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1. Generalist koinobiont parasitoids often exhibit high flexibility in their development; their larvae shorten or prolong the developmental period depending on the host quality at parasitisation. However, flexibility of the growth rate of parasitoid larvae has rarely been investigated so far. 2. This study investigated how the koinobiont parasitoid wasps Asobara japonica and Leptopilina ryukyuensis regulate their larval growth when they parasitise host Drosophila larvae with varying larval periods. 3. In both parasitoid species, the preimaginal period was longer when they parasitised 1-day-old larvae of Drosophila rufa than when they parasitised older larvae of D. rufa or when they parasitised larvae of Drosophila simulans, a species with a shorter larval period than D. rufa. After host pupariation, A. japonica accelerated its growth, thereby showing a biphasic growth curve. On the other hand, L. ryukyuensis did not accelerate its growth after host pupariation. 4. Growth retardation of parasitoid larvae in 1-day-old D. rufa larvae would contribute to avoiding excess growth before host pupariation, because the excess growth of parasitoid larvae would have negative effects on host growth. The growth rate acceleration of A. japonica after host pupariation suggests that they enhance resource utilisation in a host that has reached maximum body mass. It remains uncertain as to why L. ryukuensis does not show clear accelerated growth after host pupariation. Nonetheless, these results suggest that parasitoid larvae have the ability to detect the developmental stage of hosts in a species-specific manner.

The Drosophila parasitoid Asobara japonica Belokobylskij (Hymenoptera: Braconidae) has highly toxic venom that kills host larvae if its injection is not followed by an injection of lateral oviduct components along with egg-laying. In the present study, the venoms of seven other Drosophila parasitoids (Asobara rossica, Asobara rufescens, Asobara pleuralis, Leptopilina heterotoma, Leptopilina japonica, Leptopilina ryukyuensis, and Leptopilina victoriae) are tested against three kinds of Drosophila species (i.e. Drosophila species that are suitable as host for focal parasitoids, those that are resistant to the parasitoids, and a cosmopolitan species, Drosophila simulans). Venoms of the three Asobara species are not toxic to any of Drosophila species, whereas those of the four Leptopilina species are toxic to some Drosophila species. The toxicity of venom varies among Leptopilina species, and the susceptibility to venom also varies among host Drosophila species. Furthermore, toxicity and paralytic effects of venom are not correlated. Because the toxicity of venom is not adaptive for parasitoids, it may be an inevitable side effect of some components that play an essential role in parasitism.

Wasps of the genus Leptopilina (Hymenoptera: Figitidae) are larvo-pupal parasitoids of Drosophila species. Here we report three putative thelytokous species of this genus from Japan, with description of two new species L.tokioensis and L.tsushimaensis, which have been recorded from central Japan and Tsushima, respectively. Another thelytokous species is L.longipes occurring in northern Japan, although its European populations are assumed to be arrhenotokous. Preliminary phylogenetic analyses suggest the present three species are diversified from one another and also from the other Leptopilina species. These three thelytokous species are infected by B-supergroup Wolbachia as in European thelytokous species, L.clavipes and L.australis. The evolution of these thelytokous species is discussed based on the present and previous results, and the occurrence and distributions of Leptopilina species in Japan and surrounding regions are briefly reviewed.

International audience; Deciphering invasion routes from molecular data is crucial to understanding biological invasions, including identifying bottlenecks in population size and admixture among distinct populations. Here, we unravel the invasion routes of the invasive pest Drosophila suzukii using a multi-locus microsatellite dataset (25 loci on 23 worldwide sampling locations). To do this, we use approximate Bayesian computation (ABC), which has improved the reconstruction of invasion routes, but can be computationally expensive. We use our study to illustrate the use of a new, more efficient, ABC method, ABC random forest (ABC-RF) and compare it to a standard ABC method (ABC-LDA). We find that Japan emerges as the most probable source of the earliest recorded invasion into Hawaii. Southeast China and Hawaii together are the most probable sources of populations in western North America, which then in turn served as sources for those in eastern North America. European populations are genetically more homogeneous than North American populations, and their most probable source is northeast China, with evidence of limited gene flow from the eastern US as well. All introduced populations passed through bottlenecks, and analyses reveal five distinct admixture events. These findings can inform hypotheses concerning how this species evolved between different and independent source and invasive populations. Methodological comparisons indicate that ABC-RF and ABC-LDA show concordant results if ABC-LDA is based on a large number of simulated datasets but that ABC-RF out-performs ABC-LDA when using a comparable and more manageable number of simulated datasets, especially when analyzing complex introduction scenarios.

The diversity and host associations of parasitoids attacking mycophagous drosophilids were studied in Tokyo (a warm-temperate region) and Sapporo (a cool-temperate region) in Japan. Field collections were carried out using traps baited with mushrooms in May, June, September and October 2009 in Tokyo and in July and August 2010 in Sapporo. The major drosophilid species that emerged from mushroom baits was Drosophila bizonata in Tokyo and D.orientacea in Sapporo. In total, 13 parasitoid species emerged from drosophilids occurring in mushroom baits, and 11 of them were larval parasitoids belonging to Braconidae and Figitidae. Among the 11 larval parasitoids, 10 were collected in Tokyo, while only two were collected in Sapporo. It is not known why their diversity differed so much between these two regions. Four of the 11 larval parasitoids have also been recorded from drosophilid larvae occurring in fruit (banana). The use of these two habitats (mushrooms and fruit) by these four species seems to reflect the occurrence (i.e. resource use) of their suitable hosts. On the other hand, most larval parasitoids from Tokyo attacked D.bizonata, and two larval parasitoids from Sapporo attacked D.orientacea, suggesting that the abundance of potential hosts is one of the important factors affecting their host use.

Leafminer (Phyllonorycter, Gracillariidae, Lepidoptera) and aphid (Tuberculatus, Aphididae, Hemiptera) compositions were studied in three deciduous oak species, Quercus dentata, Q. crispula, and Q. serrata, and their hybrids in Tomakomai Experimental Forest of Hokkaido University, Hokkaido, northern Japan. Identification of trees in this forest was done mainly on the basis of discriminant analysis on leaf morphology with reference to trees in pure Q. dentata and Q. crispula stands and a Q. serrata stand mixed with Q. crispula. The results suggested that hybridization occurred in all combinations (i.e. Q. dentata-Q. crispula, Q. crispula-Q. serrata, and Q. serrata-Q. dentata) and the frequency of hybrids was about 10%. The composition of Phyllonorycter and Tuberculatus species differed between Q. dentata and Q. crispula or Q. serrata, but did not differ between Q. crispula and Q. serrata. Thus, Q. dentata would differ from Q. crispula and Q. serrata in chemical properties that determine herbivore host selection, survival and performance, possibly reflecting their eco-physiological differences or phylogenetic distances. The study insects were divided into three groups; species specialized to Q. dentata (three Phyllonorycter and one Tuberculatus species), those to Q. crispula and Q. serrata (six Phyllonorycter and two Tuberculatus species), and a species collected at least from Q. dentata and Q. crispula (one Tuberculatus species). Putative hybrid trees of Q. dentata and Q. crispula harbored both Q. dentata- and Q. crispula-specific insects.

Despite the intensive use of the Leptopilina genus and its drosophilid hosts as model-systems in the study of host-parasitoid interactions, the diversity and distribution of the species occurring in the Asian region remain elusive. Here we report the phylogeny of Japanese Leptopilina species attacking frugivorous drosophilid flies, based on COI, ITS1 and ITS2 sequences. Consistent with molecular data, hybridization experiments and morphological examination, five species were recorded in Japan. L. heterotoma, L. victoriae and three new species, two occurring in the Ryukyu archipelago, L. ryukyuensis and L. pacifica, and another species, L. japonica, distributed in Honshu and Hokkaido. L. japonica was further divided into two subspecies, L. j. japonica occurring in Japan, and L. j. formosana occurring in Taiwan. According to these results, we discuss the evolution, speciation and colonization history of Japanese Leptopilina species.

Ovipositing Asobara japonica females inject venom (containing paralysis-inducing factors) immediately after the insertion of their ovipositors into Drosophila larvae, and lay eggs a little later. Interruption of their oviposition behaviour before egg laying causes high larval mortality in host Drosophila species, while normal oviposition does not. This suggests that venom of this parasitoid is toxic to larvae of these host species but its toxicity is suppressed by factor(s) provided by parasitoid females at the time of laying egg or by parasitoid embryos developing in the hosts. On the other hand, venom does not show toxicity to larvae of non-host Drosophia species. Possible functions of venom are discussed.

Parasitoid assemblages and the rates of parasitism on tissue-feeding larvae of two Phyllonorycter leafminer species, P. persimilis and P. leucocorona, were studied from the autumn generation in 2002 to the summer generation in 2005 to understand whether parasitoids mediate interactions of the two leaf species. Fourteen species of parasitoids emerged from P. persimilis and 11 species emerged from P. leucocorona. The parasitism rate was high; i.e., 24.1-92.6% in P. persimilis and 58.9-81.7% in P. leucocorona. Thus, parasitism was a major mortality factor in the present Phyllonorycter species. The parasitoid composition was distinctly different between the two host species, although most parasitoids were able to parasitize both leafminer species. The analysis based on the quantitative parasitoid overlap revealed that the present parasitoids could mediate interactions between the present leafminer species, but their effects would be weak. This is attributable to that most parasitoids preferentially parasitize either of the leafminer species.

Flowering phenology of cleistogamous annual plants Impatiens noli-tangere and Persicaria thunbergii was studied with reference to their mortality in streamside environments in northern Japan. I. noli-tangere produced flower-buds and flowers from late June to October, whereas P. thunbergii did so from mid-August to October. Both species were often killed by rapid current and submergence after heavy rainfall, which was unpredictable in the timing and intensity. P. thunbergii was more tolerant to rapid current and submergence than I. noli-tangere. Some individuals of I. noli-tangere were killed by herbivory from sawfly larvae in summer, but P. thunbergii individuals were seldom killed by herbivory. In consequence, the percentage of individuals that survived until the end of September was higher in P. thunbergii than in I. noli-tangere. These situations are assumed to select a long reproductive life in I. noli-tangere and autumn flowering in P. thunbergii.

The endoparasitoid wasp Asobara japonica has highly poisonous venom: the host Drosophila larvae are killed by envenomation at a dose that is naturally injected by the female wasp at parasitism. This insecticidal venom is neutralized, however, because A. japonica introduces lateral oviduct components soon after venom injection at oviposition. Although the venom and lateral oviduct components of this parasitoid have been partially characterized, how the venom components favor successful development of wasp eggs and larvae in the host remains ambiguous. Here, we demonstrated that A. japonica venom did not affect host humoral immune responses, determined as expression of antimicrobial peptide (AMP) genes, but significantly diminished two cellular responses, spreading and phagocytosis, by host hemocytes. Moreover, venom components drastically elevated a serine protease-like activity 4 h after its injection. The lateral oviduct components did not negate the detrimental effects of the venom on host cellular immunities, but significantly reduced the venom-induced elevation of protease activity. Both active factors in venom and lateral oviduct components were roughly characterized as heat-labile substances with a molecular mass of at least 10 kDa. Finally, venom of A. japonica, with a wide host range, was found to be much more toxic than that of Asobara rossica, which has a limited host range. These results reveal that A. japonica venom toxicity allows exploitation of a broader range of host insects because it is essential to overcome cellular immune responses of the host for successful parasitism.