Biological properties of antimicrobial peptides (AMPs) of hemimetabolous insect are poorly characterized in innate immunity field. To investigate the biochemical properties of hemimetabolous insect’s AMPs, we purified the pyrrhocoricin-like AMP from the hemolymph of Riptortus pedestris and then named as riptocin. We successfully determined the primary protein structure and its cDNA sequence. Interestingly, the determined cDNA revealed that riptocin precursor is composed of 12 repeating units of active riptocins, which implied that riptocin precursor might require to be processed to generate active riptocins by several unidentified processing enzymes. In order to characterize the bio-processing mechanisms of riptocin precursor, we generated the antibody against active riptocin. Using quantitative PCR and Western blot analyses, we showed that gene of riptocin was started to express from the fatbody after three hours post bacterial infection. To address our hypothesis that active riptocin is generated from riptocin precursor by several processing enzymes, we need to obtain the riptocin precursor. Currently, we are expressing the recombinant riptocin precursor using in vitro translation system. Meanwhile, we investigated whether naive hemolymph (naive HL), which may contain precursor riptocin, can generate active riptocin when riptocin precursor was co-incubation with bacteria-challenged hemolymph (active HL), which may contain all processing enzymes. Actually, when naive HL was incubated with active HL, antimicrobial activity was dramatically increased, suggesting that processing enzymes in active HL may induce processing of riptocin precursor to generate active riptocins.

Riptortus pedestris possesses Burkholderia as gut symbiont in a symbiotic organ M4 midgut. To answer why Burkholderia symbionts are not eliminated by Riptortu s immune responses, we developed two hypotheses: (i) Burkholderia symbionts do not activate host innate immunity, or (ii) Burkholderia symbionts are resistant to th e host immune responses. For the first hypothesis, we compared the antimicrobial activities of the cultured Burkholderia-injected hemolymph and symbiotic Burkhol deria-injected hemolymphs. As a result, the symbiotic Burkholderia induced antim icrobial activity like the cultured Burkholderia, indicating the symbiotic cells are st ill immunogentic to host. However, when the activated hemolymph was treated to the Burkholderia cells, the symbiotic Burkholderia showed much higher susceptibi lity than the cultured Burkholderia. To understand molecular basis of these results, we purified antimicrobial peptides (AMPs) from Riptortus hemolymph. Similarly, the symbiotic Burkholderia exhibited the high susceptibility to the purified AMPs, riptocin and rip-defensin. To understand how symbiotic Burkholderia can survive in host in spite of their immuno-susceptibility, we examined the AMP expression i n the M4 midgut. Interestingly, the expression of AMPs is suppressed in the M4 mi dgut in comparison to that of the fat body. Finally, we proposed that the immuno-su sceptibility of Burkholderia symbiont helps them to retain in the symbiotic organ. Our in vivo data showing the rapid clearance of the symbiotic Burkholderia after inj ection to host Riptortus supports our proposal.

An easy and rapid resistance detection protocol for the Western flower thrips Frankliniella occidentalis was established based on the residual contact vial bioassay (RCV), in which insecticide resistance levels can be estimated at 8 h-post treatment of diagnostic doses. The RDA strain was used as a reference susceptible strain, which has been reared under laboratory conditions over 10 years without exposure to any insecticides. Seven insecticides were tested for the determination of diagnostic dose. Among them, five insecticides (chlorfenapyr, acrinathrin, spinosad, emmamectin benzoate and thiamethoxam, ranged as 0.03 ~ 0.42 μg-1cm2) were applicable to the RCV. However, two insecticides (omethoate and imidacloprid) were not able to be used for the RCV because the treated inner surface of glass vials by these insecticides were too viscous, causing non-specific mortality. The RCV detection kit was employed for the estimation of resistance levels for the five insecticides in five local populations. Almost field-collected populations revealed high levels of resistance to the four insecticides (acrinathrin, spinosad, emmamectin benzoate and thiamethoxam) by showing less than 50% mortality. The baseline resistance detection by RCV method will facilitate the selection of proper insecticides for farmers to manage insecticide resistant-populations of F. occidentalis.

Russula compacta, a wild mushroom, belongs to Russulaceae, Russulales of Basidiomycota. This study was conducted to evaluate the free radical scavenging, anti-inflammatory, anticholinesterase and anti-α-glucosidase effects from fruiting bodies of R. compacta extracted with methanol and hot water. In 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging effects, the methanol and hot water extracts showed good scavenging effects comparable with positive control, BHT. The chelating effect of methanol and hot water extracts of the mushroom were significantly higher than the positive control, BHT. The reducing power of the methanol and hot water extracts of the mushroom were lower than the positive control at the concentrations tested. In the HPLC anaysis of phenolic acids profile of the mushroom extract, 7 phenolic acids such as gallic acid, vanillin, rutin hydrate, resveratol, quercetin formononetin, and biochanin-A were detected. Nitric oxide (NO) production in lipopolysaccahride (LPS) activated RAW 264.7 cells was inhibited by 1.5-fold with the treatment of methanol extract when compared with the control. In the anti-cholinesterase activity assay, the methanol extract inhibited the acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) effects by 73.9% and 81.05% at the 1.0 mg/mL concentration, whereas galanthamine, the standard drug, inhibited the AChE and BChE activities by 97.80% and 81.12%, respectively at the same concentration. The methanol and hot water extracts of the mushroom inhibited the α-glucosidase activity by 55.44% and 62.00%, respectively at the 2.0 mg/mL concentration, while acarbose, the positive control inhibited the α-glucosidase activity by 81.81% at the 2.0 mg/mL concentration. From the experimental results, the fruiting bodies of R. compacta contained natural antioxidant, anti-inflammatory, anti-cholinesterase, and anti-diabetic substances, which might be used for health foods.

Coprinellus miaceus, belongs to Coprinaceae of Agaricales, Basidiomycota, has been used for an edible purposes in asian countries. This experiment was initiated to evaluate the free radical scavenging, free radical scavenging, anti-acetylcholinesterase, anti-inflammatory and α-glucosidase inhibitory activities of C. micaceus fruiting bodies extracted with methanol and hot water. In 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, the scavenging activities of methanol and hot water extracts were lower than that of positive control, BHT. The chelating effects of methanol and hot water extracts were significantly higher than positive control, BHT at the concentrations of 0.125-2.0 mg/mL. In the reducing power assay, methanol and hot water extracts exhibited the lower activities than the positive control, BHT at the 0.125-0.2 mg/mL concentration. In the HPLC analysis of phenolic acids profile of the mushroom fruiting bodies, 4 phenolic compounds including procatechuic acid, chlorogenic acid, (-)-epicatechin, and naringin were detected. The tyrosinase inhibitory activities of methanol and hot water extracts were 91.33% and 91.99% at 2.0 mg/mL concentration, while the inhibitory activity of kojic acid, the positive control, was 99.61%. Nitric oxide (NO) production in lipopolysaccahride (LPS) activated RAW 264.7 cells were inhibited by the methanol extract in a concentration dependent manner. In the acetylcholinesterase (AChE) inhibitory activity assay, methanol and hot water extracts of the mushroom inhibited the AChE by 94.64% and 74.19%, respectively at the 1.0 mg/mL concentration, whereas galanthamine, the standard drug inhibited the AChE activity by 97.80% at the same concentration. The methanol and hot water extracts of the mushroom inhibited the α-glucosidase activity by 62.26% and 67.59%, respectively at the 2.0 mg/mL concentration, while acarbose, the positive control inhibited the α-glucosidase activity by 81.81% at the same concentration. Therefore, it is concluded that fruiting bodies of C. micaceus contained natural antioxidant, anti-inflammatory, anti-acetylcholinesterase and α-glucosidase inhibitory substances which might be used for promoting human health.

Lentinus giganteus is a edible mushroom cultivated in Asian countries. The present study was initiated to evaluate the anti-inflammatory, anti-acetylcholinesterase, anti-α-glucosidase, and free radical scavenging activities from fruiting bodies of L. giganteus extracted with methanol and hot water. The free radical scavenging activities of methanol and hot water extracts on 1,1-diphenyl-2-picrylhydrazyl (DPPH) were 92.26% and 90.17% at 2.0 mg/mL concentration, respectively and comparable with positive control, BHT. The chelating activities of methanol and hot water extracts were significantly higher than the positive control tested. The reducing power of methanol and hot water extracts showed lower activities compared to positive control, BHT. The phenolic and flavonoid contents of hot water extract were 1.56 μg/mg and 24.35 μg/mg, respectively. Nitric oxide (NO) produced by lipopolysaccahride (LPS) activated RAW 264.7 cells were significantly inhibited by treatment of methanol and hot water extracts. The methanol extract inhibited the acetylcholinesterase (AChE) activity by 91.19% at the 2.0 mg/mL concentration, whereas galanthamine, the standard drug, inhibited the AChE activity by 97.80%. The hot water extracts inhibited the α-amylase and α-glucosidase activities by 78.86% and 80.78%, respectively at the 2.0 mg/mL concentration, while acarbose, the positive control, inhibited the activities by 89.91% and 81.81%, respectively at the same concentration. Therefore, it is concluded that fruiting bodies of L. giganteus contain antioxidant, anti-inflammatory, anti-cholinesterase, and anti-diabetic substances, which can be used for natural health food for promoting human health.

In the midst of environments with crop-mixing, there were more preferred the fruit trees by brown-winged green (Plautia stali (Scott)), and yellow-brown stink bug (Halyomorpha halys (Stål) and vegetables by Far eastern green stink bug (Nezara antennata Scott) and Bean bug (Riptortus pedestris (Thurnberg)). The major dominant species was identified as P. stali, infesting fruit trees over 99.4% among tested insects. It is severely damaged on peach of middle and late ripening, crab apple, pear and wild fruit of Actinidia arguta. In an investigation for the seasonal occurrence using aggregation pheromone trap, P. stali was peaked with three times at late-May ~ early-June, mid- and late-July, and mid-Sept. ~ Oct. And H. halys slightly peaked at mid-June, and mid- and late-July and increased the occurrence at mid-Sept. ~ mid-Oct. Using seven insecticides - dinotefuran WP, flonicamid WP, etofenprox WP, chlorpyrifos WP, cabaryl WP, chlothianidin SC, and bifenthrin WP - registered on apple, contact and residual toxicity was tested on both male and female of P. stali and H. halys. Contact toxicity of dinotefuran WP was excellent on P. stali but the others had low effect. While all insecticides except flonicamid, residual toxicity were all effective on P. stali. Dinotefuran have a excellent contact toxicity and chlorpyrifos and bifenthrin have good residual toxicity against both male and female of H. halys.

The Riptortus-Burkholderia symbiosis is a newly emerging insect-bacterium symbiotic system. This symbiosis system has a good merit as an experimental model system to produce the non-symbiotic (apo) and symbiotic (sym) host insect. In recent reported papers, the symbionts play important biological roles for the host insects. Meanwhile, juvenile hormone (JH) is one of major hormone synthesized corpora allata(CA) to control many physiology of insect. However, the study for cross-talk mechanism between symbionts and host hormones to control important physiological phenomenon of insects is almost none. In this study, we found that Riptortus speed up adult emerging and increase egg laying on presence of symbiont Burkholderia. Also we found that hexamerin proteins, which were controlled the expression by JH, were accumulated in sym-Riptortus hemolymph compare with apo-Riptortus. According as combined results, we hypothesized that the gut symbiont Burkholderia can control JH titer to conclude out beneficial effects such as development and reproduction of R. pedestris. To verify this hypothesis, we examined measurement of JH titer, expression of hexamerins as JH response genes and RNAi for hexamerin protein during whole Riptortus life on presence or absence of symbiont Burkholderia. All results demonstrated that gut symbiont controlled JH titer of Riptortus. Controlled JH amount by symbiont Burkholderia in host midgut regulated hexamerin protein expression for speeding up adult emerging and increasing egg production.

The Riptortus (stinkbug) has a specialized symbiotic organ, M4 midgut, to harboring symbiont Burkholderia. M4 midgut is located in abdomen and surrounded with insect hemolymph. Recently our group demonstrated that symbiotic Burkholderia showed different physiology after adapting in M4 gut compare with in vitro cultured Burkholderia. And population of symbiotic Burkholderia in the M4 midgut is regulated by special organ. However, the molecular mechanism to prevent spreading and migrating symbiont bacteria to other host tissues from symbiotic organ is not clear. Therefore, we assumed that symbiont Burkholderia are susceptible to host humoral immunity after established infection in M4 midgut to prevent spreading and migrating into the other host tissues through Riptortus hemolymph. To prove this assuming, we tested the susceptibility and survival rate of symbiont Burkholderia in hemolymph of Riptortus in vitro and in vivo. We also examined the susceptibility of symbiont Burkholderia using purified antimicrobial peptides (AMP), pyrrhocoricin-like, thanatin-like and defensin-like AMPs. Finally, we tested inducing ability for AMPs by systemic infection of symbiotic Burkholderia. Gene expression of purified AMPs was not different after systemic infection of both symbiont and in vitro cultured Burkholderia. Surprisingly, in vitro cultured Burkholderia resisted on bacteria injected hemolymph and purified AMPs but symbiont Burkholderia were highly susceptible in bacteria injected hemolymph and purified AMP. These results suggest that symbiont Burkholderia can't survive in the hemolymph after escaping symbiotic organ. Moreover, humoral immunity of host Riptortus is important to prevent spreading and migrating symbiont Burkholderia into the other host tissue or organ from symbiotic organ.

White-backed planthopper, Sogatella furcifera (Horvath) (Hemiptera: Delphacidae), has been a serious migratory pest in Korea. It is important to figure out the migration route and gene flow of S. furcifera. Microsatellite marker (SSR) shows high efficiency as molecular markers. Unfortunately, various microsatellite marker of S. furcifera has not been developed to see genetic diversity. S. furcifera samples were collected from Vietnam, Laos and three different sites of Bangladesh in 2012. We extracted DNA by using QIAamp DNA Mini Kit and ran next generation DNA sequencer (NGS) Roche 454 to develop a new microsatellite marker. Roughly, about 18 singleton primers and 14 contigs primers were found. We will test these primers with S. fucifera DNA samples, and figure out the accurate new microstatellite marker.

새로 조성된 간척농지의 토양개량을 위하여 새만금간척지에서 2007년부터 2011년까지 4년 동안 녹비작물 (여름철 : 세스바니아, 제주재래피, 수수×수단그라스, 겨울철 : 귀리, 호밀, 보리) 재배 및 시용이 토양이화학성에 미치는 영향을 조사하여얻은 결과는 다음과 같다.1. 녹비작물의 바이오매스 생산량은 평균 9.7~15.0ton/ha이었으며, 이 중에서 수수×수단그라스의 생산량이 가장 많았다.2. 녹비작물의 재배와 시용으로 토양유기물 함량이 1.3에서6.0g/kg로 매년 평균 1.2g/kg씩 증가하였다.3. 녹비작물의 재배와 토양환원에 의해 토양의 용적밀도가1.44에서 1.24Mg/m3로 감소하였다.4. 수수×수단그라스와 호밀의 이어짓기가 토양 용적밀도 감소에 가장 효과적이었고, 세스바니아 재배는 토양 질소함량 증가에 가장 효율적이었다.이상의 결과로 미루어 볼 때 간척지에서의 녹비작물 재배및 시용은 토양의 비옥도 요인을 일부 개선시킴으로써 간척농지 개량에 기여할 수 있을 것으로 기대된다. 앞으로 신간척지에서의 유기물과 관련된 다각적인 연구가 필요할 것으로 생각된다.

To compare the catches made using gillnets, trammel nets, and gillnets with supporting lines, several experiments were conducted with commercial vessels near Uljin and Pohang in Eastern Korea between July 2010 and May 2011. Two sets of 13 different nets were used, including 5 panels of gillnets and trammel nets each with stretched mesh sizes of 7.6, 9.1, 10.6, 12.1, and 13.6 cm and 3 panels of gillnets with a mesh size of 9.1 cm with supporting lines with different line spacing. The outer (stretched) mesh size of the trammel nets measured 51.5 cm. The target fishes of the fishing nets were various types of flounders. The catch rate of flounders was 50.7% of the total catch in weight. The total catch for all nets was 443.8 kg. The predominant species was pointhead flounder (Cleisthenes pinetorum). The total catch by trammel nets was 1.4 times that of the comparable gillnets. But more pointhead flounder were caught by gillnets than by trammel nets, though there was no significant difference. Fishermen catching the pointhead flounder in Korea said that there was no need to use trammel net to catch it; this was an unexpected finding compared to the findings of other flounder fisheries. The amounts of roughscale sole, brown sole, and blackfin flounder caught by trammel nets were greater than those caught by gillnets. The mean lengths (standard deviation) of blackfin flounder, pointhead flounder, brown sole, and roughscale sole were 21.0 (4.57), 22.9 (3.40), 24.7 (4.90), and 28.3 (5.43) cm, respectively; there were significant differences in mean length (p < 0.00001). Therefore, in order to catch flounder efficiently, the fishing nets and mesh size should be chosen according to the target species. One advantage of using supporting lines is that it prevents breakage by strengthening the material especially when utilized on a rough bottom. Catch by using gillnet with supporting lines was not greater than that by using trammel net for the conservation of fisheries resources.

Polyporus umbellatus (Syn. Grifola umbellata) is a sclerotium forming mushroom belongs to family Poly-poraceae of Polyphorales, Basidiomycota. The sclerotia of P. umbellatus have long been used for traditional medicinesin China, Korea and Japan. This study was initiated to obtain the basic data for artificial sclerotial production of P. umbel-latus. Here, we investigated the favorable conditions for mycelial growth of P. umbellatus and its symbiotic fungus Armill-aria mellea. We also evaluate the favorable carbon and nitrogen sources for sclerotial formation in dual culture betweenP. umbellatus and A. mellea. The favorable conditions for mycelial growth of P. umbellatus were 20°C and pH 4, whileoptimal conditions for mycelial growth of A. mellea were 25°C and pH 6. The carbon sources for optimal mycelial growthof P. umbellatus were fructose and glucose, while carbon sources for favorable mycelial growth of A. mellea were alsofructose and glucose. The nitrogen sources for favorable mycelial growth P. umbellatus were peptone and yeast extract,while optimal mycelial growth of A. mellea were obtained in peptone and yeast extract. When P. umbellatus and A. melleawere dual cultured on carbon sources, sclerotia were induced on basal media supplemented with glucose, fructose andmaltose at pH 4~6, while nitrogen sources inducing sclerotia were basal media supplemented with peptone and yeastextract for 60 days at 20°C under dark condition.

This study was performed to assess the current stock condition of elkhorn sculpin along the Uljin area in the East Sea of Korea. To assess the state of the stock, yield-per-recruit (YPR) and spawning biomass-perrecruit (SBPR) analyses were performed. Estimates of Fmax and F0.1 were 2.10/year and 0.48/year, respectively, and those of F35% and F40% were 0.66/year and 0.54/year, respectively. Current fishing mortality was estimated at 0.63/year and the current age at first capture was 2.41years. F40% was set as the target reference point of the stock. SBPR at F40% and current SBPR were estimated to be 41.85g and 37.77g, respectively. Estimated FOTY which is the fishing mortality for the overfished threshold yield was 0.49/year. The ratio of SBPR/SBPRMSY was calculated as 0.90 and that of F/FOTY was 1.05. The ratio of tc/tc opt was calculated as 1.15 and that of F/FOTY was 1.17. Therefore, the current stock condition of elkhorn sculpin along the Uljin area of Korea has not been overfished, however, it indicates that a light overfishing is going on this stock.

Recently, there had been reports on ethanol fermentation from mono-saccharide and disaccharide by mushroom mycelia. This experiment was conducted to study ethanol production from xylose by mycelila of mushrooms isolated from Korea. The cultures used in this study were obtained from Culture Collection and DNA Bank of Mushrooms in the Division of Life Sciences, Incheon National University. The results showed that Neolentinus lepideus, Trametes hirsuta and Cerrena unicolor produced ethanol from xylose contained media. The ethanol concentration produced in the xylose contained media ranged from 2.5∼3.8%. The highest ethanol concentration(3.8%) was obtained from fermentation of xylose by Neolentinus lepideus mycelia. All of the mushroom mycelia used in this study showed a good ability of ethanol fermentation from glucose, fructose, mannose, cellobiose and maltose.

Ganoderma applanatum is a medicinal mushroom belongs to Ganodermataceae of Polyporales, Basidiomycota. This study was conducted to evaluate the antioxidant, anti-inflammatory and anti-xanthine oxidase activities of G. applanatum fruiting bodies extracted with methanol. The antioxidant activities were performed on reducing power, 1,1-diphenyl-2-picrylhydrazyl free radical scavenging, and ferrous ion chelating activities. In addition to this, polyphenol and flavonoid contents were aslo analyzed. The methanol extract showed the good reducing power of 3.5 at the concentration of 4.0 mg/ml. The scavenging activity of methanol extract of G. applanatum 1,1-diphenyl-2-picrylhydrazy radicals was better than those of positive control, BHT and tocopherol. The ferrous ion chelating effect of methanol extract was moe effective than those of positive control, BHT and tocopherol. The antioxidant activities of the G. applanatum were increased with the increasing concentration of the extracts. The xanthine oxidase inhibitory activity of methanol extract of G. applanatum was good as positive control, allopurinol. The anti-inflammatory activity of mushroom extract was measured by carrageenan-induced hind paw edema of albino rat. The injection of 50 mg/kg of methanol and hot water extracts significantly reduced the carrageenan-induced paw edema compared with the positive control, indomethacin. The results indicated that fruiting bodies of G. applanatum could be used for natural medicine for anti-inflammatory and anti-xanthine oxidase.