蚊虫的滞育

<正> 蚊虫的滞育(diapause)是指其个体对不良环境条件长期适应的一种生理状态。滞育的发生与解除是需要一定的环境因素刺激导致体内激素参与的复杂过程。蚊虫的滞育类型主要为兼性滞育(facultative diapause),已知具有滞育现象的蚊虫多达40余种。     

昆虫滞育持续时间的影响因子及其对滞育后生物学的影响

滞育持续时间(diapause duration)是昆虫的一种生理特性,它主要由特定环境条件下的滞育强度或滞育发育速率来决定,滞育强度或滞育发育速率通常以将滞育状态的昆虫转移至解除滞育的条件(如低温处理、光周期的转换)后,滞育消除所需时间的长短来衡量。文章系统综述昆虫滞育持续时间的影响因子及其对滞育后生物学的影响。     

蜱类的滞育现象

<正> 滞育(diapause)是昆虫、蜱类等节肢动物中普遍存在的现象。对昆虫滞育现象的研究开展较早,也比较深入而对蜱类这一现象的研究则较晚。(1948)曾首次论述过硬蜱的滞育,近年来研究进展较快,已对几十种蜱类的滞育进行了研究。本文将概述蜱类滞育的表现形式,影响滞育的环境因素以及滞育期间蜱类的生理状态等问题。     

水浸对越北腹露蝗卵的影响

越北腹露蝗Fruhstorferiola tonkinensis Will.是广东北江流域河滩蝗虫的优势种。以不浸水的蝗卵为对照,研究滞育期蝗卵浸水5～95d和胚胎发育期蝗卵浸水1～30d对卵孵化率和历期的影响。结果表明,随浸水时间的增加,孵化率下降,卵发育延迟;滞育期蝗卵经受95d的浸水仍有8.01%存活,而胚胎发育期蝗卵浸水30d后全部死亡,滞育期蝗卵比胚胎发育期蝗卵具有更强的耐浸水能力。说明越北腹露蝗以卵滞育的形式发展了对水浸胁迫的适应。     

葱蝇热激蛋白DaHSP23基因的克隆及在冬滞育和夏滞育蛹中的表达分析

[目的]低分子量(12 ～43 kDa)热激蛋白(sHSPs)具有抗逆应答的功能,滞育是昆虫抵抗不良环境的特殊发育形式,但sHSPs在昆虫滞育发育过程中的作用仍不清楚.本研究克隆和特征化葱蝇Delia antiqua sHSP基因,并研究它在夏滞育和冬滞育发育过程中的表达模式,为阐明sHSPs在滞育发育上的功能奠定基础.[方法]通过RACE-PCR方法克隆了葱蝇HSP23基因,通过相似性比较分析了其特征、结构域及与双翅目代表性同源基因的系统发育关系;采用实时荧光定量PCR研究了该基因在葱蝇冬滞育蛹和夏滞育蛹发育过程中的表达情况,通过表达的差异比较揭示了该基因与滞育发育的关系.[结果]克隆出了葱蝇HSP23基因,命名为DaHSP23(GenBank登录号:HQ392521.1),其cDNA全长序列为904 bp,编码186个氨基酸,推测蛋白分子量为20.9 kDa,等电点为6.42.该基因的编码蛋白与其他双翅目昆虫的sHSPs有超过66％的氨基酸序列一致性,与已报道的其他双翅目昆虫的滞育相关HSP23基因同源.基因组测序显示该基因无内含子.DaHSP23基因在葱蝇非滞育蛹的发育过程中一直保持在较低的水平,各发育阶段间的表达量不存在显著差异.但在冬滞育和夏滞育蛹中,该基因从滞育起始期开始逐渐显著升高表达,到滞育维持期的中后期达到峰值,在滞育终止期逐渐降到较低的水平.[结论]DaHSP23基因在葱蝇冬滞育和夏滞育发育过程中明显上调表达,但存在差异,它在滞育期的调控可能是种专化的.DaHSP23可能在葱蝇两种类型的滞育上起重要作用.     

昆虫滞育研究进展

首先对昆虫滞育的若干名词概念作出解释,按照滞育过程的阶段特点划分为3个时期:(1)滞育前期(包括滞育诱导和滞育准备);(2)滞育期(包括滞育进入、滞育维持和滞育解除);(3)滞育后期。然后重点介绍这3个阶段近期分子和细胞水平上的重要工作,对一些科学问题进行讨论。     

白纹伊蚊卵滞育的实验观察

对两个地理品系的白纹伊蚊卵期滞育的实验观察表明:白纹伊蚊宜兴品系(约北纬31°N)不仅亲代受短光照作用后可诱导卵期发生滞育,而且卵期仍具有光照敏感性;海南岛品系(约北纬18°N)中,光期变化不能诱导卵期滞育.宜兴品系卵滞育率随亲代光期缩短而增高的现象只发生在一定范围内(约L:D=9:15到L:D=15:9).卵滞育还受卵期低温、干燥以及孵化时水中溶解氧的影响.卵进入深度滞育是多种环境因素作用的结果.     

茧蜂滞育的研究进展

滞育现象在多种茧蜂类天敌昆虫中存在, 通过调控茧蜂滞育, 可达到延长产品货架期、延长防控作用时间、提高产品的抗逆性和繁殖力的目标, 对茧蜂的扩繁和应用具有重要意义。本文通过总结1910年以来的2 545篇国内外滞育文献, 统计得出有明确滞育诱导或滞育解除报道的茧蜂有37种, 分属7亚科16属, 其中对滞育特征有详细描述的有23种。茧蜂大多以预蛹进入滞育, 其次为幼虫滞育, 少数为成虫滞育, 其滞育敏感阶段因种不同而异。滞育持续期相对较长, 大多可维持数月, 最长的可达14个月。光周期、温度和寄主是影响茧蜂滞育的主要因子, 湿度、亲代和寄主植物也可在一定程度上影响滞育进程。目前, 茧蜂滞育的研究仅限于生物学观察阶段, 对茧蜂滞育的生理学特征、激素调控和分子调控等尚未有研究报道, 有许多问题有待进一步探讨。     

九香虫滞育期脂肪、糖类和蛋白质含量的变化规律

【目的】研究九香虫Aspongopus chinensis Dallas滞育期体内的脂肪、糖类(总糖、糖原、海藻糖)、蛋白质含量的变化,探索体内营养物质变化与九香虫滞育的关系。【方法】本研究利用索氏提取法、蒽酮比色法和BCA蛋白定量试剂盒分别测定了九香虫滞育期(10月-次年4月)和滞育解除后(次年5月)其体内的脂肪、糖类和蛋白质含量变化。【结果】九香虫滞育期脂肪含量随着时间推移逐渐降低,到滞育后期(4月)脂肪含量最低为19.51%,与滞育初期(10月)的40.55%相比存在显著性差异(P<0.05);糖类在滞育初期(10月)开始积累,直至11月后不断减少至4月,滞育解除后(5月)糖类的含量又显著增高(P<0.05);蛋白质含量从滞育初期(10月)开始缓慢增加,直至3月开始下降,滞育解除后蛋白质含量显著低于滞育期(P<0.05)。【结论】九香虫滞育期体内脂肪、糖类和蛋白都发生了不同程度的变化。脂肪含量降低,糖类的含量先升后降,蛋白质含量先增后减是九香虫滞育过程中主要的生理特征。     

光周期对白纹伊蚊卵滞育影响的观察

过去几年在实验室饲养白纹伊蚊的过程中,作者见到每到9月中旬左右,虽平均温度仍在20℃以上,但成蚊所产的卵其孵化率大大降低,甚至完全不孵化,即使及时饲养在自然光源、25℃的恒温室中,也不能改变上述情况。近年来实验生态学的研究阐明了昼夜光照节律控制昆虫的季节发生有巨大的作用,其中最主要的是对昆虫的多型现象(polymor-phism)和滞育(diapause)形成的影响(de Wilde,1962;Andrewartha 1952)。作者受此启发,进行了光周期对白纹伊蚊产滞育卵关系的观察。     

The escape of the mink embryo from obligate diapause

The obligate embryonic diapause that characterizes gestation in mink engenders a developmental arrest at the blastocyst stage. The characteristics of escape from obligate diapause were investigated in embryos reactivated by treatment of the dams with exogenous prolactin. Protein and DNA synthesis showed marked increases within 72 h after the reinitiation of development, and embryo diameter increased thereafter. Trophoblast cells from embryos at Day 5 after activation proliferated more readily in vitro than trophoblasts from diapause or from Day 9 after activation, while in Day 9 embryos, cells from the inner cell mass (ICM) replicated comparatively more readily in vitro. There was evidence of expression of fibroblast growth factor-4 (FGF4) in both diapause and activated embryos and in ICM, but not the trophoblast. FGF receptor-2 was present in embryos from Day 5 after reactivation in both trophoblast and ICM cell lines. Trophoblast cell lines established from mink embryos proliferated in culture in the presence of FGF4 with a doubling time of 1.4 days, while in its absence, the doubling time was 4.0 days. We conclude that, during reinitiation of embryogenesis in the mink after diapause, embryo growth is characterized by gradual increases in protein synthesis, accompanied by mitosis of the trophoblast and ICM. There appears to be a pattern of differential proliferation between cells derived from these embryonic compartments, with the trophoblast phase of replication occurring mainly in the early reactivation phase, while the ICM proliferates more rapidly nearer to the time of implantation.     

The plurennial life cycles of the European Tettigoniidae (Insecta: Orthoptera)

Summary The effect of drought on embryonic development and on hatching was studied in 13 European Tettigoniidae species. Drought can affect development in three different ways: (1) Embryonic development proceeds slower than if the eggs are in contact with water; (2) it stops (for final diapause) in an earlier embryonic stage; (3) it affects maintenance and termination of the initial embryonic diapause.In many Tettigoniidae species, the initial diapause is prolonged, and may last several years. Without draught stress, between 1 and 7 cold treatments in the laboratory, and with eggs of the Tettigonia-species between 1 and 6 winters in the field were necessary to enable all eggs to complete initial diapause. In Central European species, the number of eggs maintaining initial diapause significantly increased when the eggs had no contact with water at the time when they should recover from diapause. In contrast, termination of initial diapause in Tettigonia caudata from Greece, when the environment became favorable for growth again, was highest in that group of eggs that had lost most water in a preceding period of drought. The importance of the prolonged initial diapause for the survival of unpredictable adverse conditions is discussed.     

Evidence of increased endometrial vascular permeability at the time of implantation in the short-nosed fruit bat, Cyanopterus sphinx

Early embryonic development and implantation were studied in tropical short-nosed fruit bat Cyanopterus sphinx. We report preimplantation development and embryo implantation. Different stages of cleavage were observed in embryo by direct microscopic examination of fresh embryos after retrieving them either from the oviduct or the uterus at different days, up to the day of implantation. Generally, the embryos enter the uterus at the 8-cell stage. Embryonic development continued without any delay and blastocyst were formed showing attachment to the uterine epithelium at the mesometrial side of the uterus. A distinct blue band was formed in the uterus. The site of blastocyst attachment was visualized as a blue band following intravenous injection of pontamine blue. Implantation occurred 9+/-0.7 days after mating. This study reports that bat embryonic development can be studied like other laboratory animals and that this bat shows blue dye reaction, indicating the site and exact time of implantation. This blue dye reaction can be used to accurately find post-implantational delay. We prove conclusively that this species of tropical bat does not have any type of embryonic diapause.     

Diapause in the seasonal cycle of stink bugs (Heteroptera,Pentatomidae) from the Temperate Zone

The paper reviews the data on diapause and related phenomena in stink bugs (Heteroptera: Pentatomidae). Using stink bugs as examples, the consecutive stages of the complex dynamic process of diapause (such as diapause preparation, induction, initiation, maintenance, termination, post-diapause quiescence, and resumption of direct development) are described and discussed. Out of 43 pentatomid species studied in relation to diapause in the Temperate Zone up to date, the majority (38 species) overwinter as adults, two species—as eggs, and another two species—as nymphs. Pentatoma rufipes is believed to be able to overwinter at different stages of its life cycle. Less than 5 % of pentatomid species are probably able to overwinter twice. Only five species have obligate diapause, others have the facultative one. Day-length and temperature are the main diapause inducing factors in the majority of species. The role of food in the control of seasonal development is essential in the pentatomid species feeding on plant seeds. In different species, different stages are sensitive to day-length. Some pentatomids retain sensitivity to photoperiod even after diapause, others lose it and become photo-refractory (temporarily or permanently). In Pentatomidae, such seasonal adaptations as photoperiodic control of nymphal growth rates, seasonal body colour change, migrations, and summer diapause (aestivation) are widely represented, whereas wing and/or wing muscle polymorphism has not been reported yet. In the subfamily Podopinae, induction of facultative reproductive winter diapause is under the control of photoperiod and temperature. All species feed on seeds and their seasonal development to a great extent reflects availability of food. However, the same food preferences and pattern of seasonal development are also characteristic to many species from the subfamily Pentatominae. All species of the subfamily Asopinae are predators. Among them, Picromerus bidens and Apateticus cynicus have obligate embryonic winter diapause, which is rear among true bugs. At the same time, A. cynicus and Podisus maculiventris belong to the same tribe but have different types of diapause: obligate embryonic diapause in A. cynicus and facultative adult diapause in P. maculiventris. Other Asopinae species studied up to date have facultative adult diapause controlled by photoperiod and temperature with probably only one exception: in Andrallus spinidens, adult diapause is controlled by temperature, and photoperiod plays only a secondary role. Thus, in spite of the similar habits and feeding types among Asopinae, the species of this subfamily have different types of diapause and the latter is controlled by different factors. In the subfamily Pentatominae, most species overwinter as adults and induction of their diapause is controlled by the long-day type photoperiodic response, in spite of the differences in their feeding preferences (within phytophagy). However, there are some exceptions in this subfamily, too: Palomena prasina, P. angulosa and Menida scotti have obligate diapause, which conditions univoltinism in these pentatomids. In M. scotti, only females have obligate adult diapause, whereas males remain physiologically active through the whole winter, this pattern being unusual for Heteroptera. The univoltine seasonal cycle of this species with summer diapause (aestivation) and apparent migrations is similar to that of shield bugs (Scutelleridae). According to the analysis of seasonal development, the evolution of seasonal adaptations in Pentatomidae does not directly reflect their phylogeny. However, individual genera, small tribes or even subfamilies have similar complexes of seasonal adaptations. At the same time, Pentatominae is a large and apparently collected taxon, but most of species in this subfamily have the same facultative adult diapause.     

Cloning of leukemia inhibitory factor (LIF) and its expression in the uterus during embryonic diapause and implantation in the mink (Mustela vison)

Leukemia inhibitory factor (LIF) is essential for embryo implantation in mice. Whether LIF plays a role in termination of embryonic diapause and initiation of implantation in carnivores, especially in species with obligate delayed implantation such as the mink, is not known. The objectives of this study were to clone the LIF coding sequence in the mink and determine its mRNA abundance in the uterus through embryonic diapause, implantation, and early postimplantation. We show that the mink LIF cDNA contains 609 nt encoding a deduced protein of 203 amino acids. The homologies are 80.6, 90, 88.2, 87.6, and 86.8% in coding sequence and 79.2, 90.1, 91, 90.1 and 85.4% in amino acid sequence with mouse, human, pig, cow, and sheep respectively. Glycosylation sites and disulfide bonds present in other species are generally conserved in the mink LIF sequence. Quantitation by polymerase chain reaction amplification indicates that LIF mRNA is expressed in mink uterus just prior to implantation and during the first two days after implantation, but not during diapause or later after implantation pregnancy. The abundance of LIF mRNA was significantly higher in the uterus at the embryo expansion stage (P < 0.05) than at days 1–2 of postimplantation. By immunohistochemical localization it was shown that LIF is expressed in the uterine epithelial glands at time of embryonic expansion and in early postimplantation. The coincidence of LIF expression with implantation in this species suggests that LIF is involved in the implantation process, and may be a maternal signal which terminates obligate embryonic diapause. Mol. Reprod. Dev. 51:13–21, 1998. © 1998 Wiley-Liss, Inc.     

Do ovarian steroid hormones control the resumption of embryonic growth following the period of diapause in roe deer (Capreolus capreolus)?

Embryonic diapause in the European roe deer includes a period of five months from August to December in which embryonic development is extremely decelerated. Following exit from diapause, the embryo rapidly elongates and subsequently implants. In diapausing carnivores and marsupials, resumption of embryonic growth is regulated by ovarian steroid hormones. In the roe deer, the role of steroid hormones is not known to date. In the present study, progesterone (P4), estradiol-17β (E2) and total estrogens (E_tot) were determined in blood plasma and endometrium of roe deer shot in the course of regular huntings between September and December. Steroid hormone concentrations were correlated to the corresponding size of the embryo derived from ex vivo uterine flushing and to the date of sampling. The mean plasma concentrations of P4 (5.4 ± 0.2 ng/ml, mean ± SE, N = 87), E2 (24.3 ± 2.6 pg/ml, N = 86) and E_tot (21.7 ± 2.6 pg/ml, N = 78) remained constant over the sampling period and were not correlated to embryonic size. Likewise, endometrial concentrations of P4 (66.1 ± 6.5 ng/ml), E2 (284.0 ± 24.43 pg/ml) and, E_tot (440.9 ± 24.43 pg/ml) showed no changes over time. Therefore, it was concluded that ovarian steroid hormones do not play a determining role in resumption of embryonic growth following the period of diapause in the roe deer.     

Involvement of ERK/MAPK in regulation of diapause intensity in the false melon beetle, Atrachya menetriesi

Embryonic diapause is commonly terminated by exposure to low temperature for a certain duration. Previous studies using the silkworm, Bombyx mori, showed that extracellular signal-regulated kinase (ERK), a member of the mitogen-activated protein kinase family, was activated by cold exposure and regulated diapause termination. The involvement of ERK in regulation of diapause termination was investigated in the false melon beetle, Atrachya menetriesi. Embryonic diapause of this beetle is terminated both by cold exposure and by mercury. Phospho-ERK levels remained high during the pre-diapause period but decreased after the eggs entered diapause. Exposure to 7.5 degrees C, which was effective for diapause termination, increased phospho-ERK levels, and these levels were maintained under 7.5 degrees C at least for 100 d. Incubation at 25 degrees C after the eggs were kept at 7.5 degrees C for 20 d, which intensified diapause, decreased the phospho-ERK level. An insufficient cold treatment, i.e., incubation at 0 degrees C for diapause termination did not activate ERK. However, incubation at 0 degrees C after cold treatment at 7.5 degrees C, which is effective for diapause termination, induced high phospho-ERK levels. Moreover, mercury treatment also increased phospho-ERK. Therefore, changes in the phospho-ERK level correlated well with diapause intensity. The results suggest that ERK plays a key role in the regulation of embryonic diapause.     

Effects of juvenile hormone analogs and 20-hydroxyecdysone on diapause termination in eggs of Locusta migratoria and Oxya yezoensis

To understand the hormonal control of embryonic diapause, juvenile hormone analogs (JHAs), methoprene and hydroprene, and 20-hydroxyecdysone (20E) were applied onto diapause eggs of Locusta migratoria and Oxya yezoensis. These insects enter diapause at the mid-stage of embryogenesis prior to blastokinesis. Topical application of JHAs significantly facilitated diapause termination in both species but JHA-treated embryos underwent abnormal morphogenesis, pigmentation and sclerotization without dorsal closure. The Locusta eggs immersed in the 20E solution for 24h terminated diapause in a dose-dependent manner. We also investigated phosphorylation of extracellular signal-regulated kinase (ERK), a member of mitogen-activated protein kinase (MAPK), during diapause-terminating process of Locusta migratoria and found that ERK was activated either by cold exposure or JHA treatment. The possible involvement of the hormones and ERK in embryonic diapause and the possibility of ecdysteroids synthesis by prothoracic glands of diapause embryo were proposed.