THE SYSTEMIC INSECTICIDAL ACTION OF SODIUM FLUOROACETATE AND OF THREE PHOSPHORUS COMPOUNDS ON THE EGGS AND LARVAE OF PIERIS BRASSICAE L.

Four compounds, bis dimethylaminophosphonous anhydride ( anhydride ), bis (dimethylamino)fluorophosphine oxide ( oxide ), diethyl paranitrophenyl phosphate ( E 600) and sodium fluoroacetate ( acetate ), previously shown to have systemic insecticidal activity against aphids, have now been tested against the eggs and larvae of Pieris brassicae L.
The anhydride proved to have very little toxic action on Pieris , but the other three compounds showed both contact toxicity and systemic insecticidal action when taken up by the roots of cabbage plants from solution and from soil. The acetate, but more especially the E600, also showed systemic action following application to the leaves.
In all cases the order of decreasing toxicity was E600 > acetate > oxide > anhydride. E600 is the only compound which is outstandingly toxic to Pieris eggs and larvae. It has the added interest that when watered on to the roots of cabbage plants it kills larvae in egg batches on the leaves. Death takes place at the stage when the larvae are biting through the shells. The oxide and acetate proved to be surprisingly innocuous.
Unless it is felt to be too poisonous E600 is worthy of consideration as an insecticide against Pieris eggs and larvae, since it is highly effective as a contact insecticide and also has some systemic action.     

Different responses to temperature in three closely-related sympatric cereal aphids

Cotesia glomerata L. (Hymenoptera: Braconidae) is a parasitoid of early instar larvae of Pieris brassicae L. (Lepidoptera: Pieridae). Late instars of P. brassicae can more often overcome parasitization by hemocytic encapsulation of C. glomerata eggs. Short-term hemocyte responses to parasitization were examined in third and fourth instar larvae of P. brassicae. Total and differential hemocyte counts did not differ between parasitized and unparasitized host larvae. A rapid, but temporary decrease of total hemocyte as well as plasmatocyte numbers was observed immediately after oviposition. Numbers of hemocytes adhering to tissues were shown to be the same in untreated, wounded and parasitized P. brassicae larvae by tracing hemocytes with monoclonal antibodies as markers. The in vitro spreading ability of hemocytes from unparasitized third and fourth instar larvae was lower than that of the last instar's; parasitization, however, had no influence on hemocyte spreading. We therefore suggest that the higher parasitization success of C. glomerata in earlier instars of P. brassicae is mainly due to the low spreading ability of the hemocytes. Abbreviations: ACS – anticoagulant saline; BSA – bovine serum albumin; DABCO – 1,4-diazabicyclo-[2,2,2]-octane; DHC – differential hemocyte count; FITC – fluorescein isothiocyanate; GR – granular cells; LPS – lipopolysaccharide; mAb – monoclonal antibody; OE – oenocytoids; PL – plasmatocytes; PRO – prohemocytes; PS – Pieris saline; PVP – polyvinylpyrrolidone; TBS – tris-buffered saline; THC – total hemocyte count.     

Tronchuda cabbage flavonoids uptake by Pieris brassicae

The flavonoid pattern of larvae of cabbage white butterfly (Pieris brassicae L.; Lepidoptera: Pieridae) reared on the leaves of tronchuda cabbage was analysed by HPLC-DAD-MS/MS-ESI. Twenty flavonoids were identified or characterised, namely 16 kaempferol and four quercetin derivatives. Kaempferol 3-O-sophoroside, a minor component of tronchuda cabbage, was found to be the main component in P. brassicae (15.8%). Apart from this, only two other flavonoids present in significant amounts in tronchuda cabbage (kaempferol 3-O-sophoroside-7-O-glucoside and kaempferol 3-O-sophoroside-7-O-sophoroside) were found in the larvae. The larvae have high amounts of quercetin derivatives (18.5%), which were present only in trace amounts in tronchuda cabbage extracts, suggesting that P. brassicae is able to selectively sequester these flavonoids. The occurrence of a high content of flavonoids not detectable in tronchuda cabbage extracts indicates that P. brassicae larvae are able to metabolize dietary flavonoids.     

Bird predation as a selective pressure on the immature stages of the cabbage butterflies, Pieris rapae and P. brassicae

The importance of bird predation as a selective pressure on the eggs, larvae and pupae of the cabbage butterflies, Pieris rapae L. and P. brassicae L. has been investigated in a rural garden and an allotment. Birds are the main predators of all stages in a well-tended garden whereas in a field the eggs and young larvae are mainly preyed upon by arthropods. The species of avian predators vary according to the stage of development and species of prey. Consequently the relative survival rate of P. rupae and P. brussicue varies with stage of development. This changing pattern of survival has in turn acted as a selective pressure on their two main braconid parasites. Many aspects of the size, shape, colour and behaviour of the larvae and pupae of both species of Pieris appear to be adaptations to the selective pressures exerted by bird predation.     

Relationships between parasitoid host range and host defence: a comparative study of egg encapsulation in two related parasitoid species

Abstract. Parasitoid host range may proceed from traits affecting host suitability, traits affecting parasitoid foraging behaviour, or both. We tested the hypothesis that encapsulation can be used as a reliable indicator of parasitoid host range in two closely related larval endoparasitoids of Lepidoptera. Cotesia glomerata (L.) (Hymenoptera: Braconidae) is gregarious and a generalist on several species of Pieridae, whereas C. rubecula (Marshall) is solitary and specific to Pieris rapae (L.). We determined the effects of host species ( Pieris brassicae (L.), P. napi (L.) and P. rapae ) (Lepidoptera: Pieridae) and host developmental stage (early first, second and third instar) on encapsulation of parasitoid eggs. Host species and parasitoid species, as well as the resulting interaction between these two factors had significant effects on encapsulation of Cotesia eggs. Encapsulation in Pieris hosts was much lower for C. glomerata (<34%, except for second and third instar of P. rapae ) than for C. rubecula (>32%), even when the latter was parasitizing P. rapae. Encapsulation increased with the age of the larvae, although the only significant difference was for C. glomerata. Overall, P. rapae showed a stronger encapsulation reaction than P. brassicae and P. napi. Encapsulation levels of C. glomerata corresponded well to patterns of female host species and host age preference for oviposition and parasitoid larval performance. In contrast, percentages of encapsulation of C. rubecula were not consistent with host preference and host suitability. We argue that encapsulation alone is unlikely to provide a sufficient explanation for C. glomerata and C. rubecula host range.     

Superparasitism in Cotesia glomerata: response of hosts and consequences for parasitoids

Abstract.  1. Superparasitism occurs in Cotesia glomerata (Hymenoptera: Braconidae), a gregarious endoparasitoid of Pieris spp. (Lepidoptera: Pieridae). The response of P. brassicae larvae to superparasitism and the consequences for the parasitoid were examined in order to elucidate the ecological significance of this behaviour.
2. Field surveys of a Swiss population revealed that C. glomerata brood sizes from P. brassicae larvae ranged from three to 158, and both the female ratio and the body weight of emergent wasps correlated negatively with brood size. In the laboratory, single oviposition on P. brassicae larvae did not produce any brood size larger than 62, but brood size increased with superparasitism.
3. Laboratory experiments demonstrated that both naive and experienced female wasps were willing to attack hosts that had been newly parasitised by themselves or conspecifics. Superparasitism reduced survivorship but increased food consumption and weight growth in P. brassicae larvae. Superparasitism lengthened parasitoid development and prolonged the feeding period of host larvae.
4. Despite a trade-off between maximising brood size and optimising the fitness of individual offspring, two or three ovipositions on P. brassicae larvae resulted in a greater dry female mass than did a single oviposition on the host. Thus, superparasitism might be of adaptive significance under certain circumstances, especially when host density is low and unparasitised hosts are rare in a habitat.     

Putrescine--an inhibitor of the reaction of carbon dioxide hydration in hemolymph of lepidoptera

Using high-voltage electrophoresis, thin-layer chromatography and chemical analysis tetramethylene diamine was found in hemolymph of 8 Lepidoptera species (Bombyx mori L., Ocneria dispar L., Euproctis chrysorrhoea L., Yponomeuta malinella L., Pieris brassicae L., Aporia crataegi L., Melacosoma neustria L., Mamestra brassicae L.). Its content reaches 11 mM 1 l of hemolymph. The absence of tetramethylene diamine in the intestine wall and Malpighian vessels and its presence in the fat body permits suggesting its synthesis in the fat body. Ion-selective H-electrodes were used to show the function of tetramethylene diamine as an inhibitor of the CO2 hydration reaction in the insects hemolymph.     

Chemoreception of amino acids in larvae of two species of Pieris

ABSTRACT. Specificity and sensitivity of gustatory neurones in response to twenty-two amino acids were studied in larvae of Pieris brassicae L. and Pieris rapae L. (Lepidoptera: Pieridae) using electrophysiological methods. Twelve amino acids stimulated a specific amino acid receptor cell in the lateral styloconic sensillum on the maxillary galea of both species, and a further two evoked single unit responses in the same sensillum of P.brassicae only. Histidine, phenylalanine and tryptophane were the weakest stimulants for P.brassicae , but were among the four best stimulants for P.rapae . In both species, eight amino acids were ineffective. Significant differences in stimulatory effectiveness were found between amino acids. Nutritionally essential amino acids were more effective in both species, as in five other lepidopterous species. Similarities with postulated sites for amino acid recognition in the dipteran Boettcherisca peregrina were found.
Concentration-response (C/R) relations were studied for five amino acids. Significant differences were found in saturated response levels. Parameters characterizing C/R relations were estimated using a logistic model. Comparing C/R parameters with phytochemical data on concentrations of free amino acids in a common host plant, Brassica oleracea L., shows that amino acids are effective stimuli at their natural concentrations. The amino acid chemoreceptor seems able to transmit information about concentration differences of amino acids in the plant tissue.     

The mutation R(423)S in the Bacillus thuringiensis hybrid toxin CryAAC slightly increases toxicity for Mamestra brassicae L

Bacillus thuringiensis Cry1Ac toxin is 100 times less toxic than Cry1C to Mamestra brassicae. An R(423)S mutation abolishes Cry1Ac toxin proteolysis in M. brassicae gut juice but does not increase its toxicity to this insect. The CryAAC hybrid toxin (1Ac/1Ac/1Ca) is toxic to M. brassicae but is susceptible to gut protease digestion at the R(423) residue. Accordingly we have investigated the effect of the R(423)S mutation in CryAAC on its toxicity for M. brassicae and Pieris brassicae. Bioassays demonstrated that the R(423)S mutation slightly increased the toxicity of CryAAC for M. brassicae by having a significantly inhibitory effect on the growth of surviving larvae. The mutant hybrid was still highly toxic to P. brassicae. Features of CryAACR(423)S such as, (1) stability in M. brassicae gut juice and (2) crystal solubility were investigated. Computer simulations suggest that a possible major increase in flexibility in the CryAAC loop beta7/beta8 (G(391)-P(397)) caused by the R(423)S substitution could be a reason for the increase in M. brassicae toxicity.     

Host range and virulence of five baculoviruses from lepidopterous hosts

The host range and virulence of five insect baculoviruses (two multiply-enveloped nuclear polyhedrosis viruses (MNPVs) from Agrotis segetum and Mamestra brassicae; one singly-enveloped NPV from Plusia gamma and two granulosis viruses (GVs) from A. segetum and Pieris brassicae) were studied for seven lepidopterous pests of temperate agriculture (A. segetum, Agrotis exclamationis, Lacanobia oleracea, M. brassicae, Noctua pronuba, P. gamma and Pieris rapae). None of the viruses killed all species but M. brassicae MNPV failed to infect only P. rapae. The other viruses were restricted to the homologous host, or members of its genus or subfamily. In all examples except A. segetum GV, the median lethal dose for the most susceptible host, was less than 22 virus inclusion bodies and median lethal times for all infections ranged from 5·5 to 16·6 days. The low susceptibility of A. segetum and other noctuids to GV infections is discussed in relation to the structure of inclusion bodies and the nature of the infectious unit in baculoviruses.     

A simplified method for the characterization of nuclear polyhedrosis virus genomes

A simplified restriction endonuclease analysis procedure is described which allows the characterization of baculovirus DNA obtained directly from a single larvae without purification of virus. This rapid method was used to demonstrate the genomic stability of nuclear polyhedrosis viruses (NPVs) from Agrotis segetum, Euxoa messoria and Mamestra brassicae after several passages in Euxoa scandens.     

A simplified method for the characterization of nuclear polyhedrosis virus genomes

Abstract A simplified restriction endonuclease analysis procedure is described which allows the characterization of baculovirus DNA obtained directly from a single larvae without purification of virus. This rapid method was used to demonstrate the genomic stability of nuclear polyhedrosis viruses (NPVs) from Agrotis segetum, Euxoa messoria and Mamestra brassicae after several passages in Euxoa scandens     

Parasitism of the herbivore Pieris brassicae L. (Lep., Pieridae) by Cotesia glomerata L. (Hym., Braconidae) does not benefit the host plant by reduction of herbivory

Models of tritrophic interactions often imply that attraction of herbivore natural enemies by the plant constitutes a defence. Parasitoid attack on herbivores is assumed to result in a reduction in herbivory and/or an increase in plant fitness. Coupled with the active involvement of the plant in producing signals, this can be seen as indirect mediation of wound-induced defence. The assumption that parasitism of Pieris brassicae L. by the parasitoid Cotesia glomerata L. reduces leaf consumption of cabbage is tested. The effect of parasitism on the growth and feeding rates of P. brassicae larvae compared to unparasitized larvae. Either the defensive mechanism, if it exists, is more subtle, or the natural enemy is using an incidental chemical produced as a by-product of the herbivores feeding action as a cue for host-finding, with no increaction effects. This has implications for the study of tritrophic interactions, particularly in the context of agricultural systems and the assumption that in IPM strategies, parasitoids are a plant benefit.     

大菜粉蝶颗粒体病毒对菜粉蝶幼虫取食、生长、发育的影响

菜粉蝶幼虫在一、二、三、四龄初期饲以大菜粉蝶颗粒体病毒(PbGV),食量减少99.3—38.4%;四龄末或五龄初饲毒,食量反而比健虫高出36.3%和87.2%。幼虫感病后取食期的缩短或延长是食量变化的主要原因。幼虫病死前脱皮过程先被抑制,其结果往往使得死亡时所处的龄期明显延长。幼虫在四龄末饲毒后,由于五龄期延长,食量增加,其死亡前达到的最高体重平均为357.9毫克,远高于健虫化蛹前的最高体重。PbGV对菜粉蝶化蛹也有明显影响。幼虫在五龄前饲毒,一般不能化蛹。五龄第1—3天饲毒,化蛹率26.4—87.9%,第4天饲毒对化蛹无影响。本文结果对应用PbGv防治菜粉蝶幼虫危害以及对该病毒的大量增殖均具指导意义。     

ESTABLISHMENT AND SOME CHARACTERISTICS OF A CONTINUOUS CELL LINE DERIVED FROM FAT BODYS OF THE CABBAGE ARMYWORM (LEPIDOPTERA, NOCTUIDAE)

Fat bodies from fully grown larvae of the cabbage armyworm, Mamestra brassicae , were cultivated in a synthetic medium, MGM–431. Small free cells migrated out and multiplied rapidly. The first cell passage was done after culture for 26 days, and 100 passages were performed in the following 9 months. The established cell line was designated as NIAS–MaBr–85. The cell population is morphologically heterogeneous, but most of the cells are hexaploid with 180 microchromosomes. The cells could be stored for 3 months at 5°C, or for longer when frozen at -80°C in medium containing 10% glycerol. Qualitatively the cell line requires few amino acids: only cystine, histidine, isoleucine, methionine and threonine are essential as determined by omission of individual amino acids. The cells are susceptible to Chilo iridescent virus and Autographa californica nuclear polyhedrosis virus. This cell line can grow in serum-free M-M medium.     

Response of the wasp (<Emphasis Type="Italic">Cotesia glomerata</Emphasis>) to larvae of the large white butterfly (<Emphasis Type="Italic">Pieris brassicae</Emphasis>)

The large white butterfly (Pieris brassicae L) first invaded northernmost Japan from Siberia around 1994, and after a few years, began to expand its range. The wasp, Cotesia glomerata (L) parasitizes larvae of the small white butterfly (Pieris rapae crucivora Boisduval), a usual host in the same geographic area. Some Pieris brassicae larvae in Hokkaido have been parasitized by Cotesia glomerata, but the parasitism rate of Pieris brassicae larvae tends to be lower than that of Pieris rapae. To examine the process of parasitizing Pieris brassicae larvae, we observed how the parasitoid wasp responded to the host larvae on damaged leaves. Cotesia glomerata females tended to avoid Pieris brassicae larvae, and even when female wasps inserted their ovipositors into Pieris brassicae larvae, none laid eggs. The parasitoids obtained from Pieris rapae larvae failed to parasitize Pieris brassicae during the host-acceptance step.     

Isolation of A Cytoplasmic Polyhedrosis Virus by Physical and Immunological Techniques

Procedures are described for the separation and purification of cytoplasmic polyhedrosis viruses obtained from multiply infected Heliothis armigera larvae. Separation was achieved by differential centrifugation and density gradient zone electrophoresis followed by complexing with nuclear polyhedrosis virus specific antibody. The yield of cytoplasmic polyhedrosis virus was increased by passage in larvae reared on synthetic media.     

Effectiveness of the defence mechanism of the turnip sawfly, Athalia rosae (Hymenoptera: Tenthredinidae), against predation by lizards

The turnip sawfly, Athalia rosae Linnaeus, is a pest on cruciferous crops. Larvae sequester secondary plant compounds, namely glucosinolates, in their haemolymph. When attacked, their integument is easily disrupted and a droplet of haemolymph is exuded ('easy bleeding'). This has been shown to be an effective, chemical-based, defence against invertebrate predators. The efficiency of this proposed defence was tested against a vertebrate predator, using groups of the iguanid lizard Anolis carolinensis Voigt as a model predator. Caterpillars of Pieris rapae Linnaeus and Pieris brassicae Linnaeus served as control prey species that do not sequester glucosinolates. Lizards attacked far fewer sawfly larvae than pierid caterpillars. Several of the sawfly larvae were rejected after an initial attack, demonstrating unpalatability to the lizards, while the Pieris larvae were not rejected. However, P. rapae larvae topically treated with extracts of haemolymph of A. rosae had no deterrent effect on the lizards and no avoidance learning occurred over a period of two weeks. Adult sawflies do not easy bleed but have glucosinolates carried over from the larval stage. Lizards attacked them at a higher rate than larvae and they were never rejected. The results suggest that for the defensive effectiveness of the pest sawfly species against vertebrates the chemical cue is not necessarily sufficient. Movement and colour may be important additional factors triggering the behaviour of vertebrate predators.     

Laboratory studies on the effects of pollen from Bt-maize on larvae of some butterfly species

Abstract:   Three lepidopteran species were tested to determine their susceptibility against the ingestion of pollen from genetically modified maize plants. To prove the existence of dose–response relations between the applied amount of pollen (Bt-maize) and the damages on the tested larvae, a method was developed which makes it possible to feed caterpillars with defined amounts of pollen. If their food plants were contaminated with pollen of a cultivar of the Bt-176 maize-line Pieris brassicae , Pieris rapae and Plutella xylostella -larvae fed less, grew more slowly and showed a higher mortality than caterpillars of an untreated control group. The 50% lethality (LD₅₀)-values were calculated for P.xylostella (L₄) with 19.2, for P. rapae (L₂) with 39.0 and also for P. brassicae (L₂) with 139.2 pollen of the transgenic maize Pactol CB. Studies with P. brassicae -caterpillars of different larval stages indicated, that older individuals showed a higher tolerance against pollen from Bt-maize than younger ones. It must be stated on the basis of the present studies, that ingestion of non-transgenic maize pollen has neither a positive nor a negative effect on caterpillars. It becomes clear from the information presented here that it is still difficult to make general statements about the endangering of butterflies, arising from cultivation of genetically modified maize lines. Further investigations on this issue are needed. Initially, the LD₅₀-values concerning the larvae of certain butterfly species have to be determined to anticipate the risks, and in addition the distances between habitats with caterpillar host plants and maize fields, and the abundance of these plants have to be considered.