Enhanced green fluoresce protein gene (egfp) was expressed in Beauveria bassiana ERL836 based on the Agrobacterium tumefaciens-mediated transformation (AtMT) method in this study. The ERL836 transformants were generated with pCambia-egfp binary vector. Ten transformants were randomly selected and analyzed for the T-DNA insertion and gene expression. The results revealed that 60% of the fungal putative transformants were inserted by the T-DNA fragment. Of these transformants, 33.33% (2 transformants) expressed the egfp gene. The egfp transformants showed strong green fluorescence with different expression levels. The results of this study could provide a reference for foreign protein expression in B. bassiana by using the AtMT method.

Mealworm, Tenebrio molitor L. (Coleoptera: Tenebrionidae) has high and safe protein contents, which enables it to be animal feed. However, occurrence of entomopathogenic fungi in mealworms is one of the limitations for mass production. In this work, we investigated relationships between abiotic conditions and occurrence of fungal pathogens and established an effective control method using fungicides. In virulence assay, third instar mealworm larvae were sprayed by six entomopathogenic Beauveria bassiana isolates and kept under high relative humidity; B. bassiana ERL1575 isolate had highest virulence. Under normal humidity, ERL1575 conidia showed different virulence between spray (~0% virulence) and digestion (~80% virulence) method. Furthermore, mealworms, which digested conidia, were exposed to various temperature (20-35°C) and humidity (1-3 ml distilled water spray/35 mm diam. dish) conditions for 5 days. All the treatments showed ~90% virulence except 35°C incubations (~20% virulence), but irrespective to the humidity conditions. Forty chemical fungicides were assayed against conidial germination and hyphal growth of ERL1575. Fluazinam and mancozeb showed strong inhibition of conidial germination at standard application dose (SD), 1/2 SD and 1/5 SD; besides, fluazinam showed strong inhibition of hyphal growth. When fluazinam and mancozeb were applied to the fungal conidia-inoculated wheat bran, most of mealworms were alive after 3 days post application. However, high mortality rate (~100%) were observed in the conidia-inoculated wheat bran without any fungicides. In conclusion, this work suggests that B. bassiana isolates could be pathogens at <30°C when they were digested by mealworms, and fluazinam and mancozeb would be used as effective control agents against the pathogen.

Insect cuticular melanization is regulated by the prophenoloxidase (proPO)-activating system, which is a component of innate immunity. However, the differentiation between cuticular melanization and innate immunity is not well defined. Here, we demonstrate that the proPO-activating system regulates cuticular melanization in the silkworm pupae using a different mechanism. Our results indicate that the differential and spatial regulation of key components, such as the proPO-activating factor, tyrosine hydroxylase, proPOs, and immulectin, primes the proPO-activating system for either cuticular melanization or innate immunity. This dual strategy for cuticular melanization in development and innate immunity upon infection demonstrates a two-pronged defense mechanism mediated by the priming of the proPO system.

Osmia cornifrons plays a major role in the pollination of orchards, but basic information on vitellogenin and the oocyte development is limited. To better understand vitellogenin in hymenopteran insects, we cloned a cDNA encoding vitellogenin from the hornfaced bee O. cornifrons. O. cornifrons vitellogenin cDNA contains 5477 bp with an open reading frame of 1,783 amino acid residues, and has a predicted molecular mass of approximately 200.21 kDa and a pI of 6.55. O. cornifrons vitellogenin possesses four consensus (RXXR/S) cleavage sites and has conserved DGXR and GL/ICG motifs in the C-terminus. The deduced amino acid sequence of the O. cornifrons vitellogenin cDNA showed a 66% identity with Megachile rotundata, 53% to Apis mellifera, 51% to Bombus ignitus, and 42%-30% with other hymenopteran insect vitellogenins. Phylogenetic analysis showed that O. cornifrons vitellogenin clustered with vitellogenins from Megachildae, Apidae, Vespidae, and Formicidae species but not with those from Pteromalidae, Aphelinidae or Ichneumonidae species.

Inhibitor cysteine knot (ICK) peptides exhibit ion channel blocking, insecticidal, and antimicrobial activities, but currently, no functional roles for bee-derived ICK peptides have been identified. In this study, a bee (Apis cerana) ICK peptide (AcICK) that acts as an antifungal peptide and as an insecticidal venom toxin was identified. AcICK contains an ICK fold that is expressed in the epidermis, fat body, or venom gland and is present as a 6.6-kDa peptide in bee venom. Recombinant AcICK peptide (expressed in baculovirus-infected insect cells) bound directly to Beauveria bassiana and Fusarium graminearum, but not to Escherichia coli or Bacillus thuringiensis. Consistent with these findings, AcICK showed antifungal activity, indicating that AcICK acts as an antifungal peptide. Furthermore, AcICK expression is induced in the fat body and epidermis after injection with B. bassiana. These results provide insight into the role of AcICK during the innate immune response following fungal infection. Additionally, we show that AcICK has insecticidal activity. Our results demonstrate a functional role for AcICK in bees: AcICK acts as an antifungal peptide in innate immune reactions in the body and as an insecticidal toxin in venom. The finding that the AcICK peptide functions with different mechanisms of action in the body and in venom highlights the two-pronged strategy that is possible with the bee ICK peptide.

Apolipophorin III (apoLp-III) is a multifunctional protein that is involved in lipid transport and innate immunity in insects. In this study, an apoLp-III protein that exhibits antibacterial activity was identified in honeybees (Apis cerana). A. cerana apoLp-III cDNA consists of 193 amino acids that share high protein sequence identity to other members of the hymenoptran insect apoLp-III family. A. cerana apoLp-III is expressed constitutively in the fat body, epidermis, and venom gland and is detected as a 23-kDa protein. A. cerana apoLp-III expression is induced in the fat body after injection with Escherichia coli, Bacillus thuringiensis, or Beauveria bassiana. However, recombinant A. cerana apoLp-III (expressed in baculovirus-infected insect cells) binds directly to E. coli and B. thuringiensis but not to B. bassiana. Consistent with these findings, A. cerana apoLp-III exhibited antibacterial activity against Gram-negative and Gram-positive bacteria. These results provide insight into the role of A. cerana apoLp-III during the innate immune response following bacterial infection.

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.

We investigated the insect community along altitudinal gradient to gather basic data for distributional monitoring of insect species in the forest ecosystem. The investigation area was Seon-gaksan (Mt.) in Jinan-gun, Jeollabuk-do province, where the bucket-light trap and pit-fall trap for quantification were installed in Quercus vegetation at altitude of 300m, 600m and 900m. The field collecting was performed on May, July and September 2013 respectively. ANOVA analysis was conducted to analyze the significance between insect species along altitude using the collected insect community data. Analysis of variance (ANOVA) results showed statistically significant differences among ground-beetles and ants abundance with altitude as a response variable. Although we expected a distinct cluster with the difference of altitude at each study site, nonmetric multidimensional scaling (NMS) showed distinct clusters with the moth, ground-beetles, and ant assemblage at altitudinal increase and sampling month. In the result, a total of 309 species in 18 families of nocturnal moths were collected by bucket-light trap. The insects collected in pit-fall trap were ground-beetles with 196 individuals of 26 species and ants with 11,276 individuals of 14 species respectively.

In this study, the following four species are reported for the first time in Korea: Diplopseustis perieresalis (Walker, 1859), Dolicharthria bruguieralis (Duponchel, 1833), Herpetogramma ochrimaculale (South, 1901), and Omiodes diemenalis (Guenée, 1854). All of the known species were enumerated with their available information including the collecting localities, host plants, illustrations of adults, and genitalia.

A filtration-tapingmethod was demonstrated to fabricate carbon nanotube (CNT) emitters. This method shows many good features, including high mechanical adhesion, good electrical contact, low temperature, organic-free, low cost, large size, and suitability for various CNT materials and substrates. These good features promise an advanced fieldemission performance with a turn-on fieldof 0.88 V/mm at a current density of 0.1 mA/cm2, a threshold fieldof 1.98 V/mm at a current density of 1 mA/cm2, and a good stability of over 20 h. The filtratio-taping technique is an effective way to realize low-cost, large-size, and high-performance CNT emitters.

The online game market has rapidly increased worldwide, especially in China. In 2012, the world game market equaled 111.7 billion. Many game companies compete by launching new online games in various genres. In 2010, Activision Blizzard reported that the World of Warcraft (WOW) is the world’s most popular online game. Chinese gamers constitute more than half of WOW’s 10 million users. Korea is the second largest gaming market. Thus many foreign game companies target Korea and China, in competition with domestic game companies. The purposes are 1) to understand the role of country of origin and /or brand image in game consumer behavior, 2) to find out the difference of game evaluation between game users in China and Korea, and 3) to draw strategic implication game marketers. Consumers expect products from advanced countries to provide superior performance, so they often look to country of origin in evaluating products. Recently, consumers have looked to brand image as another key element in product evaluation. In this research, we study whether gamers evaluate games on the basis of country of origin and/or brand image. In this paper, we add to the gaming research and suggest that the online game market has three dimensions. First, online games are products. Second, online games are service. Last, online games are entertainment. Data collection resulted in 355 usable responses from online game (LOL) users in Korea(166) and China(189). We conduct factor analysis and reliability analysis to check reliability and validity. Country of Origin is the key element for product evaluation of utilitarian goods but it doesn’t have significant effect on product evaluation of hedonic products which mainly provide sensual pleasure, fantasy, and fun to customers such as luxury goods or online game (Dhar & Wertenbroch, 2000).

This study explores relationship between social responsibility in advertising and brand attitude in luxury products. This study investgates how psychological constructs of attitude towards advertising affect brand attitude and purchase intention of luxury brand consumer and how it can lead the sustainable development of luxury products. Consumers no longer purchase products only but depend on quality and price of product. With globalization and rapid growth, corporate social responsibility becomes important issue. And the advertising represents corporate image and management concept. More recently, and coinciding with some major corporate ethical disasters, many companies have been including sections on governance, ethical practice, and social responsibility (David S. Waller & Roman Lanis, 2009). According to David S. Waller & Roman Lanis (2009), Corporate social responsibility (CSR) disclosure has been the subject of substantial academic accounting research (Farook and Lanis 2005; Gray, Owen, and Maunders 1987). Advertising is one of the typical means that can represent a corporate image. As defined by Lutz (1985, p. 53), attitude toward advertising in general is “a learned predisposition to respond in a consistently favorable or unfavorable manner to advertising in general.” In his framework, Lutz viewed attitude toward advertising in general as being directly influenced by general perceptions of advertising (Srinivas Durvasula et al., 1993). Authors would like to study following issues in this research. (1) How perceived social responsibility influences Attitude toward advertising. (2) How fashion consumer behavior influences Attitude toward advertising. (3) How attitude toward advertising affects brand attitude and purchase intention. (4) How proximity plays a moderating role among perceived social responsibility, attitude toward advertising and brand attitude.

Xylitol is a five-carbon sugar alcohol that inhibits the growth of oral streptococci, including Streptococcus mutans. In this study, we tested xylitol sensitivity among the oral streptococci. We also compared nucleotide homology of putative fructose phosphotransferase system (PTS) and xylitol sensitivity, since xylitol is transported via the fructose PTS. Among the tested Streptococci, S. pneumonia showed the highest resistance to xylitol while S. gordonii and S. sanguinis showed the most sensitive growth inhibition. These streptococci could be grouped according to their xylitol sensitivity. S. mutans and S. salivarius showed similar bacterial growth inhibition by xylitol. S. mitis, S. oralis, S. pneumonia, S. intermedius and S. anginosus showed relatively low sensitivity to xylitol. When the genetic homologies of five fructose PTSs were compared among the tested streptococci, closely related streptococci showed similar sensitivity to xylitol. Taken together, fructose PTSs may mediate the sensitivity to xylitol in oral streptococci.

Despite a large amount of research on reading, little attention has been paid to experimental studies on the effects of purposeful reading. This study explored the effects of reading purpose on reading comprehension and perceived difficulty. 71 high school students participated in this study and were randomly assigned to three groups – graphic organizer (Purpose 1), key sentence (Purpose 2), and control. The learners’ reading comprehension was measured through pre-, post- and delayed tests. The participants’ perceived difficulty for the texts was also checked with a sevenpoint Likert scale. The result demonstrates no immediate effect of establishing reading purpose, though immediate reading purpose effects might be found if a longer treatment period is set. The results also reveal the sustained effect of reading purpose on reading comprehension. In perceived text difficulty, the Purpose 1 group found the reading passages the easiest among the three groups. This suggests that learners establishing a reading purpose can gain greater ease in reading than those without a reading purpose.

The legume family (Fabaceae) is the third largest family of flowering plants worldwide and often damaged by stink bugs, particularly Riptortus clavatus Thunberg (Heteroptera: Alydidae). This insect selectively feeds and oviposits on these plants, resulting in >50% of damage to soybeans harvested in autumn. The most common control methods for this insect are periodic application of chemical pesticides, pheromone traps, and bait crops, but still they encounters residual problem, insect resistance or low control efficacy. The current management system needs alternative control methods, which possibly combined with the present methods. Some entomopathogenic fungi have high virulence against the nymphs and adults of R. clavatus in a favorable conditions, reaching ∼80% mortality with mycosis in 3∼5 days. The genera of Beauveria and Metarhizium are possible control agents for effective and safe management of the serious pest. Fungal infection in R. clavatus was observed using an egfp-expressing fungal transformant. Thermotolerant entomopathogenic fungal conidia can be produced in cereal substrates, which enables conidia to be stable for long times. Our considerations need to be given to the combinations with other control methods, such as chemical pesticides or pheromone trap.

Screening for antimicrobial peptide genes in the immune-induced Antheraea yamamai larvae led to the identification of a novel antifungal moricin-like peptide (MLP10) gene. The complete MLP10 cDNA is comprised of 403 bp with 174 bp open reading frame encoding a 58 amino acid precursor that contains a putative 23-residue signal peptide, a 2-residue propeptide and a 33-residue mature peptide. The deduced amino acid sequence of MLP10 has 26∼52% identity to those of moricin-related peptides from lepidopteran insects. The MLP10 was highly expressed in E. coli BL21(DE3) by fusing with ketosteroid isomerase (KSI) to avoid the cell death during induction. The resulting expressed KSI-MLP10 fusion protein was in a insoluble form. Recombinant MLP10 was released by cleavage of the fusion protein with cyanogen bromide (CNBr). Subsequently, we purified pure active MLP10 by FPLC chromatography, and 5.2mg of MLP10 was obtained from 1L culture medium. The purified MLP10 was prevented the growth of candida albicans at 6.25 uM, and was also active against gram negative and gram positive bacteria. This potent antimicrobial activity suggests that MLP10 may play a role in the immune response of A. yamamai.