Pediobius metallicus (Hymenoptera: Eulophidae): First record of a parasitoid wasp of the agromyzid fly Japanagromyza tokunagai,a serious pest of orchids

Agromyzid flies, also known as leafminer flies, are one of the most serious insect pests that infest various plants. An agromyzid fly Japanagromyza tokunagai lays eggs in the ovaries of various Japanese orchids during or shortly after anthesis. Consequently, seed predation by J. tokunagai has the potential to severely limit the reproduction of many endangered orchids in Japan. While agromyzid populations tend to be suppressed by rich communities of natural parasitoids, such parasitoids have yet to be reported. Here we report Pediobius metallicus as the first record of a parasitoid of J. tokunagai. It is highly unusual to find parasitoids infesting J. tokunagai, possibly because the fly is protected by the thick wall and three-dimensional structure of orchid capsule. Pediobius metallicus may play an important role in suppressing J. tokunagai population at least in our study site.     

First report that the wasp Gronotoma guamensis (Hymenoptera: Figitidae: Eucoilinae) parasitizes the orchid‐feeding fly Japanagromyza tokunagai in Japan

Compared with mycorrhizal relationships, the relationships between mycoheterotrophic plants and insects have rarely been studied. Here we report a previously unknown tripartite interaction among an endangered mycoheterotrophic orchid, an orchid‐feeding fly, and a parasitoid wasp. The flowers and stems of Eulophia zollingeri were heavily attacked by Japanagromyza tokunagai (Sasakawa) (Diptera: Agromyzidae), and the parasitoid wasp Gronotoma guamensis (Yoshimoto) (Hymenoptera: Figitidae) was reared from these J. tokunagai individuals. Considering the high infestation rate of J. tokunagai, G. guamensis could be important in suppressing the J. tokunagai population, and positively affect the reproductive success of E. zollingeri.     

Characterization of the damage of Spodoptera eridania (Cramer) and Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae) to structures of cotton plants

The cotton plant, Gossypium hirsutum, hosts various pests that damage different structures. Among these pests, Spodoptera cosmioides (Walker) and Spodoptera eridania (Cramer) (Lepidoptera: Noctuidae) are considered important. The objectives of this study were to characterize and to quantify the potential damage of S. eridania and S. cosmioides feeding on different structures of cotton plants. For this purpose, newly-hatched larvae were reared on the following plant parts: leaf and flower bud; leaf and boll; flower bud or boll; and leaf, flower bud and boll. The survival of S. cosmioides and S. eridania was greater than 80% and 70% for larvae fed on cotton plant parts offered separately or together, respectively. One larva of S. eridania damaged 1.7 flower buds, but did not damage bolls, while one larva of S. cosmioides damaged 5.2 flower buds and 3.0 cotton bolls. Spodoptera eridania and S. cosmioides can be considered species with potential to cause economic damage to cotton plants because they can occur throughout cotton developmental stages causing defoliation and losses of reproductive structures. Therefore, the results validate field observations that these two species of Spodoptera are potential pests for cotton.     

Confirmation by DNA analysis that Contarinia maculipennis (Diptera: Cecidomyiidae) is a polyphagous pest of orchids and other unrelated cultivated plants

The cytochrome oxidase subunit I gene in mitochondrial DNA of 53 larvae of Contarinia maculipennis Felt from flower buds of various host plants, collected from Hawaii, Japan and Thailand was analysed. Monophyly of the clade including C. maculipennis from Hawaii, Thailand and Japan was supported. There was no sequential variation within the specimens from Hawaii and Japan, which differed from one another by 6 bp (1.37%). Three haplotypes were recognized in specimens from Thailand but differences from Hawaiian and Japanese specimens were small. Overall, there were no differences in the 146 deduced amino acid residues. It is therefore concluded that C. maculipennis is a polyphagous species that can develop on plant hosts representing at least seven botanical families. This pest of Dendrobium flower buds in glasshouses is considered to have entered Hawaii, Florida and Japan from Southeast Asia, and was recently intercepted in the Netherlands. Infestations have established and spread in orchid glasshouses, causing concern about the possibility of more extensive damage to orchids and to crops, such as bitter gourd, grown in close proximity to orchid glasshouses in Japan. The potential usefulness of DNA analysis in determining host plant ranges of morphologically identical cecidomyiid species that are currently identified solely on differences of host plant is emphasized.     

Foraging behavior of Brachygastra lecheguana (Hymenoptera: Vespidae) on Banisteriopsis malifolia (Malpighiaceae): Extrafloral nectar consumption and herbivore predation in a tending ant system

In the Brazilian savanna many plant species bear regular associations with patrolling ants that are aggressive towards insect herbivores. However, not only ants but also several species of predatory wasps are attracted to plants due to the extrafloral nectaries (EFNs). Such wasps feed on both herbivores and plant exudates. In this study we describe the foraging behavior of the social Polistinae wasp Brachygastra lecheguana in the extrafloral nectaried shrub Banisteriopsis malifolia, and investigated the influence of patrolling ants Camponotus blandus on the activity of the wasp. Brachygastra lecheguana fed on the endophytic larvae of Anthonomus (Curculionidae) beetles that developed inside flower buds. The wasp lacerated the bud layers to reach the beetle larvae located at the bud core. The wasp visits to Ba. malifolia were statistically related to the abundance of flower buds and beetles. Ant exclusion experiments revealed that the hunting behavior of B. lecheguana on beetles was not related to the absence of C. blandus. However we found that wasps spent more time consuming extrafloral nectar on branches where ants were excluded. This is the first study reporting extrafloral nectar consumption by B. lecheguana, as well as the predation on herbivores in natural areas. In cerrado vegetation, ants benefit the plant by reducing insect herbivores, and our study provides evidence that the B. lecheguana – Ba. malifolia system represents a potential interaction where the wasp may also benefit the host plant. The value of this wasp species as a plant‐guard is discussed.     

Pollination of the European food-deceptive Traunsteinera globosa (Orchidaceae): the importance of nectar-producing neighbouring plants

European food-deceptive orchids generally flower early in spring and rely on naïve pollinators for their reproduction. Some species however, flower later in the summer, when many other rewarding plants species are also in bloom. In dense flowering communities, deceptive orchids may suffer from competition for pollinator resources, or might alternatively benefit from higher community attractiveness. We investigated the pollination strategy of the deceptive species Traunsteinera globosa, and more specifically whether it benefited from the presence of coflowering rewarding species. We carried out a population survey to quantify the density and reproductive success of the orchid as well as the density of all coflowering species. Our results suggest that the deceptive orchid not only benefited from the presence of coflowering species, but that interestingly the density of the species Trifolium pratense was significantly positively correlated with the orchid's reproductive success. This species might simply act as a magnet species attracting pollinators near T. globosa, or could influence the orchid reproductive fitness through a more species-specific interaction. We propose that morphological or colour similarities between the two species should be investigated in more detail to decipher this pollination facilitation effect.     

Sex-Biased Temporal Gene Expression in Male and Female Floral Buds of Seabuckthorn (Hippophae rhamnoides)

Seabuckthorn is an economically important dioecious plant in which mechanism of sex determination is unknown. The study was conducted to identify seabuckthorn homologous genes involved in floral development which may have role in sex determination. Forty four putative Genes involved in sex determination (GISD) reported in model plants were shortlisted from literature survey, and twenty nine seabuckthorn homologous sequences were identified from available seabuckthorn genomic resources. Of these, 21 genes were found to differentially express in either male or female flower bud stages. HrCRY2 was significantly expressed in female flower buds only while HrCO had significant expression in male flowers only. Among the three male and female floral development stages (FDS), male stage II had significant expression of most of the GISD. Information on these sex-specific expressed genes will help in elucidating sex determination mechanism in seabuckthorn.     

DNA barcoding identification and genetic diversity of bamboo shoot wireworms (Coleoptera: Elateridae) in South China

Wireworms are recognized as economically important oligophagous pests of Phyllostachys bamboo shoot, which have caused a huge economic loss in South China. Studies of bamboo shoot wireworm control and management were seriously hampered by the lack of reliable identification of larvae. DNA barcoding has been proved to be an important and useful tool in species identification of morphologically cryptic insects. Accurate knowledge of damage-causing species and the phylogenic structure of elaterids will provide insight into their sustainable and effective management. Here, we use interspecific variation in COI and EF1-a as a robust method of identification and consider the intra- and interspecific genetic variation of bamboo wireworms. This approach has revealed that three Melanotus larvae occurred in Phyllostachys forest. The dominance of one species in different sampling location suggests there maybe underlying difference in habit and host plant preference. These data provide a starting point for determining the distribution of wireworm species feeding on bamboo shoots in South China. The inclusions would further aid identification of wireworm species and relationships between certain wireworm species with specific environments.     

The influence of the rearing host on the response of the parasitoid Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae) to odours from Ephestia kuehniella and Plodia interpunctella in a Y-tube olfactometer

Venturia canescens (Gravenhorst) is an ichneumonid generalist parasitoid that successfully attacks the larvae of different lepidopteran pests that infest stored products. These pest species include Plodia interpunctella and Ephestia kuehniella. In this study, we aimed to evaluate the influence of the rearing host on the parasitoid’s ability to detect and respond to a new host different from the rearing species. For this reason, the trials tested the preference of parasitoids reared on P. interpunctella or E. kuehniella for products that were or were not infested with larvae of these hosts. The trials were conducted in a Y-tube olfactometer. Regardless of the rearing host species, the parasitoids showed no preference for uninfested products. The parasitoids were attracted to products infested with larvae of their rearing host in preference to uninfested products. They also showed preferential attraction to products infested with the new host over uninfested products. E. kuehniella was the preferred host, irrespectively of the parasitoid host rearing species. The results are discussed to develop a better understanding of the ecology of V. canescens for its application in biological control.     

Behavioral strategy of a lycaenid (Lepidoptera) caterpillar against aggressive ants in a Brazilian savanna

Myrmecophily is widespread in lycaenid butterflies, in which ants receive food resources and, in turn, protect caterpillars against natural enemies. This interaction ranges from obligate myrmecophily, in which immatures are invariably associated with ants and are dependent on ants for survival, and facultative myrmecophily, in which larvae are not dependent on ants for survival, but the presence of the latter may increase larvae survival. Lycaenids also include non-myrmecophilous butterflies, which do not have positive associations with ants and have developed strategies to avoid being attacked or preyed upon by them. In this study, we examined the relationship between the lycaenid Michaelus ira and two ant species associated with Distictella elongata (Bignoniaceae). This plant has extrafloral nectaries and is patrolled by Camponotus crassus and Ectatomma tuberculatum. Morphological analyses revealed that M. ira larvae have ant organs, such as dorsal nectary organs and perforated cupolas, structures associated with myrmecophily. We performed larval exposure experiments in the field, predicting that, in the absence of myrmecophily, the butterfly larva would present strategies to avoid ant attack. Results showed that larvae were attacked by both ant species. To escape ant molestation, larvae lived and fed inside silk-sealed D. elongata flower buds. We concluded that the M. ira bud-sheltering behavior was a defensive strategy against these ant species, while the dorsal nectary organs were apparently nonfunctional. Nonetheless, myrmecophily, in general, cannot be excluded in M. ira since relationships with other ant species may exist.     

Impact of the flower stalk-boring moth Neoschoenobia testacealis (Lepidoptera: Crambidae) and water-level fluctuations on the flower and fruit production of the yellow water lily Nuphar subintegerrima (Nymphaeaceae) in irrigation ponds of western Japan

The flower and fruit production of the yellow water lily Nuphar subintegerrima and the seasonal occurrence of the flower stalk-boring moth Neoschoenobia testacealis were surveyed in two irrigation ponds in western Japan. Water-level fluctuations at each pond were measured. Flowering was observed from late May to September, and fruiting from late May to October. The yellow water lily produced cumulative numbers of 189 and 181 flowers and 121 and 129 fruits in the two ponds’ quadrats. The fruit setting rates at the two ponds were 0.64 and 0.71, and decreased in the summer. The percentage of bored flower stalks was 16 and 5%. Mature larvae bored flower stalks severely during the summer. Whenever the boring larvae attacked the flower stalks, the flower buds and flowers were aborted. There was a significant negative correlation between the fruit setting rate and the boring rate per month (number of bored flower stalks/number of all flower stalks). Whenever abruptly rising water levels (about 10 cm) submerged the flowers, the flowers were aborted too and could not set fruits. The percentages of non-setting fruit due to the rising water levels at the two ponds were 6 and 8%. Overall, our results indicated that flower stalk boring was a more important factor than water-level fluctuation in the sexual reproduction of N. subintegerrima and that N. subintegerrima was well suited to small irrigation ponds.     

Consistent pollination services to Cypripedium macranthos var. rebunense (Orchidaceae) by Bombus pseudobaicalensis

Bombus pseudobaicalensis queens are known pollinators of the threatened orchid Cypripedium macranthos var. rebunense. However, whether other bumblebee species also pollinate this orchid has not yet been ascertained. A 10-year observation provided convincing evidence that B. pseudobaicalensis is the only consistent pollinator. B. beaticola moshkarareppus queens were newly found to rarely carry pollen smears of the orchid, whereas B. hypocrita sapporoensis and B. diversus queens were practically mere flower visitors. Why these three species do not equal B. pseudobaicalensis in pollination effectiveness remains unclear but differences in body size and individual abundance between B. pseudobaicalensis and the other species are considered as possible primary causes. Given the almost complete dependence of C. macranthos var. rebunense on B. pseudobaicalensis for pollination and its potential risk, conservation managers should take care not to reduce the bumblebee colonies.     

New information on host plants and distribution ranges of an invasive gall midge, <Emphasis Type="Italic">Contarinia maculipennis</Emphasis> (Diptera: Cecidomyiidae), and its congeners in Japan

Gall midges of the genus Contarinia (Diptera: Cecidomyiidae) that infest the flower buds of various plant species have been newly found in Japan in recent years. Those infesting the flower buds of Pseuderanthemum laxiflorum (A. Gray) Hubbard ex Baillon (Amaranthaceae) and Jasminum sambac (Linnaeus) Aiton (Oleaceae) in Okinawa Prefecture, and Dendrobium spp. (Orchidaceae) in Mie Prefecture were identified, on the basis of morphological features and molecular information, as an invasive gall midge, C. maculipennis Felt. C. maculipennis was recorded in Mie Prefecture for the first time, and P. laxiflorum is newly regarded as one of the host plants of C. maculipennis. Three other Contarinia gall midges that we found infesting the flower buds of Lycopersicon esculentum Miller, Capsicum annuum Linnaeus (Solanaceae), and Oxalis corniculata Linnaeus (Oxalidaceae) were not identical with C. maculipennis. Among these, the first two, which infested solanaceous plants, were identical. However, the species other than C. maculipennis could not be identified to the species level because morphological differences were obscure and DNA sequencing data of allied congeners have not yet been registered on GenBank.     

Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) eggs as alternative food for rearing of lady beetles Eriopis connexa (Germar) (Coleoptera: Coccinellidae)

Eriopis connexa (Germar) (Coleoptera: Coccinellidae) is an important predator with potential for biological control of insect pests. This research evaluated the development of E. connexa larvae fed on fresh eggs of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) without (T1) or with (T2) scales or one-day (T3) or six-month (T4) frozen, or newly-hatched larvae of S. frugiperda (T5). The percentage of E. connexa adults was higher when larvae feeding on fresh S. frugiperda eggs with or without scales, or one-day frozen eggs of this prey and lower with eggs of this Lepidoptera after frozen for six months or with newly-hatched larvae of S. frugiperda. Duration of the larval period of E. connexa was 15.7, 15.8, 16.0, 17.6, and 17.3 days, respectively, with these diets. The high survival of E. connexa fed with eggs of S. frugiperda shows the potential use of this prey in the laboratory to maintain this natural enemy.     

Identity and Specificity of Rhizoctonia-Like Fungi from Different Populations of Liparis japonica (Orchidaceae) in Northeast China

Mycorrhizal association is known to be important to orchid species, and a complete understanding of the fungi that form mycorrhizas is required for orchid ecology and conservation. Liparis japonica (Orchidaceae) is a widespread terrestrial photosynthetic orchid in Northeast China. Previously, we found the genetic diversity of this species has been reduced recent years due to habitat destruction and fragmentation, but little was known about the relationship between this orchid species and the mycorrhizal fungi. The Rhizoctonia-like fungi are the commonly accepted mycorrhizal fungi associated with orchids. In this study, the distribution, diversity and specificity of culturable Rhizoctonia-like fungi associated with L. japonica species were investigated from seven populations in Northeast China. Among the 201 endophytic fungal isolates obtained, 86 Rhizoctonia-like fungi were identified based on morphological characters and molecular methods, and the ITS sequences and phylogenetic analysis revealed that all these Rhizoctonia-like fungi fell in the same main clade and were closely related to those of Tulasnella calospora species group. These findings indicated the high mycorrhizal specificity existed in L. japonica species regardless of habitats at least in Northeast China. Our results also supported the wide distribution of this fungal partner, and implied that the decline of L. japonica in Northeast China did not result from high mycorrhizal specificity. Using culture-dependent technology, these mycorrhizal fungal isolates might be important sources for the further utilizing in orchids conservation.     

Effect of defoliation by checkerspot caterpillars (Euphydryas phaeton) and sawfly larvae (Macrophya nigra and Tenthredo grandis) on their host plants (Chelone spp.)

Summary The effect of defoliation by herbivores, checkerspot caterpillars (Euphydryas phaeton) and sawfly larvae (Macrophya nigra and Tenthredo grandis), on the reproductive output of turtlehead (Chelone spp.) was examined. Defoliation prior to development of flower buds reduced the number of reproductive stalks, flower buds, flowers and seed capsules. Severe herbivory, after flower buds appeared, decreased the final number of seed capsules and seeds per capsule. The availability of the host plants to the herbivores was a function of prior defoliation and environmental conditions. Sawfly larvae, by defoliating the plants in midsummer, forced prediapause checkerspot caterpillars to wander in search of food plants. Decimation of these perenials by postdiapause checkerspot caterpillars in a dry spring retarded growth of turtlehead and, consequently, most of the plants were not available for egg-laying by sawflies and checkerspots.