Effects of quantitative and qualitative differences in volatiles from host‐ and non‐host‐infested maize on the attraction of the larval parasitoid Cotesia kariyai

Cotesia kariyai Watanabe (Hymenoptera: Braconidae) is a specialist larval parasitoid of Mythimna separata Walker (Lepidoptera: Noctuidae). Cotesia kariyai wasps use herbivore‐induced plant volatiles (HIPVs) to locate hosts. However, complex natural habitats are full of volatiles released by both herbivorous host‐ and non‐host‐infested plants at various levels of intensity. Therefore, the presence of non‐hosts may affect parasitoid decisions while foraging. Here, the host‐finding efficiency of naive C. kariyai from HIPVs influenced by host‐ and non‐host‐infested maize [Zea mays L. (Poaceae)] plants was investigated with a four‐arm olfactometer. Ostrinia furnacalis Guenée (Lepidoptera: Crambidae) was selected as a non‐host species. One unit (1 U) of host‐ or non‐host‐infested plant was prepared by infesting a potted plant with five host or seven non‐host larvae. In two‐choice bioassays, host‐infested plants fed upon by different numbers of larvae, and various units of host‐ and non‐host‐infested plants (infestation units; 1 U, 2 U, and 3 U) were arranged to examine the effects of differences in volatile quantity and quality on the olfactory responses of C. kariyai with the assumption that volatile quantity and quality changes with differences in numbers of insects and plants. Cotesia kariyai was found to perceive quantitative differences in volatiles from host‐infested plants, preferring larger quantities of volatiles from larger numbers of larvae or plants. Also, the parasitoids discriminated between healthy plants, host‐infested plants, and non‐host‐infested plants by recognising volatiles released from those plants. Cotesia kariyai showed a reduced preference for host‐induced volatiles, when larger numbers of non‐host‐infested plants were present. Therefore, quantitative and qualitative differences in volatiles from host‐ and non‐host‐infested plants appear to affect the decision of C. kariyai during host‐habitat searching in multiple tritrophic systems.     

Temperature-sensitive eye colour mutation in the parasitoid fly Exorista japonica Townsend (Dipt.: Tachinidae)

Abstract:  An eye colour mutation was found in the tachinid fly Exorista japonica Townsend, and we established an isogenic line of the mutant. The phenotype of the mutant is red eye in adults; this can be clearly distinguished from the brown-eyed phenotype of the wild type. Crossing experiments demonstrated that the mutant phenotype is controlled by a single autosomal recessive allele. As rearing temperature during the puparial stage rose, eye colour became lighter in the mutant but did not change in the wild type, indicating that the phenotype of the mutant is temperature dependent. No differences in the percentages of egg eclosion, pupariation and adult emergence were observed between the mutant and the wild type. This mutant of E. japonica – the first in the Tachinidae to be described – could be a useful marker in the laboratory experiments to test for paternity in multiple mating and to examine intraspecific competition.     

Rhodocytin induces platelet aggregation by interacting with glycoprotein Ia/IIa (GPIa/IIa, Integrin alpha 2beta 1). Involvement of GPIa/IIa-associated src and protein tyrosine phosphorylation

Although glycoprotein Ia/IIa (GPIa/IIa, integrin alpha(2)beta(1)) has established its role as a collagen receptor, it remains unclear whether GPIa/IIa mediates activation signals. In this study, we show that rhodocytin, purified from the Calloselasma rhodostoma venom, induces platelet aggregation, which can be blocked by anti-GPIa monoclonal antibodies. Studies with rhodocytin-coupled beads and liposomes loaded with recombinant GPIa/IIa demonstrated that rhodocytin directly binds to GPIa/IIa independently of divalent cations. In vitro kinase assays and Western blotting of GPIa immunoprecipitates revealed that Src and Lyn constitutively associate with GPIa/IIa and that Src activity increases transiently after rhodocytin stimulation. Src specifically associates with p130 Crk-associated substrate (Cas) in a manner dependent upon Cas phosphorylation, suggesting that Src is responsible for Cas tyrosine phosphorylation. While all these phenomena occur early after rhodocytin stimulation in a cAMP-resistant manner, tyrosine phosphorylation of Syk and phospholipase Cgamma2, intracellular Ca(2+) mobilization, and platelet aggregation occur later in a cAMP-sensitive manner. Cytochalasin D, which interferes with actin polymerization and blocks receptor clustering, inhibits all the rhodocytin-mediated signals we examined in this study. We suggest that rhodocytin, by clustering GPIa/IIa, activates GPIa/IIa-associated Src, which then mediates downstream activation signals.     

Cytotoxic effects of β‐asarone on Sf9 insect cells

β‐Asarone is the predominant component of the essential oil of rhizomes of Acorus calamus Linn ( Sweet flag). Although rhizome extracts from this plant have long been used for insect pest control, their cytotoxic effects on insect cells are not well understood. In this study, we evaluated the potency of β‐asarone as a natural insecticide by using a Spodoptera frugiperda cell line (Sf9). To assess the cytotoxic effects of β‐asarone on Sf9 cells, we observed morphologic changes in treated cells and performed a cell proliferation assay and a DNA fragmentation assay. After 24 and 48 h of treatment with β‐asarone, the proliferation of the Sf9 cells was inhibited in a dose‐dependent manner, with IC₅₀ values of 0.558 mg/ml at 24 h and 0.253 mg/ml at 48 h. Morphologic changes in β‐asarone‐treated cells were typical of apoptosis and included loss of adhesion, cell shrinkage, and small apoptotic bodies. The DNA laddering present in β‐asarone‐treated SF9 cells and annexin V assay confirmed that this compound can induce apoptosis in insect cells. Together, these findings suggest that apoptosis induction may be one mechanism through which β‐asarone inhibits the proliferation of insect cells and thus exerts insecticidal effects.     

Visual recognition of the host in the parasitoid fly Exorista japonica

We presented the tachinid fly Exorista japonica with moving host models: a freeze-dried larva of the common armyworm Mythimna separata, a black rubber tube, and a black rubber sheet, to examine the effects of size, curvature, and velocity on visual recognition of the host. The host models were moved around the fly on a metal arm driven by motor. The size of the larva, the velocity of movement, and the length and diameter of the rubber tube were varied. During the presentation of the host model, fixation, approach, and examination behaviours of the flies were recorded. The fly fixated on, approached, and examined the black rubber tube as well as the freeze-dried larva. Furthermore, the fly detected the black rubber tube at a greater distance than the larva. The rubber tube elicited higher rates of approach and examination responses than the rubber sheet, suggesting that curvature affects the responses of the flies. The length, diameter, and velocity of host models had little effect on response rates of the flies. During host pursuit, the fly appeared to walk towards the ends of the tube. These results suggest that the flies respond to the leading or trailing edges of a moving object and ignore the length and diameter of the object.     

Production of soluble integrin alpha2beta1 heterodimer complex functionally active in vitro and in vivo.

Integrin alpha2beta1, which is a membrane protein consisting of noncovalently bound alpha2 and beta1 chains, mediates cell binding to collagen and plays a role in platelet functions. DNAs encoding the chimeric proteins in which the extracellular domains of each alpha2 and beta1 chain was fused to hinge and Fc regions of human IgG(1)gamma chain were cotransfected into CHO cells. Soluble integrin alpha2beta1 (salpha2beta1) in which alpha2 and beta1 chains were covalently bound by disulfide bonds was recovered from the culture supernatant. salpha2beta1 maintained functional characteristics of cell surface alpha2beta1 as indicated by cation-dependent binding to collagen and conformational changes induced by cations or ligand. Intravenously administered salpha2beta1 in rats colocalized with collagen in inflamed microvessels. Moreover, salpha2beta1-conjugated liposome administered intravenously reduced bleeding time of the thrombocytopenic mice. These results indicated that salpha2beta1 has pharmaceutical utilities as an agent for detecting injured vessels and a component of platelet substitute.     

Behavior of the tachinid parasitoid <Emphasis Type="Italic">Exorista japonica</Emphasis> (Diptera: Tachinidae) on herbivore-infested plants

Odors from corn plants infested with the larvae of the noctuid herbivore Mythimna separata (Walker) attract tachinid fly, Exorista japonica Townsend, females; and odors from corn plants artificially damaged also attract this fly. We investigated the responses of flies to herbivore-infested, artificially damaged, and undamaged plants before and after arrival at the target plants to measure timing of the behavioral sequence. The behavior of the flies for 10 min after takeoff from a release point was observed in a wind-tunnel bioassay. The percentage of flies attracted to the plants was higher in infested and artificially damaged plants than in undamaged plants. Latency before takeoff was longer in undamaged plants, and time from takeoff to arrival at the plant was also longer in undamaged plants. Moreover, flies stayed longer on infested and artificially damaged plants. Flies walked longer on infested plants than on artificially damaged and undamaged plants. In this paper, we summarize the behavioral data and discuss the host-searching behavior of E. japonica females.     

Delayed Response after Learning Associated with Oviposition Experience in the Larval Parasitoid,Cotesia kariyai (Hymenoptera: Braconidae)

Journal of Insect Behavior - The ability of female parasitoid wasps, Cotesia kariyai (Hymenoptera: Braconidae), to learn to recognize plant volatiles associated with an oviposition experience was...     

Laboratory rearing of Lytopylus rufipes (Hymenoptera: Braconidae: Agathidinae), a parasitoid wasp of the oriental fruit moth,Grapholita molesta (Lepidoptera: Tortricidae), using apple and a commercially available diet

Lytopylus rufipes (Nees) (Hymenoptera: Braconidae: Agathidinae) is a potential natural enemy of the oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), but there is no established method to rear this wasp continuously. In the laboratory, female wasps can produce both female and male progenies without mating (deuterotokous), but host-infested plants are necessary to trigger oviposition behavior. In this study, immature apples were used because they keep well. Grapholita molesta larvae were transferred to immature apples, and then exposed to L. rufipes females. After parasitization, these apples were transferred to blocks of artificial diet (Silkmate 2M) for further rearing. Using this transitional diet system, L. rufipes females develop in 25.1?±?1.8 (mean?±?SD) days from egg to adult, but male wasps require only 23.8?±?1.0 days. Furthermore, the longevity of female wasps was 12.2?±?7.3 (mean?±?SD) days, and the parasitism rate was 26.4 (95% Cl: 22.2–30.6). Female wasps can attack host larvae in all instars, but parasitism of first-instar larvae is more successful. Therefore, first-instar larvae of G. molesta are recommended for establishing a L. rufipes colony under laboratory conditions. This system decreases the requirement for plant material and maintains continuous production of L. rufipes.