Honey production from approximately 1.7 million colonies owned by around 21 thousand Korean beekeepers was almost 36 thousand M/T in 2014. Honey farmers have mainly obtained commodities such as honey, beeswax, pollen and royal jelly. Honeybees gather the nectars and pollens from flowers of plants or trees. Bee pollen is rich in various nutrients and called as natural complete food. Bee pollens of Quercus acutissima, Actinidia arguta, Amorpha fruticosa, Oryza sativa, Chrysanthemum morifolium and Salix koreensis were mainly collected by Apis mellifera in Korea. But stiff pollen wall hinders dissolution of polysaccharides and lowers extraction efficiency. Thus, we measured antioxidant activity as DPPH radical scavenging assay system on six kinds of pollens inoculated with fungi to confirm the husk removal effect. The antioxidant activity was evaluated by measuring the free radical scavenging activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH). Briefly, extracts prepared from different extraction conditions were mixed with freshly prepared DPPH solution. After shaking, the reaction mixtures were stand for 30 min at room temperature in dark places. The radical scavenging activity was determined by measuring the absorbance at 517 nm. The relative radical scavenging activity (%) was calculated as [1-absorbance of solution with sample and DPPH / absorbance of solution with DPPH] x 100. The scavenger activities of Quercus acutissima, Actinidia arguta, Amorpha fruticosa, Oryza sativa, Chrysanthemum morifolium and Salix koreensis were 42.73±1.25, 8.98±1.46, 4.38±3.53, 36.09±0.74, 17.28±1.95 and 62.76±2.07 at 500㎕/㎖, respectively.

Recent enactments of the Nagoya Protocol and UPOV convention precipitated international competition to secure biological resources. To address these challenges in the case of mushroom industry, collecting and preserving the genetic resources are urgently needed. Mushroom Research Division, National Institute of Horticltural and Herbal Science, Rural Development Administration, has operated mushroom resource management facilities which consist of eight separate rooms with automatic temperature and humidity controllers for the safe preservation of mycelial cultures or voucher herbarium specimens. During the past three years the liquid-nitrogen (LN) cryogenic system for the permanent/semi-permanent preservation of the strains commercially or scientifically important are successfully installed, and the computerized resource management system using QR code are adopted. Up to now, 1,586 strains of 261 species in 79 genus of edible and medicinal mushrooms were preserved.

Bee pollen is rich in various nutrients and called as natural complete food. Bee pollens of acorn (Quercus acutissima) and actinidia (Actinidia arguta) are the most collected in Korea. But stiff pollen wall hinders dissolution of polysaccharides and lowers extraction efficiency. Thus, we measured antioxidant activity as DPPH radical scavenging and the total polyphenol content of pulverized and lyophilized actinidia pollens inoculated with fungi to confirm the husk removal effect. The total polyphenol content of actinidia pollen was highest in lyophilized pollen medium inoculated with Armillaria mellea, and was lowest in pollen inoculated with Lentinula edodes. Total polyphenol content of the lyophilized pollen was higher than that of the refined pollen and the pulverized pollen in actinidia pollen germinated with A. mellea. The total polyphenol content of the lyophilized actinidia pollen germinated with A. mellea was 1.4-fold higher than that extracted with water. Measurement of antioxidant activity using the DPPH (2, 2 diphenyl-1-picrylhydrazyl) free radical scavenging method showed that the lyophilized actinidia pollen germinated with A. mellea had the highest and that germinated with L. edodes was lowest in antioxidant activities. The lyophilized actinidia pollen germinated with A. mellea was 2 to 4 times higher than that extracted with water in the antioxidant activity of DPPH free radical scavenging. Many germinated cells were formed around pore of actinidia pollen inoculated with L. edodes, while those were formed at the end of hyphae derived from actinidia pollen inoculated with A. mellea.

Differentiation of Pleurotus eryngii is laborious and time-consuming tasks especially in mycelial status. For development of a method for differentiation of P. eryngii cultivars, simple sequence repeats (SSR) from whole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSR primer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotypes. PCR with the each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism information content (PIC) values of the five markers were in range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pair-group method with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivars into 2 clusters. Cluster I and II comprised of 4 and 8 cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 and SSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivar and strains with 21 allele with a PIC value of 0.9090. These results might be useful to provide an efficient method for the identification of P. eryngii cultivars with separate PCR reactions. (This work was supported by a grant from the Gold Seed Project [Supported by a grant from the IPET (213003-04-3-WTI11), MIFAFF, Republic of Korea.]

For research and development of Silicon Carbide (SiC) mirrors, the Korea Astronomy and Space Science Institute (KASI) and National Optical Astronomy Observatory (NOAO) have agreed to cooperate and share on polishing and measuring facilities, experience and human resources for two years (2014-2015). The main goals of the SiC mirror polishing are to achieve optical surface figures of less than 20 nm rms and optical surface roughness of less than 2 nm rms. In addition, Green Optics Co., Ltd (GO) has been interested in the SiC polishing and joined the partnership with KASI. KASI will be involved in the development of the SiC polishing and the optical surface measurement using three dierent kinds of SiC materials and manufacturing processes (POCOTM, CoorsTekTM and SSGTM corporations) provided by NOAO. GO will polish the SiC substrate within requirements. Additionally, the requirements of the optical surface imperfections are given as: less than 40 um scratch and 500 um dig. In this paper, we introduce the international collaboration and interim results for SiC mirror polishing and development.

Peptidoglycan recognition proteins (PGRPs) are family of innate immune molecules that recognize bacterial peptidoglycan. PGRP-LE, a member of the PGRP family, selectively binds to diaminopimelic acid (DAP)-type peptidoglycan to activate both the immune deficiency (IMD) and proPhenoloxidase (proPO) pathways in insects. A PGRP-LE-dependent induction of autophagy to control Listeria monocytogenes has also been reported. We identified and partially characterized a novel PGRP-LE homologue, from Tenebrio molitor and analyzed its functional role in the survival of the insect against infection by a DAP-type PGN containing intracellular pathogen, L. monocytogenes. The cDNA is comprised of an open reading frame (ORF) of 990 bp and encodes a polypeptide of 329 residues. TmPGRP-LE contains one PGRP domain, but lacks critical residues for amidase activity. Quantitative RT-PCR analysis showed a broad constitutive expression of the transcript at various stages of development spanning from larva to adult. RNAi mediated knockdown of the transcripts followed by a challenge with L. monocytogenes showed a significant reduction in survival rate of the larvae, suggesting a putative role of TmPGRP-LE in sensing and control of L. monocytogenes infections in T. molitor. These results implicate PGRP-LE as a defense protein necessary for survival of T. molitor against infection by L. monocytogenes.

CD63, a member of tetraspanin membrane protein family, plays pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic ‘Cys-Cys-Gly’ motif and ‘Cys188’ residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50-56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcript was upregulated the maximum 4.5 fold in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also made significant increase in the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.

Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146 Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5’-flanking region. BLAST and phylogenetic analysis reveals that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (<26%) with other apoLp-IIIs. Homology modeling of Tm apoLp-III shows a bundle of five amphipathic helices, including a short helix 3’. The ‘helix-short helix-helix’ motif was predicted to be implicated in lipid binding interactions, through reversible conformational changes and accommodating the hydrophobic residues to the exterior for stability. Highest level of TmapoLp-III mRNA was detected at late pupal stages, albeit it is expressed in the larval and adult stages at lower levels. The tissue specific expression of the transcripts showed significantly higher numbers in larval fat body and adult integument. In addition, TmapoLp-III mRNA was found to be highly up-regulated in late stages of L. monocytogenes or E. coli challenge. These results indicate that TmapoLp-III may play an important role in innate immune responses against bacterial pathogens in T. molitor.

We have identified novel ricin-type (R-type) lectin by sequencing of random clones from cDNA library of the coleopteran beetle, T.molitor. The cDNA sequence is comprised of 495 bp encoding a protein of 164 amino acid residues and shows 49% identity with galectin of Tribolium castaneum. Bioinformatics analysis shows that the amino acid residues from 35 to 162 belong to ricin-type β-trefoil structure. The transcript was significantly upregulated after early hours of injection with peptidoglycans derived from Gram (+) and Gram (-) bacteria, beta-1, 3 glucan from fungi and an intracellular pathogen, L. monocytogenes suggesting putative function in innate immunity.

14-3-3 proteins are known to play a pivotal role in a diverse array of cellular events such as cell survival, apoptosis, and signal transduction. Numerous 14-3-3 ζ have been cloned and characterized from a host of eukaryotic organisms including human, plants, yeast, fruit fly and silkworm. However, no study on Spodoptera exigua 14-3-3ζ in conjunction with virus infection has so far been reported in insects. It appears that expression of Se14-3-3ζ was decreased starting 24 h post-SeNPV infection as SeNPV titers seemed to increase as evidenced by intense bands of SeNPV IAP3. Interestingly, confocal microscopic analysis revealed that Se14-3-3ζ is expressed at the apical side of the NPV-uninfected gut cells, whereas it was detected mainly in the nucleus of the NPV-infected cells. Thus, despite the biological significance of Se14-3-3ζ in S. exigua in conjunction with molecular interactions between SeNPV and S. exigua is unclear now, our data suggest that Se14-3-3 ζ protein plays a role to protect S. exigua from the infection or inhibit replication of SeNPV.

Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway is known to play a pivotal role in various cellular events and antiviral responses in both vertebrates and insects. In an attempt to elucidate the potential involvement of STAT on S. exigua-SeNPV interactions, the full length cDNA of SeSTAT was cloned from S. exigua. Analysis of temporal expression patterns shows that SeSTAT is expressed in all stages of life cycle such as larvae, pupae, and adult. Spatial expression analysis shows that it is highly expressed in fat body and Malpighian tubule. Interestingly, SeSTAT is induced at 24 h in response to either laminarin or LTA injection in larvae. Electrophoretic mobility shift assay (EMSA) shows that the binding of nuclear extracts from fat body cells immune-challenged with LTA to STAT5 probe was observed. In addition, SeSTAT was nuclear-translocalized in both fat body and gut cells that were challenged with LTA and laminarin, respectively. Finally, gene silencing of SeSTAT shows that SeNPV number appears to be increased. It suggests that SeSTAT may act as a negative regulator against SeNPV in midgut.