦吸浆虫滞育调控,其表达量的降低与滞育后静息阶段JH的累积有关;SmJHE在发育过程中表达量的升高可能参与幼虫到蛹的变态,表达量的降低可能与生殖发育有关。     

意大利蝗卵发育过程中血蓝蛋白基因表达分析

【目的】意大利蝗Calliptamus italicus是新疆草原的主要优势危害种,以卵在土壤中越冬。呼吸代谢可反映蝗卵的生理状态,呼吸蛋白对于呼吸系统不完善的蝗卵尤为重要。本研究旨在明确意大利蝗卵发育过程中血蓝蛋白基因的表达情况。【方法】采用实时荧光定量PCR方法检测不同发育阶段的蝗卵以及1龄蝗蝻的血蓝蛋白2个亚基基因Hc1和Hc2的表达量。【结果】根据解剖形态观察,将意大利蝗越冬卵的整个发育过程分为10个阶段,包括9个卵发育阶段(C-Ⅰ-C-Ⅹ)和1龄蝗蝻阶段(C-Ⅹ)。Hc1和Hc2在越冬蝗卵各发育阶段以及1龄蝗蝻中均有表达。其中,在蝗卵早期发育阶段(C-Ⅰ, C-Ⅱ和C-Ⅲ),Hc1表达量逐渐增加,C-Ⅲ阶段表达量显著高于C-Ⅰ和C-Ⅱ阶段;滞育阶段(C-Ⅳ, C-Ⅴ和C-Ⅵ),胚胎发育停滞,Hc1表达量较C-Ⅲ,C-Ⅶ和C-Ⅷ阶段低;滞育后发育阶段(C-Ⅶ和C-Ⅷ),蝗卵解除滞育,快速发育,Hc1表达量较早期发育阶段和滞育阶段高,其中,C-Ⅷ阶段Hc1表达量最高(212.3156±10.5470),显著高于其他所有阶段;1龄蝗蝻(C-Ⅹ)的Hc1表达量最低,为0.4017±0.1010。Hc2表达量在C-Ⅴ阶段最高(679.7511±54.5719),显著高于其他所有阶段;除C-Ⅴ阶段外,其他各阶段之间Hc2表达量差异均不显著。Hc1在蝗卵滞育后阶段高表达,而Hc2在蝗卵滞育阶段高表达。【结论】血蓝蛋白亚基基因Hc1和Hc2在整个意大利蝗卵发育过程均有表达,且具有阶段特异性。Hc1与Hc2协同作用为蝗卵发育供氧,其中,Hc1主要负责蝗卵滞育后发育期间的氧气运载,而Hc2主要维持滞育期间的氧气运载,且载氧效率较低。研究结果可为进一步探讨意大利蝗卵的抗逆机制提供科学依据。     

滞育及不同虫态下亚洲玉米螟内参基因的筛选

筛选出滞育状态下以及不同虫态下亚洲玉米螟Ostrinia furnacalis(Guenée)的实时荧光定量PCR(RT-qPCR)最稳定内参基因,本结果为研究不同虫态及滞育1个月,滞育2个月,滞育3个月和滞育4个月的亚洲玉米螟目的基因表达水平提供参考依据。本文以不同虫态及滞育状态下的亚洲玉米螟为试验材料,应用RT-qPCR技术检测β-actin,18S rRNA,EF1-α和RPS3共4个候选内参基因的表达稳定性水平,结合ge Norm,Normfinder,Best Keeper和Ref Finder软件分析各候选内参基因在不同处理下的表达稳定性。结果表明,在亚洲玉米螟不同虫态中,4个候选内参基因的稳定性大小排序为18S rRNAEF1-αβ-actinRPS3;滞育1个月,滞育2个月,滞育3个月和滞育4个月的亚洲玉米螟,稳定性大小排序为β-actinEF1-α18S rRNARPS3。18S rRNA和β-actin可分别作为不同虫态和滞育状态下的亚洲玉米螟基因表达水平分析试验中的内参基因。     

Eco-physiological phases of insect diapause

Insect diapause is a dynamic process consisting of several successive phases. The conception and naming of the phases is unsettled and, sometimes, ambiguous in the literature. In this paper, the ontogeny of diapause was reviewed and the most often used terms and the best substantiated phases were highlighted, explained and re-defined. The aim was to propose relatively simple and generally applicable terminological system. The phases of diapause induction, preparation, initiation, maintenance, termination and post-diapause quiescence were distinguished. The specific progression through diapause phases in each species, population (genotype), or even individual, is based on (thus far largely unknown) physiological processes, the actual expression of which is significantly modified by diverse environmental factors. Thus, such phases are eco-physiological in their nature.     

THE SIGNIFICANCE OF THE LIGHT AND DARK PHASES IN THE PHOTOPERIODIC CONTROL OF DIAPAUSE IN METATETRANYCHUS ULMI KOCH

The daily cycle of illumination is one of several agencies which control the onset of diapause in Metatetranychus ulmi. Both light and dark phases in the cycle are concerned in the determination process.
In general, a long light phase tends to suppress and a long dark phase to induce a diapause. In any combination, the path of development is decided by the balance between diapause-preventing (light phase) and diapause-inducing (dark phase) stimuli. However, as their effectiveness does not increase linearly with duration, the existing balance changes with the phase duration.
The effectiveness of the light phase in suppressing diapause increases most rapidly between 8 and 16 hr.; that of the dark phase rises very sharply between 8 and 12 hr. Longer dark periods of up to several days duration also induce diapause but are no more effective than a 13 hr. phase. The inclusion in the cycle of very long periods of light or darkness may also influence diapause by reducing the number of complementary phases experienced by the mite during the sensitive period of development.
M. ulmi is highly insensitive to the interruption of effective light and dark phases by short intervals of darkness or light—a further indication of the slow inception of the light-and dark-phase reactions.
These findings are discussed in terms of hypothetical mechanism involving cumulative synthesis and removal of some active substance, but the experimental results cannot yet be fully reconciled with a simple hypothesis of this kind.     

Differences in the pre‐diapause and pre‐oviposition accumulation of critical nutrients in adult females of the beetle Colaphellus bowringi

In many insect species, the differentiation of development between diapause and reproduction first becomes obvious during the diapause preparation (pre‐diapause) and pre‐oviposition phases. However, the differentiation of nutrient accumulation between these two phases remains unclear. We compared the weights of pre‐diapause and reproductive adult female Colaphellus bowringi Baly (Coleoptera: Chrysomelidae), and measured their triacylglycerol (TAG), protein, and carbohydrate content from emergence until they had fed for 4 days post‐eclosion. We also compared the ovarian development and accumulation of lipid droplets between pre‐diapause and reproductive adult females in order to determine whether we could visually detect differences in nutrient allocation. The weights of both pre‐diapause and reproductive females increased with duration of feeding. The fresh weight and water content of pre‐diapause females was significantly lower than that of reproductive females after feeding for 3 days post‐eclosion. Pre‐diapause females channeled their reserves into TAG in the fat body, whereas reproductive females converted nutrients into proteins and carbohydrates for egg development. These results quantify differences in nutrient accumulation between pre‐diapause and reproductive adult female C. bowringi, and provide clues for understanding the molecular mechanisms underlying differences in the allocation of nutrients between diapause and reproduction in insects.     

Meeting the energetic demands of insect diapause: nutrient storage and utilization

Insects in diapause characteristically feed very little or not at all, thus they are largely or totally dependent on energy reserves sequestered prior to the entry into diapause. Fats are the dominant reserve used during this period, but non-fat reserves are also important for some species, especially during certain phases of diapause. Metabolic depression, coupled with the low temperatures of winter, facilitates the economic utilization of reserves during the many months typical of most diapauses. Though many insects store additional lipid prior to the entry into diapause, our review of the literature indicates that this is not always the case. We provide evidence that interactions between nutrient storage and metabolism can influence the decision to enter diapause and determine how long to remain in diapause. In addition, the energy reserves expended during diapause have a profound effect on post-diapause fitness. Though the physiological and biochemical mechanisms that regulate nutrient homeostasis prior to and during diapause remain poorly known, we propose several mechanisms that have the potential to contribute to diapause-associated nutrient homeostasis. Potential players include insulin signaling, neuropeptide F, cGMP-kinase, AMP-activated protein kinase, and adipokinetic hormone.     

Diapause research in insects: historical review and recent work perspectives

All organisms on Earth have evolved biological rhythms to face alternation of periods of favorable and unfavorable environmental conditions, at various temporal scales. Diapause is a state of seasonal dormancy adapted to recurring periods of adverse environmental conditions and triggered by biotic and abiotic factors that precede the arrival of these conditions. Several monographs already review the mechanisms of diapause expression in arthropods, from initiation to termination phases. Rather than adding another review to the literature on this topic, this paper primarily aims to link past concepts on seasonal strategies with new perspective on diapause research in arthropods. By focusing on insects, I examine the legacy of diapause history research in terrestrial arthropods since antiquity but mostly over the past 3 centuries, its contribution to the understanding of insect seasonal ecology, and I explore some of the reasons why it is still relevant to study diapause. I highlight some of the topical issues on which current work focuses to better understand and integrate arthropod diapause with their ecology, especially in the climate change context and for the provision of ecosystem services.     

Diapause decision in the small tortoiseshell butterfly,Aglais urticae

Insects in temperate areas spend the inhospitable winter conditions in a resting stage known as diapause. In species that diapause in the larval or pupal stage, the decision whether to diapause or develop directly is customarily taken during the late instars, with long days (i.e., long light phases) and high temperatures promoting direct development. Among butterflies that overwinter as adults, data are rare and variable, but imply that the larval daylength conditions can affect the pathway decision. We studied the small tortoiseshell, Aglais urticae L. (Lepidoptera: Nymphalidae, Nymphalini), which is partially bivoltine from Central Scandinavia and southwards, and tested whether the pathway decision is taken in the larval or adult stage. We reared larvae under long‐day (L22:D2) or short‐day (L12:D12) photoperiods, and recorded the pathway taken by the eclosing adults by scoring their propensity to mate and produce eggs. We also tested whether the larval photoperiod influenced adult ability to diapause by assessing adult survival. The results clearly indicate that (1) there is no detectable effect of larval photoperiod treatment on the pathway decision taken by adults whether to enter diapause or to develop directly, (2) some individuals are obligately univoltine and insensitive to photoperiod during adulthood, whereas (3) other individuals can facultatively enter diapause or direct development, depending on the photoperiod experienced after adult eclosion.     

Oxygen consumption in relation to sorbitol utilization at the termination of diapause in eggs of the silkworm,Bombyx mori

Rates of oxygen consumption were followed throughout the entire period of diapause in eggs of Bombyx mori. In non-diapause eggs at 25 degrees C, O(2) uptake was divisible into three phases, corresponding to morphogenetic processes. In diapause eggs at 25 degrees C, O(2) uptake showed a peak (100 &mgr;l/g eggs/h) at 1 day and then suddenly dropped to reach a level of 8-10 &mgr;l/g eggs/h at 10 days and thereafter. To break diapause, eggs were exposed to 5 degrees C for varying periods. When O(2) uptake was measured at 5 degrees C, it remained at 6 &mgr;l/g eggs/h. When eggs were chilled for increasing periods and O(2) uptake was measured immediately after warming to 25 degrees C, the rates increased after a lag phase. In HCl-treated eggs, O(2) uptake increased immediately after acid-treatment. In all cases, highly increasing O(2) uptake at 25 degrees C coincided with termination of diapause. These results were discussed in relation to sorbitol utilization at the termination of diapause.     

Diapause in a Glasgow strain of the flour moth, Ephestia kuehniella

ABSTRACT. The incidence of diapause in Ephestia kuehniella Zeller from an unhealed granary in Scotland was influenced by both photoperiod and temperature. At 25°C, nearly 50% of larvae entered diapause when reared in continuous darkness (DD) and up to 30% did so in short photoperiods. Little diapause was detected around LD 14:10 but a second, smaller peak of about 20% occurred at LD 16:8 and LD 18:6, falling away again to nearly zero in continuous light. More larvae entered diapause when reared continuously at 15 or 20°C than at 25 and 30°C. However, when larvae reared from hatch at 25°C in LD 16:8 were transferred after 1 week to 15°C in LD 9:15, almost twice as many entered diapause as did those reared at 15°C throughout. The sensitive phase for diapause induction occurred near the start of the final instar. The mean duration of diapause was between 2 and 3 months in most photoperiods at 20 and 25°C, and was shorter at 15°C. However, in DD at 25°C, it lasted about 7 months. Termination of diapause was hastened in larvae reared at 25°C in DD by transferring them to LD 14:10, and also by chilling them at 7.5°C for 6 weeks before returning to 25°C in DD. In an unhealed store in southern England, viable adults emerged from May to July and originated from larvae which terminated diapause relatively late. It would appear from the results of transferring larvae back to the laboratory at various times during the winter that some phases of diapause development were completed quite early after exposure to low temperatures, although no further development took place in the store until temperatures rose again in April.