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.

ocust, Locusta migratoria (Orthoptera: Acrididae) is one of the outbreaking pests worldwide and such big occurrence was recorded in 2014, Korea, however little consideration was given to the management strategy of the pest. Herein we established a indoor locust-rearing system and constructed a locust-pathogenic fungal library to further facilitate the resources to be used as possible biological control agents. A locust colony was provided from the National Institute of Agricultural Science and Technology and reared in corn or barley plants at artificially manipulated rooms. The critical developmental stages, such as oviposition, hatching and mating were successfully proceeded. Entomopathogenic fungal granules were treated to the locust (2 g/rearing box), and in 5~7 days mycosis was observed in the membranous cuticles of head, abdomen and legs. In particular JEF-003 (Metarhizium anisopliae), JEF-186 (M. lepidiotae) and JEF-187 (Clonostachys rogersoniana) showed high virulence against the locust. A population of locust was exposed to the entomopathogenic fungal conidia-incorporated soil to investigate the possibility of the fungal isolation from natural soil, which resulted in the pathogenesis in 7~10 days in laboratory conditions. More than 80% of control efficacy was observed in the greenhouse trial of fungal granular application. This work suggests that locust rearing system was successfully established and entomopathogenic fungi can be used to control the migratory locust.

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 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. 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 effects of seed soaking treatment with the solutions of plant growth regulators IAA, GA3 and BAP on seed germination and shoot and bulb growth of Allium victorialis var. platyphyllum (Korean wild garlic) were determined. A significant variation in the seed germination rate was recorded at all treatments for various soaking periods. Maximum seed germination was obtained when seeds were soaked in IAA or GA3 solution at 200 mg L-1. The MAP treated seeds started to germinate after 3 months. Among treatments, IAA was found to be most effective in improving seed germination, but further seedling growth was not correlated to the soaking time. Seed soaking in IAA or GA3 solution enhanced further growth of seedlings compared with water control treatment. Shoot and bulb growth was highest in GA3 treatments.

In this paper, OTT (Over-The-Top) refers to voice, video and data services that arrive from a third party service provider (SP) and doesn’t require any business or technology affiliations with the network operator (Southwell 2011). Traditional telephone companies (telcos) are under big threat from OTT messaging applications which have been splitting flow of text message and voice from the former. Even though, mobile operators still enjoy some asymmetric advantages by controlling the basic telecommunications infrastructure, monitoring the network usage and distinguishing between different OTT actors (Bertin et al. 2011). From a market structure perspective, mobile telecoms industry with the entrance of OTT messaging applications is analogous to the traditional dual-channel structure. On one hand, mobile operators “sell” communication services such as SMS and voice directly to subscribers; on the other hand, they “wholesale” the network infrastructure to OTT SPs on which OTT applications distribute text, picture, video and voice for end users. Extant research has extensively examined the channel selection, channel competition and coordination in traditional sectors (Cai 2010; Cattani et al. 2006; Chiang et al. 2003; Dumrongsir et al. 2008). However, the explanation of dual channel might not be well applicable to the mobile operator and OTT issue considering some new features in the industry. 1) comparing with traditional product distribution, the mobile operators are selling “the right to use the network”, thus they could both charge the “access” and “usage” fee. 2) unlike the traditional retailing channel where consumers make payments to retailer, consumers using OTT would pay network usage fee (data fee) to the mobile operator. 3) traditional retailors earn their profits by selling products with a higher price than the wholesale price. However, OTT services aim to develop their own business after accumulation of sufficient quantities of users, such as revenues form ads, traffic guidance and so on. This paper is conducted to analyze the pricing strategy of mobile operators facing the challenge of OTT. For simplicity, we assume a monopoly market, namely one mobile operator provide network infrastructure for one OTT messaging service provider who provides communication service for consumers. Based on dynamic game theory, we show that: firstly, under non-cooperative strategy, a mobile operator should charge OTT a positive network access fee which is positively correlated to OTT platform’s future commercial value and direct communication service price, and negatively correlated to indirect communication service; secondly, under cooperative strategy, a mobile operator and OTT could create a more joint profit than that under non-cooperative strategy. The platform access fee that the joint venture charges end users is negatively correlated with OTT platform future commercial value; thirdly, despite the choice of cooperative or non-cooperative strategy, the price of direct communication products has a negative correlation with OTT platform future commercial value and a positive correlation with the platform quality; while the price of indirect communication products is positively correlated with platform future commercial value and is negatively correlated with OTT platform quality. Finally, we conclude with a discussion of the managerial implications for mobile operators and OTT SPs. With regard to policy makers, we suggest that a convenient cooperation environment should be provided, because both mobile operator and OTT SP would obtain a high profit under cooperative condition than that under non-cooperative condition and at the same time consumers could also enjoy a better welfare. This study was supported by the National Natural Science Foundation of China (71172011 and 71272160) and NCET-12-0772.

An understanding of oocyte gene expression is a necessary for the study of early female gamete development. Recently, oocyte has been used in many techniques such as somatic cell nuclear transfer, intracytoplasmic sperm injection and embryonic stem cell derivation. The purpose of this study was to investigate in the proteomes of pig oocytes and identification of differential proteins between using DIGE technique. In this experiment to overcome of limitation of 2D gel method like a low reproducibility and low sensitivity for proteome analysis of very small quantities, 2D fluorescence difference gel electrophoresis (DIGE), which enables co-detection of up to three samples on the same 2DE gels with CyDyes was used for analysis of oocyte proteins. Proteins within an isoelectric point (pI) range of 3 to 10 and a molecular weight (Mw) range of 20～100 kDa were primarily analyzed in DIGE with 2 replications of each sample. Approximately 1000 spots were detected in 2-D gel. Then, image analysis of DeCyder was performed to detect variations in protein spots between mature oocyte and parthenogenesis embryo. In the comparison of mature oocyte and parthenogenesis embryo, 11 spots were identified to be up-regulated proteins and 2 spots to be down-regulated proteins in parthenogenesis embryo, among which proteins were zona pellucida glycoprotein 4, transferrin receptor, apolipoprotein B, L-3-Hydroxyacyl Coa Dehydrogenase Revisited, cytochrome P450 2C33, similar to Monocarboxylate transporter 2, 2'-5' oligoadenylate synthetase 3, interferon alpha/ beta receptor-1, Chloride channel protein 6, pyruvate carboxylase as well as2'-5' oligoadenylate synthetase 3 using MALDI-TOF-MS. These results suggested that differential proteins were present between mature oocyte and parthenogenesis embryo.

An understanding of oocyte gene expression is a necessary for the study of biological development. Recently, Oocyte has been used in many techniques such as somatic cell nuclear transfer (SCNT), intracytoplasmic sperm injection (ICSI) and embryonic stem cell derivation. However, the molecular mechanism underlying porcine oocyte is still unclear. In this study, we present the description of the porcine oocyte proteome. Proteins within the isoelectric point ranges of 3.0 to 10.0 were analyzed separately using 2‐dimensional electrophoresis (2‐DE). About 450 spots were detected in 2‐ D gel of oocytes, stained with Coomassie blue. Subsequent excision of 227 spots from gels and MALDI‐TOF MS analysis allowed the identification of 85 proteins. Our results indicated the composite profiles of proteins in the porcine oocyte. Tubulin beta chain and meiosis‐specific nuclear structural protein 1 antibody was used to confirm those antibody expression levels in immature, mature and parthenogenetic embryo. Western blot analysis showed that expressions of those proteins increased during mature and parthenogenetic embryo. These protein profiles will make available important guides for the study of oocyte function and assist in functional analysis of the proteins.

To examine the differential expression of proteins during the cycling (70～80% confluences) and G0/G1 (full confluences) phases in porcine fetal fibroblast cells, we used a global proteomics approach by 2‐D gel electrophoresis (2‐DE) and MALDI‐TOF‐MS. Cycling cell were harvested at approximately 70% to 80% confluent state while cells in G0/G1 phase were recovered after maintenance of a confluent state for 48 hr. Cellular proteins with isoelectric points ranging between 3.0～10.0, were analyzed by 2‐DE with 2 replicates of each sample. A total of approximately 700 spots were detected by 2‐D gels stained with Coomassie brilliant blue. On comparing the cell samples obtained from the cycling and G0/G1 phases, a total of 13 spots were identified as differentially expressed proteins, of which 8 spots were up‐regulated in the cycling cell and 5 were up‐regulated in the G0/G1 phase. Differentially expressed proteins included K3 keratin, similar to serine protease 23 precursor, protein disulfide‐isomerase A3, microsomal protease ER‐60, alpha‐actinin‐2, and heat‐shock protein 90 beta. The identified proteins were grouped on the basis of their basic functions such as molecular binding, catabolic, cell growth, and transcription regulatory proteins. Our results show expression profiles of key proteins in porcine fetal fibroblast cells during different cell cycle status.

To identify DNA markers linked to a elytra polymorphism, amplified fragment length polymorphism (AFLP) analysis was performed on DNA samples from four each colour pattern individuals (2 females and males), for example, succinea 1, succinea 2, conspicua, and spectabilis. As a result of performing AFLP analysis with the restriction endonuclease combination EcoRⅠ and Mse I, total of 2,269 AFLP fragments which were specific to succinea, conspicua and spectabilis was identified using 24 different AFLP primer combinations. Among these 2,269 fragments, 16 bands which were the most specific to one color patterns were isolated, cloned and sequenced. Subsequent UPGMA cluster analysis revealed that population of H. axyridis was divided four major group and these genetic tree showed that H. axyridis elytra colour diversity was affected by genetic polymorphism. It is considered that these genetic analyses may be facilitated the understanding of molecular genetic mechanism related with the wing colour pattern formation in this species.

The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield‐the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy‐specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non‐pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non‐pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH 3.0～10.0 strip, by loading a 2‐mg milk protein sample. After the second‐dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non‐pregnant and pregnant cattle milk protein spots, using ImageMaster; this was followed by analysis with MALDI TOF‐MS. Analysis of the 2‐DE gel image resulted in a total of approximately 500～600 protein spots, of which 12 spots were differentially expressed, six spots were up‐regulated, and four spots were downregulated; two spots were identified as pregnancy‐specific proteins. These proteins were identified as lactoferrin, NADH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2‐D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

As indigenous aphid parasitoid, Aphelinus varipes kill aphids for feeding in addition to parasitization. Because of this characteristic of A. varipes, this parasitoid may have the possibility of biological control agent against aphids. So we have evaluated traits such as daily paratization, total parasization, number of aphids killed by host feeding, sex ratio, development time, pupal mortality of A. varipes parasitizing green peach aphid, Myzus persicae. At 25°C and 16L:8D, longevity, total paratization and host feeding of A. varipes female was 11.0, 25.3, and 63.3 days, respectively. And development time of male and female, sex ratio (M:F), pupa mortality of offspring of A. varipes were 12.0 days, 12.5 days, 0.88, and 11.6%, respectively. However, because these results are not enough to estimate potential of A. varipes as biological control agents/factors, other factors such as host suitability (Macrosiphum euphorbiae, Aulacorthum solani), effect of temperature, and host seeking behavior of A. varipes continually will be investigated.

As an effective generalist predator of aphids and other hemipteran pests, Harmonia axyridis has been a successful biological control agent. Interestingly, it was known that there were varied in color patterns on H. axyridis elytra. In fact, Seo & Youn (2007) reported that H. axyridis had five color patterns, for example, succinea 1, 2, conspicua, spectabilis, and axyridis. But there are uncertain that H. axyridis elytra colour patterns are regulated by genetic polymorphism. So we tried to what is the reason that color patterns are greatly variable. To identify DNA markers linked to a elytra polymorphism, amplified fragment length polymorphism (AFLP) analysis was performed on DNA samples from four female succinea, conspicua, spectabilis and Coccinella septempunctata which is another species in Coccinellidae. AFLP analysis with the restriction endonuclease combination EcoRⅠ and MseⅠwas performed. Using 12 AFLP primer pairs, nine AFLP fragments which is specific between succinea, conspicua, spectabilis was identified. These nine AFLP fragments were isolated, cloned and sequenced. Subsequent UPGMA cluster analysis revealed three major group of H. axyridis populations. These genetic tree showed that H. axyridis elytra colour diversity was affected by genetic polymorphism. For more genetically understanding elytra colour genes, different primer combinations may be need to be generate enough polymorphic markers. These genetic analyses may be facilitate the understanding of molecular mechanism behind wing colour pattern formation.