Proteomic interactions between the parasitoid Diachasmimorpha longicaudata and the oriental fruit fly,Bactrocera dorsalis during host parasitism

The oriental fruit fly, Bactrocera dorsalis, is an important agricultural pest and biological control is one of the most effective control methodologies. We conducted an investigation on the molecular response of the fruit fly to parasitism by the larval parasitoid, Diachasmimorpha longicaudata using two-dimensional gel electrophoresis and mass spectroscopy. We identified 285 differentially expressed protein spots (109 proteins) during parasitism. The molecular processes affected by parasitism varied at different time point during development. Transferrin and muscle specific protein 20 are the only two proteins differentially expressed that play a role in host immunity 24 h after parasitism. Developmental and metabolic proteins from parasitoids (transferrin and enolase) were up-regulated to ensure establishment and early development of parasitoids 48 h post parasitism. 72 h after parasitism, larval cuticle proteins, transferrin and CREG1 were overexpressed to support the survival of parasitoids while host metabolism proteins and parasitoid regulatory proteins were down-regulated. Host development slowed down while parasitoid development went up at 96 h after parasitism. All developmental, regulatory, structural, and metabolic proteins were expressed at their optimum at 120 h post parasitism. Host development was reduced, metabolism and regulatory proteins were strongly involved in the activities. The development deteriorated further at 144 h after parasitism. Enolase and CREG1 were indicators of parasitoid survival. Hexamerin and transferrin from the parasitoid was peaked at 168–216 h after parasitism, strongly indicating that parasitoid would survive. This study represents the first report that reveals the molecular players involved in the interaction between the host and parasitoid.     

Effects of a fruit and a host-derived compound on orientation and oviposition in Utetes anastrephae,a little studied opiine braconid (Hymenoptera) parasitoid of Anastrepha spp. fruit flies (Diptera: Tephritidae)

Augmentative biological control of tephritid fruit flies would benefit from: (1) synthetic attractants to monitor the survival and dispersal of released parasitoids and (2) synthetic oviposition stimulants to reduce production costs of parasitoid species that are now prohibitively costly to mass-rear. Utetes anastrephae (Viereck) is a widespread and sometimes common opiine braconid parasitoid of several pest Anastrepha spp. Despite its host range, distribution and abundance, it has attracted relatively little research and little is known of its chemical ecology. Its orientation was determined towards two chemical cues hypothesised to be useful at two spatial scales: (1) limonene derived from fruit is presumably abundant and widely dispersed and might identify from a distance patches of potentially host-containing fruit; and (2) para-ethylacetophenone (PEA), a volatile emitted by a number of tephritid larvae, presumably in relatively small amounts, and which could serve as short-range cue or oviposition stimulant. Various concentrations of limonene proved attractive to both females and males, perhaps to the later as a means of locating females accumulated in the vicinity of limonene-emitting host plants. PEA at the concentrations tested did not influence oviposition in U. anastrephae, although it did so for Diachasmimorpha longicaudata (Ashmead), another opiine tephritid parasitoid, previously known to respond to PEA and included in the experiment as a positive control. Limonene at the concentrations tested had no effect on oviposition in either species. These results advance efforts to synthesise attractants and oviposition stimulants for alternative candidates for augmentation such as U. anastrephae.     

Host discrimination and superparasitism in wild and mass-reared Diachasmimorpha longicaudata (Hym.: Braconidae) females

We compared the oviposition behavior and host discrimination ability of wild and mass-reared Diachasmimorpha longicaudata (Hymenoptera: Braconidae) females parasitizing Anastrepha ludens (Diptera: Tephritidae) larvae. Both kinds of parasitoid females were presented simultaneously with parasitized and non-parasitized larvae in choice tests, and their superparasitism performance was evaluated under a mass-rearing situation. At the time of the test, D. longicaudata had 156 generations under mass-rearing conditions. Our goal was to determine the effect of the mass-rearing process on the foraging decisions of this species. One of the primary findings was the apparent ubiquity of superparasitism by D. longicaudata females. Both types of females showed similar patterns in each of the phases of oviposition behavior evaluated. The only notable differences were among the percentages of transition between behaviors, mainly related to the intensity with which each activity was performed. Under a mass-rearing situation, both strains of females had a similar tendency to increase superparasitism (i.e., number of oviposition scars per puparium and the proportion of superparasitized larvae) over time. The mass-rearing process appears to have induced the selection of more aggressive, fertile and precocious females. Despite these observations, we concluded that the process of adaptation to mass-rearing conditions has not substantially influenced the foraging and ovipositional behaviors in this species.     

桔小实蝇本地寄生蜂长尾全裂茧蜂成虫

应用扫描电镜（SEM）对桔小实蝇本地寄生蜂长尾全裂茧蜂 Diachasmimorpha longicaudata (Ashmead) 成虫的触角感器进行观察。结果表明：长尾全裂茧蜂成虫触角由柄节、梗节和鞭节组成,鞭节共 49-52小节。在触角上共有 8种感器,分别为 Bhm鬃毛、毛形感器、刺形感器、锥形感器、板形感器、腔锥形感器、钟形感器、嗅孔。毛型感器是主要感器,数量多且分布广,其次为板形感器。除了腔锥形感器 Ⅱ外,雌雄蜂没有明显的二型现象。     

前裂长管茧蜂触角感器的扫描电镜观察

前裂长管茧蜂是许多实蝇类害虫幼虫-蛹期的重要寄生性天敌。通过扫描电镜对其触角感受器进行超微观察,结果发现,前裂长管茧蜂雌蜂触角共发现7种感受器,分别为鳞型感器、Bhm毛、毛型感器、腔型感器、栓锥型感器、钟型感器及板型感器。其中,毛型感器具有3种形态(毛型感器Ⅰ、Ⅱ、Ⅲ),锥型感器具有2种形态(锥型感器Ⅰ、Ⅱ),但在雄蜂触角上没发现锥形感器Ⅱ。毛型感器和板型感器是前裂长管茧蜂触角上的主要感器,数量较多,分布较广。     

Protein polymorphism in natural populations of Diachasmimorpha longicaudata (Hymenoptera: Braconidae) in Thailand

Diachasmimorpha longicaudata (Ashmead) is a larval parasitoid of tephritid flies and is widely used as a classical biological control agent. We have used allozyme electrophoresis to evaluate the genetic relationships of six populations of D. longicaudata in Thailand. Twelve loci were examined of which 11 were polymorphic in at least some populations, especially that of Nakornratchasima. We observed a complete lack of heterozygotes for seven of the 10 polymorphic loci in the Nakornratchisima female population, and a significant deficiency of heterozygotes at a further two loci. We discuss possible hypotheses for these findings in light of the haplo-diploid sex determination system of these wasps. This revised version was published online in July 2006 with corrections to the Cover Date.     

Suppression of Ceratitis capitata (Wied.) (Diptera: Tephritidae) populations in coffee in the Mexico–Guatemala border region through the augmentative releases of Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae)

In Guatemalan coffee plantations, Ceratitis capitata populations were suppressed by the application of augmentative releases of parasitoids. These releases significantly increased parasitism of this fruit fly over paired no-release control areas. Integrated management plus biological control presented a significant reduction in the pest population. These results support the application of biological control in C. capitata management.     

前裂长管茧蜂寄生行为对桔小实蝇幼虫生理指标的影响

通过观察测定了桔小实蝇幼虫生长发育过程中血淋巴蛋白种类和血细胞的变化以及前裂长管茧蜂的寄生行为对桔小实蝇幼虫各项生理指标的影响。结果表明:不同日龄桔小实蝇幼虫的血细胞浓度随着虫龄的增加呈显著上升趋势,由2龄的16.53×106cells/mL,到3龄的30.14×106cells/mL直至蛹前期的35.94×106cells/mL,但血淋巴蛋白种类没有明显变化。和未寄生幼虫相比,寄生后的幼虫各类型血细胞的浓度均下降,但差异均不显著;血淋巴蛋白种类无明显增减,但浓度有所变化;寄生4 h后血淋巴蛋白质浓度显著降低,接近22 h时升高,至化蛹前期浓度再次下降;寄生行为使幼虫的发育历期从8 d延长至9～11d;4日龄幼虫在被寄生后的第4 d起体重显著高于未被寄生的桔小实蝇幼虫。     

长尾潜蝇茧蜂对橘小实蝇幼虫的寄生效能

以橘小实蝇幼虫为寄主,在(25±1)℃,RH(70±10)%,光照周期14L∶10D的条件下,研究了在不同密度的寄生蜂与寄主条件下的长尾潜蝇茧蜂寄生效能。结果表明:不同密度的橘小实蝇幼虫对长尾潜蝇茧蜂的寄生作用有明显的影响,其寄生效应符合功能反应模型HollingⅡ型,方程为Na=0.7291No/(1 0.02362No)。长尾潜蝇茧蜂自身密度对寄生作用也存在干扰效应,用Hassell-Varley模型可表示为a=0.1993P-0.2966。     

Improvement of quality control methods for augmentative releases of the fruit fly parasitoids,Diachasmimorpha longicaudata and Psyttalia fletcheri (Hymenoptera: Braconidae)

Criteria were established to improve quality control methods for augmentative releases of the opiine parasitoids, Diachasmimorpha longicaudata (Ashmead) and Psyttalia fletcheri (Silvestri). These included correlating puparial weight with adult emergence and sex ratio at the Honolulu rearing facility, and determining the effect of air shipments and field releases on parasitoid emergence and subsequent mortality. There was a positive relationship between the weight of 7–10‐day‐old host puparia and percentage of emergence for both D. longicaudata and P. fletcheri. Standardization of shipping methods included placement of ice blocks in the top levels of the ice chests, prompt shipment and pick‐up of samples, and cooling of puparia before shipment to minimize metabolic heat generated in the samples, and to delay emergence of samples. Before standardization, emergence losses of up to 58% were observed for D. longicaudata and 18% for P. fletcheri. When shipping methods were standardized, emergence was no longer reduced. In addition, low emergence rates were associated with reduced flight propensity of D. longicaudata (R_s = ‐ 0.45 at Kilauea and ‐ 0.54 at Kealia). At the two release sites, 88–95% of adult D. longicaudata that did not escape the release containers were males.     

前裂长管茧蜂低温储存技术的研究

前裂长管茧蜂Diachasmimorpha longicaudata (Ashmead) 是桔小实蝇Bactrocera dorsalis (Hendel) 及多种其他实蝇害虫幼虫期重要的天敌寄生蜂。本文研究了该蜂低温储存的适宜虫期、温度和时间条件。结果表明,在5℃条件下12日龄的前裂长管茧蜂受低温影响最小,储存3 d后羽化率达94.67%,与对照的9533%无显著差异;储存6 d后蜂蛹羽化率下降到70.00%,低于对照但高于其他日龄。相同日龄的前裂长管茧蜂随着储存时间的延长,其羽化率逐渐降低,甚至不羽化。为进一步研究冷存对前裂长管茧蜂品质的影响,以储存温度(2、5、8、11和14 ℃)和储存时间(3、6、9和12 d)为参试因子,对12日龄前裂长管茧蜂冷藏后的羽化率、繁殖子代数、繁殖子代雌蜂数、子代性比、雌蜂寿命、发育历期以及种群趋势指数进行综合分析。结果表明：12日龄的前裂长管茧蜂在8℃下储存3 d,其种群趋势指数为29.32;14℃下储存6 d和9 d的种群趋势指数分别为32.85和27.04,均高于对照的25.16。因此上述组合是前裂长管茧蜂蛹低温储存的适宜条件。     

Effect of insectary rearing on host preference and oviposition behavior of the fruit fly parasitoid Diachasmimorpha longicaudata

Diachasmimorpha longicaudata (Ashmead) has been produced in the laboratory for >160 generations on the larvae of oriental fruit fly, Bactrocera dorsalis (Hendel), the propagation hosts raised routinely on a semi-synthetic wheat diet formulation. Choice tests using modified stinging units were conducted in the laboratory to investigate whether insectary rearing had altered the host seeking and oviposition behavior of female parasitoids. Results showed that fruit fly larvae that developed in papaya, Carica papaya L. var. solo, were less preferred for oviposition than fruit fly larvae that developed on wheat diet when both were exposed concurrently to naive D. longicaudata females (= females without prior oviposition experience). The substrates (pureed papaya or wheat diet) in which treatment larvae were exposed to parasitoids did not affect oviposition preference of gravid D. longicaudata for wheat diet-reared fruit fly larvae. Our study demonstrated the possibility that rearing in an insectary system may have modified the parasitization behavior of female D. longicaudata.     

Longevity of multiple species of tephritid (Diptera) fruit fly parasitoids (Hymenoptera: Braconidae: Opiinae) provided exotic and sympatric-fruit based diets

While adult parasitic Hymenoptera in general feed on floral and extrafloral nectars, hemipteran-honeydews and fluids from punctured hosts, Diachasmimorpha longicaudata (Ashmead), an Old World opiine braconid introduced to tropical/subtropical America for the biological control of Anastrepha spp. (Tephritidae), can survive on fruit juices as they seep from injured fruit. An ability to exploit fruit juice would allow such a parasitoid to efficiently forage for hosts and food sources simultaneously. Two New World opiines, Doryctobracon areolatus (Szepligeti) and Utetes anastrephae (Viereck), are also prominent Anastrepha parasitoids and are roughly sympatric. All three species were provided with: (1) pulp and juice diets derived from a highly domesticated Old World fruit (orange, Citrus sinensis L.) that is only recently sympatric with the Mexican flies and parasitoids and so offered little opportunity for the evolution of feeding-adaptations and (2) a less-domesticated New World fruit (guava, Psidium guajava L.), sympatric over evolutionary time with D. areolatus and U. anastrephae. Both sexes of D. longicaudata died when provided guava pulp or juice at a rate similar to a water-only control. D. areolatus and U. anastrephae, presumably adapted to the nutrient/chemical constituents of guava, also died at a similar rate. Survival of all three species on orange pulp and juice was greater than on water, and often equaled that obtained on a honey and water solution. In confirmatory experiments in Mexico, D. areolatus and U. anastrephae, as well as other tephritid parasitoids Doryctobracon crawfordi (Viereck) and Opius hirtus (Fisher), all died at a significantly higher rates when provided guava in comparison to a honey and water diet. Such a result is likely due to guavas being repellent, innutritious or toxic. D. longicaudata clearly consumed guava juice tagged with a colored dye. Dilutions of orange and guava juice resulted in shorter lifespans than dilutions of orange juice and water demonstrating that there while diluted orange juice provided nutrition the addition of guava created toxicity. Given the differences in fruit-food quality, adult opiine food sources would not be obtainable at all oviposition sites and in the case of guava, more additional sites and foraging for food than previously postulated may be required.     

Competitive Superiority of Early Acting Species: A Case Study of Opiine Fruit Fly Parasitoids

Koinobiont parasitoids that attack an early host stage may have an advantage in suppressing competing parasitoids that attack later stages of the same host. We examined the competitive interaction between the two most important parasitoids of tephritid fruit flies in Hawaii, Fopius arisanus (Sonan), and Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae). The former species attacks host eggs while the latter attacks host larvae, and both species emerge as adults from the host puparia. F. arisanus physiologically suppressed egg development of D. longicaudata. Over 90% of D. longicaudata eggs died in the presence of F. arisanus larvae within host larvae of either the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) or the oriental fruit fly, Bactrocera dorsalis (Hendel). D. longicaudata appeared not to discriminate against hosts previously parasitized by F. arisanus. The mechanism that F. arisanus uses to eliminate D. longicaudata is similar to that which it employs to eliminate five other larval fruit fly parasitoids so far reported in Hawaii. This suggests that there is a broad competitive superiority of the early acting species in fruit fly parasitoids. We discuss the implication of this in relation to future biological control introductions against tephritid fruit flies.