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Insect juvenile hormone: from "status quo" to high society.

Juvenile hormone (JH) exerts pleiotropic functions during insect life cycles. The regulation of JH biosynthesis by neuropeptides and biogenic amines, as well as the transport of JH by specific binding proteins is now well understood. In contrast, comprehending its mode of action on target organs is still hampered by the difficulties in isolating specific receptors. In concert with ecdysteroids, JH orchestrates molting and metamorphosis, and its modulatory function in molting processes has gained it the attribute "status quo" hormone. Whereas the metamorphic role of JH appears to have been widely conserved, its role in reproduction has been subject to many modifications. In many species, JH stimulates vitellogenin synthesis and uptake. In mosquitoes, however, this function has been transferred to ecdysteroids, and JH primes the ecdysteroid response of developing follicles. As reproduction includes a variety of specific behaviors, including migration and diapause, JH has come to function as a master regulator in insect reproduction. The peak of pleiotropy was definitely reached in insects exhibiting facultative polymorphisms. In wing-dimorphic crickets, differential activation of JH esterase determines wing length. The evolution of sociality in Isoptera and Hymenoptera has also extensively relied on JH. In primitively social wasps and bumble bees, JH integrates dominance position with reproductive status. In highly social insects, such as the honey bee, JH has lost its gonadotropic role and now regulates division of labor in the worker caste. Its metamorphic role has been extensively explored in the morphological differentiation of queens and workers, and in the generation of worker polymorphism, such as observed in ants.

Animals↗

Insect folivory in Didymopanax vinosum (Apiaceae) in a vegetation mosaic of Brazilian cerrado.

Susceptibility of Didymopanax vinosum (Apiaceae) to insect herbivores was investigated in three sites of a cerrado mosaic--composed of campo cerrado (a grassland with scattered trees and shrubs), cerradão (a tall woodland) and cerrado sensu stricto (intermediate between the two)--situated in Cerrado Pé-de-Gigante, Santa Rita do Passa Quatro, SP, Brazil. We also examined the relationship of folivory with the composition and abundance of the insect herbivore fauna, and with several nutritional and defensive plant characteristics (water, nitrogen, cellulose, lignin, tannin leaf contents, and leaf toughness). We collected insects associated with D. vinosum every month, and we measured leaf damage every three months. In general, the annual folivory differed among sites. It reached the highest rates in site 1 and site 3: 7.33 and 8.5 percent, respectively. Only 1.32 percent of annual folivory was observed in site 2. These levels resulted from the higher abundance, in sites 1 and 3, of the thrips Liothrips didymopanacis (Phlaeothripidae), the most abundant herbivore sampled, responsible for more than 90 percent of the observed damage. However, no significant relationship was found between insect activity and the chemical and physical composition of the leaves. Our findings suggest that, at least in this species, other chemical compounds or variables related to plant apparency and resource availability to herbivores (e.g. plant architecture) might play a more decisive role in the spatial variation of folivory than the nutritional and defensive traits that were analyzed.

Animals↗

Performance of transgenic corn hybrids in Missouri for insect control and yield.

The efficacy of Bacillus thuringiensis-transformed corn (Zea mays L.) hybrids compared with comparable nontransformed corn hybrids for controlling first- and second-generation European corn borer, Ostrinia nubilalis (Hübner), and second-generation southwestern corn borer, Diatraea grandiosella Dyar, was determined. Yield comparisons were obtained from the same plots of corn hybrids. Both generations of European and the second-generation of southwestern corn borer were effectively controlled, but the Bt hybrids varied in degree of control. Hybrids from Ciba Seeds, DEKALB, and Mycogen had more European corn borer tunneling than those from Novartis or Cargill, and this was generally ascribed to different transgenic events. The Bt-transformed hybrids had virtually no leaf-feeding damage and less tunneling than the non-Bt corn hybrids. Some Bt corn hybrids had no tunneling, whereas other Bt hybrids had a small amount of tunneling. All of the non-Bt hybrids had significant leaf-feeding damage and stalk tunneling from both insects. Only three live European corn borer larvae (stunted) were found in the Bt corn hybrids while splitting stalks to assess tunnel length. When insect damage was significant, and in some evaluations where damage was not significant, differences in yields among hybrids were observed. No significant insect population differences were observed for five genera of beneficial insects for Bt versus non-Bt corn hybrids. Corn hybrids that have been transformed with the Bt gene provide an effective means of control for corn borers and efforts to reduce the likelihood of development of borer resistance are warranted.

Animals↗

An in-field screen for early detection and monitoring of insect resistance to Bacillus thuringiensis in transgenic crops.

We present a field-based approach to detect and monitor insects with resistance to insecticidal toxins produced by transgenic plants. Our objective is to estimate the phenotypic frequency of resistance in a population by relating the densities of insects on genetically transformed plants to densities on nontransformed plants. We focus on European corn borer, Ostrinia nubilalis (Hübner), in sweet corn, Zea mays L., expressing Cry1Ab from Bacillus thuringiensis subsp. kurstaki Berliner to illustrate principles underlying the method. The probability of detecting one or more rare, resistant larvae depends on sample size, the density of larvae on nontransformed plants, and an assumed frequency of resistant phenotypes in a given population. Probability of detection increases with increases in sample size, background density, or the frequency of resistant individuals. Following binomial probability theory, if a frequency of 10(-4) is expected, 10(3)-10(4) samples must be collected from a B. thuringiensis (Bt) crop to have at least a 95% probability of locating one or more resistant larvae. In-field screens using transgenic crops have several advantages over traditional laboratory-based methods, including exposure to a large number of feral insects, discrimination of resistant individuals based on Bt dosages expressed in the field, incorporation of natural and Bt-induced mortality factors, simultaneous monitoring for more than one insect species, and ease of use. The approach is amenable to field survey crews working in research, extension, and within the seed corn industry. Estimates of the phenotypic frequency of resistance from the in-field screen can be useful for estimating initial frequency of resistant alleles. Bayesian statistical methods are outlined to estimate phenotype frequencies, allele frequencies, and associated confidence intervals from field data. Results of the approach are discussed relative to existing complementary methods currently available for O. nubilalis and corn earworm, Helicoverpa zea (Boddie).

Animals↗

Acoustic surveying of subterranean insect populations in citrus groves.

Subterranean insect pests cause considerable economic damage but their concealment makes detection difficult. A portable acoustic system was developed and tested for its potential to rate the likelihood that trees in citrus groves were infested with Diaprepes abbreviatus (L.) larvae. The likelihood was rated independently by a computer program and an experienced listener that distinguished insect sounds from background noises. Diaprepes abbreviatus, Phyllophaga spp., or other pest insects were excavated from all 11 sites rated at high likelihood of infestation but were absent from 20 of 25 low-rated sites. There was a significant regression between the activity rate and the number of pest organisms present at recording sites although the correlation was weaker than between activity rate and likelihood of infestation. Although the system is at an early stage of development, the success of these field tests suggests that it has considerable potential as a tool to detect and monitor hidden infestations of insects in soil.

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Functional mechanics of the plant defensive Griffonia simplicifolia lectin II: resistance to proteolysis is independent of glycoconjugate binding in the insect gut.

Griffonia simplicifolia lectin II (GSII) is a plant defensive protein that significantly delays development of the cowpea bruchid Callosobruchus maculatus (F.). Previous structure/function analysis by site-directed mutagenesis indicated that carbohydrate binding and resistance to insect gut proteolysis are required for the anti-insect activity of this lectin. However, whether there is a causal link between carbohydrate binding and resistance to insect metabolism remains unknown. Two proteases principally responsible for digestive proteolysis in third and fourth instar larvae of C. maculatus were purified by activated thiol sepharose chromatography and resolved as cathepsin L-like proteases, based on N-terminal amino acid sequence analysis. Digestion of bacterially expressed recombinant GSII (rGSII) and its mutant protein variants with the purified gut proteases indicates that carbohydrate binding, presumably to a target ligand in insect gut, and proteolytic resistance are independent properties of rGSII, and that both facilitate its efficacy as a plant defensive molecule.

Acetylglucosamine↗

Changes in Cry1Ac Bt transgenic cotton in response to two environmental factors: temperature and insect damage.

The efficacy of Cry1Ac Bacillus thuringiensis (Bt) cotton plants against field populations of Helicoverpa armigera (Hübner) has been inconsistent over the growing season. Any reduction in efficacy (where efficacy is the capacity of the plant to affect the survival of the insect) increases the opportunities for H. armigera to evolve resistance to Bt toxin. Changes in efficacy could be due to changes at the level of gene expression and/or in the physiological makeup of the plant and may be induced by environmental conditions. Two environmental factors, temperature and insect damage, were investigated. Temperature was found to affect efficacy, whether plants were grown at different temperatures continuously or were exposed to a change in temperature for a short period. Damage caused by chewing insects (H. armigera larvae) produced a dramatic increase in the efficacy of presquare Bt cotton. In contrast, damage by sucking insects (aphids) did not induce changes in efficacy. Changes in efficacy seemed to be mediated through modification of the physiological background of the plant rather than changes in the level of Cry1Ac expression or in the concentration of the Bt toxin. The impact of the non-Bt responses of plants on strains of H. armigera should be evaluated. It is possible that by enhancing existing defensive mechanisms of plants, the rate of evolution of resistance to Bt toxins could be retarded by increasing the plants overall toxicity through the additive effects of the toxins and plant defenses.

Animals↗

Evaluation of a commercially available beneficial insect habitat for management of lepidoptera pests.

A field study was conducted in 2003 and 2004 at the Center for Environmental Farming Systems in Goldsboro, NC, to evaluate the effectiveness of a commercially available beneficial insect habitat in decreasing pest caterpillar populations in organically managed tomato, Lycopersicon esculentum Mill., plots. Six pairs of tomato plots were established and a commercial beneficial insect habitat seed mix (Peaceful Valley's Good Bug Blend) transplanted around the perimeter of treatment plots, whereas a brown-top millet, Brachiaria ramose (L.) Stapf., border was planted around control plots. Egg predation, egg parasitism by trichogrammatid wasps, and larval parasitism by braconid wasps was monitored throughout the growing season to determine whether habitat increased their activity. In both years of this study, the density of Helicoverpa zea (Boddie) and Manduca spp. eggs was not significantly different between treatment and control plots. Although parasitism was the most important component of egg mortality (19-49%), parasitism was not significantly different between habitat types. Identifiable predation was a minor component (3-9%) of egg fate; it is possible that unidentified predation may be part of the approximately 35-52% of eggs that met unknown fates. Larval parasitism levels ranged from approximately 10 to 90% but was not significantly influenced by the presence of beneficial insect habitat in either year of the study. These results demonstrate that natural enemy activity in organic tomatoes was not amplified, and pest populations were not reduced by the presence of a commercially available beneficial insect habitat.

Animals↗

Standard percent DNA sequence difference for insects does not predict species boundaries.

Diagnosis and assessment of species boundaries of economically important insects are often problematic because of limited morphological and/or biological characters. DNA data can help to identify and revise species. Nonoverlapping intra- and interspecific sequence divergences are often used as evidence for species. Thus, the establishment of a standardized percent nucleotide divergence to predict species boundaries would aid in cases where species status is suspect. However, given variation in nucleotide mutation rates and species concepts, association between a standard percent sequence divergence and species is questionable. This review surveys the percent DNA sequence difference found between sister-species of economically important insects, to assess whether a standard divergence associates with all taxa. Sixty-two comparisons of intra- and interspecific pairwise DNA differences were made for mitochondrial and nuclear loci spanning families of Isoptera, Phthiraptera, Hemiptera, Coleoptera, Lepidoptera, Diptera, and Hymenoptera. Intra- and interspecific sequence divergences varied widely among insects, 0.04-26.0 and 1.0-30.7%, respectively. The ranges of intra- and interspecific sequence divergences overlapped in 28 of 62 comparisons. This implies that a standardized percent sequence divergence would fail to correctly diagnose species for 45% of the cases. Common occurrence of nonmonophyly among closely related species probably explains this observation. Nonmonophyly and overlap of intra- and interspecific divergences were significantly associated. The reviewed studies suggest that a standard percent sequence divergence does not predict species boundaries among economically important insects. DNA data can help best to predict species boundaries via its inclusion in nonphenetic phylogenetic analysis and subsequent systematic expert scrutiny.

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Improved dry-fleshed sweetpotato genotypes resistant to insect pests.

Thirty-five mostly dry-fleshed sweetpotato, Ipomoea batatas (L.) Lam. (Convolvulaceae), genotypes from the USDA-ARS/Clemson University sweetpotato breeding program were evaluated in nine field experiments at the U.S. Vegetable Laboratory, Charleston, SC, from 1998 to 2004. There were highly significant entry effects for percentage of uninjured roots; wireworm, Diabrotica, and Systena (WDS) index; percentage of roots damaged by sweetpotato weevil, Cylas formicarius elegantulus (Summers); percentage of roots damaged by sweetpotato flea beetle, Chaetocnema confinis Crotch); and percentage of roots damaged by white grub larvae (primarily Plectris aliena Chapin). The susceptible control, 'SC1149-19', had a significantly lower percentage of uninjured roots, a significantly higher WDS rating, and higher percentage infestations of flea beetle, grubs, and sweetpotato weevils than all other sweetpotato entries in this study. Twenty-seven genotypes had significantly less insect damage than 'Beauregard', the leading commercial orange-fleshed cultivar in the United States. In addition, 11 genotypes had significantly less insect injury than 'Picadito', a commercial boniato-type sweetpotato grown extensively in southern Florida. Overall, no genotypes were more resistant to soil insect pests than the resistant checks 'Sumor' and 'Regal'. Many of the advanced dry-flesh sweetpotato genotypes had high levels of resistance to soil insect pests, and they represent a useful source of advanced germplasm for use in sweetpotato breeding programs.

Animals↗

Infection of aquatic insects with trematode metacercariae carrying Ehrlichia risticii, the cause of Potomac horse fever.

We provide evidence of Ehrlichia risticii Holland, the agent of Potomac horse fever, in trematode stages found in aquatic insects collected from a pasture stream in northern California, using nested polymerase chain reaction (PCR) amplification and sequence analyses of the 16S rRNA, 51 kDa major antigen and groEL heat shock protein genes. E. risticii was detected in metacercariae found in the immatures and adults of the following insects: caddisflies (Trichoptera), mayflies (Ephemeroptera), damselflies (Odonata, Zygoptera), dragonflies (Odonata, Anisoptera), and stoneflies (Plecoptera). The prevalence of E. risticii was 31.9% (n = 454 individuals) in aquatic insects (13 of 17 species were positive). Prevalence within orders was as follows: 43.5% (n = 207) in caddisflies, 15.2% (n = 92) in mayflies, 13.9% (n = 115) in damselflies, 10.0% (n = 10) in dragonflies, and 80.0% (n = 30) in stoneflies. This study demonstrates a broad intermediate host range for trematodes that act as vector for E. risticii. Insects are likely to play an important role in the epidemiology of this disease.

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Spring succession of necrophilous insects on wildlife carcasses in Louisiana.

Seven fresh animal carcasses were monitored throughout decomposition in a mixed flatwood forest in East Baton Rouge Parish, LA from 1 April to 1 July 1999. Succession patterns of necrophilous insects were documented for the following: one Louisiana black bear (threatened species), two white-tailed deer, two alligators, and two swine as the experimental reference. Our results suggest variation in the species composition of necrophilous insects among animal carcass types. A total of 93 arthropod species, from 46 families and three classes, were manually collected from the seven carcasses. Only 19 insect species were collected on all four animal types and were represented by eight families: Coleoptera: Histeridae, Nitidulidae, Silphidae, Staphylinidae; Diptera: Calliphoridae, Muscidae, Piophilidae, Sepsidae. Eleven of the 46 families were not collected at either alligator site but were observed at bear, deer, and swine carrion: Coleoptera: Cleridae, Dermestidae, Geotrupidae, Scarabaeidae; Diptera: Micropezidae, Sarcophagidae, Syrphidae; Hymenoptera: Apidae; Lepidoptera: Nymphalidae; and Odonata: Libellulidae. Residency and succession patterns of necrophilous insects are presented for each animal type with particular emphasis on selected fly (Calliphoridae, Muscidae, Piophilidae, Stratiomyidae) and beetle species (Cleridae, Dermestidae, Histeridae, Nitidulidae, Silphidae, Staphylinidae).

Alligators and Crocodiles↗

The thyrotrophin hormone receptor of Graves' disease: overexpression of the extracellular domain in insect cells using recombinant baculovirus, immunoaffinity purification and analysis of autoantibody binding.

Since the cloning of the TSH receptor (TSH-R), the target autoantigen of Graves' disease, the receptor has been expressed in a variety of eukaryotic cells to obtain a functional molecule. Despite this success, the levels of receptor expression have been marginally higher than the extremely low levels found in thyroid cells, preventing any progress on the purification of the molecule. In this study, the large extracellular region of the TSH-R, without the membrane spanning segments, has been expressed in insect cells using recombinant baculovirus to generate substantial quantities of the receptor protein. A monoclonal antibody previously generated to a bacterial TSH-R fusion protein was used to characterize and monitor the expression of the truncated receptor in insect cells. Two polypeptides of 63 and 49 kDa were recognized as the components of the truncated recombinant receptor. The 63 kDa protein was shown to be the glycosylated form of the smaller, 49 kDa, component. Expression in different insect cell lines showed that an increase in expression of approximately tenfold was apparent in High Five cells when compared with Sf21 cells. Very small quantities of the truncated receptor were secreted by the three insect cell lines examined, with the majority of the molecule being retained within the cells. Immunoaffinity purification of milligram quantities of the truncated receptor was achieved using the monoclonal antibody. The availability of the purified TSH-R has allowed the establishment of an enzyme-linked immunosorbent assay to measure autoantibodies in the sera of patients with Graves' disease. Although the truncated receptor interacts with autoantibodies, our results show that it does not bind TSH and differs in this respect from other glycoprotein hormone receptors.

Animals↗

High-level expression of recombinant immunoreactive thyroid peroxidase in the High Five insect cell line.

Expression of a major thyroid autoantigen, thyroid peroxidase (TPO) was studied using the baculovirus-insect cell expression system. Human TPO cDNA modified so as to code for the extracellular fragment of the protein was placed under the control of the strong polyhedrin promoter in baculovirus transfer vector pBlueBacIII and cotransfected with linearized AcMNPV viral DNA. Expression in two insect cell lines Spodoptera frugiperda (Sf9) and Tricoplusia ni (High Five) was investigated and levels of recombinant TPO (rTPO) monitored by RIA and SDS-PAGE followed by Western blotting. Both insect cell lines expressed rTPO, but higher levels (30 mg/l culture medium) were obtained with High Five cells. Culture medium rTPO was purified and its glycosylation and immunoreactivity analysed. Lectin-affinity blotting and treatment with glycosidases indicated that both high mannose and complex-type sugar residues were associated with the recombinant protein. Studies with an ELISA based on biotinlabelled rTPO and an immunoprecipitation assay based on 125I-labelled rTPO indicated that the rTPO and native TPO showed similar reactivity to TPO autoantibodies (r = 0.96, P < 0.001, n = 50 and r = 0.99, P < 0.001, n = 80 respectively). In addition, rTPO expressed in High Five cells showed enzyme activity comparable with that of native TPO when the heme biosynthesis precursor delta-aminolevulinic acid was included in the culture medium. Overall, our studies indicate that the High Five insect cell line provides a useful system for the expression of relatively high levels of rTPO which should be suitable for structural analysis of TPO and TPO-TPO autoantibody complexes.

Animals↗

Phylogenetic dispersion of host use in a tropical insect herbivore community.

Theory has long predicted that insect community structure should be related to host plant phylogeny. We examined the distribution of insect herbivore associations with respect to host plant phylogeny for caterpillars (Lepidoptera), beetles (Coleoptera), and grasshoppers and relatives (orthopteroids) in a New Guinea rain forest. We collected herbivores from three lineages of closely related woody plants and from more distantly related plant lineages in the same locality to examine the phylogenetic scale at which host specificity can be detected in a community sample. By grafting molecular phylogenies inferred from three different genes into a supertree, we developed a phylogenetic hypothesis for the host community. Feeding experiments were performed on more than 100 000 live insects collected from the 62 host species. We examined patterns of host use with respect to the host plant phylogeny. As predicted, we found a negative relationship between faunal similarity, defined as the proportion of all herbivores feeding on two hosts that are shared between the hosts, and the phylogenetic distance between hosts based on DNA sequence divergence. Host phylogenetic distance explained a significant fraction of the variance (25%) in herbivore community similarity, in spite of the many ecological factors that probably influence feeding patterns. Herbivore community similarity among congeneric hosts was high (50% on average) compared to overlap among host families (20-30% on average). We confirmed this pattern using the nearest taxon index (NTI) and net relatedness index (NRI) to quantify the extent of phylogenetic clustering in particular herbivore associations and to test whether patterns are significantly different from chance expectations. We found that 40% of caterpillar species showed significant phylogenetic clustering with respect to host plant associations, somewhat more so than for beetles or orthopteroids. We interpret this as evidence that a substantial fraction of tropical forest insect herbivores are clade specialists.

Animals↗

Bioaccumulation and trophic transfer of polychlorinated biphenyls by aquatic and terrestrial insects to tree swallows (Tachycineta bicolor).

Insectivorous passerines often bioaccumulate polychlorinated biphenyls (PCBs) via trophic transfer processes. Tree swallows (Tachycineta bicolor) frequently are used for estimating PCB bioaccumulation, yet the focus on specific trophic links between contaminated sediment and bird has been limited. Bioaccumulation of PCBs from sediment to tree swallows was examined with focus on trophic pathways by simultaneously examining PCBs in emergent aquatic and terrestrial insects and gut contents of nestlings. Total PCB concentrations increased from sediment (123.65 +/- 15.93 microg/kg) to tree swallow nestlings (2,827.76 +/- 505.67 microg/kg), with emergent aquatic insects, terrestrial insects, and gut content samples having intermediate concentrations. Biota-sediment accumulation factors (BSAFs) varied among congeners for tree swallow nestlings and for male and female Chironomus spp. For nestlings, the highest BSAF was for the mono-ortho-substituted congener 118. Nestling biomagnification values were similar for gut contents and female Chironomus spp., suggesting this diet item may be the main contributor to the overall PCB transfer to nestlings. However, gut content samples were highly variable and, on a PCB congener pattern basis, may have been influenced by other taxa, such as terrestrial insects. Considering dietary plasticity of many insectivorous birds, the present study suggests that a variety of potential food items should be considered when examining PCB accumulation in insectivorous passerines.

Animals↗

Cloning of mRNA sequences for two antibacterial peptides in a hemipteran insect, Riptortus clavatus.

Escherichia coli injection rapidly induced bactericidal activity in the hemolymph of a hemipteran insect, Riptortus clavatus. This activity reached its maximum at 9 hr after injection and thereafter declined slowly. Two types of cDNA clones involved in this response were isolated by differential screening. The predominant type encoded for an open reading frame of 678 amino acids, which consisted of fourteen tandem repeats. Each repeat was rich in charged residues and had a proline-rich region which had striking sequence similarities to proline-rich antibacterial peptides from other insect species, indicating these clones encode a multipeptide precursor of antibacterial peptides. The other type encoded for a glycine-rich peptide similar to a known antibacterial peptide as well. Northern blot analyses revealed rapid induction of mRNAs corresponding to these clones after the injection. To our knowledge, this is the first report on the mRNA sequences of antibacterial peptides of hemimetabolous insects, and the second report on the occurrence of multipeptide precursor structure in insect antibacterial peptides.

Animals↗

Genome evolution in an insect cell: distinct features of an ant-bacterial partnership.

Bacteria that live exclusively within eukaryotic host cells include not only well-known pathogens, but also obligate mutualists, many of which occur in diverse insect groups such as aphids, psyllids, tsetse flies, and the ant genus Camponotus (Buchner, 1965; Douglas, 1998; Moran and Telang, 1998; Baumann et al., 2000; Moran and Baumann, 2000). In contrast to intracellular pathogens, these primary (P) endosymbionts of insects are required for the survival and reproduction of the host, exist within specialized host cells called bacteriocytes, and undergo stable maternal transmission through host lineages (Buchner, 1965; McLean and Houk, 1973). Due to their long-term host associations and close phylogenetic relationship with well-characterized enterobacteria (Fig. 1), P-endosymbionts of insects are ideal model systems to examine changes in genome content and architecture that occur in the context of beneficial, intracellular associations. Since these bacteria have not been cultured outside of the host cell, they are difficult to study with traditional genetic or physiological approaches. However, in recent years, molecular and computational approaches have provided important insights into their genetic diversity and ecological significance. This review describes some recent insights into the evolutionary genetics of obligate insect-bacteria symbioses, with a particular focus on an intriguing association between the bacterial endosymbiont Blochmannia and its ant hosts.

Animals↗