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.     

Effects of methoxyfenozide, indoxacarb, and other insecticides on the beneficial egg parasitoid Trichogramma nr. brassicae (Hymenoptera: Trichogrammatidae) under laboratory and field conditions

Trichogramma nr. brassicae is a common egg parasitoid of Helicoverpa species in Australian processing tomatoes, but its effectiveness can be severely curtailed by insecticide applications. To identify insecticides that are potentially compatible with this species, the effects of seven insecticides, including newly introduced compounds and a surfactant, were screened in laboratory and glasshouse assays for their toxicity to the wasps. Assays involved direct applications on adults, residual effects on adults, and applications on life stages still inside the host. Methoxyfenozide and indoxacarb were not toxic to Trichogramma in any assay when applied at field rates. Naled and chlorfenapyr caused 100% mortality when directly applied to adults, and 95% mortality when adults were exposed to residues of these chemicals within 24 h of application. The effects of naled residues were short lived (<48 h). Naled and chlorfenapyr were also toxic when applied to Trichogramma developing inside host eggs, reducing emergence of adults by >25%. Imidacloprid, emamectin, and tau-fluvalinate were toxic in some experiments; they caused >97% mortality in adults 1 h after direct application and in residue assays they caused 23-64% mortality during the first 24 h. In field trials, methoxyfenozide had no harmful effects on emergence from sprayed parasitized eggs, whereas indoxacarb had a small impact (<8%) on emergence. Methoxyfenozide and indoxacarb are potentially suitable for inclusion in integrated pest management strategies for management of Helicoverpa because they do not influence adult survival or development of immature stages, whereas other chemicals need to be treated cautiously.     

Are fecundity and longevity of female Aphelinus abdominalis affected by development in GNA‐dosed Macrosiphum euphorbiae?

Abstract. Snowdrop lectin ( Galanthus nivalis agglutinin, GNA) confers partial resistance to several aphid species when incorporated into an artificial diet and/or expressed in transgenic potato. First-tier laboratory-scale experiments were conducted to assess the potential effect of GNA on the longevity and fecundity of female parasitoid Aphelinus abdominalis (Dalman) that had developed in Macrosiphum euphorbiae (Thomas) fed artificial diet containing 0.1% GNA (w/v). In a previous study it was shown that GNA ingested by A. abdominalis larvae is not acutely toxic. It was also shown that GNA has a host-size mediated effect on parasitoid sex ratio and larval development, but no apparent direct effect.         In this study, we report that A. abdominalis larvae that developed in GNA-dosed aphids that were smaller than control aphids of the same age, produced smaller adults with a reduced longevity and fecundity. Aphelinus abdominalis larvae that developed in GNA-dosed aphids older than the control but of the same size, produced adults of similar size that lived as long as the control but had a reduced fecundity.         Our results suggest that GNA fed to aphids in artificial diet has both a host-mediated effect (via aphid-size) and a direct effect on adult parasitoid fecundity. It is not known how GNA affects parasitoid larval development and subsequently adult fecundity, but it is hypothesized that GNA acted as an antifeedant to parasitoid larvae, thus disturbing nutrient assimilation and conversion necessary for egg maturation.     

Toxicity of coelomic fluid of the earthworm Eisenia foetida to vertebrates but not invertebrates: probable role of sphingomyelin

The coelomic fluid (CF) of the earthworm Eisenia foetida exhibits a wide variety of biological activities. We found that the CF was not toxic to 42 species, belonging to seven invertebrate phyla, almost all in aquatic adults and larvae exposed to CF. Eleven teleostean species tested died in 0.2-1% CF mostly between 10 and 120 min and the effects were dose-dependent. Tadpoles of the toad Bufo japonicus formosus died in 0.4-2% CF between 80 and 225 min depending upon size, with larger tadpoles surviving longer. Before dying, all experimental tadpoles developed curled and shrunken tails. The Okinawa tree lizard, soft-shelled turtle, Japanese quail, mouse and rat all died after i.v. injection of CF (above 20 microl/kg). Thus, CF was not toxic to invertebrates, but toxic to vertebrates. After heating, CF lost its toxicity to fish, tadpoles and mice. Both CF and lysenin incubated with sphingomyelin-liposomes (SM-liposomes) were no longer toxic, suggesting the involvement of SM in the toxicity. Lysenin, which is a constituent of CF and known to bind specifically to sphingomyelin, exhibited toxicity similar to that of CF. Thus, lysenin in CF is probably responsible for the toxic effects of CF by binding to SM in vertebrate tissues. The bodies of invertebrates might contain little or no SM, while those of vertebrates do contain SM. The coelomic fluid of the earthworm Pheretima communissima has no toxicity to mouse.     

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.     

Evaluating the safety of Rodolia cardinalis to two species of Galapagos finch; Camarhynchus parvulus and Geospiza fuliginosa

Some species of coccinellids exude a fluid that contains an alkaloid which can be toxic to vertebrates that ingest them. Because of this, before introducing Rodolia cardinalis to the Galapagos Islands as a biological control agent for the invasive scale insect, Icerya purchasi, it was considered necessary to determine whether it is toxic to birds. In this study, experiments were carried out with two common, insectivorous finches that are endemic to the Galapagos Islands; Camarhynchus parvulus (12 individuals) and Geospiza fuliginosa (16 individuals). Finches were maintained in captivity for 12–18 days. This period was divided into three phases: adaptation, experimental, and observation. During the experimental phase, test individuals, in addition to their regular diet, were fed 0.15 g portions of larvae or adults of R. cardinalis on alternate days over four days. The control group were fed 0.15 g of Asciodes quietalis abdomens in place of R. cardinalis. Symptoms of toxicity (mortality, deterioration, critical weight loss) from feeding on R. cardinalis were not observed in either species while in captivity, but birds avoided or rejected beetles indicating that they were distasteful. Finches regurgitated a higher number of larvae than adult beetles suggesting that larvae were less palatable than adults. Although long-term monitoring was not possible, results suggest that at least two species of finch will be unaffected by the release of R. cardinalis into the Galapagos Islands.     

A novel Bacillus thuringiensis gene encoding a Spodoptera exigua-specific crystal protein

Only one of the four lepidoptera-specific crystal protein subclasses (CryIC) Bacillus thuringiensis was previously shown to be highly toxic against several Spodoptera species. By using a cryIC-derived nucleotide probe, DNA from 25 different strains of B. thuringiensis was screened for the presence of homologous sequences. A putative crystal protein gene, considerably different from the cryIC gene subclass, was identified in the DNA of strain 4F1 (serotype kenyae) and cloned in Escherichia coli. Its nucleotide sequence was determined and appeared to contain several features typical for a crystal protein gene. Furthermore, the region coding for the N-terminal part of the putative toxic fragment showed extensive homology to subclass cryIA sequences derived from gene BtII, whereas the region coding for the C-terminal part appeared to be highly homologous to the cryIC gene BtVI. With an anti-crystal protein antiserum, a polypeptide of the expected size could be demonstrated in Western immunoblots, onto which a lysate of E. coli cells harboring the putative gene, now designated as BtXI, had been transferred. Cells expressing the gene appeared to be equally toxic against larvae of Spodoptera exigua as recombinant cells expressing the BtVI (cryIC)-encoded crystal protein. However, no toxicity against larvae of Heliothis virescens, Mamestra brassicae, or Pieris brassicae could be demonstrated. The nucleotide sequence analysis and the toxicity studies showed that this novel crystal protein gene falls into a new cryl gene subclass. We propose that this subclass be referred to as cryIE.     

Laboratory evaluation of the toxicity of systemic insecticides for Control of Anoplophora glabripennis and Plectrodera scalator (Coleoptera: Cerambycidae)

Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae) is one of the most serious nonnative invasive forest insects discovered in North America in recent years. A. glabripennis is regulated by federal quarantines in the United States and Canada and is the subject of eradication programs that involve locating, cutting, and chipping all infested trees. Other control methods are needed to aid in eradication and to form an integrated management program in the event eradication fails. We conducted laboratory bioassays to determine the toxicity of two systemic insecticides, azadirachtin and imidacloprid, for potential control of A. glabripennis and the cottonwood borer, Plectrodera scalator (F.) (Coleoptera: Cerambycidae), a closely related native cerambycid. Larvae of both cerambycid species were fed artificial diet with dilutions of azadirachtin or imidacloprid for 14 wk. Both insecticides exhibited strong antifeedant effects and some toxicity against A. glabripennis and P. scalator larvae. For A. glabripennis, the highest larval mortality at the end of the bioassay was 60% for larvae fed artificial diet treated with azadirachtin (50 ppm) or imidacloprid (1.6 ppm). For P. scalator, the highest larval mortality at the end of the bioassay was 100% for larvae fed artificial diet treated with azadirachtin (50 ppm) or imidacloprid (160 ppm). At 14 wk, the LC50 values for P. scalator were 1.58 and 1.78 ppm for azadirachtin and imidacloprid, respectively. Larvae of both species gained weight when fed diet treated with formulation blanks (inert ingredients) or the water control but lost weight when fed diet treated with increasing concentrations of either azadirachtin or imidacloprid. In a separate experiment, A. glabripennis adults were fed maple twigs treated with high and low concentrations of imidacloprid. A. glabripennis adult mortality reached 100% after 13 d on twigs treated with 150 ppm imidacloprid and after 20 d on twigs treated with 15 ppm imidacloprid. There was no visible feeding by A. glabripennis adults on twigs treated at the higher imidacloprid rate, and feeding was significantly reduced for adults placed on twigs treated at the low imidacloprid rate compared with adults on untreated twigs. In summary, imidacloprid and azadirachtin had both antifeedant and toxic effects against A. glabripennis and P. scalator and have potential for use in management programs. Based on our results, the delivery of high and sustained insecticide concentrations will be needed to overcome the antifeedant effects and lengthy lethal time for both larvae and adults exposed to these insecticides.     

SYSTEMIC INSECTICIDAL ACTION OF NICOTINE AND CERTAIN OTHER ORGANIC BASES

Nicotine and nicotine salts are taken up by the roots of plants from solutions, and when 0.01–0.001 % nicotine is used the plants become toxic to Aphis fabae and to Pieris brassicae larvae and can be shown to contain nicotine. The results with Phaedon cochleariae adults and larvae are less satisfactory. No systemic action is observed when the nicotine is watered on to soil in which plants are growing and no nicotine can be detected in the plants. Apparently the nicotine is decomposed in the soil.
When applied several times to the upper surface of a bean leaf nicotine kills aphids on the underside. There is some evidence that nicotine can be translocated further through the plant following leaf applications, but the toxic action at any distance is very weak in the plants used in the present experiments and can only be produced by frequent applications of rather concentrated nicotine solutions. Leaf absorption and subsequent translocation has not been observed with nicotine salts.
The various organic bases, including some piperidine phosphonites and allied compounds tested, are of very little interest as contact or systemic insecticides against aphids.     

Distribution, Frequency, and Diversity of Bacillus thuringiensis in an Animal Feed Mill

Bacillus thuringiensis was isolated from 36 of 50 residue samples obtained from an animal feed mill (a stored-product environment). Of 710 selected colonies having Bacillus cereus-B. thuringiensis morphology isolated from the samples, 477 were classified as B. thuringiensis because of production of parasporal delta-endotoxin crystals. There was a diverse population of B. thuringiensis, as revealed by differentiation of the isolates into 36 subgroups by using (i) their spectra of toxicity to the lepidopterans Heliothis virescens, Pieris brassicae, and Spodoptera littoralis and the dipteran Aedes aegypti and (ii) their parasporal crystal morphology. A total of 55% of the isolates were not toxic to any of these insects at the concentrations used in the bioassays; 40% of all isolates were toxic to one or more of the Lepidoptera; and 20, 1, and 1% of the isolates were toxic to only P. brassicae, H. virescens, and S. littoralis, respectively. The most frequent toxicity was toxicity to P. brassicae (36% of all isolates); 18% of the isolates were toxic to A. aegypti (5% exclusively), 10% were toxic to H. virescens, and 4% were toxic to S. littoralis. Toxicity to P. brassicae was more often linked with toxicity to H. virescens than with toxicity to S. littoralis. The frequency of toxicity was significantly greater in isolates that produced bipyramidal crystals than in isolates that produced irregular pointed, irregular spherical, rectangular, or spherical crystals.     

Effects of Asobara japonica venom on larval survival of host and nonhost Drosophila species

Abstract Ovipositing Asobara japonica females inject venom (containing paralysis‐inducing factors) immediately after the insertion of their ovipositors into Drosophila larvae, and lay eggs a little later. Interruption of their oviposition behaviour before egg laying causes high larval mortality in host Drosophila species, whereas normal oviposition does not. This suggests that venom of this parasitoid is toxic to larvae of these host species but its toxicity is suppressed by factor(s) provided by parasitoid females at the time of laying egg or by parasitoid embryos developing in the hosts. On the other hand, venom does not show toxicity to larvae of nonhost Drosophia species. Possible functions of venom are discussed.     

Role of granivory and insectivory in the life cycle of the carabid beetle Amara similata

1. The cereal field carabid beetle Amara similata was selected to elucidate the role of seeds and insects as food sources. Three experiments were performed to rank different weed seeds and insects in terms of food value: (i) fecundity in relation to adult diet, (ii) larval survival in relation to diet and (iii) larval survival in relation to parental diet.
2. Seeds were found to be of high value and insects of low value both for adults and larvae. Adding insects to a seed diet gave no significant improvement. The value of single species of seeds varied within adults and larvae, but also between adults and larvae. This is the first report that Amara larvae are granivorous.
3. The value of different insects varied by species. Cereal aphid species were of the lowest value both for adult and larval beetles. On a diet of mixed insects, the adults were able to produce a low number of eggs but the larvae all died before pupation.
4. There was no simple relation between survival of the larvae and the quality of the parents' diet.     

Monoclonal Antibody Analysis and Insecticidal Spectrum of Three Types of Lepidopteran-Specific Insecticidal Crystal Proteins of Bacillus thuringiensis

We have investigated the protein composition and the insecticidal spectrum of crystals of 29 Bacillus thuringiensis strains active against lepidopteran larvae. All crystals contained proteins of 130 to 140 kilodaltons (kDa) which could be grouped into three types by the molecular weight of the protoxin and the trypsin-activated core fragment. Proteins of the three types showed a characteristic insecticidal spectrum when tested against five lepidopteran species. Type A crystal proteins were protoxins of 130 or 133 kDa, which were processed into 60-kDa toxins by trypsin. Several genes encoding crystal proteins of this type have been cloned and sequenced earlier. They are highly conserved in the N-terminal half of the toxic fragment and were previously classified in three subtypes (the 4.5-, 5.3-, and 6.6-kilobase subtypes) based on the restriction map of their genes. The present study shows that different proteins of these three subtypes were equally toxic against Manduca sexta and Pieris brassicae and had no detectable activity against Spodoptera littoralis. However, the 4.5-, 5.3-, and 6.6-kilobase subtypes differed in their toxicity against Heliothis virescens and Mamestra brassicae. Type B crystal proteins consisted of 140-kDa protoxins with a 55-kDa tryptic core fragment. These were only active against one of the five insect species tested (P. brassicae). The protoxin and the trypsin-activated toxin of type C were 135- and 63-kDa proteins, respectively. Proteins of this type were associated with high toxicity against S. littoralis and M. brassicae. A panel of 35 monoclonal antibodies was used to compare the structural characteristics of crystal proteins of the three different types and subtypes. Each type of protein could be associated with a typical epitope structure, indicating an unambiguous correlation between antigenic structure and insect specificity.     

Effects of diet on female fecundity and larval development in the carrion beetle Necrophila japonica

The carrion beetle subfamily Silphinae (Coleoptera: Silphidae) contains dominant macroinvertebrates of soil ecosystems in temperate zones. However, their feeding habits, which determine the role of each species in the ecosystem, have not been sufficiently studied. Moreover, although a diet shift from necrophagy on vertebrate carcasses to predatory feeding on invertebrates is known to occur in this subfamily, the processes and mechanisms of this shift have also been inadequately addressed. We examined female fecundity and larval development on various diets in a Silphinae species, Necrophila (Eusilpha) japonica (Motschulsky). The experimental diets included a meat diet and various invertebrate diets, which reflect the ‘ancestral’ feeding habit in Silphinae, necrophagy, and the ‘derivative’ feeding habit, predatory feeding. Female fecundity was significantly higher on the meat diet (minced beef) than on an insect larvae diet (mealworms and dipteran larvae) but did not significantly differ from that on an earthworm diet. Larval developmental performance was significantly higher on the earthworm diet than on the meat and insect larvae diets. Our results for larval development were consistent with those of previous stable isotope analyses of the same species, in which isotopic values of larval samples agree with those of hypothetical consumers that utilize earthworms. The consistency of results among different methods indicates that N. japonica larvae are most likely earthworm feeders. In contrast, our results for the female fecundity experiment differed from those of previous stable isotope analyses, in which vertebrate carcasses unlikely serve as the staple diet of adults in the field; thus, the feeding habits of N. japonica adults remain unresolved. Our observations that females and larvae performed best on the meat and earthworm diets, respectively, may indicate that, in Silphinae, the diet shift from necrophagy to predatory habits occurs earlier in larvae than in adults.     

Role of proteolysis in determining potency of Bacillus thuringiensis Cry1Ac delta-endotoxin

Bacillus thuringiensis protein delta-endotoxins are toxic to a variety of different insect species. Larvicidal potency depends on the completion of a number of steps in the mode of action of the toxin. Here, we investigated the role of proteolytic processing in determining the potency of the B. thuringiensis Cry1Ac delta-endotoxin towards Pieris brassicae (family: Pieridae) and Mamestra brassicae (family: Noctuidae). In bioassays, Cry1Ac was over 2,000 times more active against P. brassicae than against M. brassicae larvae. Using gut juice purified from both insects, we processed Cry1Ac to soluble forms that had the same N terminus and the same apparent molecular weight. However, extended proteolysis of Cry1Ac in vitro with proteases from both insects resulted in the formation of an insoluble aggregate. With proteases from P. brassicae, the Cry1Ac-susceptible insect, Cry1Ac was processed to an insoluble product with a molecular mass of approximately 56 kDa, whereas proteases from M. brassicae, the non-susceptible insect, generated products with molecular masses of approximately 58, approximately 40, and approximately 20 kDa. N-terminal sequencing of the insoluble products revealed that both insects cleaved Cry1Ac within domain I, but M. brassicae proteases also cleaved the toxin at Arg423 in domain II. A similar pattern of processing was observed in vivo. When Arg423 was replaced with Gln or Ser, the resulting mutant toxins resisted degradation by M. brassicae proteases. However, this mutation had little effect on toxicity to M. brassicae. Differential processing of membrane-bound Cry1Ac was also observed in qualitative binding experiments performed with brush border membrane vesicles from the two insects and in midguts isolated from toxin-treated insects.     

Interaction of a solitary larval endoparasitoid, Microplitis mediator, with its host, Mamestra brassicae: host acceptance and host suitability

Abstract:  Microplitis mediator (Haliday) (Hym., Braconidae) is an important parasitoid of early instar larvae of the European cabbage moth, Mamestra brassicae L. (Lep., Noctuidae). In the laboratory, we examined attack responses of female M. mediator to the first three larval instars of M. brassicae . Females were presented with M. brassicae larvae either one individual at a time in a no-choice experiment, or three individuals, one from each instar, simultaneously in a choice experiment. Whether or not there was choice, naïve female parasitoids attacked a high proportion of larvae and did not discriminate among instars. In the no-choice experiment, attacked larvae were reared, and parasitoid cocoons were produced from about 76% of larvae attacked as first and second instars, but from only 19% of larvae attacked as third instars. Dissections of attacked larvae from the choice experiment showed that about 79% of attacks on first and second instars resulted in oviposition compared with only 49% for third instars. When given choice, frequency and number of attacks on first instar larvae increased with increasing parasitoid experience. Our results suggest that first and second instar larvae of M. brassicae are suitable hosts for M. mediator , but that third instar larvae are suboptimal both because oviposition attempts were frequently unsuccessful and because immature parasitoids failed to complete development. Nevertheless, naïve attacking parasitoids exhibited minimal discrimination among instars, although experienced parasitoids most frequently attacked first instar larvae. The host selection behaviour of M. mediator is discussed in the context of optimal foraging theory and implications for biological control.     

黄玛草蛉对两种蚜虫的捕食效能

实验室条件下研究了黄玛草蛉2龄与3龄幼虫对甘蓝蚜以及玉米蚜成虫的捕食功能反应。结果表明:黄玛草蛉2龄与3龄幼虫对2种蚜虫的捕食量均随着猎物密度的增加而增加,寻找效应随着猎物密度的增加而减小。不同龄期幼虫对2种蚜虫的捕食功能反应均符合HollingⅡ模型,黄玛草蛉2龄与3龄幼虫对玉米蚜最大理论日捕食量分别为120.48头、185.18头;黄玛草蛉2龄与3龄幼虫对甘蓝蚜最大理论日捕食量分别为60.60头、86.95头。在相同猎物种类下,黄玛草蛉3龄幼虫对2种蚜虫的捕食量均高于黄玛草蛉2龄幼虫;在相同猎物密度下,黄玛草蛉2龄幼虫与3龄幼虫对玉米蚜的捕食量均高于甘蓝蚜。     

Papain protects papaya trees from herbivorous insects: role of cysteine proteases in latex

Many plants contain latex that exudes when leaves are damaged, and a number of proteins and enzymes have been found in it. The roles of those latex proteins and enzymes are as yet poorly understood. We found that papain, a cysteine protease in latex of the Papaya tree (Carica papaya, Caricaceae), is a crucial factor in the defense of the papaya tree against lepidopteran larvae such as oligophagous Samia ricini (Saturniidae) and two notorious polyphagous pests, Mamestra brassicae (Noctuidae) and Spodoptera litura (Noctuidae). Leaves of a number of laticiferous plants, including papaya and a wild fig, Ficus virgata (Moraceae), showed strong toxicity and growth inhibition against lepidopteran larvae, though no apparent toxic factors from these species have been reported. When the latex was washed off, the leaves of these lactiferous plants lost toxicity. Latexes of both papaya and the wild fig were rich in cysteine-protease activity. E-64, a cysteine protease-specific inhibitor, completely deprived the leaves of toxicity when painted on the surface of papaya and fig leaves. Cysteine proteases, such as papain, ficin, and bromelain, all showed toxicity. The results suggest that plant latex and the proteins in it, cysteine proteases in particular, provide plants with a general defense mechanism against herbivorous insects.     

Plant-toxin interactions in transgenic Bt cotton and their effect on mortality of Helicoverpa armigera (Lepidoptera: Noctuidae)

In Australia, transgenic cotton plants expressing the cry1Ac gene from Bacillus thuringiensis Berliner variety kurstaki are less toxic to first-instar Helicoverpa armigera (Hübner) after the plant is producing fruit. We developed two bioassay methods (leaf mush, leaf disk) to test if the physiological state of the plants explained changes in toxicity and a third method (diet incorporation) was developed to quantify the toxicity of Bt leaves when mixed in chickpea diet. Cry1Ac protein was less toxic to H. armigera larvae when the protein was mixed with leaves from fruiting versus presquare conventional cotton. Differences in LC50 varied from 2.4- to 726-fold, depending on the source of toxin and conventional plant material. These results suggest that plant-toxin interactions in fruiting cotton are reducing the toxicity of the Cry1Ac protein. The possible role of tannins in these changes is discussed.     

Impact of snowdrop lectin (Galanthus nivalis agglutinin; GNA) on adults of the green lacewing, Chrysoperla carnea

Based on the finding that Galanthus nivalis agglutinin (GNA) has direct negative effects on larvae of Chrysoperla carnea, laboratory experiments were conducted to investigate its toxicity to the adults. While the ingestion of GNA dissolved in an artificial diet did not affect adult longevity, there were concentration-dependent negative effects on the pre-oviposition period, daily fecundity and total fecundity (number of eggs laid). When GNA was ingested by larvae of C. carnea, it caused a significant extension of larval development time. Adults that had emerged from GNA-fed larvae did not differ from those that developed from control larvae in terms of adult fresh weight, pre-oviposition period and daily or total fecundity. However, fertility (proportion of hatching eggs) was significantly decreased in adults raised from GNA-treated larvae. Western blots revealed that GNA ingested by larvae of C. carnea was partly transferred to the adult stage and was subsequently excreted or digested within a few days. Our toxicity studies (Tier-1 tests) clearly established a hazard of GNA to adult C. carnea when administered to larvae or adults at high concentrations. Implications of these toxicity data for the non-target risk assessment of GNA-expressing transgenic crops are discussed.