Beauveria bassiana JEF-007 with strong virulence against Riptortus pedestiris was selected for the Agrobacerium tumefaciens-mediated transformation(AtMT). AtMT generated two transformants, B1-06 and C1-49, showed significantly reduced virulence against R. pedestris. To identify the virulence-related genes, thermal asymmetric interlaced(TAIL) PCR and flanking region analysis were performed. From the analysis, two genes, Complex I intermediated-associated protein 30(CIA30) and Autophagy protein 22(Atg22), possibly related virulence in B. bassiana JEF-007. For the analysis of two putative virulence-related genes in JEF-007, hairpin RNA interference (hpRNAi) is under consideration. This work can provide the functional roles of the virulence-related genes in B. bassiana JEF-007.

Chemical pesticides have been used to control persimmon pests, however the overuse of the pesticides caused insect resistance. Herein we investigate the potential of eco-friendly organic pesticides (EFOP) on the control persimmon pests, Stathmopoda masinissa and Riptortus pedestris. Eleven commercially available EFOP were sprayed on the target insects in laboratory conditions. The chemical pesticide, buprofezin+dinotefuran wettable powder served as a positive control. In the bioassay against persimmon fruit moth, the mixture of Chinese scholar tree, goosefoot and subtripinnata extracts (EFOP-2) treatment showed the highest control efficacy: 27.7% (5 days) of survival rates. In the bioassay against bean bugs, the EFOP-2 showed the highest control efficacy: 20.0% (5 days) of survival rates. These results suggest that the mixture of three plants extracts (EFOP-2) has high and multiple potential in the management of the persimmon pests.

A viral genome was assembled de novo from next-generation sequencing (NGS) data from bean bugs, Riptortus pedestris, infected with an entomopathogenic fungus, Beauveria bassiana (Bb), and was further confirmed via the RACE method. Based on the phylogenetic analysis of the RdRp sequences, RiPV-1 was clustered in the unassigned insect RNA viruses with two other viruses, APV and KFV. These three viruses were suggested to constitute a new group of insect RNA viruses. Interestingly, RiPV-1 replication was increased dramatically in bean bugs 2 to 6 days after fungal infection. In conclusion, a novel insect RNA virus was found by NGS data assembly. This virus can provide further insight into the interaction between virus, fungus and the host.

Alternative control agents, hopefully overcoming the present issues and problems in agrochemicals, should be considered and finally applied to the vector controls. Entomopathogenic fungi can used as one of the possible novel control agents and great considerations are given to the control of soil- or water-dwelling stages of vectors, such as mosquitoes and ticks. In our research group, the entomopathogenic fungal library has been constructed using the mealworm-based isolation system, which showed a variety of opportunities of their use in vector control. Important key production technologies including granular formulation have been developed to increase their industrialization. Some entomopathogenic fungal isolates showed high biological performance in the control of mosquitoes and ticks in field stands. R&D of down-stream process should be seriously considered as the key step.

Composite resins are developed as restorative materials to improve esthetics and mechanical properties. To improve the physical properties of resin material, resin filler have to be added. However, no imaging method is adopted for resin filler distribution. Optical coherence tomography (OCT) is a optical imaging technique to delineate microscopic structures within biological tissue. The OCT application to dental composites resin and its filler is not described yet. So, this new and advanced optical method is needed for clinical application for evaluation of dental composite resin. To analyze the spatial distribution of dental composite resin and to evaluate the resin restoration in cavity, frequency domain optical coherence tomography (FD-OCT) was used for their analysis. Resin restored tooth was prepared. For morphological observation, serially sectioned teeth, conventional X-ray taking and micro computed-tomography (CT) images were compared with OCT images. The experiment has done to evaluate the success of the resin restoration using 3 dimensional structure OCT image. In this research, OCT is evaluated as a new technique to image resin restoration. The evaluation of resin restored tooth was performed by OCT. Inappropriate restoration such as marginal adaptation, large porosities, internal integrity and poor contour could be detected. Resin filler also could be checked by OCT. The distribution, number, regularity and size of resin filler can be differentiated from several commercial products. Considering the characteristics of the OCT, it can be used to evaluate the defects of resin restoration, resin filler distribution, and internal integrity between resin material and tooth structure. The OCT can be considered to be a new and advanced method for the evaluation of resin restorations.

Supply and demand patterns have dynamically changed in recent years due to the increased global competition. Firms have selected quality and price as competition components and have accomplished corporate innovation in order to achieve competitive advantage. Innovation has been recognized as the way to enhance corporate competitiveness and to continuously grow in churning global competition (Homburg, Schwemmle, & Kuehnl, 2015; Moon, Miller, & Kim, 2013). Although innovation becomes a common means to improve a firm’s performance, it has the limitation of achieving a firm’s strategic goal as a long-term strategy. Thus, firms need to have more fine-grained strategies to survive in dynamically changing business environment, such as design innovation (Moon et al., 2015). For example, Apple has produced its products (e.g., iPhone, iPod and iPad) focusing on innovative product design to influence consumer purchase intention. Design can lead to a distinct competitive advantage (Bolch, 1995). Furthermore, product design can be used by firms to create amd enhance brand recognition, as well as to increase firms’ value (Mozota, 2002). Brand experience positively influences customer satisfaction and brand loyalty (Brakus, Schmitt, & Zarantonello, 2009). Thus, we assume that design experience and product innovation have a positive effect on consumers’ purchase behavior and customer value. However, while the importance of design innovation is recognized, it is not easy to apply the design innovation to marketing due to the lack of relevant research in the field. In fact, relevant research on the influence of innovative new product design and design experience on customer value is scarce. Therefore, the purpose of this study is to investigate how brand experiences and innovative product design affect customer value. Successful innovation is accomplished by identifying customer needs first and developing innovative products to satisfy their needs (Hauser et al., 2006). Design is seen as the core of innovation and the moment when a new object is conceived of, devised, and shaped in a prototype form (Landwehr, Wentzel, & Herrmann, 2013). Verganti (2008) studies the concept of "user-centered design," which describes how companies can use design to improve their relationships with users and develop a better understanding of user needs. In this research, design innovation has three dimensions: aesthetic attributes, feature attributes, and emotional attributes. First, aesthetic attributes focus on the product design itself. The aesthetic appearance of a product has a large bearing on its potential market share (Liu, 2003). Second, feature attributes focus on the product features and functional aspects that are required to satisfy customer needs. Feature attributes enable performance that can give results in the operating process (Crawford & Di Benedetto, 2007). Third, emotional attributes focus on consumers’ feeling when they purchase a new product to satisfy their needs. Emotional attributes are generated by consumers’ experience when they purchase a new product in the store. The more the product design satisfies customer's emotional needs, the more customers’ attention is attracted to the purchase of a product (Mokarian, 2007). A product satisfying the aesthetic, feature, and emotional attributes through design innovation provides a new experience to customers (Desmet & Hekkert, 2007). A consumer’s purchase decision making is affected by both direct and indirect experience of using the product and the function of product (Brakus, Schmitt, & Zarantonello, 2009). Therefore, a designer comes to design the product, taking an interest in the experience that the product gives besides its shape and function. Product experiences occur when a customer interacts with the product ̶ for example, when customers search for, examine, and evaluate products (Hoch, 2002). The product experience can be direct, i.e. when there is physical contact with the product (Hoch & Ha 1986) or indirect, i.e. when a product is presented virtually or in an advertisement (Hoch & Ha 1986; Kempf and Smith 1998). Brand experience can be split into four dimensions (sensory, affective, intellectual, and behavioral), which are differentially evoked by various brands (Brakus, Schmitt, & Zarantonello, 2009). According to previous studies, these four experience dimensions (sensory, affective, intellectual, and behavioral experience dimensions) have an effect on customer satisfaction and customer loyalty. Customer value can be defined as the trade-off between the benefits and sacrifices as a customer purchases a product or service from a supplier (Ulaga & Chacour, 2001). Ulaga and Chacour presented three dimensions of customer value: product-related components, service-related components, and promotion-related components. The researchers argued that customer value could be an important strategic marketing tool to clarify a firm's proposition to customers. Product-related components are intrinsic product characteristics. Product quality is a key factor of relationship value (Ulaga, 2003). Customer value consists of product value and service value. Customer value can be enhanced by quality, diversity, payment, and service quality and decreased by price, convenience, and risk (Jarvenpaa & Todd, 2003). Service-related components include all aspects of service associated with the product. Various service components play an important role in differentiating a supplier's offering (Narus & Anderson, 1996). Promotion-related components include all items used to promote the product to the customer. For all purchasing processes, it is necessary to assess the perceived customer value, such as service quality and promotional quality (Qualls & Rosa, 1995). In the present study, we conducted a research survey with 300 subjects and analyzed the data. In order to test the reliability of questionnaire, Cronbach's alpha was used. In order to test the validity, an exploratory factor analysis was conducted. Measurement was made for 3 variables, i.e. design innovation, design experience, and customer value. Three variables (aesthetical attributes, feature attributes, and emotional attributes) were used for design innovation. Three types of attributes were introduced for a mobile phone design innovation. Four dimensions (sensory, affective, intellectual, behavioral) were measured using a technical design experience and a humanistic design experience. Three variables (product-related customer value, service-related customer value, and promotion-related customer value) were employed for customer value. The results of the analyses demonstrate that design innovation has a positive effect on design experience, while design experience has a positive effect on customer value. Aesthetic attributes of design innovation have a positive effect on technical design experience and humanistic design experience. Feature attributes have a positive effect on the technical design experience and the humanistic design experience. Emotional attributes have an effect on the technical design experience and the humanistic design experience. The technical design experience has a positive effect on product-related customer value, service-related customer value, and promotion-related customer value. The humanistic design experience has a positive effect on product-related customer value, service-related customer value and promotion-related customer value. This study will make it possible to empirically examine how customer's experience in design innovation affects customer value. Our results will provide a theoretical foundation for examining a relationship between variables regarding how design innovation influences customer value through design experience. It is intended to give a direction as to the design innovation of firms by clarifying and presenting antecedent factors having an effect that design innovation produces on customer value. The results of the present study will inspire designers to design in consideration of design experience. Finally, our study will provide marketers with guidelines as to how design experience can influence customer satisfaction and loyalty.

A novel insect-infecting positive sense single-stranded RNA virus, Riptortus pedestris virus-1 (RiPV-1), was found in the Riptortus pedestris transcriptome data by de novo assembly and further confirmed by RACE method. The genome of RiPV-1 consists of 10,554 nucleotides (nt) excluding the poly(A) tail and contains a single large open reading frame (ORF) of 10,371 nt encoding a 3,456 aa polyprotein and flanked by 71 and 112 nt 5' and 3' noncoding regions, respectively. RiPV-1 genome contains the consensus genome organization of picorna-like RNA helicase, cysteine protease, and RNA-dependent RNA polymerase (RdRp) array in that order from the 5' to the 3' end. From the phylogenetic analysis, RiPV-1 was clustered with unassigned insect RNA viruses, APV and KFV, which suggests that these three insect picorna-like viruses might constitute a novel group of insect-infecting RNA viruses. Tissue tropism analysis revealed that RiPV-1 was relatively abundant in the thorax, abdomen, midgut and fat body. Interestingly, RiPV-1 replication was enhanced by Beauveria bassiana JEF-007 infection that was quantified using qRT-PCR. This study identified a novel insect-infecting virus and provided further insight into the relationship between virus, fungus and host.

Bean bug, Riptortus pedestris (Hemiptera: Alydidae) is an agriculturally serious pest in East Asian countries. Chemical pesticides have been contributed to the management of the pest, but nowadays insect resistance limits the use of chemical pesticides, thus alternatively new pesticides with different mode of actions such as entomopathogenic fungi are considered. Herein entomopathogenic Beauveria bassiana JEF isolates were collected, identified and assayed against bean bugs in laboratory conditions. Some isolates showed >80% virulence by contact-exposure and spray methods. The Agrobacterium tumefaciens-mediated transformation of B. bassiana JEF-007 generated random transformants and some mutants showed reduced virulence against Tenebrio molitor (Coleoptera: Tenebrionidae) larvae and R. pedestris nymph. Compared to the wild-type, the two transformants showed remarkably different morphology, conidial production, and thermotolerance. To figure out pathogenicity-related genes, thermal asymmetric interlaced (TAIL) PCR of the random transformants was performed and possibly some virulence-related genes were predicted. This work can be a strong platform for the functional genetics of bean bug-pathogenic B. bassiana.

Entomopathogenic fungi are facultative microorganisms, dwelling in soil or infecting host insects, and some of the genera have been used as biological control agents worldwide. Collection of fungal isolates should be a platform for the development of highly effective resources, thus in this work we constructed a fungal library using a mealworm (Tenebrio molitor) pathogenicity-based fungal collection method and further characterized some isolates with high virulence. A phylogenetic three was generated, and of the isolates 17 isolates’ biological features were characterized, such as morphology, spectrum of virulence, cultural characteristics, thermo-stability of fungi, production of biologically active materials, such as enzymes. This work reports an attractive entomopathogenic fungal library including the information of effective isolates in pest management.

Hypocreales entomopathogenic Beauveria bassiana and Metarhizium anisopliae are well-known biological control agents worldwide and have high potential in industrialization. However their thermo-susceptibility limits long-term storage under high temperature conditions and high insecticidal activity after application to target pests. Herein we isolated highly virulent isolates, B. bassiana JEF006 and JEF007 and M. anisopliae JEF003 and 004, and produced in three grains, as substrates for solid cultures, followed by thermotolerance assays to compare the potential of the three substrates. The JEF isolates were exposed to dry and wet heat at 50°C and overall conidia were more stable under dry heat condition rather than wet heat. Of the three grains, Italian millet was superior to the other grains in the production of thermotolerant conidia. Additionally Italian millet did not severely aggregated, which enabled air to penetrate into the substrate well compared to the sorghum and millet. JEF isolates were more thermotolerant when they were kept in oil conditions. The conidia of JEF 007 and 003 wild type and AtMT-based generating random mutants were subjected to SDS-PAGE. A significant relationship between conidial thermotoelrance and detected proteins was observed. This work suggests that Italian millet can be used as an effective substrate to produce more thermotolerant conidia, thus maintaining viability for long times under unfavorable environment and biological activity against target pests.

Bean bug, Riptortus pedestris is an agriculturally serious pest in north eastern Asian countries, damaging to several legumes and fruit trees. Chemical pesticides have been largely used to control the pest but it encounters insecticide-resistance and environmental toxicity issues. Alternatively different mode of action and environmentally sound pest management system can be found in entomopathogenic fungal insecticides. Herein we developed a platform to optimize the fungal production to express their maximum virulence against bean bug, by focusing on solid culture system for thermotolerance, formulation to select effective surfactants to carry the fungal conidia on the cuticles, and relationship between environmental abiotic factors and fungal mortality. First to produce highly thermotolerance fungal conidia, Beauveria bassiana and Metarhizium anisopliae isolates were cultured on several granular cereal substrates, which could be subjected to formulation process. Among the tested media, four media (millet, non-glutinous italian foxtail millet, glutinous italian foxtail millet, brown rice) were superior to the other grains in the spore production and thermotolerance. Next to efficiently deliver the fungal conidia on the cuticles of bean bug, total of six surfactant (CO-2.5, CO-12, LE-7, PE-61, TED-3 and Siloxane) was used to experiment. CO-12 was superior to the other surfactant in mortality of 100 ppm consistence. This work suggests that solid culture system and formulation and application should be seriously considered to reach an optimal level of mortality by inducing their maximum virulence.

Entomopathogenic fungi are facultative microorganisms, dwelling in soil or infecting host insects, and some of the genera have been used as biological control agents worldwide. Collection of fungal isolates should be a platform for the development of highly effective resources, thus in this work we constructed a fungal library using a mealworm pathogenecity-based fungal collection method and further characterized some isolates with high virulence. A phylogenetic three was generated, and of the isolates 17 isolates’ biological features were characterized, such as morphology, spectrum of virulence, cultural characteristics, thermo-stability of fungi, production of biologically active materials, such as enzymes. This work reports an attractive entomopathogenic fungal library including the information of effective isolates in pest management.

Bean bug, Riptortus pedestris is an agriculturally serious pest in East Asian countries, reducing the value of crop quality and loss of income in agribusiness. Chemical pesticides have contributed to the management of the pest, but nowadays insect resistance limits the use of chemical pesticides, thus alternatively new pesticides with different mode of actions such as entomopathogenic fungi are considered. Beauveria bassiana and Metarhizium anisopliae JEF isolates were collected, identified and assayed against bean bugs in laboratory conditions. Some isolates showed >80% virulence by spray and contact-exposure methods. Supernatant showed different level of enzyme activity including chitinase, Pr1 protease and lipase. The Agrobacterium tumefaciens-mediated transformation generated random transformants and some mutants had reduced virulence. TAIL-PCR of the random transformants revealed virulence-related genes. This work can be a strong platform for the functional genetics of bean bug-pathogenic B. bassiana.

Entomopathogenic Beauveria bassiana and Metarhizium anisopliae are well-known biological control agents worldwide and have high potential in industrialization. However their thermo-susceptibility limits long-term storage under high temperature conditions and high insecticidal activity after application to target pests. Herein we isolated highly virulent isolates, B. bassiana JEF006 and JEF007 and M. anisopliae JEF003 and JEF004, and produced in three grains, such as sorghum, millet and Italian millet as substrates for solid cultures, followed by thermotolerance assays to compare the potential of the three substrates for thermotolerance. The JEF isolates were exposed to dry and wet heat at 50°C and overall conidia were more stable under dry heat condition rather than wet heat. Of the three grains, Italian millet was superior to the other grains in the production of thermotolerant conidia. Additionally Italian millet did not severely aggregated, which enabled air to penetrate into the substrate well compared to the sorghum and millet. JEF isolates were more thermotolerant when they were kept in oil conditions as carriers of an oil-based formulation. This work suggests that Italian millet can be used as an effective substrate to produce more thermotolerant conidia, thus maintaining viability for long times under unfavorable environment and biological activity against target pests.

Bean bug, Riptortus pedestris is an agriculturally serious pest in East Asian countries, reducing the value of crop quality and loss of income in agribusiness. Chemical pesticides have contributed to the management of the pest, but nowadays insect resistance limits the use of chemical pesticides, thus alternatively new pesticides with different mode of actions such as entomopathogenic fungi are considered. Beauveria bassiana and Metarhizium anisopliae JEF isolates were collected, identified and assayed against bean bugs in laboratory conditions. Some isolates showed >80% virulence by spray and contact-exposure methods. The isolates produced high levels of pathogenesis-related enzymes, such as chitinase, Pr1 protease and lipase. The Agrobacterium tumefaciens-mediated transformation generated random transformants and some mutants had reduced virulence against bean bugs, which provided some materials to figure out pathogenicity-related genes in the fungi. Now characterization of flanking region of the integrated fragment is underway and this work may reveal some important genes in the pathogenesis. This work can be a strong platform for the functional genetics of bean bug-pathogenic B. bassiana.

The entomopathogenic fungus, Beauveria bassiana is widely used in integrated pest management (IPM), however its successful application is often limited by the little effort to explore its functions of unknown genes. In this work, egfp-expression cassette was randomly integrated into B. bassiana using Agrobacterium tumefaciensmediated transformation, and the general features of the mutants with unusual characteristics and the localization of the integrated genes were explored. To construct a transformation vector, egfp-expression cassette including gpdA promoter and trpC terminator was cut from pBARKS1-egfp using SacI and HindIII and integrated into pCAMBIA containing hygromycin B resistant hygR gene, designated as pCAMBIA-egfp. Transformed B. bassiana isolates were grown on quarter strength-Sabouraud dextrose agar containing 150 μg hygromycinB ml-1. Expression of egfp was investigated by RT-PCR and a fluorescent microscope (400×). Through the genome walking of the transformants using adaptor primers and gene specific primers, unique bands were detected on the egfp-expressing transformants, which were sequenced to figure out the flanking regions. This work provides a platform of methodology to figure out unknown functional genes of B .bassiana and possibly suggest an improved strategy to use the entomopathogen in IPM.