In this study, we tested the effect of a range of insect orders including Trichoptera as outgroups for lepidopteran phylogeny. Phylogenetic analyses performed with four different partitioning schemes using the maximum-likelihood method provided four different topologies (T1-T4) and topological test most supported T1 topology. When the means of first principle component for nucleotide frequency between A/T and G/C of PCGs was considered Trichoptera, Diptera, Coleoptera, and Orthoptera tended to result in T1 topology more frequently in the given ingroup taxa and outgroups tested. This result contradicts to the general view that the sister taxon might be the best outgroup. The T1 topology was largely consistent with a recent large molecular dataset-based lepidopteran phylogeney, presenting the relationships ((((((((((Noctuoidea + Geometroidea) + (Bombycoidea + Lasiocampoidea)) + Drepanoidea) + Mimallonoidea) + Pyraloidea) + Gelechioidea) + Papilionoidea) + Tortricoidea) + (Gracillaroidea + Yponomeutoidea)) + Hepialoidea).
Diversity and distribution of entomopathogenic nematodes were surveyed in Korea from 2010 to 2014. Soil samples were examined for the presence of steinernematid and heterorhabditid nematodes by baiting method with Galleria mellonella larvae. Out of the 189 soil samples, 47(24.9%) were positive for entomopathogenic nematodes, with the share of 7(3.7%) for Heterorhabditis and 40(21.2%) for Steinernema isolates. Morphological and molecular studies were also utilized to characterize isolates. The Heterorhabditis isolates were identified as H. megidis and Steinernema isolates as S. carpocapsae, S. monticolum and S. kraussei, respectively. H. megidis was the most common species of Heterorhabditis, which was isolated from 7 sites across the five different provinces and Steinernema monticolum was the most common species of Steinernema, which was isolated from 34 sites, respectively. S. carpocapsae was isolated from only two sites, Namwon, Jeongnam province and Iksan, Jeonbuk province, respectively. S. kraussei was the first recorded entomopathogenic nematode in Korea. Those entomopathogenic nematodes were recorded from coniferous and deciduous forests.
Some Japonica rice plant varieties show the resistance to the whitebacked planthopper (S. furcifera) and the varieties produce ovicidal compound, benzyl benzoate, to kill their eggs lying in the rice plant. We have tried to elucidate the mechanism for inducing the production of the benzyl benzoate by the Japonica rice varieties. The active 80% MeOH/H2O extract was chromatographed on ODS open column and separated into 6 fractions; 100% H2O, 20% MeOH/H2O, 60% MeOH/H2O, 80% MeOH/H2O, 100% MeOH, 100% EtOH. Of these 6fractions, 100% MeOH and 100% EtOH fractions evidently induced benzyl benzoate at 43.95μg/g of fresh rice plant(frp) and 31.04μg/gfrp, respectively. The active compounds were submitted to analyses by using NMR and GC- and LC-MS inorder to elucidate the structure. We find elicitors to induce with S. furcifera. The elicitors are in the female of S. furcifera and the structures of elicitors are species of phospholipid and glycerol.
Temperature as a major environmental factor affects on organisms on various levels including molecular, physiological, behavioral and ecological levels. Transient receptor potential channels (TRPs) are a cation channel family. Among them, thermo-TRPs are known as a thermosensor. The potential role of thermo-TRPs have been identified in the fruit fly, Drosophila melanogaster, in thermotaxis and in thermal acclimation. With RNA interference (RNAi) technique, the role of thermo-TRPs in the red flour beetle, Tribolium castaneum, was identified by measuring thermal avoidance behavior in a behavioral assay. RNAi of trpA1 reduced high temperature avoidance, 39 and 42 °C. Moreover, the effects of RNAi of thermo-TRPs on the heat-induced knockout and the death after short exposure to high temperature was measured after one minute exposure at 52 °C, either with or without a 42 °C 10-minute thermal acclimation period. Even though it was relatively short time exposure to high temperature, it was enough to induce high temperature thermal acclimation. RNAi of trpA1 made faster knockout at 52 °C. With RNAi of painless, the recovery rates from heat-induced knockout after thermal acclimation. RNAi of pyrexia reduced long-term total survivorship without thermal acclimation.
Four sympatric tortricid (Lepidoptera) species in New Zealand, Epiphyas postvittana (Walker), Planotortrix octo (Dugdale), Cydia pomonella (L.), and Cydia succedana (Denis and Schiffermüller), were studied for the morphology of antennal sensilla and the chemical communication systems using scanning electron microscopy (SEM), electro-antennogram (EAG), gas chromatography-electroantenno-detection (GC-EAD), single sensillum recording (SSR) and field tests. Antennal flagella of adult have four main types of olfactory sensilla (s.); s. trichodea, s. basiconica, s. auricillica, and s. coeloconica in the four species. The s. trichodea subtype I was longest and male-specific in all the species. The antennae of males of the four species showed characteristic higher EAG, GC-EAD and SSR responses to 12, 14-carbon acetates or alcohol. Some of the selected chemicals selected from the electrophysiological experiments caused significant antagonistic or agonistic activities to the sex pheromone of each species in the field. In this study, the four species showed distinct chemical communication systems according to sub-family, which possibly serve as a factor in species isolation.
The pea leafminer fly, Liriomyza huidobrensis Blanchard (Diptera: Agromyzidae), is an emerging pest in Korea. High population and heavy damage of this pest were observed from potato fields in the southern part of Korea in 2012. Morphological and genetic evidence confirmed the species as L. huidobrensis. The infestation was expanding throughout the country and a rich complex of parasitoid was recognized. From the exclusion study, high density infestation of leafminers reduced yield among potato varieties (Chubeak, Seohong, Goun, Dejima, and Sumi). Higher leaf damage was evident in the lower part of plants. In olfactory bioassay and feeding preference tests, leafminers choosed the Goun variety most. Given the six constant temperatures, the developmental time from egg to adult significantly decreased with increment of temperatures. From the insecticide toxicity tests, abamectin and lepimectin and spinetoram were found effective against immature stages, while cartap hydrochlorid (Neonicotinoid) was against adult insects. Overall, these studies help to pave the ways for managing this new invasive pest.
Human body and head lice are obligatory human ectoparasites. Although both body and head lice belong to a single species, Pediculus humanus, only body lice are known to be a vector of several bacterial diseases. The higher vector competence of body lice is assumed to be due to their weaker immune response than that of head lice. To test this hypothesis, immune reactions were compared between body and head lice following infections by two model bacteria, Staphylococcus aureus and Escherichia coli, and a human pathogen, Bartonella quintana. Following dermal or oral challenge, the number of these bacteria increased both in hemocoel and alimentary tract of body lice but not in head lice and the viability of the B. quintana was significantly higher in body louse feces, the major route of infection to human. In addition, body lice showed the lower basal/induced transcription level of major immune genes, cytotoxic reactive oxygen species and phagocytosis activity compared with head lice. These findings suggest that a reduced immune response may be responsible, in part, for the increased proliferation and excretion of viable bacteria which are associated with the high level of human infectivity seen in body versus head lice.
One of the overlooked points in mosquito blood feeding research is a final step before blood feeding. We provide the anatomical and chemosensory evidence that a piercing structure of the mouthpart of the mosquitoes is an essential apparatus for the penultimate stage in blood feeding in mosquitoes. Indeed, the stylet possesses a number of sensory hairs located at the tip of the stylet. These hairs house olfactory receptor neurons that express two conventional olfactory receptors of Aedes aegypti (AaOrs), AaOr8 and AaOr49, together with the odorant co-receptor (AaOrco). In vivo calcium imaging using transfected cell lines demonstrated that AaOr8 and AaOr49 were activated by volatile compounds present in blood. Taken together, we identified olfactory receptor neurons in the stylet involved in mosquito blood feeding behaviors, which in turn indicates that olfactory perception in the stylet is necessary and sufficient for mosquitoes to find host blood in order to rapidly acquire blood meals from a host animal.
A response surface methodology (RSM) was used to evaluate how pH and ionic strength (IS) affect the fate (i.e. size and colloidal stability) of an SC formulation containing the pyrethroid β-cyfluthrin. The response surfaces determined under a range of environmentally relevant conditions were then used to assess the toxicity of the SC formulation of β-cyfluthrin to D. magna. The changes in hydrodynamic diameter (HDD) and colloidal stability as determined by zeta potential measurement were closely related to either or both of the change in pH and IS with the linear factor of IS being the most significant factor affecting those changes. Thus, the concentration of SC formulation of β-cyfluthrin remaining in the water column was dependent on the pH and IS conditions and highest when the colloidal suspension contained small particles or a lack of agglomeration leading to sedimentation of the particles. The toxicity results show correspondingly higher toxicity to D. magna when exposed to the SC formulation of β-cyfluthrin when pH and IS conditions favor formation of either the smallest HDD or most stable colloidal suspensions.
In recent years, high-throughput next-generation sequencing (NGS) techniques have provided fascinating opportunities to understand the biology of non-model organisms, especially insect species. The decrease in sequencing costs and extensive sequencing services from NGS providers has brought many entomologists to be involved in genome sequencing. However, poor planning can lead to extremely fragmented genome assemblies which prevents high quality gene annotation and other desired analyses. Insect genomes can be problematic to assemble, due to combinations of high polymorphism, inability to breed for genome homozygosity, and small physical sizes limiting the quantity of DNA able to be isolated from a single individual. Given to the rapid development of host resistance to multiple classes of insecticides, it is indispensable to study the comprehensive genomic information of insects. Recent advances in sequencing technology and assembly strategies can able to fetch breakthroughs in deciphering the genetic information of insects. Here, we present the cost effective high throughput genome sequencing and assembly strategies for insect species in respects to taxonomy, evolutionary history, immune response, drug development, insect host-virus interactions and pest management etc.
Microfluidics based on nanobio sensors technologies can provide convenient and accurate diagnosis tools. In this talk, we present recent developments of nanobio sensors & diagnosis chip using microfluidics, with special emphasis on disposable plastic devices format. In detail, we overview of the common methods used in the fabrication of polymer microfluidic systems, including replica and injection mold-ing. Also we explain the different methods by which on-chip operations—such as the pumping and valving of fluid flow, the mixing of different reagents, and the separation and detection of different biochemical species implemented in a microfluidic format. Finally, a few select biotechnological applications of microfluidics are presented to illustrate both the utility of this technology and its potential applications with insect models in the near future.
Taste substances are recognized by gustatory sensory neurons that express putative seven transmembrane proteins in the gustatory receptor (Gr) family. However, the gustatory tuning of the molecular receptors encoded by these gustatory receptor genes remains unknown in honey bees. Here we first functionally characterize a gustatory receptor responding to umami taste L-amino acids in the western honey bee, Apis mellifera. Using Ca2+ imaging assay and two-voltage clamp recording, we first report that one of the gustatory receptors of honeybee, AmGr10, functions as a selectively tuned amino acid receptor in taste neurons. In addition, we report a floating electrode-based bioelectronic tongue mimicking honeybee taste systems for the detection and discrimination of umami substances. This floating electrode-based bioelectronic tongue mimicking insect taste systems can be a powerful platform for various applications such as food screening, and it also can provide valuable insights on insect taste systems.
Aedes aegypti and Aedes albopictus and Culex pipiens pallens mosquitoes transmit dengue fever and West Nile virus diseases, respectively. This study was conducted to determine the toxicity and mechanism of action of four flavonoids and seven fatty acids from Millettia pinnata (Fabaceae) seed as well as four fatty acid esters toward third instar larvae from insecticide-susceptible C. p. pallens and A. aegypti as well as wild A. albopictus. Potent toxicity was produced by karanjin, oleic acid, karanjachromene, linoleic acid, linolenic acid, pongamol, pongarotene, and elaidic acid toward C. p. pallens larvae (24 h LC50, 14.61–28.22 mg/L) and A. aegypti larvae (16.13–37.61 mg/L). Against wild A. albopictus larvae, oleic acid (LC50, 18.79 mg/L) and karanjin (35.26 mg/L) exhibited potent toxicity. Acetylcholinesterase (AChE) is the main site of action of the flavonoids, oleic acid, and palmitic acid.,Linoleic acid and linolenic acid might act on both AChE and octopaminergic receptor. Further studies will warrant possible applications of M. pinnata seed-derived products as potential larvicides for the control of mosquito populations.
Drosophila has been used as a model for studying various human diseases. Especially, it has been widely used for studying neurological disorders in humans. For example, many progressive neurological disorders such as Alzheimer's diseases, Parkinson’s disease, Amyotrophic lateral sclerosis were recapitulated in Drosophila. We have been studying DYT1 dystonais, an enigmatic movement disorder. We generated several different kinds for Drosophila models by expressing human Torsin1A genes with or without causative mutations. In addition, we employed various Omics tools to identify any genetic, proteomic, and metabolomic alterations. I will summarize our recent progression in using Drosophil as a model for studying molecular and cellular etiologies underlying DYT1 dystonia and in vivo functions of Torsin proteins.
The human β-amyloid (Aβ) cleaving enzyme (BACE-1) is a target for Alzheimer’s disease (AD) treatments. This study was conducted to determine if acacetin extracted from the whole Agastache rugosa plants had anti-BACE-1 and behavioral activities in Drosophila AD models and to determine acacetin’s mechanism of action. Acacetin (100, 300, and 500 μM) rescued amyloid precursor protein (APP)/BACE1-expressing flies and kept them from developing both eye morphology (dark deposits, ommatidial collapse and fusion, and the absence of ommatidial bristles) and behavioral (motor abnormalities) defects. The RT-PCR and Western blot analysis revealed that the protective effect of acacetin on Aβ production is mediated by transcriptional regulation of BACE-1 and APP, resulting in decreased APP protein expression and BACE-1 activity, and reduced Aβ production by interfering with BACE-1 activity and APP synthesis, resulting in a decrease in the levels of the APP carboxy terminal fragments and the APP intracellular domain, and finally, resulting in a decrease in the number of amyloid plaques.
In perennial crops virus diseases are usually caused by mixed infections rather than by individual viruses. Understanding the contribution of each virus in disease development, the interactions between viruses and how each virus spreads in the field allows for development of control measures that are targeted for disease control rather than controlling all viruses in a complex. There are multiple types of virus-vector interactions and this information can be used to inform vector control strategies to manage virus diseases. Information on virus-vector interactions and insect biology for controlling a disease caused by a virus complex in raspberry will be presented. Understanding the biology of multiple vectors as well as multiple types of virus-vector interactions for a vector of multiple viruses will be presented as a model for managing virus disease in strawberry in different environments. The goal is to describe a systems approach for controlling virus diseases in vegetatively propagated crops from developing clean plants through to fruit production.
RNA interference (RNAi) technology is a new direction for insect pest management, a biologically-based and target specific strategy. During the past decade the availability of insect genomics and computational biology has further enabled the implementation of RNAi technology to target economically important insect pests. It has shown striking results in various insect groups, suggesting that it will be a promising tool for the next generation of pest management. The mechanism of RNAi action is a specific knockdown of gene expression vie degradation of a target messenger RNA (mRNA), by double-stranded RNA (dsRNA). The applied dsRNA thus blocks target protein synthesis, leading to failure of normal physiological functions in the organism To successfully develop RNAi applications, a critical initial step is screening for appropriate candidate genes to identify RNAi targets, because the impacts of gene silencing (especially in terms of which other genes are effected) vary for different RNAi target genes and insects. The challenge with gene selection is to select suitable insect-specific target genes that provide fast-acting mortality or suppression and long-term population suppression without affecting other non-target organisms.
Even though plant protections using chemical pesticides have several advantages, non-specific toxicities to other beneficial insects and humans and rapid development of tolerance and/or resistance of target pests to chemicals are major disadvantages. Recent researches suggested that using double stranded RNA (dsRNA) could be species specific and environmental friendly pest management protocols. However, efficiency of dsRNA treatments are known to be variable according to its application methods. For example, injections of dsRNAs to pests were known to be effective in all species. However, efficiencies of oral application of dsRNAs were known to be dependent on species. Thus, development of tools that could enhance the efficacy of orally treated dsRNAs are utmost important for widening usages of dsRNAs in plant protection. Recently, we found that the efficacy of oral treated dsRNAs to target pests could be enhanced by nano-technologies. I will show how applying nano-technology to dsRNAs enhance the efficiency of dsRNAs. (This work was carried out with the support of the Cooperative Research Program for Agriculture Science & Technology Development (Project title: Studies for biological characteristics of and control methods against the migratory locust, Locusta migratoria, Project No: PJ011630042016), Rural Development Administration).
Tetranychus urticae is extremely hard to control by conventional acaricides due to its rapid development of resistance to nearly all arrays of acaricide. As an alternative control measure of acaricide-resistant mites, RNA interference (RNAi)-based method has recently been suggested. A double-stranded RNA (dsRNA) delivery method using multi-unit chambers was established and employed to screen the RNAi toxicity of 42 T. urticae genes. Among them, the dsRNA treatment of coatomer I (COPI) genes, such as coatomer subunit epsilon (COPE) and beta 2 (COPB2), resulted in high mortality [median lethal time (LT50) = 89.7 and 120.3 h, respectively]. The transcript level of the COPE gene was significantly (F3,9 = 16.2, P = 0.001) reduced up to 24% following dsRNA treatment, suggesting that the toxicity was likely mediated by the RNAi of the target gene. To identify the deferentially expressed gene upon dsRNA ingestion, RNA-seq was employed to compare the transcriptional profiles between mites fed dsEGFP and dsCOPB2. Approximately 928 of genes were up- or down-regulated significantly (P < 0.05) compared to control and 182 genes were commonly responded to the treatment of both dsRNAs. Those dsRNA-responsible genes were mainly categorized into metabolic enzymes, transporters and secretory proteins. Further study would be necessary to elucidate the roles of dsRNA-responsible genes in mite’s dsRNA uptake and defense.
Polydnaviruses (PDVs) are a group of insect viruses and symbiotic to some endoparasitoid wasps. Genome analyses of different PDVs provide a number of genes putatively associated with alteration of host insect physiological processes. Especially, PDV gene products assist host wasp development by suppressing immune responses and delaying larval development of parasitized lepidopteran hosts. Thus, PDV genes can be applied to control insect pests by incorporating them into crop genomes. This talk illustrates two examples of PDV genes: CpBV-CST1 and CpBV-ELP1. CpBV-CST1 has been known to inhibit insect cysteine proteases to suppress immune and development, while CpBV-ELP1 exhibits a high cytotoxicity to insect cells. Transgenic tobaccos expressing CpBV-CST1 or CpBV-ELP1 were constructed by using Agrobacterium-mediated transgenic system and exhibited antixenosis against chewing and sucking insect pests on tobacco.