Propagation of the Colorado potato beetle [Col.: Chrysomelidae] egg parasiteEdovum puttleri, [Hym.: Eulophidae]

The parasitic wasp,Edovum puttleri Grissell, was successfully reared onLeptinotarsa decemlineata (Say) in the laboratory and increased in sufficient numbers for inoculative release studies in the spring and summer. The effects of temperature on parasite development, and host-parasite ratios in the laboratory are presented. The sex ratio of parasites reared in the laboratory are compared to those released and recovered in the field.     

Parasitism ofLeptinotarsa decemlineata [Coleoptera: Chrysomelidae] byEdovum puttleri [Hymenoptera: Eulophidae] in different cultivars of eggplant

Experiments were conducted to quantify parasitism of Colorado potato beetle,Leptinotarsa decemlineata (Say), by the egg parasitoid,Edovum puttleri Grissell, on 3 different cultivars of eggplant,Solanum melongena L. Levels of parasitism were higher (P<0.05) on ‘Black Pride’ than on other cultivars. The percentage of egg masses that were parasitized was 1.2-fold higher (P<0.05) on ‘Black Pride’ than on ‘Harris Special’ and ‘White’. The number of eggs per mass that were parasitized was 1.3- and 1.4- fold greater (P<0.05) on ‘Black Pride’ than on ‘Harris Special’ and ‘White’, respectively. The percentage of eggs that were parasitized per mass and percentage of emerged adult parasitoids did not differ (P>0.05) among cultivars; between 2.1- to 2.6- fold more females than males emerged from eggs on all cultivars during the growing season.Edovum puttleri suppressed the 2nd generation ofL. decemlineata on ‘Black Pride’ and ‘Harris Special’, but did not suppress populations on ‘White’.      

Edovum puttleri [Hym.: Eulophidae]: Biological activity and responses to normal and frozen eggs ofLeptinotarsa decemlineata [Col.: Chrysomelidae]

Biological control of eggplant pests in general, and (CPB)Leptinotarsa decemlineata (Say) in particular, is highly problematic.Edovum puttleri Grissell, an exotic egg parasitoid which has been found potentially effective, is being reared to this end. Frozen CPB eggs (?18°C) proved to be accepted and suitable for parasitization byE. puttleri. However, when given a choice between 20 frozen and 20 normal eggs every 2 days, the females exhibited a significant preference for the latter. Parasitization and host predation patterns related to female age are described by 2 opposing fourth-order polynomial functions. Parasitization increased up to day 10, followed by progressive decline. Average life span per female was 31.10±.58, with a recorded maximum of 56 days. High predatory behaviour of older females, which showed a decrease in parasitization, was observed in these experimental conditions.     

Incidence of natural enemies of the Colorado potato beetle,Leptinotarsa decemlineata [Coleoptera: Chrysomelidae] on a native host in Mexico

In central Mexico, the center of origin forLeptinotarsa decemlineata (Say) and its principal host plants,L. decemlineata is associated with a diverse complex of natural enemies. Observations during 2 years in Morelos, Mexico revealed 4 species of asopine pentatomids, 7 species of foliar searching carabids, and 2 coccinellids among the predators ofL. decemlineata. Parasitoids include 3 species of tachinids andEdovum puttleri Grissel, an eulophid egg parasitoid. These natural enemies include several predators that have not previously been reported attackingL. decemlineata. University of Maine Agriculture Experiment Station No. 1553     

Reproductive biology of two biotypes of Edovum puttleri, a parasitoid of Colorado potato beetle eggs

Emergence of adults of Edovum puttleri Grissell (Hymenoptera: Eulophidae) has a diel periodicity: most emergence occurred within 2 h of lights on (16L:8D, 24±1°C). Males emerged 2.0±3.0 h prior to females, and soon thereafter mated with emerging females. Courtship behavior did not differ between the Colombian and Mexican biotypes and was similar to that of members of the eulophid subfamily Tetrastichinae. The preoviposition period and oviposition period did not differ between biotypes. Males and females of both biotypes were long-lived, with the longevity of females of the Mexican biotype being significantly longer than that of Colombian females. males of both biotypes lived equally long. Mexican females produced more offspring than Colombian females, although this difference was not significant. After approximately 3 weeks, production of female offspring declined in both biotypes. A single mating was sufficient for females to produce a full complement of female offspring. Head capsule width of females correlated positively with fecundity.

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Résumé L'émergence des adultes de E. puttleri Grissell (Hymen.: Eulophidae) présente une périodicité quotidienne, la plupart de émergences ayant lieu dans les deux premières heures de la photophase (16L/8D, 24°±1°C). Les mâles apparaissent 2±3 h avant les femelles, et s'accouplant immédiatement avec les femelles émergentes. Les biotypes mexicains et columbiens ont le même comportement de cour, qui correspond à celui décrit pour les Tetrastichini et pour les Entedontinae. Il en est de même pour la durée, d'une part des périodes précédent la ponte (3,7 ±2,8 j), et d'autre part de la ponte elle-même (25,1 ±4,5 j). Les adults des deux biotypes vivent longtemps, mais les femelles mexicaines vivent plus longtemps (57,4±18 j) que les colombiennes (46,2±14,8 j); les longévités des mâles sont identiques (45,7 ±18 j). Les femelles mexicaines produisent apparemment plus d'oeufs (221,2±29,5) que les colombiennes (166,6±58,1), mais ces différences ne sont pas significatives. Après trois semaines environ la production de descendants régresse chez les deux types. Un seul accouplement suffit à produire le contingent total de filles. La largeur de la capsule céphalique des femelles est liée positivement à leur fécondité.

     

Intraspecific mitochondrial DNA variation in the Colorado potato beetle,Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

Restriction endonuclease analyses of mitochondrial DNA (mtDNA) were used to examine genetic variability and population structure inLeptinotarsa decemlineata (Say). A group of three enzymes, EcoRI, HpaI, and PstI, was used to reveal polymorphism both within and among some of the 10 populations tested, yielding 16 haplotypes in combination. The frequencies of these 16 haplotypes differed significantly across geographic regions, indicating some partitioning of mtDNA haplotypes. Estimates of mtDNA sequence divergence (δ) between haplotypes ranged from 0.016 to 0.135%, suggesting local differentiation of mtDNA in some populations. Analysis of these data suggests that Texas was colonized by more than one mtDNA lineage, most likely originating in Mexico. We hypothesize that a larger founder size for the initial introductions or high levels of variability in the parent population at the edge of the CPB expanding range led to the initial partitioning of haplotypes observed in samples from Texas.     

Oviposition behavior of Edovum puttleri, reared on two hosts, Leptinotarsa decemlineata and L. texana

Female Edovum puttleri Grissell [Hymenoptera: Eulophidae], reared from eggs of Leptinotarsa decemlineata (Say) or Leptinotarsa texana Schaeffer [Coleoptera: Chrysomelidae], were videotaped as they attacked egg masses of L. decemlineata containing 20 host eggs. We identified 15 components of ovipositional behavior. Parasitoids reared on L. texana attacked and oviposited in significantly more host eggs than did females reared on L. decemlineata. Ethometric analyses of behavioral transitions and a clustering analysis of 34 behavioral parameters showed that females reared on L. texana attacked the host egg mass in a different manner than those reared from L. decemlineata. It was concluded that differences were associated with the host species upon which they were reared. Contrary to previous reports, mortality of unparasitized hosts was caused by an ovipositor probe of short duration, which was not related to host-feeding.     

Sperm utilization and post-copulatory female-guarding in the Colorado potato beetle, Leptinotarsa decemlineata

Crosses between white and black color morphs of the Colorado potato beetle, Leptinotarsa decemlineata Say, used as genetic markers, showed that male beetles ride and guard females to achieve the three copulations required to fill the spermatheca. Multiple matings are necessary for the females to realize their full reproductive potential. Sperm mixing occurs in the spermatheca providing partial sperm precedence. The data support the competitive mate searching theory.

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Résumé Différentes combinaisons d'accouplements entre des mutants noirs et blancs du doryphore de la pomme de terre, Leptinotarsa decemlineata, servirent à étudier le comportement de guarde de la femelle par le mâle après un premier accouplement. Les résultats indiquent que le sperme de deux mâles accouplant la même femelle se mélange dans la spermathèque assurant ainsi un certain niveau de précédence du sperme du dernier mâle.La masse de sperme dans la spermathèque était inférieure à sa capacité chez les femelles ayant été accouplées une ou deux fois. Le doryphore mâle aurait donc avantage à demeurer sur la femelle après un premier accouplement afin d'obtenir le minimum de trois accouplements requis pour remplir la spermathèque avant que les autres mâles puissent accoupler cette femelle. Les autres mâles ont avantage à essayer de dérober cette femelle pour remplir eux mêmes la spermathèque ou tout au moins profiter d'une certaine précédence du sperme du dernier mâle pour fertiliser une partie des oeufs.Sans accouplements multiples, la femelle ne possède pas suffisamment de sperme pour féconder tous ses oeufs. II semble aussi que les accouplements répétés puissent augmenter sa fécondité.Les résultats supportent une stratégie reproductive de compétition plutôt qu'une simple maximisation du nombre d'accouplements.

     

Host-habitat conditioning in the eulophid egg parasitoid, Edovum puttleri: effects of herbivore's host and its food plant

The egg parasitoid, Edovum puttleri Grissell, was cultured from eggs of two Leptinotarsa hosts, L. decemlineata (Say) and L. texana Schaeffer, and experienced on eggs from three L. decemlineata food plants, namely potato, Solanum tuberosum tuberosum L., eggplant, S. melongena L., and tomato, Lycopersicon esculentum Mill. Host preference of the various cultures was determined using choice tests. parasitoid choices were influenced by the Leptinotarsa rearing host. Wasps, reared from L. decemlineata eggs from a mixture of all three food plants and experienced on egg masses from all three food plants, preferred to parasitize egg masses from potato. Wasps, reared from L. texana eggs from silver leaf nightshade, S. elaeagnifolium L., and experienced on egg masses from all three food plants, preferred to parasitize egg masses from tomato. Choices were also influenced by the herbivore's food plant that wasps were conditioned to. Wasps, reared from L. texana and experienced on egg masses from potato, preferred to parasitize egg masses from potato. Wasps, reared from L. decemlineata on potato and experienced on egg masses from potato, preferred to parasitize egg masses from potato. Wasps, reared from L. decemlineata on tomato and experienced on egg masses from tomato, did not display a distinct host preference. Within-egg mass parasitism (i.e., number of eggs per mass that were parasitized, probed, etc.) did not consistently differ among the three food plants. The implications of these data in parasitoid ecology and biological control programs for L. decemlineata are discussed.

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Résumé L'oophage, E. puttleri a été élevé à partir des oeufs de Leptinotarsa decemlineata et L. texana. Il a été mis en expérience sur l'une des 3 plantes consommées par L. decemlineata: Solanum tuberosum tuberosum, S. melongena et Lycopersicon esculentum. Des expériences de choix ont permis de déterminer ses préférences pour les différentes catégories d'oeufs. Le choix de l'oophage est influencé par l'espèce de Leptinotarsa sur laquelle il a été élevé. E. puttleri élevé sur oeufs de L. decemlineata récoltés sur un mélange des 3 solanées, et mis en présence d'un choix de groupes d'oeufs provenant de ces 3 plantes, a parasité préférentiellement les oeufs récoltés sur S. t. tuberosum; élevé à partir d'oeufs de L. texana récoltés sur S. elaeagnifolium, et mis en présence d'un mélange de pontes des 3 solanées, il a préféré parasiter les oeufs provenant de L. esculentum. Son choix a été influencé aussi par la plante qui l'avait conditionné. E. puttleri, élevé à partir de L. texana et mis en expérience sur des pontes provenant de S. t. tuberosum a préféré pondre sur les oeufs provenant de cette solanée. De même, élevé à partir d'oeufs de L. decemlineata pondus sur S. t. tuberosum, il a préféré pondre sur les oeufs provenant de cette plante. Cependant, les oophages élevés à partir d'oeufs de L. decemlineata provenant de L. esculentum, et mis en présence de pontes provenant de L. esculentum, n'ont pas préféré significativement ces oeufs au détriment de ceux provenant des autres solanées, bien que les pontes récoltées sur L. esculentum aient été parasitées plus fréquemment quand 2 des 3 pontes (une de chaque plante) avaient été sondées ou parasitées. Le parasitisme à l'intérieur d'une ponte (nombre d'oeufs de la ponte parasités, sondés, etc) ne varie pas significativement avec la solanée d'origine. La discussion a porté sur les conséquences de ces observations sur l'écologie de l'oophage et sur la lutte biologique contre L. decemlineata.

     

Ultrastructure of the neurosecretory system of the Colorado potato beetle,Leptinotarsa decemlineata (Say)

Summary The ultrastructure of seven types of neurosecretory cells (NSC) in the medial and lateral groups of the protocerebrum is described. The differences among cell types established earlier by light microscopy parallel differences in size and appearance of the neurosecretory particles observed in electron micrographs. No relationship was found between the affinity for Gomori's paraldehyde fuchsin stain and the nature of the particles.The secretions of the A-, A₁-, and C-types of NSC of the medial group are characterized by electron-dense neurosecretory granules of 1250 Å dia., medium-dense granules of 2100 Å, and electron-lucent vesicles of 1700 Å, respectively. The L-type NSCof the lateral group contain smaller (1300 Å) or larger (1700 Å) neurosecretory granules. The medial B- and E-types of NSC and the lateral L_B-type contain granulated vesicles (1200 Å) of the same appearance. These cell types differ in other respects and most likely have separate functions.The author wishes to thank the Laboratory of Virology of the Agricultural University for the use of the electron microscope, Mr. J. Groenewegen and Miss J. van Rinsum for technical assistance, and Professor J. Lattin for correcting the English text. Part of the work has been done while the author was in the service of the Netherlands Organization for the Advancement of Pure Research (ZWO, grant 942-48), and the National Council for Agricultural Research (TNO).     

Antifeedant and toxic effects of drimanes on Colorado potato beetle larvae

Among the drimane compounds tested, the dialdehydes polygodial and warburganal were the most active as antifeedants against Colorado potato beetle larvae, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae), in a dual-choice assay with potato, Solanum tuberosum L., leaf discs. Lactones were less effective. Direct observations showed that decreased feeding on leaf discs treated with polygodial and warburganal was accompanied by increased locomotry activity. Topical application of these two compounds on the insect's cuticle decreased food intake of untreated leaf discs, indicating that besides deterrent effects, toxic properties of these molecules influence feeding behaviour.     

A comparison of food-related behaviour between geographic populations of the Colorado potato beetle (Coleoptera, Chrysomelidae), on six solanaceous plant species

Some components of the food-related behaviour (biting, feeding, walking on the plant or walking elsewhere, and resting) of two North American and three European Colorado potato beetle (CPB) populations, on six solanaceous plant species were monitored with the help of a microcomputer-based program, testing the hypothesis whether populations showed differences at the behavioural level to alternative host plants (S. melongena, S. dulcamara, andLycopersicon esculentum), as well as to non-host plants occasionally encountered (S. luteum, S. nigrum.). The few significant differences found were among populations of the same continent in 1) the number of activities monitored, 2) the duration of feeding periods, 3) the number of bites taken to the onset of feeding, 4) the ‘amount of behavioural plasticity’, and 5) the ‘behavioural diversity’. On the basis of two-way ANOVA analyses, no plant-by-population interactions were detected indicating that the CPB populations studied did not have specific relationships with any of the plant species tested. The lack of substantial differences in food-related behaviour was unexpected since considerable isolation in space and time exists between the populations occurring on the two continents.     

Structure-activity studies of modified citrus limonoids as antifeedants for Colorado potato beetle larvae, Leptinotarsa decemlineata

Limonin and ten structurally modified limonins were evaluated as antifeedants against 4th instar larvae of the Colorado potato beetle, Leptinotarsa decemlineata Say, in no-choice leaf dics assays. The epoxide and furan groups were shown to be essential structural requirements for high antifeedant activity.

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Résumé Le pouvoir phagodissuasif de la limonine et de 10 limonines dont la structure avait été modifiée a été évalué sur des disques de feuilles de Solanum tuberosum offerts à des larves du 4ème stade de L. decemlineata. Aucune activité dissuasive n'ayant été observée avec le tétrahydrolimonine ou le déoxylimonine, l'anneau furane et le groupe époxy sont donc nécessaires pour obtenir cette activité. Ni la réduction de la fonction 7-kéto, ni la rupture de l'anneau A de la limonine n'ont eu d'effêt sur cette activité. La réduction des fonctions 16-carbonyl et 7-kéto de la limonine ont diminué l'activité, mettant en évidence une action possible de l'anneau D-lactone dans l'activité phagodissuasive. Tandis que la déépoxydation de la limonine entraîne une perte d'activité totale, une restauration partielle de cette activité a pu être obtenu par une réduction ultérieure de la fonction 7-kéto.

     

Overwintering biology ofMyiopharus aberrans andMyiopharus doryphorae (Dipt.: Tachinidae) larval parasitoids of the colorado potato beetle (Col.: Chrysomelidae)

The overwintering biology ofMyiopharus aberrans (Townsend) andMyiopharus doryphorae (Riley), tachinid parasitoids of the Colorado potato beetle, was investigated. Sixty seven overwintered Colorado potato beetle adults were dug from the soil before beetle emergence in the spring of 1990 from a field that had been planted to potatoes the previous year. Five adultM. aberrans and two adultM. doryphorae flies were reared from seven beetles. This is the first record of howM. aberrans andM. doryphorae flies overwinter.      

A proctolin-like peptide and its immunocytochemical localization in the Colorado potato beetle,Leptinotarsa decemlineata

Summary Evidence is presented that neurons in the adult Colorado potato beetle contain a proctolin-like substance. By use of immunocytochemical methods the location of immunoreactive neurons in the central and stomatogastric nervous systems is described. No such neurons were found in the proto- and deutocerebrum or optic lobe. Few immunoreactive neurons are present in the tritocerebrum and numerous proctolin-immunoreactive neurons occur in all ventral ganglia and in the frontal ganglion. Two groups of neurosecretory cells in the suboesophageal ganglion contain a proctolin-immunoreactive substance. In these cells this material is co-localized with a bovine pancreatic polypeptide/FMRF amide-like substance and with a vasopressin/vasotocin/oxytocin-like substance. Proctolin-immunoreactive axon terminals were found on the musculature of the fore- and hindgut and of the vas deferens, and on some segmental muscles. Furthermore, proctolin-immunoreactive neurosecretory axon terminals were found in the corpus cardiacum. The proctolin-like substance may therefore function both as a neurotransmitter/neuromodulator and as a neurohormone. The presence of a proctolin-like substance was also demonstrated with a sensitive bioassay. On fractionation of extracts of the nervous systems of Leptinotarsa decemlineata with high performance liquid chromatography most of the proctolin-like bioactive material comigrated with authentic proctolin. This shows that a proctolin-like substance in this insect is very similar to, if not identical with, the known pentapeptide proctolin.     

Suppression of larval Colorado potato beetle growth and development by digestive proteinase inhibitors

Evidence is presented that the Colorado potato beetle (CPB) (Leptinotarsa decemlineata (Say)) depends, at least partially, on cysteine proteinases for protein digestion. Midgut homogenates of CPB larvae have a mildly acidic pH and exhibit major proteolytic activity in the mildly acidic pH range. This proteolytic activity is activated by reducing agents, is inhibited by E-64 (a specific cysteine proteinase inhibitor), and is not inhibited by serine proteinase inhibitors. In addition, consumption of E-64 treated potato leaves by CPB larvae at rates as low as 0.8 g/cm² of leaf tissue has a deleterious effect on larval growth and development.

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Résumé Les protéinases intestinales des larves de Leptinotarsa decemlineata sont en partie caractérisées par les pH limitant leur activité, et par l'effet d'inhibiteurs de protéinases de spécificités connues. Les protéinases ont été testées avec la méthémoglobine tritiée comme substrat et nous avons déterminé les taux relatifs de peptides radioactives TCA solubles, libérées, dans des conditions précises, pour des pH compris entre 2,0 et 12,0. Le taux le plus élevé d'activité protéolytique a été observé pour des pH 5,0 à 6,0, bien qu'il y ait eu une activité appréciable aux pH 4,0 et 8,0. Parmi les inhibiteurs examinés, le E-64, inhibiteur protéinase cystéine très sélectif, a été le plus efficace sur l'activité protéinase. La pepstatine, inhibiteur protéinase aspartique, a été actif, mais seulement dans une gamme plus moin de pH. Les inhibiteurs protéinase sérine, ont été pratiquement inactifs sur l'activité protéinase.Chez des larves de doryphores nourries de feuilles de pommes de terre traitées avec différentes concentrations de E-64 ou de pepstatine, la consommation de E-64 a retardé fortement la croissance larvaire et le développement, la pepstatine a provoqué un retard du développement plus limitée.Nos résultats suggèrent que le doryphore dépend, tout au moins partiellement, des protéinases cystéine pour l'assimilation des protéines.

     

Survival of the Colorado potato beetle on virus-infected tomato in relation to plant nitrogen and alkaloid content

Infection by tobacco mosaic virus improved the suitability of tomato, Lycopersicon esculentum Mill. for survival of Colorado potato beetle, Leptinotarsa decemlineata (Say), larvae. This improvement was due, at least in part, to the increase in total nitrogen content of virus-infected plants. The simultaneous increase in tomatine content had no discernable effect on L. decemlineata survival. Our results are consistent with the suggestions that virus infection may improve the suitability of partially resistant or non-preferred hosts, and that virus infection may facilitate the adaption of phytophagous insects to such marginal host plant species.

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Résumé Pour vérifier l'hypothèse selon laquelle une contamination par un virs végétal peut accroître la consommation de plantes délaissées ou partiellement résistantes, nous avons examiné les interactions entre Leptinotarsa decemlineata Say, Lycopersicon esculentum Mill., et le virus de la mosaïque du tabac (TMV). La survie des larves de L. decemlineata a augmenté avec la contamination par TMV; les teneurs en azote et en tomatine étaient toutes deux plus élevées dans les plantes contaminées par le virus. La survie a augmenté linéairement avec la teneur en azote, mais sans dépendre de la teneur en tomatine. L'influence positive pour L. decemlineata de la contamination par le virus peut être attribuée au moins en partie à la teneur en azote plus élevée. Nos résultats correspondent à l'hypothèse selon laquelle la consommation de plantes délaissées ou partiellement résistantes peut être augmentée par la contamination virale, et que cette contamination peut faciliter l'adaptation d'insectes phytophages sur des plantes marginalement consommables.

     

Effects of selected physical and chemical treatments of Colorado potato beetle eggs on host acceptance and development of the parasitic wasp, Edovum puttleri

Effects of various physical and chemical treatments of Colorado potato beetle [Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)] eggs on parasitization and development of the egg parasitoid Edovum puttleri (Hymenoptera: Eulophidae) were investigated. UV irradiation did not affect host acceptance but reduced host suitability for UV exposure times 90 min. Susceptibility of host eggs to UV irradiation varied with host age; eggs were most vulnerable to damage from irradiation at 12, 18, and 24 h post-oviposition. The rate of parasitization also was influenced by host age. Percent parasitization was greatest in freshly laid eggs and 24–30 h old eggs. Seventy-seven percent of host eggs frozen at –20 °C (5 min) were parasitized by E. puttleri, but extended exposure of eggs to –20 °C reduced both acceptance and suitability. Host eggs that had been washed with hexane (removal of kairomone and sticky layer) also were parasitized. After 5 min of washing, application of kairomone significantly increased the rate of parasitism (from 74.7% to 88.2%), but with longer periods of washing, kairomone application had no significant effect on percent parasitism. Thus, the sticky material(s) coating the egg did not appear to be essential for parasitization to occur. Our results provide effective methods and times for treating Colorado potato beetle eggs to maximize parasitization and development of E. puttleri.