A Trichogramma Species Showing a Better Adaptation to High Temperature than its Symbionts

High temperature cures Wolbachia endocytobionts in their symbiotic Trichogramma hosts reproducing by thelytokous parthenogenesis, and restores a bisexual reproduction. This restoration occurs at a lower temperature in T. cordubensis than in T. pretiosum. High temperature affects host biological characteristics to a lesser extent in T. cordubensis than in T. pretiosum. Therefore, adaptive differences to thermal conditions not only exist among Wolbachia symbionts but also among their Trichogramma hosts, and these differences appear to be independent in the two partners.     

How does infection with parthenogenesis-inducing Wolbachia reduce the fitness of Trichogramma?

We analyzed the survival rate of the immature stages of Trichogramma species and lines that differed in their mode of reproduction. Specifically, we compared the mortality of arrhenotokous (W(-)), irrevertable thelytokous (W(-)), and Wolbachia-associated thelytokous (W(+)) forms. The embryonic mortality of the W(+) strains was significantly higher than that of the W(-) lines. The embryonic mortality was negligible for the arrhenotokous Trichogramma evanescens and the thelytokous Trichogramma cacoeciae, which is not infected with Wolbachia. Only 30% of the eggs of the Wolbachia-infected strains developed to adults, while the emergence rate of the Wolbachia-free strains was more than 78%, irrespective of the origin of the strains. More than 78% of the overall mortality in W(+) strains happened during the early stages of development. About 35% of embryos of W(+) strains remain in the mitotic stage even 48 h after oviposition. Most embryos of W(-) strains had already developed to cellular blastoderm after 6 h, regardless of strains. The mortality of immature stages in W(+) strains was mainly caused by the failure of the mitotic divisions.     

Wolbachia感染对拟澳洲赤眼蜂寿命、生殖力和嗅觉反应的影响

研究了通过共享同一寄主卵,短管赤眼蜂Trichogramma pretiosum体内共生的Wolbachia被水平传递到拟澳洲赤眼蜂T. confusum体内后,Wolbachia对新宿主拟澳洲赤眼蜂的影响。结果表明： Wolbachia的侵染能使拟澳洲赤眼蜂进行不完全的产雌孤雌生殖,增加拟澳洲赤眼蜂子代雌性比例,但却导致了雌蜂寿命缩短和繁殖力降低的生理损失。Wolbachia感染的当代处女蜂及其建立的种群连续5代(F₁～F₅),其子代雌蜂百分率分别为79.17%、76.60%、68.66%、72.58%、68.15%和64.06%,基本上呈现出逐代降低的趋势,并越来越接近对照的63.85%。处女蜂及F₁～F₅代雌蜂的平均寿命分别为4.33、5.50、5.60、6.68、7.32和7.50天,而未感染交配雌蜂寿命为7.59 天;处女蜂及F₁～F₅代雌蜂平均产卵量分别为11.33、70.00、86.41、93.90、102.92和124.38粒/雌,除F₅代外,均显著低于未感染交配雌蜂的产卵量134.81粒/雌。用四臂嗅觉仪测定了Wolbachia新宿主拟澳洲赤眼蜂对寄主小菜蛾的嗅觉反应,结果表明Wolbachia的侵染具轻微干扰拟澳洲赤眼蜂嗅觉反应的负面影响。未感染Wolbachia的拟澳洲赤眼蜂及Wolbachia供体短管赤眼蜂对寄主小菜蛾具较强的嗅觉反应,其雌蜂在小菜蛾腹部鳞片正己烷提取液和小菜蛾卵表正己烷提取液处理区内滞留时间显著或极显著长于对照区。而感染Wolbachia的拟澳洲赤眼蜂F₂代和F₃代雌蜂,尽管在小菜蛾腹部鳞片正己烷提取液处理区内滞留时间比对照区长,但没有达到显著水平;其F₂代雌蜂在小菜蛾卵表正己烷提取液处理区内滞留时间与对照区相比,也没有达到显著水平。随Wolbachia在拟澳洲赤眼蜂种群中垂直感染代数的增加,拟澳洲赤眼蜂对寄主小菜蛾的嗅觉反应恢复正常,在Wolbachia感染后的 F₄～F₆代,雌蜂在两种提取液处理区内的滞留时间均显著或极显著长于对照区。     

Biological Control Potential of Wolbachia -infected Versus Uninfected Wasps: Laboratory and Greenhouse Evaluation of Trichogramma cordubensis and T. deion Strains

The effects of thelytoky-inducing Wolbachia ( &#102 -proteobacteria) on Trichogramma cordubensis and T. deion (Hymenoptera, Trichogrammatidae) were studied in laboratory and greenhouse conditions. One infected (thelytokous, all female) line of each wasp species was compared with its conspecific uninfected (arrhenotokous, sexual) counterpart for several fecundity and dispersal traits. Arrhenotokous lines had a higher fecundity than their thelytokous counterparts, which suggests that Wolbachia negatively affect the fecundity of Trichogramma females. The arrhenotokous females dispersed more in the laboratory than their thelytokous counterparts. In the greenhouse, the opposite effect or no difference between lines was found, indicating that the laboratory set-up used to measure dispersal is not useful to predict relative dispersal of the females in the greenhouse. Calculations show that by releasing 100 adult wasps of both lines, thus including arrhenotokous males in the sexual line, more eggs are parasitized by the thelytokous wasps. Therefore, in spite of their lower individual female fecundity, thelytokous lines have a better potential for biological control than their arrhenotokous counterparts.     

安荔赤眼蜂在中国野外首次发现及其体内Wolbachia的检测

【目的】为丰富赤眼蜂Trichogramma的种类资源,明确野外新采集的一种赤眼蜂的种类,探明该赤眼蜂所感染Wolbachia的类型。【方法】采用挂米蛾Corcyra cephalonica卵卡法在华南农业大学树木园诱集到两批赤眼蜂,通过形态鉴定和PCR扩增ITS2序列并测序分析的分子鉴定手段对野外采集的赤眼蜂材料进行种类鉴定;通过PCR扩增Wolbachia的外膜蛋白基因(wsp)序列,检测赤眼蜂体内Wolbachia的感染情况;通过PCR扩增wsp序列和多位点序列分型(multilocus sequence typing, MLST)对检测到的赤眼蜂体内的Wolbachia进行同源性分析。【结果】所诱集到的两批赤眼蜂均被鉴定为安荔赤眼蜂Trichogramma oleae Voegelé & Pointel,体内Wolbachia的感染率达100%。该Wolbachia株系与安荔赤眼蜂(前南斯拉夫品系)、短管赤眼蜂T. pretiosum(乌拉圭品系)以及T. deion(荷兰品系)体内Wolbachia亲缘关系较近,属于B超组Sib亚组,对应MLST序列型为ST486。【结论】安荔赤眼蜂T. oleae为中国野外首次发现,是完全感染Wolbachia的产雌孤雌生殖品系。本研究为害虫生物防治提供了一种新的天敌种类资源,并为进一步探明Wolbachia与赤眼蜂的互作提供了研究材料。     

Wolbachia在我国广赤眼蜂种群内的感染

Wolbachia是广泛分布于节肢动物生殖组织内的一类细胞内共生菌,它属于原细菌的α亚类,能够通过调控寄主的生殖活动而促进其在寄主种群中的扩散。通过对wsp 基因的克隆及PCR-RFLP分析确定了Wolbachia在我国广赤眼蜂种群内的存在,并发现有2种Wolbachia 菌系的感染,命名为wEvaA和wEvaB。经过克隆分离得到了这2种Wolbachia的wsp基因序列,在GenBank的登录号为AY390279和 AY390280 ,并由基于wsp基因的聚类树中发现,这两种Wolbachia菌系均属于A组。     

Comparative Analysis of the Reproductive Attributes of Three Commercially-Produced Trichogramma Species (Hymenoptera: Trichogrammatidae)

The reproductive attributes of commercially-produced Trichogramma platneri, T. minutum and T. pretiosum reared from eggs of both Ephestia kuehniella and Sitotroga cerealella were monitored at 25 C. The age-specific fecundity, longevity and progeny sex ratio and adult size were determined for individual females from all six combinations of Trichogramma and rearing host, using E. kuehniella as the experimental host. Rearing host had a significant influence on the lifetime fecundity, 3-day fecundity and longevity of all three Trichogramma species. In general, the performance of T. minutum and T. pretiosum was better when reared from S. cerealella, but that of T. platneri was superior when reared from E. kuehniella. The lifetime fecundity of the Trichogramma species was linearly related to longevity and the ranking between species was T. pretiosum > T. minutum > T. platneri. The age-specific pattern of oviposition for T. platneri was distinctly precocious, with 40% of its lifetime fecundity oviposited on the first day, in contrast to 17-24% for the other two species. Progeny sex ratio over the lifetime of the Trichogramma females was slightly male biased and differed significantly from 0.5 for T. minutum and T. platneri. Daily sex ratio for parasitoids reared from the most productive rearing host was female biased only for the first day of oviposition for T. platneri in contrast to the first 5-6 days of oviposition for the other two species. There was little evidence that any of the reproductive attributes of these Trichogramma species, reared from small host eggs, was dependent on the size of the adult females.     

几种赤眼蜂对灰白蚕蛾寄生能力的影响

通过室内功能反应试验,测定松毛虫赤眼蜂T.dendrolimiMatsumura、短管赤眼蜂T.pretiosumRiley、暗黑赤眼蜂T.pintoiVoegele、玉米螟赤眼蜂T.ostriniaePangetChen、卷蛾分索赤眼蜂Trichogrammatoidea bactraeNagaraja、拟澳洲赤眼蜂T.confusumViggiani、舟蛾赤眼蜂T.closteraePangetChen、广赤眼蜂T.evanescensWestwood对广州地区重要园林害虫灰白蚕蛾的寄生能力。结果表明,除卷蛾分索赤眼蜂外,其它7种赤眼蜂对灰白蚕蛾的寄生作用均能用HollingⅡ型功能反应模型拟合。松毛虫赤眼蜂对灰白蚕蛾的寄生能力为最强,短管赤眼蜂和暗黑赤眼蜂次之,最大寄生卵量分别为25.8粒/雌、19.0粒/雌和18.6粒/雌,且在灰白蚕蛾卵内都能发育成功和正常羽化,可进一步进行筛选。虽然玉米螟赤眼蜂和舟蛾赤眼蜂对灰白蚕蛾有较强的寄生能力,但其后代雌蜂率都较低,且玉米螟赤眼蜂不能正常羽化,因此不适宜用来防治灰白蚕蛾。卷蛾分索赤眼蜂、广赤眼蜂和拟澳洲赤眼蜂的最大寄生量都较低,均不超过15.0粒,对灰白蚕蛾卵的寄生潜能不大。     

Effects of the Host Species and the Number of Parasitoids per Host on the Size of Some Trichogramma Species (Hymenoptera: Trichogrammatidae)

The size of some Trichogramma spp. adults and especially the ovipositor length depends on the species, but is also related to the host species and to the number of parasitoids per host. The length is greater in T. evanescens than in T. pretiosum itself greater than in T. exiguum, but the width is similar in the three species. For T. evanescens, the size obtained in Mamestra brassicae host when three or four insects emerged is similar to that obtained in Ephestia kuehniella host when singly parasitized. The size of the ovipositor is important because it may influence the possibility of in vitro egg laying in artificial host eggs. A shorter or a narrower ovipositor could cause difficulties in egg-laying into artificial host eggs composed of a membrane of unsuitable thickness.     

Cytology of Wolbachia-induced parthenogenesis in Leptopilina clavipes (Hymenoptera: Figitidae).

Parthenogenesis induced by cytoplasmatically inherited Wolbachia bacteria has been found in a number of arthropod species, mainly Hymenoptera. Previously, two different forms of diploidy restoration have been reported to underlie parthenogenesis induction in Hymenoptera by Wolbachia. Both are a form of gamete duplication, but each differs in their timing. We investigated the cytology of the early embryonic development of a Wolbachia-infected strain of the parasitoid wasp Leptopilina clavipes and compared it with that of an uninfected sexual strain. Both strains have a similar meiosis. In the infected parthenogenetic strain, diploidy is restored by anaphase restitution during the first somatic mitosis, similar to Trichogramma, but not to Muscidifurax. Our results confirm the occurrence of different cytological mechanisms of diploidy restoration associated with parthenogenesis-inducing Wolbachia in the order Hymenoptera.     

Wolbachia在玉米螟赤眼蜂内的三重感染

Wolbachia是一类广泛存在于节肢动物体内的共生菌。玉米螟赤眼蜂Trichogramma ostriniae是我国玉米田间的优势赤眼蜂种, 据报道, 赤眼蜂种内有Wolbachia感染。本文利用Wolbachia的16s rDNA和wsp基因引物通过PCR方法对玉米螟赤眼蜂的野生种群进行了调查, 发现以wsp基因为鉴定依据, 检测的所有个体都感染了3种Wolbachia ［wOstGDAa (GenBank accession no. EU157103), wOstGDAb (GenBank accession no. EU157104) 和 wOstGDB (GenBank accession no. EU157105)］。本文首次报道了野生赤眼蜂种群内Wolbachia的三重感染率几乎为100%。根据本研究的结果, 可以推测当不同种赤眼蜂寄生同一寄主时, Wolbachia可能会在不同赤眼蜂种间进行横向传播。     

Addition of wsp Sequences to the Wolbachia Phylogenetic Tree and Stability of the Classification

Wolbachia are symbiotic bacteria altering reproductive characters of numerous arthropods. Their most recent phylogeny and classification are based on sequences of the wsp gene. We sequenced wsp gene from six Wolbachia strains infecting six Trichogramma species that live as egg parasitoids on many insects. This allows us to test the effect of the addition of sequences on the Wolbachia phylogeny and to check the classification of Wolbachia infecting Trichogramma. The six Wolbachia studied are classified in the B supergroup. They confirm the monophyletic structure of the B Wolbachia in Trichogramma but introduce small differences in the Wolbachia classification. Modifications include the definition of a new group, Sem, for Wolbachia of T. semblidis and the merging of the two closely related groups, Sib and Kay. Specific primers were determined and tested for the Sem group.     

Collection of Trichogramma Westwood (Hymenoptera: Trichogrammatidae) from tropical northern Australia: a survey of egg parasitoids for potential pest insect biological control in regions of proposed agricultural expansion

Abstract  Trichogramma limit pest damage to crops by killing the developing embryo of their insect host at the egg stage. Their impact on the potentially insecticide resistant species, Helicoverpa armigera (Hübner), is considered integral to the resistance management plan for transgenic cotton production in the Ord River Irrigation Area in northern Western Australia. The dominant species of egg parasitoid in Ord River Irrigation Area cotton crops is the introduced Trichogramma pretiosum (Riley). Surveys of similar northern Australian regions earmarked for agricultural expansion revealed that T. pretiosum has been introduced, or has adventitiously dispersed, to all developing agricultural regions of northern Australia. Several previously unsequenced species were collected during surveys in more pristine habitats. Trichogramma pretiosum demonstrates an apparent ability to supersede native Trichogramma species and is perhaps favoured by agricultural expansion.     

Comparison of Hemolymph and Holotissues of Different Species of Insects as Diet Components forin VitroRearing ofTrichogramma galloiZucchi andT. pretiosumRiley

The aim of this work was to compare pupal holotissues (Diatraea saccharalis, Anticarsia gemmatalis,andBombyx mori) and larval hemolymph of several moth species (Helicoverpa zeaandBombyx mori) commonly produced in Brazilian laboratories as components of artificial diets forin vitrorearing ofTrichogramma galloiandTrichogramma pretiosum.Diets based on larval hemolymph ofH. zeaproduced the best development of both natural enemies, although a diet with pupal holotissues ofB. moriallowed full development ofT. pretiosum.Amino acid analysis showed qualitative differences among the insect derivates and eggs of the natural and alternative hosts, but with a quantitative similarity. Electrophoretic analysis resulting in a diverse protein-stained bands evidenced the qualitative differences among these components.     

Studies on three species of Trichogramma. I. Foraging behaviour for food or hosts

The foraging strategies for both food or hosts of three trichogrammatid species: Trichogramma evanescens, Trichogramma cacoecia and Trichogramma dendrolimi were compared using artificial patches of Sitotroga cerealella eggs. In all experiments, inexperienced and newly emerged wasps of < 1 h old were singly tested. When the females were allowed to land upon the hosts, the decision-making process for initial acceptance or rejection was species-dependent. The initial search in T. evanescens was for food, whereas T. dendrolimi or T. cacoeciae start to oviposit immediately after their emergence. When honey-deprived or undeprived females were each exposed to single patches for 20 or 60 min, variations in mean number of probing females 'drilling and ovipositing' were also species-dependent. Acceptance of host eggs by honey-deprived females and subsequent egg deposition were higher in both T. cacoeciae and T. dendrolimi than in T. evanescens . For all species, the probing of honey-undeprived females was higher than that of deprived ones. When the exposure period was prolonged to 24 or 48 h and the number of patches per female increased to three, most of the honey-deprived or undeprived T. evanescens females attacked one patch, and only a few of them attacked two patches. Foraging activity of honey-deprived females of T. cacoeciae was restricted to single patches, whereas most undeprived ones attacked more than one patch. In contrast, honey-deprived or undeprived T. dendrolimi females attacked more than one patch. The experiments showed that T. dendrolimi females have more powerful ovipositing urges than looking for food and that the opposite was the case for T. evanescens , with T. cacoeciae being intermediate.     

Opposite sex-specific effects of Wolbachia and interference with the sex determination of its host Ostrinia scapulalis

In the adzuki bean borer, Ostrinia scapulalis, the sex ratio in most progenies is 1 : 1. Females from Wolbachia-infected matrilines, however, give rise to all-female broods when infected and to all-male broods when cured of the infection. These observations had been interpreted as Wolbachia-induced feminization of genetic males into functional females. Here, we show that the interpretation is incorrect. Females from both lines have a female karyotype with a WZ sex-chromosome constitution while males are ZZ. At the time of hatching from eggs, WZ and ZZ individuals are present at a 1 : 1 ratio in broods from uninfected, infected and cured females. In broods from Wolbachia-infected females, ZZ individuals die during larval development, whereas in those from cured females, WZ individuals die. Hence, development of ZZ individuals is impaired by Wolbachia but development of WZ females may require the presence of Wolbachia in infected matrilines. Sexual mosaics generated (i) by transfection of uninfected eggs and (ii) by tetracycline treatment of Wolbachia-infected mothers prior to oviposition were ZZ in all tissues, including typically female organs. We conclude that: (i) Wolbachia acts by manipulating the sex determination of its host; and (ii) although sexual mosaics can survive, development of a normal female is incompatible with a ZZ genotype.     

赤眼蜂防治小菜蛾适宜蜂种或品系的筛选(英文)

以控制小菜蛾为目的,在实验室对２９种赤眼蜂或品系进行了筛选。筛选评价根据５项指标：（１）能在小菜蛾卵上产卵的雌蜂比例,（２）每雌蜂寄生卵量,（３）子峰在每粒寄主上的存活率,（４）羽化率,和（５）子代性比。研究结果表明：有２３种赤眼蜂或品系能够寄生小菜蛾卵,但无论在种间还是品系间均存在很大差异。根据５项筛选指标综合评价,２９种中较好的为：前苏联的Ｔｒｉｃｈｏｇｒａｍｍａ ｔｒｊａｐｉｔｚｎｉ,美国的Ｔ．ｐｒｅｔｉｏｓｕｍ,法国的Ｔ．ｎａｇａｒｋａｔｔｉｉ,中国台湾的Ｔｒｉｃｈｏｇａｒａｍｍａｔｏｉｄｅａ ｂａｃｔｒａｅ,中国广东的Ｔ．ｃｈｉｌｏｎｉｓ和法国的一个种Ｔ．ｓｐ。其中美国的Ｔ．ｐｒｅｔｉｏｓｕｍ,中国台湾的Ｔｒｉｃｈｏｇａｒａｍｍａｔｏｉｄｅａ ｂａｃｔｒａｅ,中国广东的Ｔ．ｃｈｉｌｏｎｉｓ可以考虑作为大田防治小菜蛾的首选蜂种。田间筛选及大田验证试验正在进行之中。     

SELECTION OF TRICHOGRAMMA SPECIES FOR CONTROLLING THE DIAMONDBACK MOTH PLUTELLA XYLOSTELLA(L.)

Abstract  The parasitization of twenty-nine species or strains of Trichogramma and Trichogrammatoidea on the eggs of diamondback moth (DBM), Plutella xylostella (L.), was tested in the laboratory. Five indexes were evaluated: proportion of female wasps of egg-laying, the number of parasitized host eggs by each female wasp, survival ratio of progeny in each host-egg, emergence ratio and sex ratio of offsprings. The results showed that 23 species or strains were able to parasitize the eggs of DBM and significant difference was found not only between species but also between strains. According to the five indexes, T. trijapitzni (from former USSR), T. pretiosum (from USA), T. nagarkattii (from France), Trichogrammatoidea bactrae( from Taiwan, China), T. chilonis (from Guangdong, China), and T . sp. (from France) showed better performance, among which T. pretiosum, Trichogrammatoidea bactrae and T. chilonis (Guangdong)were considered as suitable candidates to control DBM in the field.     

Host-foraging success of three species of Trichogramma (Hymenoptera: Trichogrammatidae) in a simulated retail environment

Three species of trichogrammatid egg parasitoids (Trichogramma deion Pinto & Oatman, Trichogramma ostriniae Pang & Chen, and Trichogramma pretiosum Riley) (Hymenoptera: Trichogrammatidae) were evaluated under laboratory conditions as potential biological control agents for the Indianmeal moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae), on retail shelves. A single shelving unit was used in each trial and a grid of sentinel egg disks was used to evaluate foraging success. The shelving consisted of pallet units with five shelves that were either bare or stocked with empty cereal boxes. In each replicate, approximately 500 female Trichogramma were released at the center of the shelving unit and allowed to forage for 48 h. Percentage of egg parasitism and percentage of host egg mortality were recorded after 7 d. Foraging success as well as the spatial pattern of parasitism differed significantly among the three Trichogramma species. Percentage of egg parasitism was approximately 4 times greater for T. deion than for T. ostriniae or T. pretiosum. The vertical distribution of parasitism by T. deion was also more uniform than for the other two species. In addition, the presence of packaging affected the foraging efficiency of T. ostriniae and T. pretiosum but not T. deion. Based on these findings, Trichogramma deion may be the best-suited candidate for augmentative biological control of P. interpunctella in retail stores, and a central release point of T. deion will likely provide adequate coverage of products on pallet-type shelving.     

Diversity of Wolbachia endosymbionts in heteropteran bugs

An extensive survey of Wolbachia endosymbionts in Japanese terrestrial heteropteran bugs was performed by PCR detection with universal primers for wsp and ftsZ genes of Wolbachia, cloning of the PCR products, restriction fragment length polymorphism analysis of infecting Wolbachia types, and molecular phylogenetic characterization of all the detected Wolbachia strains. Of 134 heteropteran species from 19 families examined, Wolbachia infection was detected in 47 species from 13 families. From the 47 species, 59 Wolbachia strains were identified. Of the 59 strains, 16 and 43 were assigned to A group and B group in the Wolbachia phylogeny, respectively. The 47 species of Wolbachia-infected bugs were classified into 8 species with A infection, 28 species with B infection, 2 species with AA infection, 3 species with AB infection, 5 species with BB infection, and 1 species with ABB infection. Molecular phylogenetic analysis showed little congruence between Wolbachia phylogeny and host systematics, suggesting frequent horizontal transfers of Wolbachia in the evolutionary course of the Heteroptera. The phylogenetic analysis also revealed several novel lineages of Wolbachia. Based on statistical analyses of the multiple infections, we propose a hypothetical view that, in the heteropteran bugs, interactions between coinfecting Wolbachia strains are generally not intense and that Wolbachia coinfections have been established through a stochastic process probably depending on occasional horizontal transfers.