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.

We have used bulked segregant analysis to screen the strain-specific DNA marker associated thermophilic strain of Pleurotus eryngii. Bulked genomic DNAs of Pleurotus eryngii were amplified by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S primers to screen the strain-specific DNA marker. A unique DNA fragment of 550 bp was amplified with OP-S07 primer from the thermophilic strain and sequenced. A sequence characterized amplified region (SCAR) marker was designed on the basis of the determined sequence and named as OP-S07-1. The PCR analysis with the OP-S07-1 primer showed that this SCAR marker clearly distinguish the thermophilic strains from the control strains.

Insect-derived Kazal-type serine protease inhibitors exhibit thrombin, elastase, plasmin, proteinase K, or subtilisin A inhibition activity, but so far, no functional roles for bee-derived Kazal-type serine protease inhibitors have been identified. In this study, a bee (Apis cerana) venom Kazal-type serine protease inhibitor (AcKTSPI) that acts as a microbial serine protease inhibitor was identified. AcKTSPI contained a single Kazal domain that displayed six conserved cysteine residues and a P1 threonine residue. AcKTSPI was expressed in the venom gland and was present as a 10-kDa peptide in bee venom. Recombinant AcKTSPI Kazal domain (AcKTSPI-Kd) expressed in baculovirus-infected insect cells demonstrated inhibitory activity against subtilisin A (Ki 67.03 nM) and proteinase K (Ki 91.53 nM), but not against α-chymotrypsin or typsin, which implies a role for AcKTSPI as a microbial serine protease inhibitor. However, AcKTSPI-Kd exhibited no detectable inhibitory effects on factor Xa, thrombin, tissue plasminogen activator, or elastase. Additionally, AcKTSPI-Kd bound directly to Bacillus subtilis, B. thuringiensis, Beauveria bassiana, and Fusarium graminearum but not to Escherichia coli. Consistent with these findings, AcKTSPI-Kd showed antibacterial activity against Gram-positive bacteria and antifungal activity against both plant-pathogenic and entomopathogenic fungi. These findings constitute molecular evidence that AcKTSPI acts as an inhibitor of microbial serine proteases. This paper provides a novel view of the antimicrobial functions of a bee venom Kazal-type serine protease inhibitor.

The pear psylla, Cacopsylla pyricola Foerster (Homoptera: Psyllidae), is a serious insect pest of commercial pear crops. The species, which resides on pear trees throughout its life cycle, is rapidly spreading in some regions of the world. Given the life cycle, it is unclear how such a rapid spread has been facilitated. Presently, the population genetic structure of the species including genetic diversity and gene flow was studied to understand the nature of dispersal and field ecology of the species. Pear psylla was collected from several pear orchards in Korea. The 658-bp region of mitochondrial COI gene and the 716-bp long complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA were sequenced. Unlikely other previously studied insect pests, the COI-based genetic diversity of the pear psylla was extremely low (maximum sequence divergence of 0.15%). This finding allowed us to conclude that the species may have been introduced in Korea relatively recently, possibly with the phenomenon of genetic bottlenecks. ITS2 sequence-based analyses of phylogeny, population differentiation, gene flow, and hierarchical population structure all concordantly suggested that the pear psylla populations in Korea are neither genetically isolated nor hampered for gene flow. These genetic data are concordant with the dispersal of an overwintering winterform morph outside the non-pear habitat in the fall and the possibility of subsequently longer distant dispersal.

To control pest of Brassicaceae leafy vegetable(leaf broccoli, Red Mustard Leaf, Tatsoi) which grows at vinyl house in IKSAN, Jeollabuk-do, the non-treatment was set as negative control, and treatment was divided into Parasitic natural treatment group and general treatment group. And incidence density of pest was surveyed and control effect was analyzed. As a result, in case of spring plants, the leaf damage ratio was decreased by 31% in Leaf broccoli, 30% in Red mustard leaf and 27% in Tatsoi compared to untreatment. In case of autumn plants, it was decreased by 32%, 41% and 17% respectively. The key pests were Plutella xylostella, Pieris rapae, Myus persicae Sulzer, Thrips palmi and Striped cabbage flea-beetle. Compared with the untreatment, the incidence density of each was significantly controlled. Other pests include Spodoptera exigua, Macdunnoughia purissima, Macdunnoughia purissima which showed high incidental density sometimes. In case of spring plants, the number by treatment was increased by 117% in Leaf broccoli, 85% in Red mustard leaf and 1,000% in Tatsoi. In autumn plants, it was increased by 132%, 257% and 1,077% respectively. The used Parasitic natural and eco-friendly materials were Cotesia glomerata, lacewing, ladyburg, Orius laevigatus, Encarsia formosa, Entomopathogenic nematode, Chungjigi and Togkaki. During early development of each pest, the Parasitic naturals were grazed 2~3 times at the interval of 7~10days. During the peak time, eco-friendly materials were sprayed 1~2 times. Based on this, comprehensive management model was drafted by period for each Brassicaceae vegetables pest.

To control Thrips tabaci in Korean leek and green onion which grow at vinyl house in IKSAN, Jeollabuk-do, the non-treatment was set as negative control. In general treatment group, 120 mesh gauze was installed on the side window after planting(May, 2nd) and, in spring, Orius strigicollis (1.0/m2) was grazed 3 times every 7 days from mid of May which is early development stage and In the mid of June, which is the peak stage, plant extracts were sprayed one time. In autumn, O. strigicollis was grazed 3 times every 7 days from mid of September which is early development stage, and in the mid of October, which is the peak stage, plant extracts were sprayed one time. The result shows that the leaf damage ratio was decreased by 22% in Korean leek and by 27% in green onion compared to the untreatment. And the control value of Thrips tabaci shows 78.7% in korean leak and 90.6% in green onion. The density control effect of Thrips tabaci was significantly controlled under max 6.2/plant in general treatment group compared max 25/plant in the untreatment and this result was similar in green onion. The yields by general treatment was increased by 85% in Korean leek and 56% in green onion, compared with non-treatment yields, which was 900kg/10a in Korean leek and 1,287kg/10a in green onion.

This study was conducted for establishment of Economic injury levels(EILs) of the tea red spider mite, Tetranychus kanzawai on Rubus coreanus. T. kanzawai was innoculated on May 7 in differently 0, 5, 10, 20 and 40 adults per plant. And pesticide was treated in late April, early May, middle May and late May, respectively. After inoculated of T. kanzawai, the density was increased until the mid-June and decreased gradually in all plots. And higher inoculation density were increased higher than lower inoculation density. Growth variables were not different among experimental plots except number of fruit set, but the number of fruit set and yields were decreased with increasing initial mite density. Densities of T. kanzawai on treated time of pesticide was lower in later treatment time than early treatment time. The late April treatments were not effective of pesticide in harvesting season. The rates of yield loss increased with increasing initial mite density. The relationship between initial T. kanzawai densities and yield losses was well described by a linear regression, Y = 0.6545X + 3.0425, R2=0.9313. Based on the relationship, the number of adults per plant which can cause 5% loss of yield was estimated to be approximately 3.0. And EILs was estimated to be approximately 8.3 adults/leaf in mid-May.

We analyzed a portion of mitochondrial COI gene sequences (658 bp) to investigate the genetic diversity and geographic variation of the swallowtail butterfly, Papilioxuthus L., and the cabbage butterfly, Pieris rapae (Lepidoptera: Papilionidae). P. xuthus showed a moderate level of sequence divergence (0.91% at maximum) in 15 haplotypes, whereas P. rapae showed a moderate to high level of sequence divergence (1.67% at maximum) in 30 haplotypes, compared with other relevant studies. Analyses of population genetic structure showed that most populations are not genetically differentiated in both species. The distribution pattern of both species appears to be consistent with category IV of the phylogeographic pattern sensu Avise (Avise et al. 1987): a phylogenetic continuity, an absence of regional isolation of mtDNA clones, and extensive distribution of close clones. The observed pattern of genetic diversity and geographic variation of the two butterfly species seems to reflect the abundant habitats, abundant host plants, and flying abilities in connection with the lack of historical biogeographic barriers.

The 15,338-bp long complete mitochondrial genome (mitogenome) of the Japanese oak silkmoth, Antheraeayamamai (Lepidoptera: Saturniidae) was determined. This genome has a gene arrangement identical to those of all other sequenced lepidopteran insects, but differs from the most common type, as the result of the movement of tRNAMet to a position 5’-upstream of tRNAIle. No typical start codon of the A. yamamai COI gene is available. Instead, a tetranucleotide, TTAG, which is found at the beginning context of all sequenced lepidopteran insects was tentatively designated as the start codon for A. yamamai COI gene. Three of the 13 protein-coding genes (PCGs) harbor the incomplete termination codon, T or TA. All tRNAs formed stable stem-and-loop structures, with the exception of tRNASer(AGN), the DHU arm of which formed a simple loop as has been observed in many other metazoan mt tRNASer(AGN). The 334-bp long A+T-rich region is noteworthy in that it harbors tRNA-likestructures, as has also been seen in the A+T-rich regions of other insect mitogenomes. Phylogenetic analyses of the available species of Bombycoidea, Pyraloidea, and Tortricidea bolstered the current morphology-based hypothesis that Bombycoidea and Pyraloidea are monophyletic (Obtectomera). As has been previously suggested, Bombycidae (Bombyxmori and B.mandarina) and Saturniidae (A.yamamai and Caligula boisduvalii) formed a reciprocal monophyletic group.

We determined the complete mitochondrial genome (mitogenome) of the jewel beetle, Chrysochroa fulgidissima (Coleoptera: Buprestidae) from two overlapping fragments and subsequent sub fragments. The 15,592-bp long C. fulgidissima mitogenome contains gene arrangement and content identical to the most common arrangement found in insects. Most individual C. fulgidissima mitochondrial (mt) genes were well within the range found in the respective genes of other insects. The 875-bp A+T-rich region is shortest among the coleopteran mitogenomes sequenced in their entirety. The region is interesting in that it contains several stem-and-loop structures and tRNA-like structure found in the A+T-rich regions of other insect mitogenomes. As seen in other insect motogenomes the start codon of C. fulgidissima COI gene also is unusual because no typical start codon is available. Three of the 13 protein-coding genes have incomplete termination codon T or TA. All tRNA formed stable stem-and-loop structure, except for tRNASer(AGN), the DHU arm of which formed a simple loop as seen in many other metazoan mt tRNASer(AGN).

본 연구는 국내 특산식물이며 신 관상식물로 기대되는 산꼬리풀[Veronica rotunda var. subintegra (Nakai) T.Yamaz.]의 효과적인 재배법을 구명하고자 수행되었다. 플러그 육묘는 파종용기와 파종량을 달리하였다. 재배는 묘의 소질, 식재용기, 토양종류 및 차광정도를 달리하여 처리하였다. 묘의 소질은 162, 200 및 288구 트레이를 이용하여 육묘한 묘를 사용였으며, 파종량은 1, 2, 4 및 6립을 파종하여 생산된 묘를 사용하였다. 본 연구에서 산꼬리풀의 육묘 시 트레이 종류는 162구가 적절하였으며, 셀 당 1립씩 파종할 경우 각 개체의 생육이 증가하였으나 4립 파종 시에는 전체 식물의 생육에 유리하였다. 묘의 소질에 따른 실험은 162구 트레이에서 생산된 묘와 4립씩 파종하여 생산된 묘에서 각기 우수한 생육을 보였다. 산꼬리풀 재배 시 식재용기의 용적량이 커질수록 지상부 및 지하부의 생육이 증가 하는 경향이었으며, 용기의 재질에 따른 유의적인 차이는 나타나지 않았다. 토양조건별 생육은 원예상토에서 가장 왕성하였으며, 혼용토에서는 피트모스에 비해 마사토의 함량이 높은 조건에서 양호한 생육을 나타냈다.

Present study aimed to determine the effect of ‘bitter melon’, a popularly used fruit in Bangladesh and several other Asian countries, on high-fat-diet-induced type 2 diabetes. To investigate the effect, ethanol extract from bitter melon (BME) as a dietary supplement with mouse chow was used. BME was found to significantly attenuate the high-fat diet (HFD) -induced body weight and total fat mass. BME also effectively reduced the insulin resistance induced by the HFD. Furthermore, dietary supplementation of BME was highly effective in increasing insulin sensitivity and reducing hepatic fat and obesity. These results indicate that BME could be effective in attenuating type 2 diabetes and could therefore be a preventive measure against type 2 diabetes.

Background: Mahonia Nepalensis DC. (Hoang lien o ro), the specie of the family Berberidaceae, is widely distributed in the high mountainous areas at altitudes 1700 – 1900 m of Vietnam. It is found that the stem of Mahonia nepalensis indicated anti-inflammatory, anti-bacterial and antifungal activities and they are used particularly for the treatment of eczema, psoriasis, and other skin conditions. However, no study on the antioxidant and anti-cancer activities of Mahonia Nepalensis stem has been previously reported. The aim of this study was to evaluate anti-oxidant and anti-cancer activities of Mahonia Nepalensis stem. Methods and Results: The stem pieces of Mahonia Nepalensis were dried and extracted three times with 100% methanol. After that, the extract was suspended in distilled water and then partitioned with n-hexane, ethyl-acetate (EtOAc) and butanol (water saturated BuOH) fractions were then evaporated using a vacuum rotary evaporator. Evaluation of the anti-oxidative activity of Mahonia Nepalensis was carried out using a DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-producing system. The results revealed that the ethyl acetate fraction of M. nepalensis possessed higher potential DPPH radical scavenging activity (IC50, 81.88 ± 1.33㎍/㎖) than other fractions as well as BHT (2,6-Di-tert-Butyl-4-methylphenol) (IC50, 250.49 ± 1.60㎍/㎖). The reducing power assay was also investigated and EtOAc fraction showed higher absorbance values than other fractions. At 1.0 mg/ml concentration, EtOAc fraction showed absorbance of 1.72, be higher than Ascorbic acid. Cell viability was evaluated according to the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyl tetrazolium Bromide) assay. By MTT assay, all fractions showed a significant reduction in cell viability on COLO 205 (Human colon carcinoma cell) at the highest concentration tested (200㎍/ ㎖) with over 70% decrease in cell viability was obtained, and the highest significantly inhibiting effect occurred in butanol fraction with approximately 90% reduction in cell viability. Conclusion: We demonstrated that Mahonia Nepalensis stem extract has highly potential in anti-cancer activity. Further studies are necessary in order to explore the variety of Mahonia Nepalensis stem to be applied as a valuable natural material.

Background : Sorghum is a major cereal food crop used in many parts of the world. It has been grown on a subsistence level by farmers, under various conditions of environmental stresses in the semi-arid tropics of Africa and Asia. This crop has received significant attention because of its ability to lower cholesterol in the blood, and its anti-dementia, antioxidant, and antimicrobial properties. It is possible to develop a functional and commercially viable sorghum variety by using superior cultivars of sorghum. The objective of this study was to construct a database of superior sorghum accession. Methods and Results : We used 250 sorghum accessions collected from different geographical bioregion in Korea. We determined various growth characters including germination rate and ear length of these accessions. To determine the antioxidant capacity, we measured the DPPH radical scavenging activity, ABTS scavenging activity, total phenolic contents, and total flavonoid contents. 189 accession showed higher germination (> 80%) than other accessions. Higher DPPH radical scavenging activity was observed in 11-SB-078 (RC50: 1.89 ± 2.88 ㎍/ ㎖), and higher ABTS scavenging activity was recorded in 11-SB-116 (RC50: 35.48 ± 2.42 ㎍/㎖) than other accessions. The ear length ranged from 15 ㎝ to 48 ㎝, the total phenolic contents ranged from 3 ㎎·GAE/g to 77 ㎎·GAE/g, and total flavonoid contents ranged from 0.09 ㎎·QE/g to 1.07 ㎎·QE/g. Conclusion : Among 250 sorghum accessions, we selected 10 with both superior agronomic characters and higher functional food quality.

Background : Haskap berries commonly refer to fruits of Lonicera caerulea L., recognized by the Japanese aborigines as the “The elixir of life.”. Due to their recent arrival on the North American market, haskap berries have not yet been positioned among other berries and compared in terms of their phytochemical content. And haskap berries have higher ascorbic acid and anthocyanin content than other berries known for their health-promoting benefits, such as blueberries. However, no study has reported on the antioxidant and anti-cancer activity of Lonicera caerulea stem. The purpose of this study is to present the current research on the chemical content, antioxidant and anti-cancer activities of Lonicera caerulea stem. Methods and Results : The stem of Lonicera caerulea L. ware dried in the shade at room temperature and extracted with 100% methanol. The extract was suspended in deionized water and partitioned sequentially with n-hexane, chloroform, ethyl-acetate and butanol (water saturated BuOH) fractions. Antioxidant activities were measured by determination of antioxidants, DPPH (2,2-diphenyl-1-picrylhydrazyl). Cell viability was determined by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. All cell lines were purchased from the Korean Cell Line Bank (Seoul, Korea). All results were performed with three replications were processed statistically. By DPPH assay, the Lonicera caerulea L. the highest activity was obtained from the ethyl-acetate fraction (IC50=15.46 ㎍/㎖). By MTT assay, the chloroform fraction showed a significant growth inhibiting effect on MCF-7 (Human breast cancer, IC50=225.91 ㎍/㎖), COLO 205 (Human colon cancer, IC50=179.55 ㎍/㎖), but on AGS (Human stomach cancer) and other fractions it did not show effect. Conclusion : We demonstrated that Lonicera caerulea L. stem extract and fractions has antioxidant and antiproliferation activity in vitro. Further studies should identify the active constituents in Lonicera caerulea L stem to evaluate the potential in vitro antioxidant and antiproliferation activities of the extract.

Background : Astilboides tabularis (Hemsl.) Engl. is a perennial herbaceous plant, distributed in the northern high mountains of the Korean peninsula and China. It is an excellent ornamental plant currently at risk of overharvesting and therefore, is designated as an endangered wild plant Class II by the Ministry of Environment. Physiological research on A. tabularis has not be reported. Therefore, in this study, using A. tabulari extracts, antioxidant and Anti-inflammatory effects were determined. Methods and Results : The antioxidant and free radical scavenging activities of A. tabularis extracts were evaluated using DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. The results showed that the ethyl acetate fraction of A. tabularis possesses potent DPPH radical scavenging activity (2.90±0.08㎍/㎖), similar to the scavenging activity of ascorbic acid (2.19±0.06㎍/㎖), and better than the powerful antioxidant α-tocopherol (10.60±0.40㎍/㎖) as well as BHA (butylatedhydroxy anisole)(6.12±0.27㎍/㎖). The ethyl acetate fraction possessed a significantly higher concentration of total phenolic (549.70±2.72㎎GAE/g) and flavonolic content (154.58±1.04㎎QE/g). It was also found that the ethyl acetate fraction exhibited high reducing power and inhibition of ROS (Reactive Oxygen Species) formation. Different fractions of A. tabularis were tested for anti-inflammatory activity using LPS stimulated Raw 264.7 cells. The n-hexane and ethyl acetate fractions exhibited a high inhibitory effect on NO (Nitrite oxide) production (22.43±1.06%, 19.30±0.45%, respectively) at 200㎍/㎖ concentration. The mRNA of IL-1β, iNOS and COX-2 gene expression was decreased by treatment with the ethyl acetate fraction. These results showed that A. tabularis extracts can be used as natural substances to control inflammation. Conclusion : These result showed that A. tabularis extracts can be used in a variety of antioxidant and other functional product research and development processes as valuable natural materials.

Background : Temperature is major factor for growth plant. Recently, because of global warming, abnormal temperature included drought, deluge, sudden temperature change and heavy snow damaged crops in the world. In Korea, crops have been sensitive to low temperature on early growth stage, e.g. fruit tree and ginseng, were damaged owing to sudden heavy snow and cold on Spring. Therefore, recently interest in cold resistance crops were increased in demand rapidly. This study was performed to establish transgenic Nicotiana benthamiana by transforming cold resistant gene related to cold tolerance S-adenosylmethionine synthetase (SAMS) isolated from Miscanthus sinensis. Methods and Results : Total RNA was extracted from leaves of M. sinensis using Trizol assay and isolated MsSAMS. Isolated MsSAMS was insert into SacⅠ- XbaⅠ sites of pMBP1 vector. The vector was transformed to Agrobacterium tumefaciens strain LBA4404 by DH5α. A. tumefaciens with binary plasmid were selected at YEP medium supplemented with kanamycin. Cut leaves of tobacco were co-cultured with selected A. tumefaciens. Co-cultured leaves was grown on regeneration medium for a month at dark condition, and transferred to at light condition. Regeneration shoot from callus were excised and transferred to root-induction medium. Approximately, 58% of leaves explant produced callus. Nearly, 30% of callus had shoot and approximately, 94% of shoots were rooted in root-induction medium. Conclusion : We established an efficient transformation system of N. benthamiana transformed by using MsSAMS gene related to cold tolerance isolated from M. sinensis. We may use the produced transgenic plants to prevent damages carried by cold.

Background : Perilla frutescens L. is a bisexual, annual plant belonging to the Labiatae. Dormancy period of Perilla seed usually ranged from 85-200 days. The germination enhancing effects in a reduced seed vigor during the GA3 treatment has been reported. Objectives of the present study was to evaluate the effect of GA3 on germination rate and the expression of germination-related genes expression by using perilla seeds. Methods and Results : Three different cultivars (Saeyeopsil, Okdong) and accession (line 141) of perilla were used in the experiment and treated with GA3 at different concentration (50, 100, 300, 500 μM) for one day at dark condition. Germination test for perilla seed was carried out by using 100 perilla seeds treated with GA3 at 25℃. Rate of germination were evaluated after 10 days and the experiments were repeated three times in similar condition. Samples were collected from each cultivar and accessions for RNA extraction. After cDNA synthesis quantification, templates were subjected to RT-PCR using actin primers. EST blast performed for sequencing the RT-PCR products. Conclusion : Result showed that 100μM GA3 treated seeds of three perilla cultivars and accessions showed the highest germination rate. However, concentration of GA3 over 100 μM and lower than 100 μM resulted into reduced germination rate. Furthermore, expression of germination-related genes from 100 μM treated GA3 was higher compared to untreated sample by using 100 μM GA3 treated sample by using pC12, pJ14 primers.