Hideyuki Mitsui

Spatial distributions and clutch sizes of Drosophila species ovipositing on cherry fruits of different stages

In the aggregation theory, aggregation of eggs is one of important conditions for the coexistence of species. However, aggregation of eggs by clutch laying does not always promote coexistence, whereas aggregation of eggs by aggregated distributions of ovipositing females always has a significant contribution to the coexistence. In this study, spatial distributions of three Drosophila species across naturally occurring cherry fruits were studied with relation to their clutch sizes. Drosophila suzukii oviposited eggs mainly on fresh fruits on trees, and its eggs were randomly distributed across cherry fruits. The emergence data also indicated random spatial distributions of this species. Random egg distributions of this species are explained by random visits of females to fruits and the production of clutches of mostly single eggs. On the other hand, D. lutescens and D. rufa oviposited on fallen fruits, showed aggregated distributions in the emergence data, and frequently produced clutches of a few eggs. In these species, the degree of aggregation was usually significantly lower than the expectation based on random visits of females to fruits and their clutch sizes observed in the present experiments, indicating that their aggregation is unlikely to arise from aggregated distributions of ovipositing females. Thus, the spatial aggregation of these species does not necessarily lead to their coexistence.

Seasonal life cycles and resource uses of flower- and fruit-feeding drosophilid flies (Diptera: Drosophilidae) in central Japan

Seasonal life cycles and resource uses of flower‐ and fruit‐feeding drosophilids (Diptera: Drosophilidae) were studied from low to high altitudes in central Japan to understand their adaptation to seasonal changes of environmental conditions. Drosophila unipectinata and D. oshimai specialized to flowers, D. suzukii and D. subpulchrella depended almost on fruits, while D. lutescens, D. rufa, D. auraria, D. biauraria and D. sternopleuralis used both of them. It was assumed that D. unipectinata moved from low to high altitudes in June while D. oshimai, D. suzukii and D. subpulchrella in July. Migration of D. unipectinata is considered as a means to avoid summer heat or exploit early‐summer resources at high altitudes. On the other hand, D. oshimai, D. suzukii and D. subpulchrella have the capacity to pass the summer at low altitudes, and therefore their migration is assumed as a means to escape from resource‐poor conditions in summer at low altitudes or exploit resources at high altitudes. The generalist species, D. lutescens, D. rufa, D. auraria, D. biauraria and D. sternopleuralis, would not perform such extensive movements between low and high altitudes. They may pass the summer at low or mid altitudes depending on accidentally fallen immature fruits and/or some other resources such as decayed leaves.

Geographical distributions and host associations of larval parasitoids of frugivorous Drosophilidae in Japan

In Japan, dominant parasitoids attacking frugivorous Drosophilidae species were Asobara (Braconidae, Alysiinae), Leptopilina, and Ganaspis species (Figitidae, Eucoilinae). Asobara japonica was found throughout Japan, and its populations in the main islands of Japan were parthenogenetic whereas those in the subtropical islands were sexually reproducing. Other parasitoids showed rather restricted distributions; A.tabida, A. rossica, A. rufescens, and Leptopilina heterotoma occurred mainly in northern to central parts of the main islands, Ganaspis xanthopoda from central to southern parts of the main islands, A. leveri in a southern part of the main islands, and A. pleuralis, L. victoriae, and Ganaspis sp. mainly in the subtropical islands. Their major hosts were species of the D. melanogaster species group in the main islands, and species of the D. melanogaster, immigrans, and polychaeta species groups in the subtropical islands. Host use considerably varied among parasitoid species, especially in the subtropical islands.

Host range of Asobara japonica (Hymenoptera: Braconidae), a larval parasitoid of drosophilid flies

We studied the host range of Asobara japonica, a larval‐pupal parasitoid of drosophilid flies. Habitat selection was found to be an important determinant of host range in this parasitoid; it attacked drosophilid larvae breeding on banana and mushrooms, but seldom attacked those breeding on decayed leaves. This parasitoid was able to use diverse drosophilid taxa as hosts. Attack by A. japonica sometimes killed hosts at the larval stage, and therefore parasitoid larvae also died. Drosophila elegans and D. busckii suffered particularly high larval mortality due to the attack by A. japonica (in the latter species only when young larvae were attacked). Many individuals of D. subpulchrella also died at the pupal stage without producing parasitoids when they were parasitized at the late larval stage. In contrast, D. bipectinata, D. ficusphila, D. immigrans, D. formosana and D. albomicans were resistant to attack: large proportions of the larvae of these drosophilid species grew to adulthood, even in the presence of parasitoids. On the basis of phylogenetic information, we concluded that phylogenetic position has only limited importance as a factor determining whether a species is suitable as a host for A. japonica, at least within the genus Drosophila.

Ecological, morphological and molecular studies on Ganaspis individuals (Hymenoptera: Figitidae) attacking Drosophila suzukii (Diptera: Drosophilidae).

Ganaspis individuals parasitizing Drosophila suzukii (Matsumura), a pest of fruit crops, were examined for host use and molecular and morphological differences from those attacking D. lutescens Okada and some other Drosophila species that breed on fermenting fruits. Wild cherry fruits were collected in the suburbs of Tokyo, and drosophilid pupae obtained from these fruits were examined for parasitism. Drosophila suzukii was the only drosophilid species infesting fresh wild cherry fruits, and Ganaspis individuals were the major parasitoids attacking D. suzukii in wild cherry fruits. In parasitism experiments, these Ganaspis individuals parasitized D. suzukii larvae in fresh cherry fruits, but did not parasitize those in Drosophila medium. In addition, they did not parasitize larvae of some other fruit-feeding Drosophila species even when these occurred in fresh cherry fruit. These Ganaspis individuals parasitizing D. suzukii were different from those parasitizing D. lutescens and some other drosophilids in nucleotide sequences of the COI gene, as well as in ITS1 and ITS2. They were also different in forewing and antenna morphology, although they showed some overlap in morphological traits. They are tentatively assigned as the suzukii- and lutescens-associated types of G. xanthopoda Ashmead. In the present field survey, Leptopilina japonica Novković & Kimura and some Asobara species were also observed to attack D. suzukii larvae in wild cherry fruit.

Capacity of Japanese Asobara species (Hymenoptera; Braconidae) to parasitize a fruit pest Drosophila suzukii (Diptera; Drosophilidae)

Drosophila suzukii is a pest of soft fruit such as cherry, strawberry or blueberry and recently colonized North America and Europe from Asia. In this study, we assessed the utility of Japanese Asobara species as agents for the biological control of this pest species by examining their host use in nature and their capacity to parasitize this pest species. From Japan, eight Asobara species including putative three undescribed species were found; Asobara japonica was recorded from all over Japan, Asobara tabida from high altitude areas in central and northern Japan, Asobara rossica and Asobara rufescens from central to northern Japan, three undescribed species from central to western or southern Japan and Asobara pleuralis from a subtropical island of Japan. Among them, an undescribed species Asobara sp. TK1 may be useful as an agent for the biological control because it was recorded only from D. suzukii. All of the remaining seven species were generalists mainly using drosophilid species associated with fermenting fruits, and four of them had no or very low abilities to parasitize D. suzukii, indicating that these seven species are not or less appropriate as agents for biological control.

Taxonomy and phylogeny of Leptopilina species (Hymenoptera: Cynipoidea: Figitidae) attacking frugivorous drosophilid flies in Japan, with description of three new species

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: Leptopilina 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. Leptopilina japonica is 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.

Coexistence of drosophilid flies: Aggregation, patch size diversity and parasitism

We carried out field experiments to investigate the coexistence of Drosophila species in domestic and forest areas on the basis of the aggregation model. Three cosmopolitan species Drosophila simulans Sturtevant, Drosophila melanogaster Meigen and Drosophila immigrans Sturtevant, and a native species, Drosophila auraria Peng, emerged abundantly from banana placed at the domestic station, while Drosophila immigrans and five native species, Drosophila lutescens Okada, Drosophila rufa Kikkawa and Peng, Drosophila bizonata Kikkawa and Peng, Drosophila sternopleuralis Okada and Kurokawa and Scaptodrosophila coracina (Kikkawa and Peng), were abundant at the forest station. The present analysis suggests that their coexistence was facilitated by the aggregation mechanism. In the cosmopolitan species, the density of individuals that emerged from patches increased with the increase of patch size, but the relationship between fly density and patch size was not clear in the native species. This difference in distribution patterns between the cosmopolitan and native species is likely to be due to the difference in the female visiting behavior. In the present analysis, however, it was not clear whether patch size diversity facilitated their coexistence or not. The effect of patch size diversity may have been masked, because the effect of aggregation was more prominent. The rate of parasitism by wasps was high in October at the domestic station, and in May and June at the forest station. The present result suggests that the rate of parasitism was density-dependent, at least at the domestic station, and therefore parasitism facilitates the coexistence of drosophilid species in domestic areas.

Genetic differentiation of Ganaspis brasiliensis (Hymenoptera: Figitidae) from East and Southeast Asia

Ganaspis brasiliensis (Ihering) (Hymenoptera: Figitidae: Eucoilinae) is a Drosophila parasitoid that has often been misidentified as G. xanthopoda (Ashmead) in recent studies. This study aims to clarify genetic differentiation of G. brasiliensis based on the nucleotide sequences of the mitochondrial cytochrome oxidase subunit 1 (CO1) gene and three nuclear DNA regions, the inter-transcribed spacers 1 and 2 (ITS1 and ITS2) and putative 60S ribosomal protein L37 (RpL37), as well as crossing experiments. Four lineages are recognized in individuals assigned as G. basiliensis by morphology, (1) individuals occurring in Japan and probably South Korea, (2) individuals from a small subtropical island of Japan, Iriomote-jima, (3) individuals from temperate lowlands of Japan and high altitude areas of Southeast Asia, and (4) individuals occurring widely in Asia, America, Hawaii and Africa. The first lineage is a specialist of Drosophila suzukii (Matsumura), a pest of fresh fruit, and also the fourth lineage has a capacity to parasitize this pest species. The first, third and fourth lineages occur sympatrically at least in Tokyo. The third and fourth lineages differed in mate choice and host use to some extent, but post-mating isolation between them was almost absent.

Diversity and host associations of parasitoids attacking mycophagous drosophilids (Diptera: Drosophilidae) in northern and central Japan

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.