丽蝇蛹集金小蜂的微生态环境及其对个体发育的影响

丽蝇蛹集金小蜂(Nasonia vitripennis walker)是膜翅目的一种寄生性昆虫,其幼期阶段寄生于蝇蛹体内,取食蛹内组织,起到灭蝇作用。该蜂在寄主蝇蛹内进行胚胎期、幼虫期和蛹期各阶段的发育。蜂卵孵化出幼虫,经过三个龄期、预蛹与蛹再羽化出成蜂啮破蝇蛹壳而出,整个过程都是在寄主蝇蛹内进行。羽化出的成蜂再交配、寻找寄主、产卵寄生。因此,作者认为丽蝇蛹集金小蜂的寄主蝇蛹就是该蜂生长发育的微生态环境(Microen vironment)。蜂幼并在寄主蝇蛹内越冬,所以,蝇蛹自然成为蜂幼虫越冬的微生态环境。关于丽蝇蛹集金小蜂的生物学与生态习性的研究报告很多,但对该蜂的微生态环境及其对个体发育的影响报告很少。对该蜂微生态环境及其对个体发育的影响研究有助于其生物学与生产实践利用的探讨。作者于1984年4月至1985年12月对该蜂的微生态环境及其对个体发育的影响进行了初步研究,现报告如下。     

去除Wolbachia 对丽蝇蛹集金小蜂繁殖适合度和成蜂寿命的影响

【目的】明确内共生菌 Wolbachia 对丽蝇蛹集金小蜂 Nasonia vitripennis 繁殖适合度和成蜂寿命的影响。【方法】通过给自然感染 Wolbachia 的丽蝇蛹集金小蜂成蜂喂食不同浓度的利福平来消除其体内的 Wolbachia,然后进行10个世代的连续饲养,探究不同浓度利福平对丽蝇蛹集金小蜂体内 Wolbachia 的去除效果和去除 Wolbachia 后对丽蝇蛹集金小蜂繁殖力、性比（雌蜂占子代数量的比值）和成蜂寿命的影响。【结果】低浓度利福平（0.1~0.5 mg/mL）对丽蝇蛹集金小蜂的毒害作用较小,而高浓度利福平（0.7~10.0 mg/mL）对丽蝇蛹集金小蜂的毒害作用较大,但二者均能去除丽蝇蛹集金小蜂体内的 Wolbachia;去除 Wolbachia 后丽蝇蛹集金小蜂的出蜂量显著下降（P <0.01）,子代中性比显著下降（P <0.01）,但寿命无明显差异。【结论】不同浓度利福平均能去除丽蝇蛹集金小蜂体内Wolbachia,但效果不一致;Wolbachia 对丽蝇蛹集金小蜂的出蜂量和子代性比均有显著影响,对成蜂寿命无显著影响。     

丽蝇蛹集金小蜂的大量繁殖与释放方法

丽蝇蛹集金小蜂Nasonia vitripennis Wa-lker是膜翅目的一种拟寄生性昆虫(Parasi-toid),其幼期寄生于蝇蛹内,取食蛹内组织,致使蝇蛹死亡。在利用蝇蛹寄生蜂防治蝇类     

金小蜂基因组学研究前沿

金小蜂不仅是重要的昆虫天敌资源,还是理想的模式生物。2010年1月15日,3种金小蜂(丽蝇蛹集金小蜂Nasonia vitripennis、吉氏金小蜂N.giraulti和长角金小蜂N.longicornis)的基因组在《科学》上发表。这一事件标志着金小蜂研究"后基因组时代"的开始。金小蜂基因组测序完成后,科学家们就利用生物信息学、比较基因组学、功能基因组学等方法,基于基因组平台,在进化遗传学、发育生物学、神经生物学、行为学等领域开展了系列研究,取得了重要进展,国际著名杂志《昆虫分子生物学》和《遗传》还以特刊的形式进行刊载。本文就金小蜂基因组学相关研究取得的进展予以扼要概述,并探讨其研究方向和发展前景。     

丽蝇蛹集金小蜂气味结合蛋白基因的克隆与序列分析

利用RACE技术,首次从丽蝇蛹集金小蜂Nasonia vitripennis中克隆获得一个气味结合蛋白全长cDNA序列.该基因全长553 bp,开放阅读框411 bp,3'和5'端非编码序列分别为13 bp和129 bp.其推导的氨基酸序列编码136个氨基酸,推测编码蛋白质的分子量为15.4 kDa,等电点为8.76.同源性比对分析发现,丽蝇蛹集金小蜂气味结合蛋白基因与现已报道的其它昆虫气味结合蛋白基因在氨基酸水平上的相似性均低于30%,拥有6个保守半胱氨酸位点等气味结合蛋白所具有的典型特征.系统进化树分析表明,丽蝇蛹集金小蜂气味结合蛋白与意大利蜜蜂Apis mellifera气味结合蛋白2,3,4,5,6,7,8和12聚为同一族,与意大利蜜蜂气味结合蛋白5,6和8的进化程度最近.RT-PCR分析表明,丽蝇蛹集金小蜂气味结合蛋白基因不仅在雌、雄成虫触角中高度表达,而且在头部和足中有微弱表达.     

取食行为对丽蝇蛹集金小蜂雌雄成虫体内可培养真菌的影响

【目的】本研究旨在阐明取食行为对丽蝇蛹集金小蜂Nasonia vitripennis(Walker)雌雄虫体内可培养真菌群落结构的影响,对其体内可培养真菌进行多样性研究。【方法】采用不同浓度、不同培养基通过分离培养法研究丽蝇蛹集金小蜂体内真菌,将培养所得真菌采用形态和分子方法(ITS基因序列)进行鉴定。【结果】取食前后丽蝇蛹集金小蜂雌、雄虫体内共分离得到49种真菌,隶属子囊菌门,盘菌亚门的座囊菌纲、散囊菌纲、锤舌菌纲、子囊菌纲,共10个属,其中以链格孢属(Alternaria)、枝孢属(Clacdosporium)、青霉属(Penicillium)为优势属。【结论】丽蝇蛹集金小蜂体内可培养真菌物种数目,雄虫多于雌虫,取食后多于取食前。本研究结果说明食物、性别是丽蝇蛹集金小蜂体内可培养真菌群落结构产生差异的重要因素。     

家蝇蛹和果蝇蛹繁育的蝇蛹金小蜂对果蝇蛹的寄生比较

【目的】蝇蛹金小蜂Pachycrepoideus vindemmiae(Rondani)是杨梅园等果园果蝇类害虫蛹期常见寄生蜂种类,在对果蝇类害虫的生物防治上具有重要价值。本文旨在探讨使用家蝇蝇蛹为替代寄主繁育蝇蛹金小蜂的方法。【方法】探讨分别以家蝇蛹和果蝇蛹繁育的蝇蛹金小蜂对家蝇和果蝇蝇蛹的选择性,并比较了在两种寄主上繁育的蝇蛹金小蜂在大小、寿命、产卵期、后代产量和性比等方面的差异。【结果】结果表明与果蝇蛹相比,家蝇蛹明显较大,在家蝇蛹上发育的蝇蛹金小蜂后代个体也明显较大;家蝇蛹和果蝇蛹发育的寄生蜂雌蜂寿命为(13.4±4.11)和(3.94±2.49)d、产卵期分别为(11.4±4.11)和(3.13±2.42)d、单头雌蜂后代雌蜂数量分别为(34.31±31.83)和(7.88±3.58)头,在家蝇蛹上繁育的寄生蜂明显具有较长的寿命和产卵期、更多的雌雄蜂后代数量;在对家蝇蛹和果蝇蛹的选择上,繁育自家蝇和果蝇的蝇蛹金小蜂雌蜂选择频率的差异不大。【结论】利用家蝇蛹繁殖的蝇蛹金小蜂在寄生果蝇蛹时具有更大优势,在繁殖蝇蛹金小蜂控制杨梅园等果蝇的为害时,可以选择家蝇蛹作为替代寄主。     

云南省斑翅果蝇寄生性天敌昆虫种类调查

斑翅果蝇作为危害蓝莓、杨梅、葡萄、樱桃等软皮水果的重要害虫之一,已经引起国内外广泛关注,明确斑翅果蝇寄生性天敌昆虫的种类、分布和自然寄生状况,可以为斑翅果蝇天敌的保护利用提供科学依据。本文通过在云南省斑翅果蝇适生区采集斑翅果蝇的栽培寄主和野生浆果果实,带回实验室培养5~8d,解剖果实并挑出斑翅果蝇蛹,收集其中羽化的斑翅果蝇寄生性天敌昆虫,并记录其种类和数量,计算寄生率。本研究共采集调查了45种植物果实,有15种植物果实可被斑翅果蝇危害,其中杨梅中的斑翅果蝇种群数量最大,达到96.03头/百果;共收集到5种斑翅果蝇的寄生蜂,其中幼虫寄生蜂有丽盾瘿蜂Ganaspis brasiliensis、细毛瘿蜂Leptopilina japonica和反颚茧蜂Asobara sp.,蛹寄生蜂有果蝇锤角细蜂Trichopria drosophilae和蝇蛹金小蜂Pachycrepoideus vindemmiae,寄生蜂的自然寄生率最高可达27.81%。结果表明,丽盾瘿蜂的虫口数量最多,分布最广,为斑翅果蝇的田间优势种寄生蜂。斑翅果蝇的寄生性天敌昆虫在斑翅果蝇化蛹后的1~2周达到羽化高峰期,对斑翅果蝇具有较好的自然控制作用。     

抑制性消减杂交技术在昆虫滞育中的利用和展望

滞育是多数天敌昆虫固有的遗传属性,掌握天敌昆虫滞育诱导、维持和解除技术,能及时将扩繁的天敌昆虫以滞育状态进行储存,在释放前再打破滞育使其进入活跃状态以防控害虫。开展天敌昆虫滞育研究,还有助于掌握其发育特点与发生动态,提高害虫防治效率,加深对天敌昆虫发育机制的认识,探寻昆虫对环境适应机制及进化途径。因此,滞育调控及机理研究成为近年来昆虫学研究的一个重要领域。抑制性消减杂交技术（Suppression Subtractive Hybridization, SSH）是近年来发明的用于挖掘差异性调控或特异表达cDNA探针和构建文库的一种方法。本文在总结抑制性消减杂交技术的测试原理及其优越性的基础上,归纳了该技术在昆虫滞育研究中的应用,概述了通过构建抑制性消减文库,在鞘翅目、直翅目、鳞翅目、双翅目等昆虫中探明的滞育差异表达基因,进一步探讨了差异表达基因与滞育的关系。随着昆虫全基因组测序和转录组测序的信息增加,抑制性消减杂交技术将更广泛的应用于昆虫学各个领域的差异基因的筛选与挖掘,促进昆虫分子生物学科学体系的发展。     

小蜂滞育的研究进展

滞育现象在多种小蜂类天敌昆虫中存在,通过研究小蜂滞育技术,可实现蜂种的长期贮存、延长防控作用时间、提高产品的抗逆性,对小蜂工厂化生产及应用具有重要意义。本文在分析国内外小蜂总科昆虫滞育文献的基础上,总结了已开展滞育研究的69种小蜂类昆虫的滞育虫态、滞育持续期、主要诱导因子以及亲代效应等,分属小蜂科、赤眼蜂科、姬小蜂科、跳小蜂科、金小蜂科、蚜小蜂科、旋小蜂、长尾小蜂科、广肩小蜂、四节小蜂科10科。小蜂多以幼虫或预蛹滞育,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。一种寄生麦红吸浆虫的金小蜂Macroglenes penetrans在2.5℃的土壤中,其滞育持续期可达16个月。低温、短日照和寄主是影响多数小蜂滞育的主要因子;但也有少数小蜂进行夏滞育,如普金姬小蜂Chrysocharis pubicornis、Aphelinus flavus、车轴草广肩小蜂Bruchophagus platypterus等。另外,亲代也可对小蜂滞育产生一定影响。目前,对小蜂滞育后发育生物学评价的研究报道较少,尚待进一步探索研究。     

Water relationships in the ectoparasitoid Nasonia vitripennis during larval diapause

Abstract. Final instar larvae of the ectoparasitoid Nasonia vitripennis overwinter in diapause within the puparium of the host fly. Wasp larvae that are in diapause have a different set of water relations than larvae that are not in diapause. Diapausing larvae contain less water, lose water at a lower rate, and have the capacity to absorb water vapour. Water conservation in larvae of N. vitripennis is also enhanced considerably by the puparium of its fly host.     

Cytonuclear genic incompatibilities cause increased mortality in male F2 hybrids of Nasonia giraulti and N. vitripennis

The haplodiploid wasp genus Nasonia is a promising model for studying the evolution of genic incompatibilities due to the existence of interfertile species and haploid males. The latter allows for significantly reducing the sample size required to detect and map recessive dysfunctional genic interactions. We exploited these features to study the genetics of intrinsic hybrid inviability in male F2 hybrids of Nasonia giraulti and N. vitripennis. Analyzing marker segregation in 225 hybrid embryos, we inferred a linkage map with 38 framework markers. The markers were tested for marker transmission ratio distortion (MTRD) and interchromosomal linkage disequilibrium in populations of embryonic and adult hybrids. We found evidence for four transmission ratio distorting loci (TRDL). Three TRDL showed a deficit of the N. giraulti allele in hybrids with N. vitripennis cytoplasm. A separate TRDL exhibited a deficiency of the N. vitripennis allele in hybrids with N. giraulti cytoplasm. We ascribe the observed MTRD in adult hybrids to cytonuclear genic incompatibilities causing differential mortality during development since hybrid embryos did not show MTRD. The identified cytonuclear genic incompatibilities in F2 hybrids with N. vitripennis cytoplasm account for most of the intrinsic hybrid inviability in this cross. The high mortality rate in F2 hybrids with N. giraulti cytoplasm cannot be explained by the single identified TRDL alone, however.     

Chromosomal anchoring of linkage groups and identification of wing size QTL using markers and FISH probes derived from microdissected chromosomes in Nasonia (Pteromalidae: Hymenoptera)

Nasonia vitripennis is a small parasitic hymenopteran with a 50-year history of genetic work including linkage mapping with mutant and molecular markers. For the first time we are now able to anchor linkage groups to specific chromosomes. Two linkage maps based on a hybrid cross (N. vitripennis x N. longicornis) were constructed using STS, RAPD and microsatellite markers, where 17 of the linked STS markers were developed from single microdissected banded chromosomes. Based on these microdissections we anchored all linkage groups to the five chromosomes of N. vitripennis. We also verified the chromosomal specificity of the microdissection through in situ hybridization and linkage analyses. This information and technique will allow us in the future to locate genes or QTL detected in different mapping populations efficiently and fast on homologous chromosomes or even chromosomal regions. To test this approach we asked whether QTL responsible for the wing size in two different hybrid crosses (N. vitripennis x N. longicornis and N. vitripennis x N.giraulti) map to the same location. One QTL with a major effect was found to map to the centromere region of chromosome 3 in both crosses. This could indicate that indeed the same gene/s is involved in the reduction of wing in N. vitripennis and N. longicornis.     

Female dispersal and isolation-by-distance of Nasonia vitripennis populations in a local mate competition context

Dispersal behavior directly influences the level of inbreeding, but the effect of inbreeding avoidance on dispersal is less well studied. The parasitoid wasp Nasonia vitripennis (Walker) (Hymenoptera: Chalcidoidea: Pteromalidae) is known to mate exclusively on the natal patch, and females are the only dispersing sex. A previous study has shown that foundresses on a patch are typically unrelated, implying that females disperse for a considerable distance from their natal patch after mating. We investigated dispersal of N. vitripennis on two scales. On a local scale we used a mark-release-recapture experiment, and on the larger scale we investigated isolation by distance using a population genetic approach. We found that N. vitripennis females are long-distance dispersers, capable of covering at least 2 km in 48 h. Populations within a range of 100 km showed no substructure, but larger distances or major geographical barriers restricted gene flow and led to significant population structure. The results provide a basis for future research on dispersal of parasitoids and are discussed in the context of dispersal abilities and inbreeding avoidance in Nasonia .     

The jewel wasp Nasonia: querying the genome with haplo-diploid genetics

The jewel wasp Nasonia vitripennis is considered the "Drosophila melanogaster of the Hymenoptera." This diminutive wasp offers insect geneticists a means for applying haplo-diploid genetics to the analysis of developmental processes. As in bees, haploid males develop from unfertilized eggs, while diploid females develop from fertilized eggs. Nasonia's advantageous combination of haplo-diploid genetics and ease of handling in the laboratory facilitates screening the entire genome for recessive mutations affecting a developmental process of interest. This approach is currently directed toward understanding the evolution of embryonic pattern formation by comparing Nasonia embryogenesis to that of Drosophila. Haplo-diploid genetics also facilitates developing molecular maps and mapping polygenic traits. Moreover, Nasonia embryos are also proving amenable to cell biological analysis. These capabilities are being exploited to understand a variety of behavioral, developmental, and evolutionary processes, ranging from cytoplasmic incompatibility to the evolution of wing morphology.     

多组学分析在昆虫滞育中的应用

组学技术将生物的相关问题分别展现在基因、蛋白质和代谢物等不同层次水平上，已成为解读生命过程的重要工具。本文分别从转录组学、蛋白质组学、代谢组学以及组学间的联合应用等方面概括总结了组学技术在昆虫滞育研究中的应用情况，阐述了以转录组学、蛋白质组学和代谢组学为代表的多组学技术在昆虫滞育调控分子机制中取得的重要成果，并针对当前研究现状，对昆虫滞育中组学技术应用的前景和局限性进行了总结和展望，以期为昆虫滞育调控分子机制的研究提供参考依据。     

Species-diagnostic single-nucleotide polymorphism and sequence-tagged site markers for the parasitic wasp genus Nasonia (Hymenoptera: Pteromalidae)

Wasps of the genus Nasonia are important biological control agents of house flies and related filth flies, which are major vectors of human pathogens. Species of Nasonia (Hymenoptera: Pteromalidae) are not easily differentiated from one another by morphological characters, and molecular markers for their reliable identification have been missing so far. Here, we report eight single-nucleotide polymorphism and three sequence-tagged site markers derived from expressed sequenced tag libraries for the two closely related and regionally sympatric species N. giraulti and N. vitripennis. We studied variation of these markers in natural populations of the two species, and we mapped them in the Nasonia genome. The markers are species-diagnostic and evenly spread over all five chromosomes. They are ideal for rapid species identification and hybrid recognition, and they can be used to map economically relevant quantitative trait loci in the Nasonia genome.     

Shifts in metabolomic profiles of the parasitoid Nasonia vitripennis associated with elevated cold tolerance induced by the parasitoid's diapause,host diapause and host diet augmented with proline

The ectoparasitoid wasp, Nasonia vitripennis can enhance its cold tolerance by exploiting a maternally-induced larval diapause. A simple manipulation of the fly host diapause status and supplementation of the host diet with proline also dramatically increase cold tolerance in the parasitoid. In this study, we used a metabolomics approach to define alterations in metabolite profiles of N. vitripennis caused by diapause in the parasitoid, diapause of the host, and augmentation of the host's diet with proline. Metabolic profiles of diapausing and nondiapausing parasitoid were significantly differentiated, with pronounced distinctions in levels of multiple cryoprotectants, amino acids, and carbohydrates. The dynamic nature of diapause was underscored by a shift in the wasp's metabolomic profile as the duration of diapause increased, a feature especially evident for increased concentrations of a suite of cryoprotectants. Metabolic pathways involved in amino acid and carbohydrate metabolism were distinctly enriched during diapause in the parasitoid. Host diapause status also elicited a pronounced effect on metabolic signatures of the parasitoid, noted by higher cryoprotectants and elevated compounds derived from glycolysis. Proline supplementation of the host diet did not translate directly into elevated proline in the parasitoid but resulted in an alteration in the abundance of many other metabolites, including elevated concentrations of essential amino acids, and reduction in metabolites linked to energy utilization, lipid and amino acid metabolism. Thus, the enhanced cold tolerance of N. vitripennis associated with proline augmentation of the host diet appears to be an indirect effect caused by the metabolic perturbations associated with diet supplementation.     

Genetics of cuticular hydrocarbon differences between males of the parasitoid wasps Nasonia giraulti and Nasonia vitripennis

Many insects rely on cuticular hydrocarbons (CHCs) as major recognition signals between individuals. Previous research on the genetics of CHCs has focused on Drosophila in which the roles of three desaturases and one elongase were highlighted. Comparable studies in other insect taxa have not been conducted so far. Here, we explore the genetics of CHCs in hybrids of the jewel wasps Nasonia giraulti and Nasonia vitripennis. We analyzed the CHC profiles of pure strain and of F(2) hybrid males using gas chromatography coupled with mass spectrometry and distinguished 54 peaks, of which we identified 52 as straight-chain, monounsaturated, or methyl-branched CHCs. The latter compound class proved to be particularly abundant and diverse in Nasonia. Quantitative trait locus (QTL) analysis suggests fixed genetic differences between the two strains in 42 of the 54 studied traits, making Nasonia a promising genetic model for identifying genes involved in CHC biosynthesis. QTL for methyl-branched CHCs partly clustered in genomic regions with high recombination rate: a possible indication for pleiotropic genes that control their biosynthesis, which is largely unexplored so far. Finally, we identified and mapped genes in the Nasonia genome with high similarity to genes that have been implicated in alkene biosynthesis in Drosophila and discuss those that match in their position with predicted QTL for alkenes.