Parasitism of Lacanobia oleracea (Lepidoptera,noctuidae) by the ectoparasitic wasp Eulophus pennicornis,results in the appearance of a 27 kDa parasitism-specific protein in host plasma

Little is known about the effects of ectoparasitoids and their secretions on the plasma protein profiles of their insect hosts. To address this deficit, a study has been made of the interactions between an ectoparasitic wasp, Eulophus pennicornis, and its host, the tomato moth, Lacanobia oleracea. In particular, the quantitative and qualitative effects of parasitism or the experimental injection of wasp venom on host plasma proteins were investigated. Results demonstrated that both treatments caused an initial increase in L. oleracea total plasma protein concentration up to day 5 of treatment, but whereas the protein concentration remained high in the experimentally envenomated group, a decrease towards day 8 occurred in parasitized insects. Parasitism was also associated with the appearance of a protein with an estimated molecular weight of 27 kDa. This protein first appeared on day 3 after parasitization and its levels subsequently increased. The protein was not detected in any of the unparasitized larvae (including all the various control groups) or in experimentally envenomated L. oleracea larvae. In addition, the appearance of this protein was not a non-specific result of nutritional deprivation, nor was it a general injury, stress, or infection induced protein. Its appearance was strictly associated with parasitism of L. oleracea by E. pennicornis and thus, it may be described as a parasitism-specific protein (PSP). The PSP has been partially purified using whole gel elution. Gel filtration and SDS PAGE indicated that it has a native molecular weight of 27 kDa and that it does not appear to aggregate to produce higher molecular weight molecules, nor dissociate into lower molecular weight subunits held together by disulphide or covalent bonds. The precise site of synthesis of the 27 kDa PSP is not yet known but some evidence leads us to speculate that it may be synthesised by the feeding E. pennicornis larvae and introduced into their host. This possibility is discussed in relation to previous work detailing the effects of parasitism on L. oleracea haemocyte morphology, function and viability, and the effects of endoparasitoids on host plasma proteins.     

Proteins synthesized and secreted by larvae of the ectoparasitic wasp, Eulophus pennicornis

A microscopic examination of Eulophus pennicornis larvae on their host Lacanobia oleracea, revealed that peristaltic waves travelled from the anterior to posterior end of the feeding wasp larvae, and vice versa. In addition, when wasp larvae were immersed in PBS in vitro, they released a variety of proteins, with molecular weights ranging from (at least) 14 to 200 kDa. Amongst these was a protein with an estimated molecular weight similar to that of the 27 kDa parasitism-specific protein (PSP) detected in plasma from parasitized L. oleracea [Richards and Edwards, Insect Biochem Mol Biol 29:557-569 (1999)]. Similar results were obtained when the wasp larvae were incubated on balls of cotton wool soaked in tissue culture medium or sucrose, i.e., conditions that resemble their natural feeding behaviour. These results (and others) indicate that the wasp larvae release proteins, putatively through their mouth. Protein synthesis studies using (35)S-methionine indicated that the wasp larvae synthesize and secrete a variety of proteins in vitro, including one with a molecular weight corresponding to that of the L. oleracea 27 kDa PSP. As expected, only a portion of the total proteins synthesized by the parasitoid larvae were subsequently secreted. In addition, the autoradiogram of secreted proteins contained significantly fewer bands than silver-stained SDS gels of proteins released into PBS or onto cotton wool. Thus, some of the additional bands detected on the latter gels are thought to represent proteins that were not of wasp origin. Instead, these proteins released by the wasp larvae are speculated to be derived from their gut and, as such, probably represent proteins derived from host haemolymph and ingested during feeding. This possibility was supported by an electrophoretic analysis of homogenate supernatants prepared from wasp larvae with or without their gut contents. These studies indicated that the gut contents of the larval parasitoid contributes several distinct bands to the total protein profile. The ability of E. pennicornis larvae to synthesize, secrete, and release proteins is discussed with reference to those produced by endoparasitoid larvae. Published 2001 Wiley-Liss, Inc.     

Host regulation and release of parasitism-specific proteins in the system Toxoneuron nigriceps-Heliothis virescens

The braconid wasp Toxoneuron nigriceps induced qualitative and quantitative changes in the protein composition of the moth Heliothis virescens host hemolymph. Total protein concentration was found to be higher in parasitized host 4 days after parasitism as compared to control hosts, mainly due to changes in a particular group of proteins. Host proteins with a molecular mass of 173 and 72 kDa were found in higher levels in the hemolymph of parasitized larvae as control hosts approached pupation, while an 80 kDa peptide was found in reduced concentration in the hemolymph of parasitized hosts. Levels of these three peptides were maintained throughout parasitoid development, while two of them (173 and 72 kDa) were cleared from the host hemolymph close to pupation. Besides the regulation of host proteins, three parasitism-specific proteins (PSPs) were released into the host hemolymph. Two of them (PSP1-MW=116 kDa, pI=6.3; PSP2-MW=114 kDa, pI=6.2) first appeared in the hemolymph of parasitized hosts soon after pupation of control host and increased in concentration as the parasitoid developed. The third PSP (PSP3-MW=56 kDa, pI=5.8) was produced towards the end of parasitoid larval development, close to parasitoid egression. Database searches based on the amino acid composition and amino terminal sequence of PSP1 and PSP2 did not produce any significant matches, while PSP3 was identified as a putative chitinase. Incubation of host derived tissues, parasitoid larvae and teratocytes in 35S conditioned media suggested PSPs were a product of teratocytes. The role of the regulation of host proteins and release of PSPs by teratocytes for the successful development of T. nigriceps are discussed.     

The effect of snowdrop lectin (GNA) delivered via artificial diet and transgenic plants on Eulophus pennicornis (Hymenoptera: Eulophidae), a parasitoid of the tomato moth Lacanobia oleracea (Lepidoptera: Noctuidae)

Snowdrop lectin (Galanthus nivalis agglutinin, GNA) has previously been shown to confer significant levels of protection against the lepidopteran pest Lacanobia oleracea when expressed in transgenic potato. The effect of GNA on the parasitism of L. oleracea by the gregarious ectoparasitoid Eulophus pennicornis was investigated. Maize-based, and potato leaf-based diets containing GNA, and excised transgenic potato leaves expressing GNA, were fed to L. oleracea larvae from the beginning of either the third or fourth larval instar. Lacanobia oleracea larvae were individually exposed to single mated adult female E. pennicornis parasitoids from the fifth instar onwards.The success of the wasp was not reduced by the presence of GNA in any of the diets, or by the length of feeding of the host prior to parasitism. However, the mean number of wasps that developed on L. oleracea reared from the third instar on the GNA-containing maize diet was significantly higher than on the controls (20.6 and 9.3 adults/host respectively). In all other cases differences were not significant. Eulophus pennicornis progeny that developed on L. oleracea reared on GNA-containing diets showed little or no alteration in size, longevity, egg load and fecundity when compared with wasps that had developed on hosts fed the respective control diets.The results suggest that expression of GNA in transgenic crops to confer resistance to lepidopteran pests will not adversely affect the ability of the ectoparasitoid E. pennicornis to utilise the pest species as a host.     

Parasitism of Lacanobia oleracea (Lepidoptera) by the ectoparasitoid, Eulophus pennicornis, is associated with a reduction in host haemolymph phenoloxidase activity

When haemolymph from fifth instar Lacanobia oleracea was incubated in vitro, rapid melanization occurred. Similar levels of melanization occurred in haemolymph from larvae that had been experimentally injected with venom from the ectoparasitic wasp, Eulophus pennicornis. In contrast, haemolymph from larvae parasitized by this wasp melanized more slowly and less extensively. Phenoloxidase assays indicated that enzyme activity was present in haemocyte lysate supernatants, serum and plasma from L. oleracea and that on day 5 post-parasitization, fractions prepared from parasitized larvae had significantly less phenoloxidase activity than similar fractions from untreated or experimentally envenomated larvae. In addition, no PO activity was detectable in wasp venom, and the venom had no effect on L. oleracea plasma phenoloxidase activity in vitro. These results indicate that parasitism of L. oleracea by E. pennicornis suppresses host haemolymph phenoloxidase activity and that this suppression is not induced by adult wasp venom. The results are discussed with reference to the survival advantages of suppressing the activity of this host enzyme, and to the possible source(s) of putative suppressive factors.     

Expression of a parasitism-specific protein in lepidopteran hosts of Chelonus sp.

The hemolymph of each noctuid species successfully parasitized by Chelonus near curvimaculatus possessed a parasitism-specific protein (PSP) previously identified in host T. ni (Insect Biochem. 19:445; 21:845). Expression of PSP occurred in a stage-specific manner in the stadium during which the host undergoes precocious metamorphosis. The appearance of the protein was not due to nutritional stress associated with parasitism of hosts, since starved nonparasitized larvae did not produce the protein, or to low juvenile hormone titers occurring in precociously metamorphosing hosts, but rather was dependent on the presence of the endoparasite larva. Results of in vivo incorporation experiments with [³⁵S]-methionine showed that synthesis and subsequent appearance of the protein in the hemolymph of parasitized hosts was abrogated by prior surgical removal of endoparasite. Immunoprecipitation analysis of proteins from C. near curvimaculatus larvae cultured in vitro using antibodies specific to PSP indicated that the source of the protein was the endoparasite. Synthesis of PSP by the endoparasitic larvae with its subsequent secretion into the hemocoel of hosts was specific to the advanced stages of parasite development prior to its egression from the host. © 1993 Wiley-Liss, Inc.     

Effects of Galanthus nivalis agglutinin (GNA) expressed in tomato leaves on larvae of the tomato moth Lacanobia oleracea (Lepidoptera: Noctuidae) and the effect of GNA on the development of the endoparasitoid Meteorus gyrator (Hymenoptera: Braconidae)

The effect of ingestion of transgenic tomato leaves expressing the plant lectin Galanthus nivalis agglutinin (GNA) on development of larvae of Lacanobia oleracea (Linnaeus) was studied under laboratory conditions. When L. oleracea larvae were fed on tomato line 14.1H, expressing approximately 2.0% GNA, significant increases in the mean larval weight and in the amount of food consumed were found. This resulted in an overall reduction in the mean development time to the pupal stage of approximately 7 days. A significant increase in the percentage survival to the adult moth was also recorded when newly hatched larvae were reared on transgenic tomato leaves (72%) compared to larvae reared on untransformed leaves (40%). The effects of ingestion of GNA by L. oleracea larvae, via artificial diet or the leaves of transgenic tomato or potato plants, on the subsequent development of its solitary endoparasitoid Meteorus gyrator (Thunberg) was also studied. No significant effects on the life cycle parameters of M. gyrator developing in L. oleracea fed on GNA-containing diets were observed. Experiments with transgenic potato plants indicated that the stadium of the host larvae at parasitism had a greater influence on M. gyrator development than the presence of GNA. Potential GNA-binding glycoproteins were detected in the gut and body tissues of larval M. gyrator. Despite detection in host tissues, GNA could not be detected in adult M. gyrator and therefore it is likely that at the time of pupation M. gyrator are able to void the GNA in the meconial pellet.     

Effect of microsporidian infection in Lacanobia oleracea (Lep., Noctuidae) on prey selection and consumption by the spined soldier bug Podisus maculiventris (Het., Pentatomidae)

Abstract:  The predatory behaviour of Podisus maculiventris was investigated when this bug was presented with Lacanobia oleracea larvae infected with the microsporidian pathogen Vairimorpha necatrix . In choice tests, adult predatory bugs attacked V. necatrix -infected L. oleracea prey in similar numbers to uninfected larvae. Exposure to infected prey during nymphal development increased the rate at which adult bugs attacked diseased L. oleracea larvae. Fifth instar P. maculiventris nymphs, however, attacked infected prey in the majority of cases (>80% of occasions). Consumption of healthy and infected prey was measured for both adult and nymphal bugs. Over the course of 1 week, the mean number of V. necatrix -infected prey eaten by P. maculiventris adults (7.0 ± 0.82) was approximately twice the number of uninfected prey consumed (3.8 ± 0.42). Similarly, the number of prey larvae attacked by the bug over the course of the final nymphal stadium was also increased, with 2.9 ± 0.42 uninfected larvae eaten as opposed to 4.9 ± 0.27 V. necatrix -infected prey. However, small-scale investigations into the rate of P. maculiventris reduced small populations of L. oleracea indicated that the combination of the predator and pathogen would produce, at best, an additive effect.     

Parasitism of Lacanobia oleracea (lepidoptera) by the ectoparasitic wasp, Eulophus pennicornis, disrupts the cytoskeleton of host haemocytes and suppresses encapsulation in vivo

Parasitism of Lacanobia oleracea larvae by the ectoparasitic wasp Eulophus pennicornis suppressed host haemocyte-mediated encapsulation of Sephadex DEAE A-25 beads in vivo. Beads dissected out of parasitized larvae had fewer haemocytes associated with them. Moreover, those haemocytes that were associated with the beads tended to retain a rounded configuration and rarely flattened. Similar results were obtained using in vitro encapsulation assays. SDS PAGE indicated that for parasitized and PBS injected larvae, there were some differences in the plasma proteins that bound to Sephadex DEAE A-25 beads, suggesting that parasitism-mediated changes to host plasma proteins might contribute to the differences in the encapsulation response occurring in these larvae. However, in vitro encapsulation assays using beads that had been pre-incubated in plasma from parasitized and unparasitized larvae, demonstrated that major differences in the extent of encapsulation did not occur. These results, plus in vitro haemocyte attachment and spreading assays, suggest that parasitism-mediated suppression of encapsulation is primarily due to reductions in the ability of host haemocytes to attach to (i.e., recognize) and flatten over non-self surfaces and other haemocytes. This proposal is corroborated by staining of actin in the haemocyte cytoskeleton by FITC-labelled phalloidin, which indicated that parasitism disrupts the formation of stress fibers and focal adhesions in plasmatocytes. By contrast, experimental injection of adult female wasp venom into unparasitized L. oleracea larvae had no significant effect on in vivo encapsulation responses or the haemocyte cytoskeleton. Arch. Insect Biochem. Physiol. 49:108-124, 2002. Published 2002 Wiley-Liss, Inc.     

Seasonal phenology and stage-specific parasitism of the apple ermine moth, Yponomeuta malinellus Zeller, in Korea

The apple ermine moth, Yponomeuta malinellus Zeller (Lepidoptera: Yponomeutidae), is a tent caterpillar that feeds on Malus spp. in Korea. Populations of the moth in native areas appeared to be regulated by the assemblage of parasitoids. Phenological associations between host stages and parasitoids, susceptible stage(s) of the host for each parasitoid, and stage‐specific parasitism were studied. The egg larval parasitoid Ageniaspis fuscicollis (Dalman) had highest parasitism of first instar larvae (24%), with 14% parasitism of other larval stages. Dolichogenidea delecta (Haliday) was recovered from all larval instars with the highest parasitism rate of second instar larvae (20.1%), followed by 19.9% parasitism of mid‐larval hosts. Herpestomus brunicornis Gravenhorst was reared from second instar larvae through to pupal collection, and had the highest parasitism rate (29.9%) at the pupal stage. The larval pupal parasitoid Zenillia dolosa (Meigen) was recovered from mid‐larval to pupal stages with the highest parasitism rate (5.5%) occurring in third to fourth instar larvae. The host stages for developing A. fuscicollis completely overlap with those of D. delecta, and with those of H. brunicornis to some degree. A statistically significant negative correlation exists between A. fuscicollis and these dominant parasitoids, indicating competitive interaction within the host.     

The effect of host stage and temperature on selected developmental parameters of the solitary endoparasitoid Meteorus gyrator (Thun.) (Hym., Braconidae)

Abstract:  The development of the solitary endoparasitoid Meteorus gyrator was compared in the six larval stages of its host, the tomato moth Lacanobia oleracea , and at five constant temperatures. The host instar at the time of parasitism had a marked effect on the larval developmental period of the parasitoid, such that larvae derived from eggs oviposited in first instar hosts took approximately 18 days to egress, whilst those derived from eggs oviposited in sixth instar hosts took <10 days. The weight of cocoons was greatest when oviposition was into final instar hosts, where female cocoons averaged 12.8 mg, and lowest in those derived from eggs oviposited into first instars (9.2 mg). The parasitoid's larval development rate in third instar hosts increased with temperature increments in a linear fashion up to 25°C , after which development times were only marginally increased. At 10°C, the mean larval development time was approximately 90 days and pupal development 35–40 days, whilst at 25°C development times were 10–11 days for larvae and 6–7 days for the pupae. In the majority of cases, overall development times were marginally longer (<1 day) in females than in males.     

Gypsy moth parasitoids in the declining outbreak in Lithuania

A 3 year study was conducted on the parasitoids of gypsy moth larvae in two reducing outbreak areas in Lithuania. Overall parasitism of 25.0 ± 2.0% in the first post-culmination year was significantly lower than the 36.3 ± 1.4 and 35.2 ± 1.4% parasitism in the two subsequent years. When analysed in terms of the life stage at which the host was collected, the total parasitism over 3 years was constantly increasing from 3.1 ± 0.8 in the first to 72.5 ± 2.9% in the sixth instar. Parasetigena silvestris R.-D. dominated causing 48.7 ± 1.5% parasitism and 16.7 ± 0.6% larval mortality preferably in late instars. Phobocampe disparis Vier. contributed to 21.9 ± 1.2% parasitism and 7.5 ± 0.5% mortality recovering from early instar larvae. Meteorus pulchricornis Wes. parasitized 4.3 ± 0.6% gypsy moth larvae causing 1.5 ± 0.2% mortality and a few Apanteles species provided 2.8 ± 0.6% parasitism and 1.0 ± 0.2% mortality. The gypsy moth in Lithuania was reported to act as host for Rogas sp. (Hym., Braconidae) and Siphona boreata Mes. (Dipt., Tachinidae).     

Differential parasitism of Plutella xylostella (Lepidoptera: Plutellidae) larvae by the parasitoid Cotesia plutellae (Hymenoptera: Braconidae) on two host plant species

Laboratory experiments were conducted to examine host selection by Cotesia plutellae Kurdjumov when larvae of its host, Plutella xylostella (Linnaeus), fed on Chinese cabbage, Brassica campestris L. ssp. pekinensis and those fed on common cabbage, Brassica oleracea L. var. capitata were provided simultaneously, and to investigate the roles of plant and host volatiles in mediating host selection. When C. plutellae were provided with equal numbers of host larvae on plants of the two species in one arena, the parasitoid parasitized 4- to 15-fold more host larvae on Chinese cabbage than on common cabbage. This preference changed little with host density. However, an experience of searching coupled with an oviposition in a host larva on a leaf of the less-preferred plant, common cabbage, significantly increased the preference for parasitizing host larvae on this plant and resulted in twice as many host larvae parasitized on this plant than on Chinese cabbage. Dual choice tests with a Y-tube olfactometer showed that plant volatiles from Chinese cabbage were more attractive to female C. plutellae than those from common cabbage when plants of both species were either intact or infested. In parallel to the increased parasitism on common cabbage following experience, oviposition in a host larva on this less-preferred plant significantly increased the response to volatiles emanating from that plant. These results indicate that host plants may strongly influence the foraging behaviour of C. plutellae, but their differential attractiveness to the parasitoid may be altered by experience of the parasitoid.     

卵液成分改变及卵表涂施引诱剂对玉米螟赤眼蜂产卵发育的影响

试验通过部分改变人造卵卵液成分和在人造卵表涂施引诱剂2种途径来试图提高人造卵繁育玉米螟赤眼蜂Trichogramma ostriniae Panget Chen的成功率。结果表明,不同比例的亚洲玉米螟OstriniafurnacalisGuenée幼虫和蛹匀浆替代部分柞蚕蛹匀浆后,对赤眼蜂寄生率有明显提高,但赤眼蜂在人造卵内的化蛹率和羽化率均较低,赤眼蜂发育至老熟幼虫因卵液过剩而被淹死;在原卵液卵表涂施亚洲玉米螟卵和鳞片正己烷提取液后,评价繁蜂成功率的各项参数都有明显提高,赤眼蜂可以完成发育,羽化成蜂,但繁蜂成功率还较低。综合上述试验结果,应用人造卵繁殖玉米螟赤眼蜂研究重点要解决卵液过剩以及其它相关的营养学问题。     

Changes in haemolymph proteins of the fall armyworm, Spodoptera frugiperda (J. E. Smith), associated with parasitism by the braconid parasitoid Cotesia marginiventris (Cresson)

Changes in haemolymph proteins of the fall armyworm, Spodoptera frugiperda, associated with parasitism by the parasitoid Cotesia (= Apanteles) marginiventris were monitored by sodium dodecyl sulphate polyacrylamide gel electrophoresis. As early as hour 4 after parasitization treatment, several electrophoretically slow-migrating, high-molecular-weight proteins were detected in the host's haemolymph. These proteins were detected earlier in haemolymph from parasitized larvae than in haemolymph from control larvae, and their concentrations were higher in heavily parasitized host larvae (≥ 3 eggs/host) than in lightly parasitized larvae (1 egg/host). Additionally, unique proteins that migrated electrophoretically with bovine serum albumin appeared in the haemolymph of parasitized larvae at hour 8 after parasitization treatment and were evident in haemolymph collected through to hour 64.     

Alteration of hemolymph polypeptides in Manduca sexta larvae parasitized by Cotesia congregata: A two-dimensional electrophoretic analysis and comparison with major bacteria-induced proteins

Analyses using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) previously demonstrated that parasitization by the braconid wasp Cotesia congregata significantly alters the normal hemolymph polypeptide profile of host Manduca sexta larvae. In the present study two-dimensional gel analyses corroborated our earlier findings and provided additional evidence that multiple parasitism-specific polypeptides were induced, which varied according to the stage of development of the wasps. Parasitization additionally elicited changes in the total protein concentration detected in the blood. Initially an elevation was observed, with newly parasitized larvae exhibiting a twofold elevation in hemolymph protein concentration by 12–24 h postoviposition. In contrast, terminal-stage hosts with second instar parasites had significantly less protein in the hemolymph, likely due to reduced growth and inhibition of arylphorin synthesis by the fat body during the final stages of parasitism. Comparison of the array of hemolymph polypeptides produced in unparasitized larvae injected with 10⁶cells of the gram-negative bacterium Enterobacter cloacae with those proteins induced by parasitization indicated the two classes are different. Our findings confirm that the hostresponse to parasitism is a specific one, and not mimicked by bacterial challenge. Duringshort-term in vitro culture of wasp larvae dissected from the host hemocoel, several proteins were detected in the medium using SDS - PAGE, with their appearance in vitro suggestive of secretion by the wasps in vivo. Moreover, hemolymph from the parasites had significant amounts of putative host proteins, including an arylphorin - like polypeptide and a protein with a mobility similar to that of insecticyanin. Thus, a dynamic interchange of proteins may occur, with the parasites accumulating host proteins while simultaneously secreting a variety of factors into the host hemocoel.     

Effects of host-plant shift on immune and other key life-history traits of an eruptive Geometrid, Epirrita autumnata (Borkhausen)

Abstract.  1. Population density of Epirrita autumnata (Lepidoptera: Geometridae) reaches outbreak densities regularly in northernmost Scandinavia. During these outbreak years, the most abundant host species, the mountain birch ( Betula pubescens ssp. czerepanovii ), is regularly exhausted, although larvae may rescue themselves from starvation by using alternative host species.
2. In this paper, the effects of the shift of host species on the immune defence and other life-history traits of E. autumnata were investigated, and possible consequences for population dynamics were briefly discussed. Moth larvae were reared on the leaves of the main host, mountain birch, until larvae reached their third instar. After this, larvae were allocated randomly to five treatments: larvae were either allowed to finish larval stage on the mountain birch or were shifted onto four alternative host species that are typical species for the area.
3. As expected, the host species had a major effect on fitness traits: body weight, development, and survival rate of the moths. The pupal weight was lower and development rates slower on the three alternative host species, Salix myrsinifolia Salisb., Vaccinium uliginosum L., and Betula nana L., than on the main host, mountain birch.
4. The immunity was, however, the same or better on the alternative hosts than on the main host. The immunity and pupal weights were negatively related, suggesting a trade-off between body size and immunocompetence.
5. The decreased body size and fecundity of E. autumnata during outbreak years may be partly due to the shift to alternative host species whereas the host-plant species probably does not affect markedly the rate of parasitism.     

Influence of host plant nitrogen fertilization on hemolymph protein profiles of herbivore Spodoptera exigua and development of its endoparasitoid Cotesia marginiventris

Nitrogen has complex effects on plant–herbivore–parasitoid tritrophic interactions. The negative effects of low nitrogen fertilization in host plants on insect herbivores can be amplified to the higher trophic levels. In the present study, we examined the impact of varying nitrogen fertilization (42, 112, 196, and 280 ppm) of cotton plants (Gossypium hirsutum L.) on the interactions between the beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), and the hymenopteran endoparasitoid Cotesia marginiventris (Cresson) (Hymenoptera: Braconidae). We predicted that the development and fitness of C. marginiventris would be adversely affected by low host plant nitrogen fertilization through the herbivore S. exigua. The percentage of C. marginiventris offspring developing to emerge and spin a cocoon, and total mortality of parasitized S. exigua larvae were unaffected by nitrogen level. The developmental time of C. marginiventris larvae in S. exigua larvae feeding on low (42 ppm) nitrogen cotton plants was approximately 30% longer than that of those feeding on higher (112, 196, and 280 ppm) nitrogen plants. Parasitoid size (length of right metathoracic tibia), a proxy for fitness, of C. marginiventris males was positively affected by nitrogen level. Total amounts of S. exigua hemolymph proteins were not affected by nitrogen level, but were reduced by parasitism by C. marginiventris. Two proteins with molecular weights of ca. 84 and 170 kDa dominated the S. exigua larval hemolymph proteins. Concentrations of the 170 kDa hemolymph protein were unaffected by nitrogen treatment, but parasitism reduced concentrations of the 170 kDa protein. Concentrations of the 84 kDa protein, on the other hand, were interactively affected by parasitism and nitrogen treatment: higher nitrogen fertilization (112, 196, and 280 ppm) increased protein concentrations relative to the 42 ppm treatment for unparasitized S. exigua larvae, whereas nitrogen treatment had no effects on parasitized larvae. For S. exigua larvae feeding on 42 ppm nitrogen plants, parasitism increased concentration of the 84 kDa protein, while for those feeding on 112, 196, and 280 ppm nitrogen plants, parasitism decreased concentrations of the protein. Possible mechanisms and ecological consequences for the extended development of C. marginiventris on S. exigua hosts grown on low-nitrogen plants are discussed.     

Bottom-up cascading effects in a tritrophic system: interactions between plant quality and host-parasitoid immune responses

Abstract.  1. Little is known about underlying mechanisms by which plants indirectly affect parasitism success in hymenopteran endoparasitoids. The hypothesis that host-plant effects can challenge the innate immune system of an insect host was experimentally tested in this study using a model tritrophic, crucifer – lepidopteran [ Plutella xylostella (L.)] – parasitoid [ Cotesia plutellae (Kurdjumov)], system.
2. The effects of host-plant suitability on herbivore performance and parasitism were examined. The bottom-up effect of plant suitability on host-parasitoid immune responses was then evaluated using measures of cellular and humoral effectors.
3. Host-plant quality showed a significant effect on the encapsulation response of P. xylostella to first instar but not to second instar parasitoid larvae. Encapsulation was never sufficient to prevent parasitoid emergence.
4. Poor host-plant suitability suppressed phenoloxidase activity in the absence of the parasitoid. The suppressive effect of C. plutellae on phenoloxidase activity was much greater and no plant effects were detectable after insects had been parasitized.
5. Despite strong plant effects on parasitism, those on immune effectors of the host were transitory or overwhelmed by the effect of the parasitoid.
6. These results demonstrated that plant-mediated variation in parasitism success by C. plutellae were not as a result of plant nutritional status or other attributes affecting the immune function of P. xylostella , nor to host-plant effects on superparasitism.
7. In these experiments, P. xylostella was a fully permissive host to C. plutellae and host-plant-mediated effects on the innate immune response appeared to play no part in parasitoid survival within hosts.     

Molecular cloning and characterization of a cDNA encoding a novel cuticle protein from the Chinese oak Silkmoth, Antheraea pernyi.

In our research to identify gene involved in the cuticle protein, we cloned a novel cuticle protein gene, ApCP13, from the Chinese oak silkmoth, Antheraea pernyi, larvae cDNA library. The ApCP13 gene encodes a 120 amino acid polypeptide with a predicted molecular mass of 13 kDa and a pI of 4.01, and is intron-less gene. The ApCP13 contained a type-specific consensus sequence identifiable in other insect cuticle proteins and the deduced amino acid sequence of the ApCP13 cDNA is most homologous to another wild silkmoth, A. yamamai CP12 (86% protein sequence identity), followed by Bombyx mori LCP18 (35% protein sequence identity). Northern blot analysis revealed that the ApCP13 showed the epidermis-specific expression. This is the first report of cuticle protein gene in the wild silkmoth, A. pernyi.