Admirandus multicavus Belogurov and Belogurova, 1979, a new record of free-living marine nematode species belonging to family Oncholaimidae is reported based on the specimens collected from the intertidal sediments of Korea. Admirandus multicavus is characterized by the following unique characteristics: cephalic setae (1-2 μm long), spicules (71-126 μm long) and gubernaculum (15-31 μm long), midventral preanal supplementary organ, three glandular duct openings of the Demanian system, and stoma length measuring twice the width. A comparative analysis of the biogeographical and ecological characteristics of Admirandus species is presented. DIC (Differential interference contrast) photomicrographs of the species are also provided herein. This report represents the first taxonomic analysis of the genus Admirandus Belogurov and Belogurova, 1979 discovered from the Korean seawaters.

South Korea has over 0.38 million of managed honey bee (Apis cerana) colonies before 2009 years ago, which produce the highest quantity of honey in the Korea; however, almost colony (90%) were collapsed by Korean Sacbrood Virus (KSBV) in South Korea. Korean Sacbrood Virus (KSBV) is the pathogen of A. cerana Sacbrood disease, which poses a serious threat to honeybee A. cerana, and tends to cause bee colony and even the whole apiary collapse. Colony collapse of A. cerana was first reported on the Pyeong-Chang of the South Korea in 2009. Several scientists and governments has been tried research for cure the sacbrood disease in A. cerana colony by medicines and management techniques. Unfortunately, The sacbrood disease dosen’t improve. So, we were developed a better breed of A. cerana for resistance of sacbrood virus by selection and then artificial insemination. A. cerana breeding technique was first successful applied with A. cerana in Korean. Queens was grafted from sacbrood resistance line and then it was growing in sacbrood disease colony that was survived 100%. Altogether selected 18 queens were artificially inseminated and 2,000 drones of A. cerana in Korea was used to evaluate amount of semen collection. We are select two scabrood resistance A. cerana line (R and H). R line be used for rearing the Queen. Drone was reared in H line colony. The RH hybrid were not infected sacbrood virus even spread sacbrood virus (2×106). RH colonies have very excellent hygienic behavior, brood, and sacbrood disease resistance activity.

The objectives of the present study were to examine the antioxidant activity of fractions with different isoelectric points from salmon enzymatic hydrolysates and obtain peptide fractions of sufficient amounts with higher antioxidant peptide fraction, which could be applied to the food and animal model systems. The salmon enzymatic hydrolysates were fractionated on the basis of the amphoteric nature of sample peptides by preparative isoelectric focusing without toxic solvents and reagents, which is termed autofocusing. Acidic and basic fractions showed higher 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity than the other fractions. The basic fractions showed higher hydroxyl (OH) radical scavenging activity. The weak acidic and weak basic fractions showed higher oxygen radical absorbance capacity (ORAC) values than the acidic and basic fractions. The acidic fraction showed higher metal chelating activity than other fractions. The acidic fraction suppressed lipid oxidation in the cooked patties to a greater extent than other fractions. These results suggest that peptides fractions from salmon enzymatic hydrolysate are effective antioxidant, and that autofocusing could be useful to increase antioxidant activity for application to food and animal model systems.

In vitro culture of murine embryos is an important step for in vitro production systems including in vitro fertilization and generations of genetically engineered mice. M16 is widely used commercialized culture media for the murine embryos. Compared to other media such as potassium simplex optimization medium, commercial M16 (Sigma) media lacks of amino acid, glutamine and antibiotics. In the present study, we optimized M16 based embryo culture system using commercialized antibiotics-glutamine or amino acids supplements. In vivo derived murine zygote were M16 media were supplemented with commercial Penicillin-Streptomycin-Glutamine solution (PSG; Gibco) or MEM Non- Essential Amino Acids solution (NEAA; Gibco) as experimental design. Addition of PSG did not improved cleavage and blastocyst rates. On the other hand, cleavage rate is not different between control and NEAA treated group, however, blastocyst formation is significantly (P<0.05) improved in NEAA treated group. Developmental competence between PSG and NEAA treated groups were also compared. Between two groups, cleavage rate was similar. However, blastocyst formation rate is significantly improved in NEAA treated group. Taken together, beneficial effect of NEAA on murine embryos development was confirmed. Effect of antibiotics and glutamine addition to M16 media is still not clear in the study.

KO mice provide an excellent tool to determine roles of specific genes in biomedical filed. Traditionally, knockout mice were generated by homologous recombination in embryonic stem cells. Recently, engineered nucleases, such as zinc finger nuclease, transcription activator-like effector nuclease and clustered regularly interspaced short palindromic repeats (CRISPR), were used to produce knockout mice. This new technology is useful because of high efficiency and ability to generate biallelic mutation in founder mice. Until now, most of knockout mice produced using engineered nucleases were C57BL/6 strain. In the present study we used CRISPR-Cas9 system to generate knockout mice in FVB strain. We designed and synthesized single guide RNA (sgRNA) of CRISPR system for targeting gene, Abtb2. Mouse zygote were obtained from superovulated FVB female mice at 8-10 weeks of age. The sgRNA was injected into pronuclear of the mouse zygote with recombinant Cas9 protein. The microinjected zygotes were cultured for an additional day and only cleaved embryos were selected. The selected embryos were surgically transferred to oviduct of surrogate mother and offsprings were obtained. Genomic DNA were isolated from the offsprings and the target sequence was amplified using PCR. In T7E1 assay, 46.7% among the offsprings were founded as mutants. The PCR products were purified and sequences were analyzed. Most of the mutations were founded as deletion of few sequences at the target site, however, not identical among the each offspring. In conclusion, we found that CRISPR system is very efficient to generate knockout mice in FVB strain.

Lentinus lepideus is an edible mushroom, belongs to the family Tricholomaceteae and order Agaricales. The purpose of this study was to evaluate the antioxidant activities, tyrosinase inhibitory effects on the fruiting bodies of L. lepideus extracted with acetone, methanol and hot water. The antioxidant activities were performed on β-carotene-linoleic acid, reducing power, 1,1-diphenyl-2-picrylhydrazyl free radical scavenging, and ferrous chelating abilities. In addition to this, phenolic acid and flavonoids contents were also analyzed. Hot water extract of L. lepideus showed the strongest β-carotene-linoleic acid inhibition as compare to others extracts. At 8 mg/ml, methanolic extract showed a high reducing power of 1.21. The scavenging effects on 1,1-diphenyl-2-picrylhydrazyl radicals, acetonic and methanolic extracts were effective than hot water extract. The strongest chelating effect (87.50%) was obtained from the methanolic extract at 1.0 mg/ml concentration. Antioxidant activities of the extracts from the fruiting bodies of L. lepideus were increased with the increasing concentration. After application of reverse phase high performance liquid chromatography, coupled to a diode array detector and electrospray ionisation mass spectra, six phenolic compounds namely, chlorogenic acid, vanillin, naringin, naringenin, formononetin and biochanin were identified from acetonic extract. Tyrosinase inhibition of acetonic, methanolic, and hot water extracts of L. lepideus were increased with the increasing of concentration. Results revealed that acetonic and methanolic extract showed good, while hot water showed moderate activities of the tyrosinase inhibition at the concentration tested. This study suggests that fruiting bodies of L. lepideus can potentially be used as a readily accessible source of natural antioxidants.

Lentinus edodes is a popular edible mushroom in South-East Asia. This study was initiated to evaluate the antioxidant activities, tyrosinase inhibitory effects on the fruiting bodies of L. edodes extracted with acetone, methanol and hot water. The antioxidant activities were performed on β-carotene-linoleic acid, reducing power, 1,1-diphenyl-2-picrylhydrazyl free radical scavenging, and ferrous chelating abilities. In addition to this, phenolic acid and flavonoids contents were also analyzed. Methanolic extract of L. edodes showed the strongest β-carotene-linoleic acid inhibition as compare to others extracts. At 8 mg/ml, hot water extract showed a high reducing power of 0.96. The scavenging effects on 1,1-diphenyl-2-picrylhydrazyl radicals, acetonic extract was effective than other extracts. The strongest chelating effect (86.45%) was obtained from the acetonic extract at 1.0 mg/ml concentration. Antioxidant activities of the extracts from the fruiting bodies of L. edodes were increased with the increasing concentration. After application of reverse phase high performance liquid chromatography, coupled to a diode array detector and electrospray ionisation mass spectra, four phenolic compounds namely, naringenin, hesperetin, formononetin and biochanin were identified from acetonic extract. Tyrosinase inhibition of acetonic, methanolic, and hot water extracts of L. edodes were increased with the increasing of concentration. Results revealed that acetonic and methanolic extracts showed good, while hot water showed moderate activities of the tyrosinase inhibition at the concentration tested. This study suggests that fruiting bodies of L. edodes can potentially be used as a readily accessible source of natural antioxidants.

Slow seedling growth rate and nodulation failure of white clover (Trifolium repens L.) has been limited its good establishment to pastures. The experiment was done to determine the effect of removal of cotyledon and unifoliolate on the shoot, root growth,

Background : Cudrania tricuspidata Bureau is a widely used medicinal perennial woody plant. Obtaining information about the genetic diversity of plant populations is highly important for conservation and germplasm utilization. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific ecotypes of C. tricuspidata via amplification refractory mutation system (ARMS)-PCR analyses. We performed molecular authentication of twelve C. tricuspidata ecotypes from different regions using DNA sequences in the chloroplast TrnL-F intergenic region. Methods and Results : SNPs were identified based on the results of nucleotide sequence for the intergenic region of TrnL-TrnF gene (chloroplast). Molecular markers were designed for those SNPs with additional mutations on the second base from SNPs for amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). HRM pattern analyses were performed using the Mx3005P QPCR System (Agilent Technologies, CA, USA). Conclusion : We collected 12 individual lines of C. tricuspidata from various region in South Korea and China. Based on the nucleotide sequence in the trnL-trnF intergenic region of these lines, six SNPs and a deletion of 12 bps were identified and 12 individual lines were able to be grouped in one Korean ecotype and two different ecotypes of chinese lines, chinese line 1 and 2. The SNP markers developed in this study are useful for rapidly identifying these specific C. tricuspidata ecotypes collected from different regions.

Background : Schizandra chinensis Baillon have five tastes and lately it is using a beverage broadly. Schizandra chinensis is one of the top producing medicinal plant in Korea. Mungyeong of Gyongbuk province produce almost of Schizandra chinensis. Maturity of Schizandra chinensis get 3 years and proliferation of Schizandra chinensis was not a manual. It is needed that a new cultivar has a big fruit and high quality chracteristics using processed food and beverage. Methods and Results : 105 lines of Schizandra chinensis were collected on Mungyeong, Yeongwol, Jinan. It were studied it’ characteristics especially it’s fruit trait. Fruit traits of Schizandra chinensis were researched on fruit length, fruit weight, maturity, number of fruit, male and female ratio, powdery mildew. Fruit length of Schizandra chinensis is relation of fruit weight. It were founded 15 lines of long fruit length. 5 lines were studied high fruit weight and it’s weight were 32 to 41g. Number of fruit has relation with fruit weight and high fruit weight gets many fruits. it’s numer of fruits were 3 to 41. Male and female ratio were very impotant characteristic. High level of female ratio has quantity of fruit. High level of female ratio were founded 2 lines. Finally It was selected 3 good breed lines of Schizandra chinensis. Conclusion : 105 lines of Schizandra chinensis Baillons were collected on Mungyeong, Yeongwol, Jinan. It were founded 15 lines of long fruit length and 5 lines were studied high fruit weight. High level of female ratio were founded 2 lines. 3 good breed lines of Schizandra chinensis were selected.

Watermelon (Citrullus lanatus) is one of the most economically important cucurbitaceous crop over the world. Anthracnose disease caused by Colletotrichum orbiculare, result to severe damage to cucurbits worldwide. Anthracnose is a typical plant disease which significantly affecting the yield of cucurbit crop. Pathogeneis-related (PR) proteins are well-known plant defense proteins against pathogens. Therefore, we observed PR genes expression patterns when watermelon got anthracnose disease. We did RT-PCR experiment to evaluate differences of PR genes expression pattern among Au-Producer(R), one of the representative of resistance watermelon varieties against anthracnose, and 920533(S), one of the representative of susceptible watermelon varieties against anthracnose. As a result, there were differences of the expression of several PR genes between the R and S watermelon. Analysis of the function of these genes is expected to perform in the future.

Watermelon (Citrullus lanatus) is one of the most economically important cucurbitaceous crop over the world. Screening of proper reference genes was needed to reverse transcription quantitative real-time PCR (qRT-PCR), and it is bausic step of many researches including gene expression analysis. However, the reference genes on watermelon has not yet been reported systematically. Therefore, eight candidates of reference genes were selected with reference to Arabidopsis or cucumber papers. They are β-Actin, elongation factor 1-α, glyceraldehy-3-phosphate-dehydrogenase, NADP-isocitrate dehydrogenase, leunig, polypyrimidine tract-binding protein1, ubiquitin-conjugating eznyme E2, and 18S ribosomal RNA. The expression levels of genes were evaluated by qRT-PCR under biotic stress (Colletotrichum orbiculare treatment), plant hormone treatment (100 μM ABA), and abiotic stresses such as drought, cold (4℃), salt (250 mM NaCl) stresses. We founded appropriate reference genes which did not induce or reduce gene expression levels under broad spectrum of stresses by qRT-PCR analysis. These results may provide proper information for the use of appropriate reference genes for gene expression studies in watermelon qRT-PCR analysis.