Biochemical and immunological properties of different electrophoretic forms of juvenile hormone esterase from Trichoplusia ni (Hübner)

The two major electrophoretic forms (pI 5.5, 5.3) of juvenile hormone esterase were independently isolated from hemolymph of larval Trichoplusia ni. A simple and rapid preparation procedure of poly(ethylene glycol) precipitation, Sephadex gel filtration and chromatofocusing is described. Analytical isoelectric focusing showed only one peak of juvenile hormone esterase activity in the respective purified samples, whereas there were four (two major) such peaks in the hemolymph. The amino acid composition of the two forms was similar. The comparison of peptides obtained after protein fragmentation by cyanogen bromide showed that juvenile hormone esterases A and B were very similar, although definitely not identical, in amino acid sequence. The immunological comparisons of juvenile hormone esterases suggested that the number of polyclonal antibody binding sites on both forms was the same. There were no detected differences between immunoreactive properties of juvenile hormone esterase from the hemolymph of different stages of larval maturation. The influence of the active site of the enzyme on its antigenic properties was studied by immunocompetition. The inactive, heat-denatured juvenile hormone esterase can only partially protect against inhibition of its activity by the antibodies, whereas an organophosphate inhibitor which covalently binds to the catalytic center of the enzyme did not change the immunoreactive properties in comparison to active juvenile hormone esterase from hemolymph. These data show that heat-denatured juvenile hormone esterase has lost at least one or more epitopes, but the catalytic site of the enzyme is distinct from the epitopes.     

Effects of juvenile hormone I and the anti-juvenile hormone fluoromevalono-lactone on development and juvenile hormone esterase activity in post-feeding last-stadium larvae of Trichoplusia ni (Hübner)

Treatment of post-feeding (early day 3; wandering phase) last-stadium larvae of the cabbage looper, Trichoplusia ni, with the anti-juvenile hormone, fluoromevalonolactone, prevented the normal ecdysis to the pupa. It caused the formation of larval-pupal intermediates, a dose-dependent delay in the time of tanning, and a decrease in juvenile hormone esterase activity at the time of the prepupal juvenile hormone esterase peak. Fluoromevalonolactone was inactive as juvenile hormone esterase inhibitor in vitro. Conversely, juvenile hormone I accelerated the time of tanning, induced the early appearance of juvenile hormone esterase activity, and prevented adult eclosion. Although most of the larvae that were treated with fluoromevalonolactone immediately after the prepupal burst of juvenile hormone (late on day 3; post-spinning phase) still became larval-pupal intermediates, the time of tanning and juvenile hormone esterase activity were close to normal. Topical treatment of day-3 larvae with radiolabelled juvenile hormone I resulted in the rapid appearance and decline of radiolabelled juvenile hormone I in the haemolymph which was associated with the increased production of juvenile hormone I acid and the induced appearance of juvenile hormone esterase activity. Thus, in post-feeding last-stadium larvae of T. ni, juvenile hormone seems to be necessary for the proper formation of the pupa. Juvenile hormone is also involved in determining the time of pupation, and it appears to induce its own degradation.     

The evaluation of anti juvenile hormones using last stadium larvae of the cabbage looper,Trichoplusia ni (Hübner)

Treatment of prepupae of the cabbage looper, Trichoplusia ni (Hübner), with the anti juvenile hormone (AJH) fluoromevalonolactone (FMev) reduces juvenile hormone esterase (JHE) activity and either delays or inhibits the normal larval-pupal ecdysis in a dose-dependent manner, resulting in the formation of pupae with a suite of characteristic abnormalities. A variety of putative AJHs were evaluated using these teratogenic effects and changes in JHE activity as indicators of AJH activity. Like FMev, the compactin analog L-643,049-01K01 and, at very high doses, 3,3-dichloro-2-propenyl hexanoate (DPH) behaved as AJHs. FMev and L-643,049-01K01 were most effective when larvae were treated at gut purge, while DPH was the most effective after the initiation of spinning behavior.     

Effect of the insect parasite Hyposoter exiguae (Viereck) on the carbohydrate metabolism of its host,Trichoplusia ni (Hübner)

Parasitization of Trichoplusia ni by Hyposoter exiguae resulted in elevations in the haemolymph trehalose concentration and fat body glycogen level. These increases in tissue carbohydrate reserves were accompanied by an elevation in the maximal velocities of fat body phosphofructokinase and fructose 1,6 bisphosphatase. The activity of the latter, however, was several-fold greater than that of phosphofructokinase and the potential gluconeogenic flux through the fructose phosphate step was, therefore, markedly increased following parasitization. The changes in adenylate levels in parasitized fat body, specifically an increase in AMP and decrease in ATP, were consistent with an elevation in phosphofructokinase activity in vivo but not in fructose 1,6 bisphosphatase activity. The latter enzyme, however, was shown to be less sensitive to both AMP and fructose 2,6 bisphosphate inhibition in fat body of parasitized individuals. Decreased haemolymph glucogenic amino acid concentrations and an increase in faecal uric acid level accompanied the above effects. It was concluded that parasitization of T. ni by H. exiguae caused a stress-induced hyperglycemia that may result from an alteration in the hormonal regulation over carbohydrate synthesis.     

External antennal morphometry of Trichoplusia Ni (Hübner) (Lepidoptera: Noctuidae)

A detailed morphometric analysis of the antennae of male and female Trichoplusia ni (Hübner) is presented, based on light and scanning electron microscopic observations. These measurements of general antennal dimensions and the number, distribution, dimensions, and surface features of the antennal sensilla are applied in the consideration of olfactory transduction processes and in the derivation of a mathematical representation of the antennal surface. The surface area and the number of sensilla on the antennal flagellum are found to vary with the weight of the pupa and adult moth. The long Type 1 sensilla trichodea are hypothesized to be innervated by pheromone-sensitive olfactory cells on the basis of morphological comparisons with sensilla of other insects.     

Influence of a haemolymph protein fraction on the binding of juvenile hormone in homogenates of insect epidermis (Plodia interpunctella (Hübner))

Summary A carrier protein fraction (CPF) from larval haemolymph was found to influence binding and catabolism of tritiated juvenile hormone (JH) in homogenates of larval epidermis. The CPF reduced binding of tritiated JH in all of the particulate fractions but did not alter the relative binding pattern when compared with JH alone. The CPF also protected the hormone from degradative enzymes in the membrane vesicle and microsomal + cytosol fractions but not in the nuclear and mitochondrial fractions. Preliminary evidence exists for high-affinity binding sites for JH in the nuclear and mitochondrial fractions. We conclude that the CPF influences catabolism of the tritiated JH but does not participate in subcellular recognition of JH in homogenized target tissue.Mention of a proprietary product in this paper does not constitute an endorsement of that product by the U.S. Department of Agriculture     

Does PBAN play an alternative role of controlling pheromone emission in the cabbage looper moth,Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae)?

There is an active process by which sex pheromone reserves of female cabbage looper moths, Trichoplusia ni, are transported to the gland's surface during the nocturnal period of calling. We hypothesized that this mobilization was controlled by a head factor, possibly related to the pheromone biosynthesis activating neuropeptides (PBAN) that in other species stimulate pheromone synthesis. We evaluated the impact of head extracts of T. ni on pheromone emission and glandular content of pheromone. During the photophase injected head extracts stimulated an increased pheromone emission rate in females, but glandular content of pheromone was not affected. Head extracts of H. virescens, a species with known PBAN activity, and synthetic PBAN stimulated an increased pheromone emission rate in T. ni. There was some specificity of the response of female T. ni to PBAN, in that several other unrelated polypeptides did not stimulate this type of response. Previously it had been determined that brain factors do not play a role in stimulating pheromone biosynthesis in T. ni. Our results indicate that there may be additional avenues by which PBAN or related neuropeptides control pheromone emission, including transport of pheromone reserves to the surface of the sex pheromone gland.     

Binding of Insecticidal Crystal Proteins of Bacillus thuringiensis to the Midgut Brush Border of the Cabbage Looper, Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae), and Selection for Resistance to One of the Crystal Proteins

The susceptibility of Trichoplusia ni larvae to several Bacillus thuringiensis insecticidal crystal proteins (ICPs) was tested. Neonatal larvae proved to be susceptible to solubilized trypsin-treated CryIA(a), CryIA(b), and CryIA(c) (50% lethal concentrations [LC(50)s], 570, 480, and 320 ng/cm, respectively) but showed little susceptibility to CryIB and CryID (LC(50)s, 5,640 and 2,530 ng/cm, respectively). The toxicity of ICPs was correlated to binding to the epithelial brush border of the midgut, as revealed by immunocytochemical staining with monoclonal antibodies. In vitro binding experiments with iodinated ICPs and brush border membrane vesicles indicated that CryIA(b) and CryIA(c) share the same high-affinity binding site, whereas CryIA(a) binds to a different one. The affinities of CryIA(b) and CryIA(c) for the binding site were similar (K(d) = 3.6 and 4.7 nM, respectively), and the mean binding-site concentration was 0.71 pmol/mg of vesicle protein. Selection of a population with increasing concentrations of CryIA(b) produced 31-fold resistance in seven generations. The realized heritability (h) was 0.19. The increase of homozygosity (for resistance factors) as selection proceeded was reflected in the increase in the slopes of the dose-mortality curves. Resistance was specific for CryIA(b) and did not extend to CryIA(a) or even to CryIA(c). This result was not predicted by the binding-site model, in which CryIA(b) and CryIA(c) bind to the same high-affinity binding site. This result may suggest a more complicated relationship between in vitro binding of ICPs to specific sites in the epithelial membrane of the midgut and the in vivo toxic effect.     

DNA synthesis in the imaginal wing discs of the American bollwormHelicoverpa armigera (Hübner)

The effect of two insect growth regulators of plant origin viz. plumbagin and azadirachtin and the ecdysteroids 20-hydroxyecdysone, makisterone A and a phytoecdysteroid on DNA synthesis in imaginal wing discs of day 4 final instarHelicoverpa armigera larvae was studied. DNA synthesis increased with increase in time of incubation up to 8 h and decreased later without the addition of moulting hormone. Addition of 20-hydroxyecdysone supported long term acquisition of competence for DNA synthesis in the wing discs. Both DNA synthesis and protein content were drastically reduced in plumbagin and azadirachtin-treated insects. Underin vitro conditions, plumbagin had a more pronounced inhibitory effect than azadirachtin. All the ecdysteroids tested, viz. makisterone A, 20-hydroxyecdysone and the ecdysteroidal fraction from the silver fernCheilanthes farinosa enhanced DNA synthesis     

Nutrition of the cabbage looper,Trichoplusia ni (Hübn.)—I. Some requirements for larval growth and wing development

The cabbage looper, Trichoplusia ni (Hübner), was reared on a meridic diet containing wheat germ (oil and solids) as the only plant adjuvant. Some attempts were made to determine the specific nutrients contained in wheat germ which are essential for Trichoplusia. Wheat-germ residue after chloroform: methanol extraction contained a factor required for normal larval growth, and the lipid fraction contained a factor required both for larval growth and normal wing development. Lack of the wing development factor resulted in various gradations of wing deformity or in failure of the moths to emerge from the pupal case. The factor was contained in the saponifiable portion of wheatgerm oil. A dietary supplement of methyl linoleate promoted larval growth and normal pupation, but did not produce normal wings.Ascorbic acid was also demonstrated to be an essential dietary requirement for Trichoplusia. Deficiency of ascorbic acid resulted in complete larval mortality at an immature stage.     

A taxonomic review of the genus Coleophora Hübner (Lepidoptera: Coleophoridae) in Korea

Twenty-seven species of the genus Coleophora Hübner (Lepidoptera, Coleophoridae) are recognized in the Korean peninsula, including seven species that are new to Korea: C. albicans Zeller, C. falkovitshella Vives, C. honshuella Baldizzone & Oku, C. japonicella Oku, C. kononenkoi Baldizzone & Savenkov, C. rectimarginalis Li, and C. vestianella (Linnaeus). For all known species, bibliographies, collecting localities, distribution, biological information, and some taxonomic notes are provided, with illustrations of the male or the female genitalia.     

Fecundity and life table parameters of Campoletis sonorensis (Hymenoptera: Ichneumonidae), an endoparasitoid of the Cabbage Looper Trichoplusia ni Hübner (Lepidoptera: Noctuidae), under laboratory conditions

The fecundity and life table parameters of Campoletis sonorensis females were measured using thirty 4-day-old Trichoplusia ni larvae daily at 24°C, 60% RH and a photoperiod of 12 h L:12 h D. The mean longevity was 34.5±2.8 days, the mean oviposition period was 22.7±1.9 days, and the mean constant oviposition period and the mean post-oviposition period were 15.9±1.3 and 11.9±2.2 days, respectively. The mean realised fecundity and the mean fertility differed significantly at 66.9±7.8 and 60.4±7.8 parasitoids per female, respectively. The mean sex ratio for the mean oviposition period (23 days) was 0.13±0.07, indicating a highly female biased ratio. The life table parameters were: intrinsic rate of natural increase (r _m), 0.135 female/day; gross reproductive rate (GRR), 50.30; net reproductive rate (R _o), 49.96; finite capacity for increase (λ), 1.14 female/day; mean generation time (T), 28.97 days; doubling time (DT), 5.13; capacity for increase (r _c), 0.33; and cohort generation time (T _c), 11.69. Campoletis sonorensis may be a suitable candidate for a biocontrol program of T. ni populations mostly because the primary selection criterion, r_m , obtained for this parasitoid can be similar to or larger than the rate obtained for T. ni.