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Extracts of botanical origin naturally contain a complex mixture of chemicals considered effective in managing lepidopteran pests. Chemical screening of the ethanolic leaf extracts of two fig tree species, Ficus lyrata and Ficus auriculata, delivered 12 and 15 phyto‐compounds with relatively high peak area percentages in phytol and flavone, respectively. Larvicidal activity against Spodoptera litura yielded higher mortality rates at maximum‐concentration treatment (600 ppm) with Ficus lyrata (91.3%) and Ficus auriculata (98.5%) extracts during the second instar. Sub‐lethal dosages (300 ppm) of both Ficus lyrata and Ficus auriculata extracts impeded the development and reproduction of lepidopteran pests. The enzyme‐inhibition activity of both fig extracts elicited a significant reduction in the major digestive enzymes alkaline phosphatase, acid phosphatase, and lactate dehydrogenase dose‐responsively. Mid‐gut histological screening of Ficus lyrata and Ficus auriculata extracts displayed gut lumen disruptions, shape alterations of columnar cells, and brush‐border membrane damage. Further, the non‐target toxicity of fig extracts against the red worm Eisenia foetida was minimal compared with that of the chemical temephos. Overall, the Ficus extracts proved to be eco‐friendly strategies for managing the polyphagous pest Spodoptera litura and are potentially more sustainable and less harmful to non‐target earthworms. Nonetheless, in silico predictions suggest that the active compounds in fig extracts are predominantly toxic against honeybees (16 compounds) and violate TICE rules (10 compounds). Therefore, the biological actions of fig extracts' individual novel chemistries need to be examined on target and non‐target species to develop better pest management strategies. [ABSTRACT FROM AUTHOR]

Abstract Two genes, Distal-less (Dll) and spalt (sal), are known to be involved in establishing nymphalid butterfly wing patterns. They function in several ways: in the differentiation of the eyespot’s central signalling cells, or foci; in the differentiation of the surrounding black disc; in overall scale melanisation (Dll); and in elaborating marginal patterns, such as parafocal elements. However, little is known about the functions of these genes in the development of wing patterns in other butterfly families. Here, we study the expression and function of Dll and sal in the development of spots and other melanic wing patterns of the Indian cabbage white, Pieris canidia, a pierid butterfly. In P. canidia, both Dll and Sal proteins are expressed in the scale-building cells at the wing tips, in chevron patterns along the pupal wing margins, and in areas of future scale melanisation. Additionally, Sal alone is expressed in the future black spots. CRISPR knockouts of Dll and sal showed that each gene is required for the development of melanic wing pattern elements, and repressing pteridine granule formation, in the areas where they are expressed. We conclude that both genes likely play ancestral roles in organising distal butterfly wing patterns, across pierid and nymphalid butterflies, but are unlikely to be differentiating signalling centres in pierids black spots. The genetic and developmental mechanisms that set up the location of spots and eyespots are likely distinct in each lineage.

Agricultural intensification has led to a dramatic impoverishment of biodiversity. Moths are not an exception: rapid declines of common and widespread species have been reported and it is still not clear which conservation measures should be applied to restore their populations.We investigated how much of the variation in moth assemblages inhabiting extensively managed meadows in a region of high‐intensity agriculture is explained by surrounding woody landscape features, meadow management and vegetation composition, and which specific variables are the most influential.Results show that the percentage of forest and the length of hedgerows within a 250 m radius were especially important for forest macromoths, explaining 42% of variation in abundance and 23% of species richness, whereas meadow management played a relatively marginal role. The abundance of grassland macromoths was also positively influenced by the length of hedgerows in the landscape (combined with elevation it explained 10% of the variance), while it responded negatively to uncut refuge and to delayed mowing (7%). Regarding grassland macromoth species richness, the annual harvesting frequency was the main predictor (4%). In contrast, micromoth abundance was more influenced by meadow management, notably the date of the first cut (4%) and vegetation composition (8%, though not conclusive due to a single outlier), whereas landscape features explained nothing.Altogether, these results demonstrate the importance of both woody features within farmed landscapes and biodiversity‐friendly meadow management for the maintenance of integral moth communities. [ABSTRACT FROM AUTHOR]

Two genes, Distal-less (Dll) and spalt (sal), are known to be involved in establishing nymphalid butterfly wing patterns. They function in several ways: in the differentiation of the eyespot's central signalling cells, or foci; in the differentiation of the surrounding black disc; in overall scale melanisation (Dll); and in elaborating marginal patterns, such as parafocal elements. However, little is known about the functions of these genes in the development of wing patterns in other butterfly families. Here, we study the expression and function of Dll and sal in the development of spots and other melanic wing patterns of the Indian cabbage white, Pieris canidia, a pierid butterfly. In P. canidia, both Dll and Sal proteins are expressed in the scale-building cells at the wing tips, in chevron patterns along the pupal wing margins, and in areas of future scale melanisation. Additionally, Sal alone is expressed in the future black spots. CRISPR knockouts of Dll and sal showed that each gene is required for the development of melanic wing pattern elements, and repressing pteridine granule formation, in the areas where they are expressed. We conclude that both genes likely play ancestral roles in organising distal butterfly wing patterns, across pierid and nymphalid butterflies, but are unlikely to be differentiating signalling centres in pierids black spots. The genetic and developmental mechanisms that set up the location of spots and eyespots are likely distinct in each lineage. [ABSTRACT FROM AUTHOR]

Green pesticides, derived from natural sources, have gained wider attention as an alternative to synthetic pesticides for managing polyphagous pests, such as Spodoptera litura. In this study, the methanolic flower extract of Nyctanthes arbor-tristis (Mx-Na-t) was subjected to chemical screening, and 3-hydroxy-1,2-dimethyl-4(1H)-pyridone (3H-dp) and tyrosol (Ty-ol) were identified as the major derivatives. The toxic effects of Mx-Na-t (500 ppm) were highest in third-instar S. litura larvae (96.4%), while those of 3H-dp and Ty-ol (5 ppm) were highest in second-instar larvae (76.5% and 81.4%, respectively). The growth and development of S. litura larvae and pupae were significantly reduced by all three treatments. Fecundity rates were also reduced by all treatments [from 1020 eggs (control) to 540 eggs by Mx-Na-t treatment, 741 eggs by 3H-dp treatment, and 721 eggs by Ty-ol treatment]. The extract and its active constituents decreased adult emergence and slowed total larval development in a dose-dependent manner. A decrease was noted in the major gut enzymes of young S. litura larvae exposed to Mx-Na-t, 3H-dp, and Ty-ol. Moreover, midgut tissues of fourth-instar larvae were severely damaged by Mx-Na-t (250 ppm), 3H-dp (2.5 ppm), and Ty-ol (2.5 ppm); the treatments induced structural damage to the epithelial cells and gut lumen. The earthworm Eisenia fetida was used to assess nontarget toxicity. Compared with cypermethrin, the phytochemicals exhibited minimal effects on the earthworm's detoxifying enzymes superoxide dismutase and catalase after 14 days of treatment. Moreover, in silico predictions using BeeTox and ProTox-II indicated little or no toxicity of 3H-dp and Ty-ol toward honey bees and other nontarget species. • Toxicity against S. litura larvae of Mx-Na-t (at 500 ppm) was highest in 3rd instars. • Phytometabolites of Mx-Nat delivered significant inhibition of S. litura midgut enzymes. • Growth and development of S. litura larvae and pupae were significantly reduced. • The nontarget toxicty of phytometabolites delivered harmless actions against earthworm. [ABSTRACT FROM AUTHOR]

Entomopathogenic fungi, particularly Metarhizium and Beauveria species, are emerging as effective biocontrol agents for combating lepidopteran pests in agricultural and forestry settings. This review provides a comprehensive overview of their modes of action, secondary metabolites, extracellular enzymes, and infection mechanisms. These fungi employ various strategies, including the secretion of proteolytic enzymes, chitinolytic enzymes, esterases, and lipases, to penetrate the insect cuticle and initiate infection. The process involves spore recognition, adhesion, germination, and differentiation into infective structures. The impact of fungal strains on the insect immune system and the commercial availability of fungal pesticides are also discussed. Furthermore, the advancements in genetically engineered mycotoxins and the key challenges facing their implementation are addressed, in addition to listing future research directions. This review offers valuable insights for researchers involved in the development and application of entomopathogenic fungi for sustainable pest management practices. [Display omitted] • Exploration of the mode of actions of Metarhizium and Beauveria fungi, for targeting lepidopteran pests. • Examination of secondary metabolites contributing to the pathogenicity of entomopathogenic fungi. • Analyze the mechanism of extracellular enzymes, in the degradation of insect cuticles. • Insights into the infection mechanisms employed by mycotoxins to invade lepidopteran hosts. [ABSTRACT FROM AUTHOR]

BACKGROUND Broflanilide has been registered in China for the control of Lepidoptera and Coleoptera pests, and is widely used to control the target pests at lethal and sublethal levels. The lethal and sublethal effects of broflanilide on the common cutworm (CCW) Spodoptera litura Fabricius, a representative Lepidopteran pest in agricultural crops, were examined to explore its ecological influence on pests. RESULTS: In F0, broflanilide had little influence on the hatchability of eggs, but significantly reduced the neonate survival rate. The lethal activity of broflanilide towards third‐instar larvae and adults was 0.13 mg kg–1 (LD50) and 3.59 mg L–1 (LC50) respectively at 48 h. After being treated with a sublethal dose (LD10 and LD30) of broflanilide, the duration of third‐ to sixth‐instar larvae and the mean fecundity of reproductive females were significantly increased, but pupation rate, weight of pupae and life‐cycle rate were significantly decreased. In F1, the duration of F1 larvae and the doubling time were prolonged, whereas the rates of pupation and the life cycle were decreased by 14.92% and 18.00%, respectively. The intrinsic rate of increase, finite rate of increase and net reproductive rate in the sublethal group were lower than in the control group. The relative fitness of F1 treated by LD10 and LD30 was 0.81 and 0.66, respectively. CONSLUSION: Broflanilide not only has highly lethal activity, but also suppresses the population growth and progeny of CCW, as a critical factor for guidelines of its usage in the field. [ABSTRACT FROM AUTHOR]

RNA interference (RNAi) is a method of gene silencing where dsRNA is digested into small interfering RNA (siRNA) in the presence of enzymes. These siRNAs then target homologous mRNA sequences aided by the RNA-induced silencing complex (RISC). The mechanism of dsRNA uptake has been well studied and established across many living organisms including insects. In insects, RNAi is a novel and potential tool to develop future pest management means targeting various classes of insects including dipterans, coleopterans, hemipterans, lepidopterans, hymenopterans and isopterans. However, the extent of RNAi in individual class varies due to underlying mechanisms. The present review focuses on three major insect classes viz hemipterans, lepidopterans and coleopterans and the rationale behind this lies in the fact that studies pertaining to RNAi has been extensively performed in these groups. Additionally, these classes harbour major agriculturally important pest species which require due attention. Interestingly, all the three classes exhibit varying levels of RNAi efficiencies with the coleopterans exhibiting maximum response, while hemipterans are relatively inefficient. Lepidopterans on the other hand, show minimum response to RNAi. This has been attributed to many facts and few important being endosomal escape, high activity dsRNA-specific nucleases, and highly alkaline gut environment which renders the dsRNA unstable. Various methods have been established to ensure safe delivery of dsRNA into the biological system of the insect. The most common method for dsRNA administration is supplementing the diet of insects via spraying onto leaves and other commonly eaten parts of the plant. This method is environment-friendly and superior to the hazardous effects of pesticides. Another method involves submergence of root systems in dsRNA solutions and subsequent uptake by the phloem. Additionally, more recent techniques are nanoparticle- and Agrobacterium -mediated delivery systems. However, due to the novelty of these biotechnological methods and recalcitrant nature of certain crops, further optimization is required. This review emphasizes on RNAi developments in agriculturally important insect species and the major hurdles for efficient RNAi in these groups. The review also discusses in detail the development of new techniques to enhance RNAi efficiency using liposomes and nanoparticles, transplastomics, microbial-mediated delivery and chemical methods. [ABSTRACT FROM AUTHOR]

RNA interference (RNAi) is a method of gene silencing where dsRNA is digested into small interfering RNA (siRNA) in the presence of enzymes. These siRNAs then target homologous mRNA sequences aided by the RNA-induced silencing complex (RISC). The mechanism of dsRNA uptake has been well studied and established across many living organisms including insects. In insects, RNAi is a novel and potential tool to develop future pest management means targeting various classes of insects including dipterans, coleopterans, hemipterans, lepidopterans, hymenopterans and isopterans. However, the extent of RNAi in individual class varies due to underlying mechanisms. The present review focuses on three major insect classes viz hemipterans, lepidopterans and coleopterans and the rationale behind this lies in the fact that studies pertaining to RNAi has been extensively performed in these groups. Additionally, these classes harbour major agriculturally important pest species which require due attention. Interestingly, all the three classes exhibit varying levels of RNAi efficiencies with the coleopterans exhibiting maximum response, while hemipterans are relatively inefficient. Lepidopterans on the other hand, show minimum response to RNAi. This has been attributed to many facts and few important being endosomal escape, high activity dsRNA-specific nucleases, and highly alkaline gut environment which renders the dsRNA unstable. Various methods have been established to ensure safe delivery of dsRNA into the biological system of the insect. The most common method for dsRNA administration is supplementing the diet of insects via spraying onto leaves and other commonly eaten parts of the plant. This method is environment-friendly and superior to the hazardous effects of pesticides. Another method involves submergence of root systems in dsRNA solutions and subsequent uptake by the phloem. Additionally, more recent techniques are nanoparticle- and Agrobacterium-mediated delivery systems. However, due to the novelty of these biotechnological methods and recalcitrant nature of certain crops, further optimization is required. This review emphasizes on RNAi developments in agriculturally important insect species and the major hurdles for efficient RNAi in these groups. The review also discusses in detail the development of new techniques to enhance RNAi efficiency using liposomes and nanoparticles, transplastomics, microbial-mediated delivery and chemical methods.

Plants have intrinsic structural, chemical, and proteomic defense mechanisms against pests that can be developed as natural biopesticides as an environmentally friendly and safe alternative. This review investigates advances in biopesticides, focusing on protease inhibitors (PIs) that interfere with the digestion of proteins in insect guts, adversely affecting insect nutrition, growth, and metabolism. One type of PI trypsin inhibitor [TI] was assessed from seeds of 56 plants belonging to 8 families for insecticidal and fungicidal activity with up to 90% effectiveness. Lepidopterans were the main insects causing the most damage to important cultivated crops, and TIs from Momordica cochinchinensis showed up to 80% inhibitory activity. They were selected for further assessment for their potential as biopesticides for pest management. TIs from M. cochinchinensis have been well characterized, and their small sizes, high potency, and genetic simplicity make them ideal candidates for development and commercialization as natural biopesticides. [ABSTRACT FROM AUTHOR]

We detected enzymatic activity that generates 20-nucleotide (nt) RNA from double-stranded RNAs (dsRNAs) in crude extracts prepared from various silkworm (Bombyx mori) organs. The result using knocked-down cultured cells indicated that this dicing activity originated from B. mori Dicer-2 (BmDcr2). Biochemical analyses revealed that BmDcr2 preferentially cleaves 5'-phosphorylated dsRNAs at the 20-nt site-counted from the 5'-phosphorylated end-and required ATP and magnesium ions for the dicing reaction. This is the first report of the biochemical characterization of Dicer-2 in lepidopteran insects. This enzymatic property of BmDcr2 in vitro is consistent with the in vivo small interfering RNA profile in virus-infected silkworm cells., (© 2024 Wiley Periodicals LLC.)

Low tunnels covered with spun-bonded fabric (row covers) provide season extension for vegetable production and also afford a physical barrier against airborne insects and other non-soil pests. Brussels sprouts, Brassica oleracea L. group Gemmifera (Brassicaceae), is a popular vegetable in local markets in Virginia; however, unprotected field production is severely affected by insect pest infestation. This study's objective was to determine the level of protection low tunnels provide against insect infestation and leaf herbivory injury. The experiment was conducted at the Virginia Tech Eastern Shore Agricultural Research and Extension Center in Painter, Virginia. The experimental design was split-plot with polyethylene soil mulches (white or black) as whole plot factors and production systems (low tunnel or open field) as subplot factors. In this study, low tunnels reduced insect infestation and chewing herbivory leaf injury to Brussels sprouts. Compared to an unprotected open field, infestations of lepidopteran insects and harlequin bug, Murgantia histrionica (Hahn) (Hemiptera: Pentatomidae) were reduced on plants under low tunnels. However, aphids (Hemiptera: Aphidae) infestation occurred under low tunnels in fall. There was no effect of color mulches (white or black) and no interaction between tunnel and mulch color on insect infestation and chewing injury. Fewer insect infestations and feeding injury indicate that low tunnels can be an effective management tool for sustainable vegetable production. [ABSTRACT FROM AUTHOR]

Pest insects pose a heavy burden on global agricultural industries with small molecule insecticides being predominantly used for their control. Unwanted side effects and resistance development plagues most small molecule insecticides such as the neonicotinoids, which have been reported to be harmful to honeybees. Bioinsecticides like Bacillus thuringiensis (Bt) toxins can be used as environmentally-friendly alternatives. Arachnid venoms comprise another promising source of bioinsecticides, containing a multitude of selective and potent insecticidal toxins. Unfortunately, no standardised insect models are currently available to assess the suitability of insecticidal agents under laboratory conditions. Thus, we aimed to develop a laboratory model that closely mimics field conditions by employing a leaf disk assay (LDA) for oral application of insecticidal agents in a bioassay tray format. Neonate larvae of the cotton bollworm (Helicoverpa armigera) were fed with soybean (Glycine max) leaves that were treated with different insecticidal agents. We observed dose-dependent insecticidal effects for Bt toxin and the neonicotinoid insecticide imidacloprid, with imidacloprid exhibiting a faster response. Furthermore, we identified several insecticidal arachnid venoms that were active when co-applied with sub-lethal doses of Bt toxin. We propose the H. armigera LDA as a suitable tool for assessing the insecticidal effects of insecticidal agents against lepidopterans. [Display omitted] • Oral activity is often overlooked when characterising insecticidal compounds. • We established a tray-based leaf disc assay (LDA) that mimics lepidopteran larval feeding in nature. • Our LDA confirmed the oral activities of existing commercial insecticides imidacloprid and Bt toxin. • We found that several arachnid venoms caused oral toxicity in the presence of sublethal Bt toxin doses. [ABSTRACT FROM AUTHOR]

The present research investigated the chemical characterization and insecticidal activity of n-Hexane extracts of Epaltes divaricata (NH-EDx) along with their chief derivatives n-Hexadecanoic acid (n-HDa) and n-Octadecanoic acid (n-ODa) against the dengue vector Aedes aegypti and lepidopteran pest Spodoptera litura. Chemical screening of NH-EDx through GC–MS analysis delivered nine major derivatives, and the maximum peak area percentage was observed in n-Hexadecanoic acid (14.63%) followed by n-Octadecadienoic acid (6.73%). The larvicidal activity of NH-EDx (1000 ppm), n-HDa (5 ppm), and n-ODa (5 ppm) against the A. aegypti and S. litura larvae showed significant mortality rate in a dose-dependent way across all the instars. The larvicidal activity was profound in the A. aegypti as compared to the S. litura across all the larval instars. The sublethal dosages of NH-EDx (500 ppm), n-HDa (2.5 ppm), and n-ODa (2.5 ppm) also showed alterations in the larval/pupal durations and adult longevity in both the insect pests. The enzyme activity revealed that the α- and β-carboxylesterase levels were decreased significantly in both the insect pests, whereas the levels of GST and CYP450 uplifted in a dose-dependent manner of NH-EDx, n-HDa, and n-ODa. Correspondingly, midgut tissues such as the epithelial layer (EL), gut lumen (GL), peritrophic matrix (Pm), and brush border membrane (BBM) were significantly altered in their morphology across both A. aegypti and S. litura against the NH-EDx and their bioactive metabolites. NH-EDx and their bioactive metabolites n-HDa and n-ODa showed significant larvicidal, growth retardant, enzyme inhibition, and midgut toxicity effects against two crucial agriculturally and medically challenging insect pest of ecological importance.

The objectives of this research are to identify the Lepidoptera and parasitoid that associated on C. odorata. The research was conducted in Laboratory of Pest and Disease of Crop of Agriculture Faculty UNS and in C. odorata plantation in Banyudono, Boyolali Regency, Central Java. Observation was done at seedling and crop yield. Lepidoptera (larvae, pupae, adult) and parasitoids that found were taken to be identified in laboratory. Larvae and pupae which infected by parasitoid or pathogen placed in separated jar to be kept and waited for parasitoid emerged. Collection of Lepidopteras and parasitoids were identified by comparing with picture, using key of determination, and combination both of them. Identifying was done at family and several of them at species.In this research was found eight species of Lepidopteras and two parasitoids. Eight species of Lepidopteras consist of species of family Lymantridae, Nymphalidae, Papilionidae, Satyridae, Psychidae, and Saturniidae (Cricula trifenestrata and Attacus atlas). Two parasitoid were Apanteles sp. and Pteromalus sp.)

Humans have drastically increased the availability of many once-limited nutrients, resulting in potential attraction of animals to now toxic, novel nutritional conditions. For instance, herbivores are attracted to sodium in many terrestrial ecosystems because sodium occurs in much lower concentrations in plants than in animals. However, sodium availability has increased due to crop irrigation or distribution of de-icing salts along roads. While moderate increases in once-limited sodium may be beneficial, large increases are potentially toxic. Here, we investigate whether increased sodium along roadsides is functioning as an ecological trap, drawing butterflies to oviposit and forage on plants with toxic levels of sodium. As in many other animals, butterflies exhibit sodium-specific foraging behaviour – males will 'puddle' for minerals on mud, dung or carrion; yet sodium-seeking behaviours at other life stages remain unexplored. We present data from a series of field- and laboratory-based sodium preference trials using both caterpillars and ovipositing adult females. Female monarch and cabbage white butterflies did not show any preference for laying eggs on sodium-enriched plants, either at moderate levels that are likely to be physiologically beneficial, or at potentially toxic levels typical of high-traffic roads. Monarch caterpillars somewhat avoided the plants highest in sodium, but this behaviour is unlikely to compensate for the failure of ovipositing females to avoid toxic high-sodium plants. These results suggest that while butterflies frequently use sodium-enriched roadside habitats, they are unlikely to be seeking out or avoiding these areas due to elevated sodium. • Butterflies did not alter oviposition with respect to sodium levels in plants. • Caterpillars did not avoid plants containing moderate levels of sodium. • Caterpillars somewhat avoided plants with toxic levels. • Butterflies are neither attracted to nor repelled by elevated sodium along roadsides. [ABSTRACT FROM AUTHOR]

Neuropeptides are small signalling molecules acting as neurohormones, neurotransmitters and neuromodulators. Being part of the chemical communication system between cells within an organism, they are involved in the regulation of different aspects of animal physiology and behaviour such as feeding, reproduction, development and locomotion. Transcriptomic data from larval and adult tissues have been obtained and mined to generate a comprehensive neuropeptidome for the polyphagous insect pest Spodoptera exigua. Sixty‐three neuropeptides have been identified and described based on their tissue specificity and their regulation in response to different abiotic perturbations. Expression analyses have identified those neuropeptides involved in ingestive and digestive behaviour of S. exigua larvae and revealed a general pattern of upregulation in the midgut during larval starvation. Our results represent a comprehensive neuropeptidome of a lepidopteran species that will be highly relevant to future studies and provide novel information of the insect's perception of its environment. [ABSTRACT FROM AUTHOR]

Highlights • Silkworm moths can learn. • Oviposition decisions are mediated by Pavlovian conditioning. • The Hopkins Host Selection Principle is replicated in the silkworm. • This is evidence for the important adaptive role of Pavlovian conditioning. Abstract One of the most relevant behaviors in the life of some insects is oviposition, because reproductive fitness largely depends on the choice of egg-laying site. We report one experiment that explored the influence of Pavlovian conditioning on oviposition choices of the silkworm. Our results show that moths that jointly experienced an odor (conditioned stimulus) and mulberry leaves (the preferred oviposition place for the moths) preferred to lay their eggs near the odor when it was present, whereas moths in which the odor and the mulberry leaves never appeared together showed no preference. This result provides evidence of the important role that a psychological process such as Pavlovian conditioning has for the survival of this species. [ABSTRACT FROM AUTHOR]

Evaluation of monocrotophos and Azadirachta indica oil, herein referred to as ‘Vijayneem’, were combined in ratios of 10% (10: 90), 20% (20: 80), 30% (30: 70), 40% (40: 60) and 50% (50: 50) [Monocrotophos: Vijayneem] under laboratory conditions to determine efficacy for properties of pesticidal, repellency, attraction, stimulant of consumption, development and reproduction. Asa biopesticide, Vijayneem oil (LC 50 = 10.619%), caused greater mortality in dermal toxicity bioassays (LC 50 = 18.045%) over ingestion. The greatest repellent activity was observed in third (Access Proportion Index = −0.25), fourth (API = −0.52) and fifth instar larvae (API = −0.67) in sprays of cotton leaves with monocrotophos. The detoxification enzymes like esterase (0.3, 11.1 μl/min/mg) and lactate dehydrogenase (6.8, 13.8 μl/min/mg) levels increased postingestion of treatments. Insects which ingested treated leaves showed an increase in carbohydrate (27.0, 18.0%), protein (4.7, 7.7%) and lipid (4.0, 10.4%) content. The greatest deformation of larvae, pupae and adults occurred in monocrotophos alone treatments, while the least deformities were recorded for monocrotophos + Vijayneem treatments. The male and female ratio and percentage of adult emergence was high in control than in A. indica oil alone treated S. litura . The greatest fecundity was observed in control (1620/4 female). In conclusion, the synergistic effects of A. indica oil + monocrotophos can be used for use the management of S. litura . [ABSTRACT FROM AUTHOR]

The velvetbean caterpillar, Anticarsia gemmatalis is one of the most important pests of soybean crops in tropical America. By feeding on leaves, significant defoliation occurs resulting in reduced photosynthetic capacity required for plants’ maintenance and growth, which subsequently can lead to crop losses and reduced agricultural productivity. Many studies have sought to look for compounds that have insecticidal effects. One class of compounds is phenolics, which are produced by plants and have been found to influence the behavior and development of defoliators, representing an important alternative approach to many synthetic insecticides. Particularly, Araucaria angustifolia is a plant rich in polyphenols, which are compounds able to alter cellular dynamics through modulating redox status. In this study, A. angustifolia extract (AAE) was added to the artificial diet of A. gemmatalis. The results demonstrated that AAE was able to reduce larval viability by inducing morphological changes and a delay in the insect’s development. In addition, AAE was found to induce oxidative damage to lipids and proteins, as well as increased nitric oxide levels in A. gemmatalis larvae. AAE treatments also decreased the antioxidant defense systems, leading to a redox imbalance. The reduction in viability in A. gemmatalis was positively correlated with oxidative markers, suggesting that redox imbalance can lead to larvae’s death. These results suggest that AAE possess insecticidal potential through the mechanisms of action of altering cellular redox state. Though further studies are required to confirm this, our study nevertheless contributes to a better understanding of AAE’s mechanisms of action as potential biopesticides in pest management, opening new perspectives on the development of compounds with insecticidal action.

Spodoptera frugiperda (SMITH, 1797) (Lepidoptera: Noctuidae) affects diverse crops of great economic interest, for instance, it can cause severe yield losses in maize, rice and sorghum. In this study, a selection and characterization of Bacillus thuringiensis (BERLINER, 1911) isolates with a high insecticidal activity against S. frugiperda was performed. Fifty-two crystal-forming B. thuringiensis isolates that were identified from 3384 Bacillus-like colonies were examined and screened by PCR for the presence cry genes (cry1, cry1Aa, cry1Ab, cry1Ac, cry1D, cry2 and cry2Ab). Four isolates that showed high toxicity towards S. frugiperda were shown to harbor cry2 genes. The crystals were analyzed by electron microscopy and showed bipyramidal and cuboidal shapes. Furthermore, these four isolates had lethal concentration (LC50) values of 44.5 ng/cm2 (SUFT01), 74.0 ng/cm2 (SUFT02), 89.0 ng/cm2 (SUFT03) and 108 ng/cm2 (SUFT 04) to neonate S. frugiperda larvae. An ultrastructural analysis of midgut cells from S. frugiperda incubated with the SUFT01 spore-crystal complex showed disruptions in cellular integrity and in the microvilli of the midgut columnar cells. The isolates characterized in this work are good candidates for the control of S. frugiperda, and could  be used for the formulation of new bioinsecticides.

Various lepidopteran insects are responsible for major crop losses worldwide. Although crop plant varieties developed to express Bacillus thuringiensis (Bt) proteins are effective at controlling damage from key lepidopteran pests, some insect populations have evolved to be insensitive to certain Bt proteins. Here, we report the discovery of a family of homologous proteins, two of which we have designated IPD083Aa and IPD083Cb, which are from Adiantum spp. Both proteins share no known peptide domains, sequence motifs, or signatures with other proteins. Transgenic soybean or corn plants expressing either IPD083Aa or IPD083Cb, respectively, show protection from feeding damage by several key pests under field conditions. The results from comparative studies with major Bt proteins currently deployed in transgenic crops indicate that the IPD083 proteins function by binding to different target sites. These results indicate that IPD083Aa and IPD083Cb can serve as alternatives to traditional Bt-based insect control traits with potential to counter insect resistance to Bt proteins.

Antimicrobial peptides (AMPs) are small-molecule peptides that play crucial roles in insect innate immune responses. To better understand the function of AMPs in Plutella xylostella, one of the main pests of cruciferous vegetables, three full-length cDNAs encoding moricins were cloned from Pl. xylostella. Two variants of the moricin named PxMor2 and PxMor3 were heterologously expressed and purified. A secondary structure analysis using circular dichroism demonstrated that the two peptides adopted an α-helical structure in the membrane-like environment, but in aqueous solution, they were present in random coiled conformation. Antimicrobial activity assays demonstrated that PxMor2 exhibited high activity against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli; however, PxMor3 only demonstrated high activity against E. coli. Scanning electron microscopy and confocal laser-scanning microscopy analyses suggest that PxMors can lead to the disruption of bacterial membrane, which might be the mechanism by which PxMors inhibit bacterial growth. This study contributes to the understanding of Pl. xylostella AMPs and immune responses, and also enriches the knowledge of insect moricin. [ABSTRACT FROM AUTHOR]

BACKGROUND An insecticide screening effort identified N-(4-bromophenyl)-4,6-bis(2,2,2-trifluoroethoxy)-1,3,5-triazine-2-amine as having weak potency against two lepidopteran species, Helicoverpa zea and Spodoptera exigua. A structure-activity relationship study about the trifluoroethoxy substituents and the aniline of this compound was carried out in an effort to improve insecticidal potency. RESULTS Initially, a series of analogs bearing various substituents on the aniline were prepared, and the insecticidal potency was evaluated against H. zea and S. exigua in greenhouse diet feeding assays. The results showed that electron-withdrawing substituents, such as Cl, Br and CF3, were preferred over electron-donating substituents, such as methoxy, and that potency was significantly better when the substituent was in the para-position. Additional investigations showed that bis(anilino)trifluoroethoxytriazines were more potent. Replacement of the remaining trifluoroethyl group in the bis(anilino)triazine series with an alkyl amine lead to compounds of equal or superior efficacy. CONCLUSION The work presented showed that electron-withdrawing substituents in the para-position of the aniline ring of the initial hit delivered the best levels of insecticidal potency against the two insect species tested. Further investigations showed that potency could be improved by replacing one of the two trifluoroethoxy groups with additional 4-substituted aniline. This level of potency was maintained or further improved when the remaining trifluoroethoxy was replaced with a substituted amine. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Using RNA-seq digital difference expression profiling methods, we have assessed the gene expression profiles of hemocytes harvested from Heliothis virescens that were challenged with Helicoverpa zea single nucleopolyhedrovirus (HzSNPV). A reference transcriptome of hemocyte-expressed transcripts was assembled from 202 million 42-base tags by combining the sequence data of all samples, and the assembled sequences were then subject to BLASTx analysis to determine gene identities. We used the fully sequenced HzSNPV reference genome to align 477,264 Illumina sequence tags from infected hemocytes in order to document expression of HzSNPV genes at early points during infection. A comparison of expression profiles of control insects to those lethally infected with HzSNPV revealed differential expression of key cellular stress response genes and genes involved in lipid metabolism. Transcriptional regulation of specific insect hormones in baculovirus-infected insects was also altered. A number of transcripts bearing homology to retroviral elements that were detected add to a growing body of evidence for extensive invasion of errantiviruses into the insect genome. Using this method, we completed the first and most comprehensive gene expression survey of both baculoviral infection and host immune defense in lepidopteran larvae.

Dwindling resources might be expected to induce a gradual decline in immune function. However, food limitation has complex and seemingly paradoxical effects on the immune system. Examining these changes from an immune system network perspective may help illuminate the purpose of these fluctuations. We found that food limitation lowered long-term (i.e. lipid) and short-term (i.e. sugars) energy stores in the caterpillar Manduca sexta. Food limitation also: altered immune gene expression, changed the activity of key immune enzymes, depressed the concentration of a major antioxidant (glutathione), reduced resistance to oxidative stress, reduced resistance to bacteria (Grampositive and -negative bacteria) but appeared to have less effect on resistance to a fungus. These results provide evidence that food limitation led to a restructuring of the immune system network. In severely food-limited caterpillars, some immune functions were enhanced. As resources dwindled within the caterpillar, the immune response shifted its emphasis away from inducible immune defenses (i.e. those responses that are activated during an immune challenge) and increased emphasis on constitutive defenses (i.e. immune components that are produced consistently). We also found changes suggesting that the activation threshold for some immune responses (e.g. phenoloxidase) was lowered. Changes in the configuration of the immune system network will lead to different immunological strengths and vulnerabilities for the organism. [ABSTRACT FROM AUTHOR]

Study and research of Bt (Bacillus thuringiensis) transgenic plants have opened new ways to combat insect pests. Over the decades, however, insect pests, especially the Lepidopteran, have developed tolerance against Bt delta-endotoxins. Such issues can be addressed through the development of novel toxins with greater toxicity and affinity against a broad range of insect receptors. In this computational study, functional domains of Bacillus thuringiensis crystal delta-endotoxin (Cry1Ac) insecticidal protein and vegetative insecticidal protein (Vip3Aa) have been fused to develop a broad-range Vip3Aa-Cry1Ac fusion protein. Cry1Ac and Vip3Aa are non-homologous insecticidal proteins possessing receptors against different targets within the midgut of insects. The insecticidal proteins were fused to broaden the insecticidal activity. Molecular docking analysis of the fusion protein against aminopeptidase-N (APN) and cadherin receptors of five Lepidopteran insects (Agrotis ipsilon, Helicoverpa armigera, Pectinophora gossypiella, Spodoptera exigua, and Spodoptera litura) revealed that the Ser290, Ser293, Leu337, Thr340, and Arg437 residues of the fusion protein are involved in the interaction with insect receptors. The Helicoverpa armigera cadherin receptor, however, showed no interaction, which might be due to either loss or burial of interactive residues inside the fusion protein. These findings revealed that the Vip3Aa-Cry1Ac fusion protein has a strong affinity against Lepidopteran insect receptors and hence has a potential to be an efficient broad-range insecticidal protein. [ABSTRACT FROM AUTHOR]

Southern blotting experiments performed on Mamestra brassicae genomic DNA after digestion withmethylation-sensitive restriction enzymes indicated that defensin gene is methylated at CpG targets inthe promoter region. However, defensin gene is actively transcribed despite the presence ofmethylation. Experiments performed by genome demethylation indicated that demethylated defensingene resulted in an altered expression after bacterial induction. In particular, no increase in geneexpression was observed after induction with Gram positive bacteria if defensin gene wasdemethylated. The present results are intriguing since they indicate that in M. brassicae DNAmethylation is not involved in gene silencing, but also that DNA methylation could be essential toassure the expression of specific genes. On the whole, data here presented argue against an unifyingand evolutionary conserved role of cytosine methylation from invertebrates to vertebrates.

Lepidopteran silks number in the thousands and display a vast diversity of structures, properties and industrial potential. To map this remarkable biochemical diversity, we present an identification and screening method based on the infrared spectra of native silk feedstock and cocoons. Multivariate analysis of over 1214 infrared spectra obtained from 35 species allowed us to group silks into distinct hierarchies and a classification that agrees well with current phylogenetic data and taxonomies. This approach also provides information on the relative content of sericin, calcium oxalate, phenolic compounds, poly-alanine and poly(alanine-glycine) β- sheets. It emerged that the domesticated mulberry silkmoth Bombyx mori represents an outlier compared with other silkmoth taxa in terms of spectral properties. Interestingly, Epiphora bauhiniae was found to contain the highest amount of β-sheets reported to date for any wild silkmoth. We conclude that our approach provides a new route to determine cocoon chemical composition and in turn a novel, biological as well as material, classification of silks. [ABSTRACT FROM AUTHOR]

We describe a simple and straightforward procedure for the purification and separation of apyrene and eupyrene forms of lepidopteran sperm. The procedure is generally applicable to both butterfly and moth species with results varying according to the relative amounts of sperm produced and size of sperm storage organs. The technique relies upon inherent differences between eupyene sperm bundles and free apyrene sperm morphology. These differences allow for separation of the sperm morphs by repeated “panning” of sperm bundles into the center of a plastic dish. The purified eupyrene sperm bundles can then be removed and apyrene sperm collected from the supernatant by centrifugation. Efficacy of the purification process was confirmed by light microscopy and gel electrophoresis of the resulting fractions. Both one- and two-dimensional gel electrophoresis identified significant protein differences between the fractions further suggesting that the panning procedure effectively separated eurpyrene from apyrene sperm. The panning procedure should provide a convenient and accessible technique for further studies of sperm biology in lepidopterans. [ABSTRACT FROM AUTHOR]

The moth Heortia vitessoides Moore (Lepidoptera: Crambidae) is a major pest of ecologically, commercially and culturally important agarwood-producing trees in the genus Aquilaria. In particular, H. vitessoides is one of the most destructive defoliating pests of the incense tree Aquilaria sinesis , which produces a valuable fragrant wood used as incense and in traditional Chinese medicine [33]. Nevertheless, a genomic resource for H. vitessoides is lacking. Here, we present a chromosomal-level assembly for H. vitessoides , consisting of a 517 megabase (Mb) genome assembly with high physical contiguity (scaffold N50 of 18.2 Mb) and high completeness (97.9% complete BUSCO score). To aid gene annotation, 8 messenger RNA transcriptomes from different developmental stages were generated, and a total of 16,421 gene models were predicted. Expansion of gene families involved in xenobiotic metabolism and development were detected, including duplications of cytosolic sulfotransferase (SULT) genes shared among lepidopterans. In addition, small RNA sequencing of 5 developmental stages of H. vitessoides facilitated the identification of 85 lepidopteran conserved microRNAs, 94 lineage-specific microRNAs, as well as several microRNA clusters. A large proportion of the H. vitessoides genome consists of repeats, with a 29.12% total genomic contribution from transposable elements, of which long interspersed nuclear elements (LINEs) are the dominant component (17.41%). A sharp decrease in the genome-wide percentage of LINEs with lower levels of genetic distance to family consensus sequences suggests that LINE activity has peaked in H. vitessoides. In contrast, opposing patterns suggest a substantial recent increase in DNA and LTR element activity. Together with annotations of essential sesquiterpenoid hormonal pathways, neuropeptides, microRNAs and transposable elements, the high-quality genomic and transcriptomic resources we provide for the economically important moth H. vitessoides provide a platform for the development of genomic approaches to pest management, and contribute to addressing fundamental research questions in Lepidoptera. • Heortia vitessoides is a major pest of the lign aloes Aquilaria malaccensis and Aquilaria sinensis in Malaysia and southern China. • A high-quality chromosomal level genome of Heortia vitessoides is assembled by Illumina short-read, 10× Genomics linked-read, and Hi-C sequencing data. • Transcriptomic resources (mRNA and small RNAs) have facilitated predicting 16,421 gene models and identifying 85 conserved and 94 novel microRNAs. • Gene family expansions have been detected in H. vitessoides , including duplications of cytosolic sulfotransferase (SULT) genes shared among lepidopterans. • Transposable elements account for 29.12% total genomic contribution, with long interspersed nuclear elements (LINEs) identified as the dominant component (17.41%). [ABSTRACT FROM AUTHOR]

The natural influences of American bison have been virtually eliminated from the North American landscape, making it difficult to study native grassland ecosystems. The Sandhill Wildlife Area provides a unique opportunity to study this once-natural phenomenon in an oak savanna landscape. This area is home to the largest known population of the federally endangered Karner blue butterfly. We evaluated lepidopteran and lupine distribution relative to cover types and generated statistical models to predict occupancy and abundance relative to in situ groundcover variables as well as American bison related habitat variables. Overall variance of lepidopteran populations was best explained by increasing nectar and forb cover (29.4 % occurrence explained), while lepidopterans excluding Karner blues were further associated with decreasing cover of shrubs/trees (25.8 % occurrence, 59.8 % abundance explained) and were more likely to occur in areas of disturbance cover type. Karner blue occurrence was best explained (41.6 %) by increasing nectar and forb cover and an increasing number of bison chips (indicative of the recent presence of bison). Karner blue females, specifically, were more likely to be present in areas of wallow cover type and were best explained by increasing nectar plant cover (33.4 % occurrence, 60.1 % abundance explained) and increasing shrub cover. Females were also associated with an increasing number of bison chips, number of wallows and total size of wallows. Karner blue male occupancy were best explained (40.7 %) by increasing nectar cover and increasing number of bison chips. Lupine was significantly less likely to occur in areas of shrubs/trees cover. Lupine was explained (55.7 % occupancy and 88.2 % abundance), by the decreasing cover of shrubs/trees, and by the cover of nectar, forbs, and grass, and total size of wallows. We conclude that the activities caused by bison reduce the overall woody growth and produce improved habitat for nectar plants and especially for disturbance-dependent lupine, and subsequently enhance habitat for lepidopterans. Karner blues were significantly related to bison activities, suggesting that improved management techniques should consider mimicking megaherbivore activities through increased disturbance frequency and increase mineral-soil disturbance. [ABSTRACT FROM AUTHOR]

Abstract: The invertebrate octopaminergic system affects many diverse processes and represents the counterpart to the vertebrate adrenergic/noradrenergic system with the classes of octopamine receptor (OAR) being homologous to those of vertebrate adrenergic receptors. However, there is still little information on the OARs present in different insect species, and the levels and distribution of these receptors throughout the body. cDNAs sharing high similarity with known insect OARs were cloned in three lepidopteran species: the cabbage looper, Trichoplusia ni; the true armyworm, Pseudaletia unipuncta; and the cabbage white, Pieris rapae. Seven major larval tissues and one adult tissue were examined in T. ni using quantitative real-time PCR to determine the relative expression levels of each receptor transcript across different tissues, as well as of all receptor transcripts within individual tissues. A subset of these tissues was also examined in P. unipuncta and P. rapae. All receptor transcripts were expressed in the nervous system of all three species, however, the distribution of the different receptor types varied between species. In all tissues, the OARbeta2 transcript was the most highly expressed, except in the Malpighian tubules where OARbeta1 was highest, and the midgut where there was no significant difference in receptor transcript levels. [Copyright &y& Elsevier]

Garlic ( Allium sativum) lectins are promising candidate molecules for the protection against chewing (lepidopteran) as well as sap sucking (homopteran) insect pests. Molecular mechanism of toxicity and interaction of lectins with midgut receptor proteins has been described in many reports. Lectins show its effect right from sensory receptors of mouth parts by disrupting the membrane integrity and food detection ability. Subsequently, enter into the gut lumen and interact with midgut glycosylated proteins like alkaline phosphatase (ALP), aminopeptidase-N (APN), cadherin-like proteins, polycalins, sucrase, symbionin and others. These proteins play critical role in life cycle of insect directly or indirectly. Lectins interfere with the activity of these proteins and causes physiological disorders leading to the death of insects. Lectins further transported across the insect gut, accumulated in various body parts (like haemolymph and ovary) and interact with intracellular proteins like symbionin and cytochrome p450. Binding with cytochrome p450 (which involve in ecdysone synthesis) might interfere in the development of insects, which results in growth retardation and pre-mature death. [ABSTRACT FROM AUTHOR]

Abstract: An antimicrobial peptide (AMP) of the cecropin family was isolated by HPLC from plasma of the insect pest, Spodoptera frugiperda. Its molecular mass is 3910.9Da as determined by mass spectrometry. Thanks to the EST database Spodobase, we were able to describe 13 cDNAs encoding six different cecropins which belong to the sub-families CecA, CecB, CecC and CecD. The purified peptide identified as CecB1 was chemically synthesized (syCecB1). It was shown to be active against Gram-positive and Gram-negative bacteria as well as fungi. Two closely related entomopathogenic bacteria, Xenorhabdus nematophila F1 and Xenorhabdus mauleonii VC01T showed different susceptibility to syCecB1. Indeed, X. nematophila was sensitive to syCecB1 whereas X. mauleonii had a minimal inhibitory concentration (MIC) eight times higher. Interestingly, injection of live X. nematophila into insects did not induce the expression of AMPs in hemolymph. This effect was not observed when this bacterium was heat-killed before injection. On the opposite, both live and heat-killed X. mauleonii induced the expression of AMPs in the hemolymph of S. frugiperda. The same phenomenon was observed for another immune-related protein lacking antimicrobial activity. Altogether, our data suggest that Xenorhabdus strains have developed different strategies to supplant the humoral defense mechanisms of S. frugiperda, either by increasing their resistance to AMPs or by preventing their expression during such host-pathogen interaction. [Copyright &y& Elsevier]

The reversible mitochondrial transhydrogenase in Manduca sexta has been characterized and is involved with endocrine-mediated post-embryonic larval development in this model insect. While biochemical in vivo and in vitro studies have been accomplished, robust molecular studies of the transhydrogenase have not been possible due to deicient genomic data. In the present study, using a combination of degenerate oligonucleotide primers and raw genomic data, we have determined the structure and sequence of the transhydrogenase gene from the model insect Manduca sexta. The encoded protein is highly similar to other transhydrogenase proteins and this sequence is the irst lepidopteran sequence reported to date. [ABSTRACT FROM AUTHOR]

The cotton bollworm Helicoverpa armigera and the oriental tobacco budworm H. assulta are sibling species, both being important agricultural pests. Morphologically, the two insects are almost indistinguishable at the egg, larval and pupal stages. One of the big challenges in the study of these insects, in particular in integrated pest management, is a timely and dependable identification of these insects at their early stages of development. Here, we report a H. armigera-specific nuclear DNA marker, and demonstrate that it can be employed to reliably discriminate between H. armigera and H. assulta by simple polymerase chain reaction amplification experiment. [ABSTRACT FROM AUTHOR]

Using RNA-seq digital difference expression profiling methods, we have assessed the gene expression profiles of hemocytes harvested from Heliothis virescens that were challenged with Helicoverpa zea single nucleopolyhedrovirus (HzSNPV). A reference transcriptome of hemocyte-expressed transcripts was assembled from 202 million 42-base tags by combining the sequence data of all samples, and the assembled sequences were then subject to BLASTx analysis to determine gene identities. We used the fully sequenced HzSNPV reference genome to align 477,264 Illumina sequence tags from infected hemocytes in order to document expression of HzSNPV genes at early points during infection. A comparison of expression profiles of control insects to those lethally infected with HzSNPV revealed differential expression of key cellular stress response genes and genes involved in lipid metabolism. Transcriptional regulation of specific insect hormones in baculovirus-infected insects was also altered. A number of transcripts bearing homology to retroviral elements that were detected add to a growing body of evidence for extensive invasion of errantiviruses into the insect genome. Using this method, we completed the first and most comprehensive gene expression survey of both baculoviral infection and host immune defense in lepidopteran larvae. [ABSTRACT FROM AUTHOR]

The process of uptake of hexamerins during metamorphosis from insect haemolymph by fat body cells is reminiscent of receptor-mediated endocytosis. Previously, we had identified a hexamerin-binding protein (HBP) and reported for the first time that uptake of hexamerins is dependent on the phosphorylation of HBP partly by a tyrosine kinase, which is, in turn, activated by 20-hydroxyecdysone (20E). However, the exact nature of HBP and the mechanism of interaction are still unknown. Here we report the possibility of HBP being a GPI-anchored protein in the fat body of Achaea janata and its role in the tyrosine-kinase-mediated phosphorylation signalling. Digestion of fat body membrane preparation with bacterial phosphatidylinositol-specific phospholipase C (PI-PLC), and the subsequent recognition by antibodies specific for the cross-reacting determinant (CRD), revealed that HBP is glycosylphosphatidylinositol (GPI)-anchored protein and, further, that the hexamerin binding to HBP was inhibited after digestion. Hexamerin overlay assay (HOA) of co-immunoprecipitated in vitro phosphorylated HBP showed exclusive binding to ~120 kDa protein. Lectin-binding analysis of hexamerins revealed the presence of N-acetylgalactosamine (GalNAc) and N-acetylglucosamine (GluNAc), whereas HBP showed the presence of GalNac alone. Mild chemical deglycosylation studies and binding interaction in the presence of sugars revealed that glycan moieties are possibly not involved in the interaction between HBP and hexamerins. Taken together, these results suggest that HBP may be a GPI-anchored protein, and interaction and activation of HBP is through lipid-linked non-receptor src tyrosine kinases. However, additional studies are needed to prove that HBP is a GPI-anchored protein. [ABSTRACT FROM AUTHOR]

The present paper describes the purification, characterization and determination of the partial primary structure of the first trypsin inhibitor isolated from the family Sapindaceae. A highly stable, potent trypsin inhibitor (SSTI) was purified to homogeneity. SDS-PAGE analysis revealed that the protein consists of a two-polypeptide chain with molecular masses of approximately 15 and 3 kDa. The purified inhibitor inhibited bovine trypsin at a 1:1 M ratio. Kinetic analysis revealed that the protein is a competitive inhibitor with an equilibrium dissociation constant of 10 M for trypsin. The partial NH- terminal sequence of 36 amino acids in SSTI indicates homology with other members of the trypsin-inhibitor family from different sources. This inhibitor is highly stable in the presence of denaturing agents. SSTI showed significant inhibitory activity against trypsin-like proteases present in the larval midgut on Anagasta kuehniella, Corcyra cephalonica, Diatreae saccharalis and Anticarsia gemmatalis. [ABSTRACT FROM AUTHOR]