The honey bee soluble acetylcholinesterase 1 (AmAChE1) is overexpressed under the overwintering and brood rearing-suppressed conditions. To investigate the role of AmAChE1 in regulating acetylcholine (ACh) titer, ACh concentrations both in the head (neuronal) and abdomen (non-neuronal) were analyzed. ACh titer was significantly lower in both tissues of worker bees under the overwintering and brood rearing-suppressed conditions compared to control bees. The expression levels of another two factors that regulate ACh titer, choline acetyltransferase (AmAChT) and acetylcholinesterase 2 (AmAChE2), were not altered as judged by qPCR and native PAGE, suggesting that the lower ACh titer was mainly regulated by AmAChE1. For precise verification of AmAChE1 as an ACh titer regulator, honey bees were put under brood rearing-suppressed condition to induce AmAChE1 and injected AmAChE1 dsRNA to knock down the gene. The ACh titer of AmAChE1-knocked down honey bees was 1.9 and 2.6 folds higher than that of control bees in head and abdomen, respectively. Taken together, in spite of its extremely low catalytic activity, the overexpression of AmAChE1 is likely to be related with the low level of ACh homeostasis, perhaps via ACh sequestration, under brood rearingsuppressed condition, and likely induce metabolic changes through ACh receptors-related pathways.

Small hive beetle (Aethina tumida) (SHB) is an invasive species to most northern hemisphere countries, including Korea. In an attempt to obtain basic information for efficient management of SHB, genes encoding conventional insecticide targets [voltage-sensitive sodium channel α-subunit (VSSC) and acetylcholinesterase (AChE)] were annotated and characterized following the analysis of whole transcriptomes of adults and larvae. A single VSSC gene was identified but no apparent mutations associated with pyrethroid resistance were detected. Genes encoding two AChEs (AtAChE1 and AtAChE2) were identified from the SHB transcriptome. AtAChE1 was determined to be the main catalytic enzyme, thereby being a toxicologically more relevant target. No apparent mutations associated with resistance to organophosphorus and carbamate insecticides was identified in the AtAChE1 gene, whereas the S238G mutation, originally identified from the Colorado potato beetle, was detected in the AtAChE2 gene.

The honey bee soluble acetylcholinesterase 1 (AmAChE1) is overexpressed under the overwintering and brood rearing-suppressed conditions. To investigate the role of AmAChE1 in regulating acetylcholine (ACh) titer, ACh concentrations in both the head (central nervous system) and abdomen (peripheral nervous system) were analyzed. ACh titer was significantly lower in both tissues of worker bees under the overwintering and brood rearing-suppressed conditions compared to control bees. Interestingly, the expression levels of choline acetyltransferase (AmChAT) and molecular marker genes of immune systems were significantly reduced in honey bee head under the same conditions. Taken together, ACh titer appears to be reduced via a cooperative interaction of the AmAChE1 overexpression and AmChAT underexpression and to be linked to reduced inmmune responses under the overwintering and brood rearing-suppressed conditions. The roles of AmAChE1 (with little catalytic activity) and AmChAT in the ACh homeostasis and signaling was discussed in the contexts of immune response and longevity regulation in honey bees.

Among two different acetylcholinesterase (AmAChE1 and AmAChE2) of the western honey bee, the soluble AmAChE1might be related with a stress response as judged from its over-expression in honey bee workers when brood rearingwas suppressed. In this study, to ensure the nature of AmAChE1 responding to stress factors, the expression patternsof AmAChE1 were investigated following various treatments, including varroa mite infestation, bacterial challenge, broodrearing suppression, thermal stresses, chemical treatments, ultraviolet B irradiation, starvation, water restriction and crowdingstress. In addition, transcription profiles of four heat shock protein genes known as general stress markers and vitellogeningene, which is induced in several stress conditions, were tested as positive references. In every tested condition, onlybrood rearing suppression and heat shock were related with the expression of AmAChE1.

There are two different types of acetylcholinesterase (AChE1 and AChE2) in the western honeybee as in most of insects. It is suggested that soluble AmAChE1 might be related with a stress response as judged from its elevated expression level in honey bee workers when brood rearing was suppressed. In this study, to ensure the nature of AmAChE1 responding to stress factors, the expression patterns of AmAChE1 following heat shock, brood rearing suppression and chemical treatments (Imidacloprid and fluvalinate) were investigated. Also, several heat shock protein (hsp) genes (hsp10, hsp60, hsp70 and hsp90) known as general stress markers were tested as positive references. Heat shock induced expression of every tested hsp along with AmAChE1. In brood rearing-suppressed worker bees, 7 days old bees showed much higher expression level of AmAChE1 and hsp90 compared to control honey bees. However, treatment of imidacloprid and fluvalinate did not induce any apparent overexpression of these genes. These results confirm that both HSP and AmAChE1 genes generally respond to temperature and brood rearing suppression and further suggest that AmAChE1 can serve as a potential biomarker along with hsps for the detection of stress in honey bee colonies.

Two different types of acetylcholinesterae (AChE1 and AChE2) are present in majority of insects, including the Western honey bee. Out of the two honey bee AChEs (AmAChEs), the soluble AmAChE1 with little catalytic activity is widely distributed in both neuronal and non-neuronal tissues, including fat body. In this study, to identify stresss factors that can induce AmAChE expression, we tested various conditions that honey bees can encounter in natural setting, including heat shock, cold shock, bacterial challenge (Escherichia coli and Staphylococcus aureus) and Varroa mite infestations, and evaluated their effects on AmAChE expression. Among the stress factors tested, only heat shock condition induced AmAChE expression in a dose dependet manner. This finding suggests that one function of AmAChE1 is related with thermoregulations, especially against heat shock stress in honey bees.

Acetylcholinesterase 1 (AmAChE1) has low catalytic activity and is abundantly expressed in both neuronal and non-neuronal tissues. In previous experiments, we observed that AmAChE1 is rarely expressed in summer while highly expressed in winter. Through additional experiments, the expression of AmAChE1 was suggested to be associated with brood rearing status. Under the assumption that abnormal suppression of brood rearing activity may result in stressful condition in honey bee social community, it was further suggested that AmAChE1 is likely involved in stress management particularly during winter. We hypothesized that the increased docility usually observed in overwintering bees is likely an outcome of stress management in colony, which is mediated by AmAChE1 expression. To verify this, worker bees expressing abundant AmAChE1 were collected in early winter and injected with Amace1 dsRNA to knockdown Amace1. Then, the behavioral activity of the bees was investigated using the EthoVison video tracking system. Honey bees injected with Amace1 dsRNA showed significantly increased motility, which was strongly correlated with the suppressed expression level of AmAChE1 in the abdomen. No apparent reduced expression of AmAChE1 in the head was observed perhaps due to the limited efficacy of RNA interference in the blood-brain-barrier. Our finding suggests that behavioral activity can be regulated, at least, by AmAChE1 expression level in non-neuronal tissue (i.e., fatbody) perhaps via metabolic alteration.

We tested the identification ability of DNA barcodes comparing with morphological data using the Korean butterflies. The 921 samples (4.6 samples per species) for 202 resident Korean species except migratory species were used. The obtained samples were morphologically identified based on wing patterns. In a result, genetic divergence to the nearest-neighbouring taxon varied from 0 to 28.2%, with an average of 13.4 per cent. The neighbour joining (NJ) tree profile showed that sequence data for 185 of the 202 species formed distinct barcode clusters. Thus, our results indicated that 91.6 percent of the species were possible to allow the reliable identification using DNA barcoding. The rest 17 species (8.4%) consist of following four cases: clustering separated from each species by less than 1% branch length (two species pairs), paraphyletic clustering (two species pairs and one triple species pair), polyphyletic clustering with sharing barcodes (three species pairs), and clustering separated from existing species by the deep branch divergence (four clusters). However, it was not easy to interpret these ambiguous cases only using our current taxonomic evidences. Therefore, we are performing integrative taxonomy on these cases using other additional evidences such as examination on male genitalia and analysis of other gene regions.

Background : Angelica gigas is a monocarpic perennial plant. A. gigas, also called DangGui or Korean Angelica, is a major medicinal herb used in Asian countries such as Korea, Japan and China. In Korea, we are using the roots of A. gigas. but, Chinese using Angelica sinensis and Japanese using Angelica acutiloba with the same name 'DangGui'. The biggest problem in the use of A. gigas is the confusion with A. acutiloba or A. sinensis. This confusion can cause an medical accident or lack of pharmacological ingredients. In this study, we developed chloroplast InDel markers that can distinguish A. gigas, A. acutiloba or A. sinensis. Methods and Results : We collected 14 Angelica plant samples including A. gigas, A. acutiloba and A. sinensis and extrated DNA using CTAB method. The DNA was diluted to 10 ng/㎕ and kept -20℃. We designed the primer sets using CLC Main Workbench based on chloroplast DNA InDel region of between A. gigas and A. acutiloba. PCR were performed on the 14 Angelica plant samples including A. gigas, A. acutiloba and A. sinensis (5 repeats each). Electrophoresis was performed using fragment analyzer automated CE system. We designed 6 InDel primer sets and the primer sets amplified the amplicons effectively. Three of the 6 primer sets showed polymorphism. Conclusion : We could distinguish A. gigas, A. acutiloba, and A. sinensis using 2 newly developed InDel markers.

This study was to verity that the uptake inhibition and accumulation of nitrogen in different potassium levels. Lettuce was used as model plant in this study and grown in pot of 10cm's in diameter and depth with mixture media of vermiculite and perlite under supply of different culture solution for three weeks. Nitrogen absorption at root was inhibited by increased potassium concentration in nutrient solution, and nitrate accumulation of plant was depended on absorption of nitrogen because nitrate content of 0 K level was 4-5 times higher than that of 2 K level, Concentration of ascorbic acid was decreased by increasing the nitrogen absorption, since ascorbic acid (AsA) content of 2K level was higher than those of OK level in both of old leaf and flesh leaf