A parasitoid wasp of Drosophila employs preemptive and reactive strategies to deplete its host’s blood cells

The wasps Leptopilina heterotoma parasitize and ingest their Drosophila hosts. They produce extracellular vesicles (EVs) in the venom that are packed with proteins, some of which perform immune suppressive functions. EV interactions with blood cells of host larvae are linked to hematopoietic depletion, immune suppression, and parasite success. But how EVs disperse within the host, enter and kill hematopoietic cells is not well understood. Using an antibody marker for L. heterotoma EVs, we show that these parasite-derived structures are readily distributed within the hosts’ hemolymphatic system. EVs converge around the tightly clustered cells of the posterior signaling center (PSC) of the larval lymph gland, a small hematopoietic organ in Drosophila. The PSC serves as a source of developmental signals in naïve animals. In wasp-infected animals, the PSC directs the differentiation of lymph gland progenitors into lamellocytes. These lamellocytes are needed to encapsulate the wasp egg and block parasite development. We found that L. heterotoma infection disassembles the PSC and PSC cells disperse into the disintegrating lymph gland lobes. Genetically manipulated PSC-less lymph glands remain non-responsive and largely intact in the face of L. heterotoma infection. We also show that the larval lymph gland progenitors use the endocytic machinery to internalize EVs. Once inside, L. heterotoma EVs damage the Rab7- and LAMP-positive late endocytic and phagolysosomal compartments. Rab5 maintains hematopoietic and immune quiescence as Rab5 knockdown results in hematopoietic over-proliferation and ectopic lamellocyte differentiation. Thus, both aspects of anti-parasite immunity, i.e., (a) phagocytosis of the wasp’s immune-suppressive EVs, and (b) progenitor differentiation for wasp egg encapsulation reside in the lymph gland. These results help explain why the lymph gland is specifically and precisely targeted for destruction. The parasite’s simultaneous and multipronged approach to block cellular immunity not only eliminates blood cells, but also tactically blocks the genetic programming needed for supplementary hematopoietic differentiation necessary for host success. In addition to its known functions in hematopoiesis, our results highlight a previously unrecognized phagocytic role of the lymph gland in cellular immunity. EV-mediated virulence strategies described for L. heterotoma are likely to be shared by other parasitoid wasps; their understanding can improve the design and development of novel therapeutics and biopesticides as well as help protect biodiversity.     

Effects of sugar feeding on carbohydrate and lipid metabolism in a parasitoid wasp

Lifetime patterns of carbohydrate and lipid metabolism were compared in starved and sucrose‐fed adults of the parasitoid Macrocentrus grandii (Goidanich) (Hymenoptera: Braconidae). As expected, sucrose‐fed individuals lived longer than did starved individuals. Macrocentrus grandii males and females eclosed with levels of simple storage sugars (presumably primarily trehalose) and glycogen that were below maximum levels recorded from sucrose‐fed parasitoids. Both of these nutrients dropped to very low levels in starved individuals within 4 days post‐emergence and were maintained at high levels in sucrose‐fed individuals throughout their lives. Lipid reserves at emergence represented the highest lipid levels for both sexes in the two diet treatments, with levels declining over the lifetimes of males and females from both diet treatments. Our results therefore suggest that dietary sucrose is used to synthesize trehalose and glycogen, but not lipids in M. grandii. Also, in contrast to the patterns observed for the simple sugars and glycogen, lipid levels in starved individuals did not drop below levels observed in sugar‐fed individuals. The average number of mature eggs carried by females at emergence was 33 and increased to approximately 85 in sucrose‐fed and 130 in starved females by the age of 5 d in the absence of hosts. The egg maturation rate was therefore higher in starved than in sugar‐fed females. Potential explanations for this unexpected result are discussed.     

Effects of spontaneous mutation accumulation on sex ratio traits in a parasitoid wasp

Sex allocation theory has proved extremely successful at predicting when individuals should adjust the sex of their offspring in response to environmental conditions. However, we know rather little about the underlying genetics of sex ratio or how genetic architecture might constrain adaptive sex-ratio behavior. We examined how mutation influenced genetic variation in the sex ratios produced by the parasitoid wasp Nasonia vitripennis. In a mutation accumulation experiment, we determined the mutability of sex ratio, and compared this with the amount of genetic variation observed in natural populations. We found that the mutability (h(2)(m)) ranges from 0.001 to 0.002, similar to estimates for life-history traits in other organisms. These estimates suggest one mutation every 5-60 generations, which shift the sex ratio by approximately 0.01 (proportion males). In this and other studies, the genetic variation in N. vitripennis sex ratio ranged from 0.02 to 0.17 (broad-sense heritability, H(2)). If sex ratio is maintained by mutation-selection balance, a higher genetic variance would be expected given our mutational parameters. Instead, the observed genetic variance perhaps suggests additional selection against sex-ratio mutations with deleterious effects on other fitness traits as well as sex ratio (i.e., pleiotropy), as has been argued to be the case more generally.     

Host-size-dependent sex ratio in a parasitoid wasp

Charnov's host-size model explains parasitoid host-size-dependent sex ratio as an adaptive consequence when there is a differential effect of host size on the offspring fitness of parasitoid males versus females. This article tests the predictions and the assumptions of the host-size model. The parasitoid wasp Pimpla nipponica Uchida (Hymenoptera: Ichneumonidae) laid more female eggs in larger or fresher host pupae when choice among hosts of different sizes or ages was allowed. Then, whether an asymmetrical effect of host size and age on the fitness of females versus males existed in P. nipponica was examined. Larger or fresher host pupae yielded larger wasps. Larger females lived longer, whereas male size did not influence male longevity. Large males mated successfully with relatively large females but failed with small females, whereas small males could mate successfully either with small or with large females. Thus, small-male advantages were found, and this held true even under male–male competition. Ovariole and egg numbers at any one time did not differ among females of different sizes. Larger females attained higher oviposition success and spent less time and energy for oviposition in hosts. Larger females produced more eggs from a single host meal. Taken together, females gained more, and males lost more, by being large. Host size and age thus asymmetrically affected the fitness of offspring males versus females through the relationships between host size or hast age and wasp size, which means the basic assumption of the host-size model was satisfied. Therefore, sex ratio control by P. nipponica in response to host size and age is adaptive. Received: November 13, 1998 / Accepted: January 18, 1999     

Genetic dimension of the coevolution of virulence-resistance in Drosophila -- parasitoid wasp relationships

Variations observed in parasite virulence and host resistance may be the outcome of coevolutionary processes. Recent theoretical developments have led to a 'geographic mosaic theory' of coevolution according to which there are some localities where reciprocal selection occurs (hot spots) and others where it is strongly reduced (cold spots). Studies of host-parasitoid systems back this up, revealing a geographical variation of traits subjected to antagonistic selection governed by variations in the strength of the ecological interactions. A more detailed analysis of the genetic basis of these geographic variations in a model system -- the interaction between Drosophila melanogaster and its specific parasitoid Leptopilina boulardi -- suggests that cold spots and hot spots are also driven by the amount of genetic variation available for the trait considered. Our approach, based on isolating reference strains, has been found to predict the result of sympatric interactions and it will be helpful in identifying the selective forces responsible for the coevolution. In this model, host resistance to a standardised reference strain is a weak predictor of the outcome of interactions in the field, and the main parameter accounting for the geographic variations is the number of host species available, with less parasitoid virulence towards D. melanogaster being found in areas displaying a more diversified host community.     

Fitness consequences of ovicide in a parasitoid wasp

Ovicide, superparasitism and host rejection are alternative reproductive tactics facing female parasitoid wasps encountering an already-parasitized host. Superparasitism is simply the addition of an egg or a clutch of eggs by the secondary parasitoid, but under ovicide the primary clutch is removed or destroyed. Host rejection occurs if the wasp leaves without laying a clutch. The ectoparasitoid Laelius pedatus (Say) (Hymenoptera: Bethylidae) performs ovicide in this situation. Clutch manipulation experiments show that secondary clutches suffer high mortality in competition with primary clutches, which increases with increasing time delay between clutches. Primary clutches however suffer little in competition with secondary clutches, even if there is minimal time delay between clutches. These data suggest that the offspring of ovicidal females are substantially fitter than the offspring of superparasitizing females. Handling time and clutch size do not differ significantly between first (sole) parasitoids and second (ovicidal) parasitoids. The same is true for offspring survival and development time. However, offspring of second females are slightly smaller. This suggests that parasitized and unparasitized hosts are resources of similar quality when ovicide is performed. These data strongly support the predictions of evolutionary models of ovicide. They may also give some insight into the taxonomic distribution of ovicide in parasitoids.     

Mixed sex allocation strategies in a parasitoid wasp

The sex allocation strategy of the parasitoid Laelius pedatus (Hymenoptera: Bethylidae) on different-sized hosts was investigated. The wasp lays from one to five eggs, and clutch size increases with host size. On the smallest hosts, single male eggs are laid, while on slightly larger hosts single female eggs are laid. On still larger hosts, gregarious clutches are laid which nearly always consist of a single male and one or more female eggs. The sex ratio strategy of the wasp appears to be influenced by a combination of local mate competition and conditional sex expression based on host quality. Received: 6 June 1996 / Accepted: 13 October 1996     

Visualization of polydnavirus sequences in a parasitoid wasp chromosome

Polydnaviruses, obligatorily associated with endoparasitoid wasps, are unique in that their segmented genome is composed of multiple double-stranded DNA circles. We present here the first cytological evidence that virus segments are integrated in the wasp genome, obtained by using in situ hybridization of virus probes with viral sequences in the chromosomes of a wasp from the braconid family of hymenopterans.     

Spread of the wasp parasitoid

The measurement of parasitism rates of wasp nests;lt Pelorus Bridge, New Zealand, at different distances from the initial release point suggests that the mean displacement of the parasitoid has increased by 1—1.5 km y(-1) from 1988 to 1993. Since average parasitism rates within this radius at any given site show little trend over time, this suggests an approximate 3-fold increase in the total parasitoid population each year, two-thirds of which is devoted to dispersal and one-third to maintaining local populations.     

Effects of age and repeated mating on male sperm supply and paternity in a parasitoid wasp

Post-copulatory paternity biases after female multiple mating are major constraints on both male and female reproductive systems. The outcome of paternity in certain situations is only controlled directly by male sperm stock. This was tested experimentally in the parasitoid wasp Anisopteromalus calandrae (Howard) (Hymenoptera: Pteromalidae), in which sperm stocks are small (several hundred) and the fertilizing efficiency of stored sperm is high (the ratio of sperm stored/fertilized eggs is about 0.75). Sperm in seminal vesicles and paternity of males of different status (virgin young, virgin old, or young previously mated) were measured after female single and double mating. The amount of sperm in the seminal vesicle differed according to male status (increasing from previously mated males to old males), but there was no difference in sperm stored by females after a single mating. In double mating experiments with two males of different status, paternity increased linearly with the relative amount of sperm in seminal vesicles. Paternity distribution conforms to 'a fair raffle' of sperm from both donors following complete mixing of sperm prior to fertilization. Thus, in a female multiple mating context, male fitness depends principally on their sperm stock, which in turn depends on life history parameters, such as age and previous mating.     

Mapping candidate genes for Drosophila melanogaster resistance to the parasitoid wasp Leptopilina boulardi

Drosophila melanogaster resistance against the parasitoid wasp Leptopilina boulardi is under the control of a single gene (Rlb), with two alleles, the resistant one being dominant. Using strains bearing deletions, we previously demonstrated that the 55E2-E6; 55F3 region on chromosome 2R is involved in the resistance phenomenon. In this paper, we first restricted the Rlb containing region by mapping at the molecular level the breakpoints of the Df(2R)Pc66, Df(2R)P34 and Df(2R)Pc4 deficiencies, using both chromosomal in situ hybridization and Southern analyses. The resistance gene was localized in a 100 kb fragment, predicted to contain about 10 different genes. Male recombination genetic experiments were then performed, leading to identification of two possible candidates for the Rlb gene. Potential involvement of one of this genes, edl/mae, is discussed.     

枸杞瘿螨姬小蜂的生物学特性

枸杞瘿螨姬小蜂Cirrospilus eniophyesiYang(新种)是枸杞瘿螨Aceria pallidaKeifer的重要天敌,以幼虫在虫瘿内取食枸杞瘿螨的成虫和若虫。在(25±1)℃下,该蜂完成1个世代需16~21d。成虫寿命及产卵量与补充营养有关,取食10%蜂蜜水的雌虫寿命为(6.94±0.67)d,雄虫为(5.14±0.49)d。不进行补充营养的雌雄成虫寿命分别为(1.96±0.27)d和(1.06±0.29)d;补充10%蜂蜜水和水可显著提高产卵量,每雌平均产卵量分别为(29.13±6.12)粒和(23.14±3.16)粒,产卵高峰期在羽化后的第2天。成虫羽化时间多集中在6:00~8:00am和4:00~6:00pm,羽化当天即可交尾。枸杞瘿螨姬小蜂自然种群消长与寄主虫口密度相关,在6月底7月初寄生率达到83.3%。     

Kin discrimination and sex ratios in a parasitoid wasp

Sex ratio theory provides a clear and simple way to test if nonsocial haplodiploid wasps can discriminate between kin and nonkin. Specifically, if females can discriminate siblings from nonrelatives, then they are expected to produce a higher proportion of daughters if they mate with a sibling. This prediction arises because in haplodiploids, inbreeding (sib-mating) causes a mother to be relatively more related to her daughters than her sons. Here we formally model this prediction for when multiple females lay eggs in a patch, and test it with the parasitoid wasp Nasonia vitripennis. Our results show that females do not adjust their sex ratio behaviour dependent upon whether they mate with a sibling or nonrelative, in response to either direct genetic or a range of indirect environmental cues. This suggests that females of N. vitripennis cannot discriminate between kin and nonkin. The implications of our results for the understanding of sex ratio and social evolution are discussed.     

Factors affecting refuge from parasitoid attack in a cynipid wasp, Aphelonyx glanduliferae

We examined seasonal patterns of gall morphology, growth, and survivorship of the agamic generation of a cynipid wasp, Aphelonyx glanduliferae, and discussed its mortality factors, especially from the point of view of refuge from parasitoid attack. Although the initiation period varied greatly among individual galls, the larvae of A. glanduliferae grew rapidly and reached their maximum size within 3 weeks before pupating in late September to early October. This growth period corresponded to the period when the gall walls became thinner. Parasitoid attack, which was the principal factor in the mortality of A. glanduliferae in the tree crown, was concentrated around the pupation period of the cynipid. Gall walls were significantly thinner in galls attacked by parasitoids than in those still containing a living cynipid. Therefore, the period available to parasitoids seems to be limited by both gall wall thickness and cynipid size. Thus, the growth pattern of A. glanduliferae larvae can have significance in that it narrows the window of vulnerability to parasitoids to a particular period. Although delaying gall initiation will also shorten the exposure period to parasitoid attacks, it was likely to increase the risk of death from gall abortion caused by seasonal degradation in the quality of host plant tissues. Although many cynipids were killed by disease in the galls that fell to the ground, the falling of mature galls to the ground may be another way to a parasitoid-free space. It is thus suggested that a trade-off among life history traits against multiple factors operates in the refuge of A. glanduliferae from parasitoid attack. Received: May 15, 2001 / Accepted: February 1, 2002     

Sperm stock and mating of males in a parasitoid wasp

Hymenoptera are haplodiploid insects, consequently sex ratio depends on female's sperm management which itself arises from the reproductive capacity of neighbouring males. To study the influence of ageing on male reproductive potential, laboratory experiments were conducted on Dinarmus basalis (Hymenoptera, Pteromalidae) males, a tropical wasp in which sperm counts are known to constrain sex ratio. Two groups of virgin males were compared: 1-day and 30-days old. Parameters recorded were sperm quantity and viability in seminal vesicles, shape of testis, mating ability in both individual and competitive situations and sperm stored by females after male multiple mating. Older males had twice as much sperm as young males, but their reproductive capacities did not differ. They were able to copulate with 20 successive virgin females in a short period. Sperm stored in spermathecae decreased with female mating order. In competition, old and young males had the same access to females. The difference between old and young males was visible at the level of reproductive tract: young males have functional testis and old males have empty non-functional testis. Spermatozoa are kept viable in male seminal vesicles for long periods. In this species, the reproductive potential of males is not altered by ageing. At the population level this may represent an adaptation for maintaining continuous reserves of sperm at the disposal of females.     

Single-locus recessive inheritance of asexual reproduction in a parasitoid wasp

The evolutionary maintenance of sex is one of the big unresolved puzzles in biology. All else being equal, all-female asexual populations should enjoy a two-fold reproductive advantage over sexual relatives consisting of male and female individuals. However, the "all else being equal" assumption rarely holds in real organisms because asexuality tends to be confounded with altered genomic constitutions such as hybridization and polyploidization or to be associated with parthenogenesis-inducing microbes. This limits the ability to draw general conclusions from any particular system. Here we describe a new system that permits unbiased comparisons of sexual and asexual reproduction: the parasitic wasp Lysiphlebus fabarum. Crossing experiments demonstrated that asexual reproduction has a simple genetic basis in this species and is consistently inherited as a single-locus recessive trait. We further show that the asexuality-inducing allele exhibits complete linkage to a specific allele at a microsatellite marker: all asexual lines in the field were homozygous for this allele, and the allele cosegregated perfectly with asexual reproduction in our experimental crossings. This novel system of contagious asexuality allows the production of closely related individuals with different reproductive modes, as well as the monitoring of the asexuality-inducing allele in natural and experimental populations.     

Potential fitness consequences of associative learning in a parasitoid wasp

Several studies have documented associative learning in insects,but the adaptive value of such learning is not yet well understood.To evaluate this issue, we quantified long-term fitness consequencesof associative learning in the parasitoid wasp, Biosteres arisanus.We compared individual wasps that were allowed to choose hostsubstrate based on experience ("learning" wasps) to wasps thatcould only make random substrate choice ("random" wasps) inan environment where only one out of two substrates containedhost eggs. In two experiments, the average number of host eggsparasitized and offspring produced were significantly largerfor learning than for random wasps. Our results allow detailedexamination of the conditions under which learning would havepositive fitness effects in ecological systems similar to ours.These conditions include relatively long search duration forhosts; the ability to remember a learned preference over extendedperiods of interfering activities; and large mean differencesbetween alternatives, and small variances, which together allowrapid evaluation of alternatives and long duration of exploitingthe superior one.     

变温贮藏僵蚜对烟蚜茧蜂耐寒能力的影响

为明确变温贮藏以麦二叉蚜为寄主的僵蚜对烟蚜茧蜂耐寒能力的影响,探究其体内的生化物质变化规律,测定了变温处理后羽化的烟蚜茧蜂雌雄成虫过冷却点、结冰点、体内含水量、脂肪、蛋白质和总糖含量的变化.结果表明:与对照(20℃)相比,经4℃22 h/20℃2h和4 ℃ 46 h/20℃2h处理1周后烟蚜茧蜂的耐寒能力显著增强.经变温处理后,烟蚜茧蜂雌雄个体的过冷却点、结冰点均出现不同程度的下降,雌蜂经4℃22 h/20℃2h处理后过冷却点和结冰点最低,分别为-26.38和-25.51 ℃;雄蜂经4 ℃ 46 h/20℃2h处理后过冷却点和结冰点最低,分别为-26.82和-26.38℃.经变温处理后烟蚜茧蜂僵蚜雌雄个体体内糖和蛋白质含量上升而脂肪和体内含水量下降,尤以经4℃22 h/20 ℃ 2 h和4℃46h/20℃2h处理后的变化最为明显.变温可以提高烟蚜茧蜂僵蚜的低温抵抗能力,且其耐寒能力的增加与其体内生化物质含量的变化密切相关.僵蚜经4℃22h/20℃2h和4℃46h/20℃2h贮藏1周后更有利于烟蚜茧蜂的生存和实践应用.