Background : The research is designed to investigate the optimal cultivation technology and the growth of above-ground and below-ground sections as well as the photosynthetic characteristics for new ginseng variety “K-1” by differentiating the planting density under the conditions of transplanting and direct seedling. Methods and Results : The K-1 variety and hybrid variety (Jakyungjong) were selected for the research and the ginseng varieties were transplanted and directly sown in Yeoncheon area in 2013. The transplanting was made in the form of 5 lines × 9 rows (45 plants), 6 lines x 9 rows (54 plants), 7 lines × 9 rows (63 plants) and 8 lines × 9 rows (72 plants) in each lot (1.65㎡) while the direct seedling for testing was conducted three times in randomly blocked design in the form of 11 lines × 14 rows (154 plants), 12 lines × 14 rows (168 plants), 13 lines × 14 rows (182 plants). Various measures were collected from the 4-year transplanted ginseng and 3-year direct seedling ginseng in 2015 to find out the growth features and photosynthesis of above-ground section (rate of germination, leaf length, leaf width, stem length and leaf area index (LAI)) and the below-ground section (length, diameter, weight and class of roots). Conclusion : After the planting of the ginseng, the germination rate of K-1 for the transplanting was 85.1 ~ 92.0% across different plantation densities while that for the direct seedling was 67.7% ~ 77.9% across plantation densities, thus showing no significant difference between the two planting methods. LAI was higher for the higher planation density for both transplanting and direct seedling. As for the photosynthesis speed, the form of 6 lines × 9 rows showed the higher speed in transplanting while the form of 12 lines x 14 rows showed the higher speed in direct seedling. The photosynthesis of K-1 was higher than that of Jakyungjong. In the 4-year ginseng cultivated under the transplanting, diameter of roots, number of branch roots and weight of raw ginseng were the highest in the plantation density of 5 lines × 9 rows. The distribution of root weight was high with 23.3% and 20.0% for the 51~70g group and the 71g or above group, respectively, for the 4 year transplanted plants in the form of 5 lines × 9 rows. The growth for above-ground and below-ground sections for K-1 was better than that for Jakyungjong. As a result, it was found that the proper plantation density for the 4-year root in the transplanted K-1 was 5 lines × 9 rows considering the growth of the above-ground section, quantity and distribution of root weight.

Background : Lycium ruthenicum Murr. was known as black wolfberry. It belongs to Solanaceae. In order to elucidate biological activities and functional substances of Lycium ruthenicum Murr., the methanol extract of Lycium ruthenicum Murr. harvested from Qaidam Besin. area was prepared and was studied. Methods and Results : Antioxidant, anticancer, anti-inflammatory and antidiabetic activities of the methanol extract of Lycium ruthenicum Murr. were examined using diverse in vitro bioassays and functional substances were analyzed by HPLC and LC-MS/MS. The methanol extract of Lycium ruthenicum Murr. showed strong DPPH and ABTS radical scavenging activities and also showed strong SOD-like activity. To examine the effects of the methanol extract of Lycium ruthenicum Murr. for the expression of antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), RT-PCR was performed. SOD expression was strongly increased by the methanol extract of Lycium ruthenicum Murr. CAT expression was also increased with dose-dependent manner. The methanol extract of Lycium ruthenicum Murr. showed inhibitory effects against α-glucosidase and α-amylase, and also inhibited NO production. Anticancer effects of the methanol extract of Lycium ruthenicum Murr. assay were examined with five human cancer cell lines. The potent anticancer activity exhibited in HepG2 hepatoma cells. As a result of main component analysis by HPLC, chlorogenic acid and rutin were identified as the major flavonoids of Lycium ruthenicum Murr.. In addition, by LC-MS/MS, petunidine-3-O-rutinoside (trans-p-coumaroyl)-5-O-glucoside was determined as a major anthocyanin of Lycium ruthenicum Murr.. Conclusion : Lycium ruthenicum Murr. showed various biological activities and could be used as a food and a functional material to enhance human health. Further study to identify an unknown flavonoids of Lycium ruthenicum Murr. would be needed.

Safflower (Carthamus tinctorius L.) seeds have long been clinically used in Korea to promote bone formation and prevent osteoporosis. In addition, the safflower buds (SB) were found to have more useful functional ingredients than safflower seed. Thus, we investigated the preventive effects of SB diet in ovariectomized (OVX) rats. The rats were divided into five groups; sham operated group, OVX alone group, OVX plus 17β-estradiol (E2 10 ㎍/㎏, i.p.) and OVX plus SB diet feeding group (0.3% or 1%). Feeding of SB diet (0.3% or 3%) to OVX rats markedly increased trabecular formation in femur compared to OVX rats. Feeding of SB diet (0.3% or 3%) to OVX rats also decreased TRAP activity compared to OVX rats. These results suggest that SB diets have bone sparing effects by the decrease of osteoclast activity. We also observed that OVX rats fed with SB diet (0.3% or 3%) exhibited the decrease of calcium and phosphorus in serum compared to OVX-induced rats. Therefore, SB may be beneficial for the patients of osteoporosis, especially in postmenopausal women.

Zinc finger nucleases (ZFNs) have been used for targeted mutagenesis in eukaryotic cells. Custom-designed ZFNs can induce double-strand breaks (DSBs) at a specific locus. Our custom ZFN dimer was designed 3-finger of left and 4-finger of right with 2 kb size using 2A. A Ti-plasmid vector, pTA7002 containing the target site of SSS4A gene for a ZFN pair, that was shown to be active in yeast, was integrated in the rice genome. This promising technique for genome engineering was induced into 4 exon region of SSS4A gene in rice genome using Agrobacterium-mediated transformation. The SSS4A full-length cDNA was 5,070 bp consisting of a 318 bp 5′-untranslated region (UTR), a complete ORF of 2,928 bp encoding a polypeptide of 975 amino acids and a 3′-UTR of 1,824 bp. The vector is based on glucocorticoid receptor inducible gene expression system. Thus, SSS4A::ZFN expression was tightly controlled and the phenotype in low concentrations 10uM of the glucocorticoid hormone dexamethasone (DEX). In plant cells, transient ZFN expression is achieved by direct gene transfer into the target cells. For an alternative, ZFN delivery and production of mutant plants using a tobacco transient expression system for indirect transient delivery of ZFNs into a variety of tissues and cells of plants. ZFN activity was determined by PCR and sequence analysis of the target site. ZFN induced plants were obtained in up to 2% of the PCR products, consisting of deletions ranging between 1and 100 bp and insertions ranging between 1 and 10 bp. Our results describe an alternative to direct gene transfer for ZFN delivery and for the production of mutated rice.

Plant breeding requires genetic diversity of useful traits for crop improvement. EMS-induced mutation is practiced to generate mutations at loci regulating economically important traits and/or to knock out the genes to elucidate their functions. The present study was aimed to induce mutations in a Korean local land race Capsicum annuum ‘Yuwol-cho’. This accession is pungent and also has advantage to mature early. A total of about 1,500 M2 families were screened and three non-pungent mutants were identified and crossed with wild type ‘Yuwol-cho’. After phenotyping of F2 population for pungency, MutMap approach will be used to identify the genes controlling the pungency in mutants. In addition to this, another C. annuum accession “Micro-Pep” was used to develop a mutant population. Micro-Pep is a small, pungent pepper generally used as ornamental purpose. Having compact growth habit, and small size, it has advantage to handle and utilize easily in mutation study and molecular research. On the basis of preliminary experiment 1.3% of mutagen was used for treatment of pepper seeds and 30% less germination percentage was observed in EMS treated seeds in comparison to control seeds. A total of 4,674 M1 plants are grown under greenhouse condition and M2 population will be studied for characterization of phenotypic variation including fruit color and pungency. Newly constructed mutant populations will be valuable assets for identification of functional genes and molecular breeding of pepper.

Superjami is a new rice breed resulted from crossing ‘C3GHi (has high amount of Cyanidin 3-glucoside, and was developed from a cross between ‘Heugjinjubyeo’ and ‘Suweon 425’) and ‘Daeribbyeo 1’. Superjami has 10.9 times higher C3G content compared with ‘Heugjinjubyeo’. It also contains the highest essential amino acids of all kinds (except tryptophan content). This study was done to investigate the effects of extracts from superjami bran on the in-vitro antioxidant metabolism, in-vivo antioxidant metabolism and bone metabolism on menopause- induced condition in experimental rats. Overall, extract from superjami bran was confirmed of improving antioxidant and bone metabolism which can be considered as a good dietary supplement.

GWAS (Genome-wide association study) provides a useful to associate phenotypic variation to genetic variation. It has emerged as a powerful approach for identifying genes underlying complex diseases or morphological traits at an unprecedented rate. Despite benefits, there are only a few examples applied in crop plants due to lack of effective genotyping techniques and well prepared resources for developing high density haplotype maps. In this study, 350 core accessions selected from almost 5,000 Capsicum accessions were used for GWAS. We are planning to construct a high-density haplotype map using GBS platform and perform GWAS for various agronomic traits including fruit traits and metabolites related to pungency to identify genes controlling the traits. These results will not only provide a list of candidate loci but also a powerful tools for finding genetic variants that can be directly used for crop improvement and deciphering the genetic architecture of complex traits.

The ovariectomized Sprague-Dawley female rats were randomly assigned to Sham-Control, OVX-Control, OVX-Superjami (extract) groups. The results showed that the activity of glucokinase to keep the blood sugar constant is increased by increasing insulin secretion from pancreatic β- cells and the homeostatic regulation of glucose. Meanwhile the glyconeogenesis which is involved in the actions of the enzymes glucose-6-phosphatase and phosphoenolpyruvate carboxykinase showed that the glucose level is decreased. It was confirmed that these enzymes regulate the carbohydrate metabolism. On the other hand, results of the measurement of the lipid metabolism in the fat tissue and liver tissue, effect of β-oxidation enzymes and carnitine palmitoyl transferase which is involved in fatty acid oxidation for energy generation is increased. Moreover, the activity of fatty acid synthase, glucose-6-phosphate dehydrogenase and malic enzyme have been reduced, therefore, it was confirmed that these enzymes regulate the lipid metabolism.

Pepper (Capsicum spp.) germplasm shows diverse phenotypic variations including fruit size, color, pungency, and many other horticultural traits. Traditional markers including SSR, AFLP, and RFLP have been used to construct genetic maps using biparental populations. However to assess the genetic diversity of large number of germplasm, a robust and rapid marker development and genotyping approach is needed. We used six pepper accessions including C. annuum, C. chinense, C. baccatum and C. frutescens and performed genotyping-by-sequencing (GBS). To select the most appropriate condition, eight different 2 bp selective nucleotides were used to make GBS libraries. Selective nucleotide ‘OO’ showed the largest number of reads in all samples, and 11,026 to 47,957 high-quality SNPs were called in six accessions. When C. annuum ‘CM334’ genome sequence was used as a reference, C. annuum showed the smallest number of SNPs, while C. baccatum which was known to be a different Capsicum clade showed the largest number of SNPs. Pepper core collection chosen to represent the genetic diversity of whole germplasm will be genotyped by high-density SNPs developed from GBS. We will perform genome-wide association study (GWAS) using genetic and phenotypic variation to identify the functional genetic loci controlling horticultural traits.

Capsicum annuum ‘Bukang’ is a resistant variety to Cucumber mosaic virus isolate-P0 (CMV-P0), CMV-P1 can overcome the CMV resistance of ‘Bukang’ due to mutations in Helicase (Hel) domain of CMV RNA1. To identify host factors involved in CMV-P1 infection, a yeast two-hybrid system derived from C. annuum ‘Bukang’ cDNA library was used. A total of 156 potential clones interacting with the CMV-P1 RNA helicase domain were isolated. These clones were confirmed by β-galactosidase filter lift assay, PCR screening and sequence analysis. Then, we narrowed the ten candidate host genes which are related to virus infection, replication or virus movement. To elucidate functions of these candidate genes, each gene was silenced by virus induced gene silencing in Nicotiana benthamiana. The silenced plants were then inoculated with green fluorescent protein (GFP) tagged CMV-P1. Virus accumulations in silenced plants were assessed by monitoring GFP fluorescence and enzyme-linked immunosorbent assay (ELISA). Among ten genes, silencing of formate dehydrogenase (FDH) or calreticulin-3 (CRT3) resulted in weak GFP signals of CMV-P1 in the inoculated or upper leaves. These results suggested that FDH and CRT3 are essential for CMV infection in plants. The importance of FDH and CRT3 in CMV-P1 accumulation was also validated by the accumulation level of CMV coat protein confirmed by ELISA. Altogether, these results demonstrate that FDH and CRT3 are required for CMV-P1 infection in plants.

Carotenoids are vital pigments responsible for yellow, orange and red color in plants. In Capsicum, capsanthin-capsorubin synthase (CCS), phytoene synthase (PSY), β-Carotene hydroxylase (CRTZ-2) and lycopene β-cyclase (LCYB) were identified to be involved in the carotenoids synthesis pathway. Previously molecular markers based on the CCS and PSY genes have been developed to distinguish fruit colors in pepper. However these markers can distinguish fruit colors of limited pepper genotypes. Therefore, there is need of developing additional markers for accurate prediction of fruit colors using molecular markers. In this study carotenoids contents of 16 pepper accessions were analyzed and the CCS, PSY, CRTZ-2, LCYB genes were sequenced to identify the genes affecting the fruit color. Among all the analyzed carotenoids, capsanthin was accumulated in much higher amount in red and orange fruits (1100-2500 mAU·min and 30-500 mAU·min respectively) while violaxanthin (20-1200 mAU·min) was accumulated more in yellow fruits. Sequence analysis revealed that deletions and two frame shift mutations in CCS gene for yellow accessions. Frame shift mutations of the PSY gene were detected in two orange accessions. These results show that mutations in CCS and PSY genes affect the fruit colors of pepper, and markers can be developed using mutations of these genes.

Powdery mildew disease caused by Leveillula taurica is a serious fungal threat to greenhouse pepper production. In contrast to most epiphytic powdery mildew species, L. taurica is an endophytic fungus which colonizes in the mesophyll tissues of the leaf. In the genus Capsicum, several studies have been conducted to identify resistance sources to L. taurica. In previous studies, five quantitative trait loci (QTLs) for powdery mildew resistance have been identified. An F2 population derived from self-pollination of the commercial cultivar Capsicum annuum ‘PM Singang’ was used for genetic analysis of powdery mildew resistance. Resistance of the F2 plants was tested under the natural environmental conditions. Sporulation intensity on infected leaves was used as a disease scale to assign resistance levels to plants, where 0-5% is Resistant, 6-15% Moderate resistant and 16-100% Susceptible. A total of 83 F2 plants were evaluated for resistance. The results showed that 59 plants were resistant, 10 susceptible and 14 moderately resistant. If we consider MR as S, segregation ratio fitted to a single dominant resistance gene model. In the future study, closely linked molecular marker will be developed and tested to locate this gene. The developed marker will be used to identify the powdery mildew resistance gene.

The transposable element is a DNA sequence that can be changed its position within the genome, sometimes it can create or reverse mutations and altering the cell's genome size. Target region amplification polymorphism (TRAP) is a rapid and efficient PCR-based marker technique, which uses bioinformatics tools and expressed sequence tag (EST) database information to generate polymorphic markers around targeted candidate gene sequences. TE-TRAP is a new marker system which used terminal inverted repeat (TIR) instead of targeted candidate gene sequences. Sorghum holds a good potential plant organism for transposon tagging due to its small genome size, low amount of repetitive DNA and co-linearity with other cereal genomes, which allows the use of information derived from sorghum in other cereal grasses. IS2868 of sorghum accession was treated Gamma irradiation on seed. To define availability and utilization of TE-TRAP, twenty-one accessions were used to evaluate the genetic diversity and underlying relationships. One-thousand thirty-three TE-TRAP markers were amplified by thirty-one primer combination. Altogether, 712 (62.8%) markers were observed polymorphic segregation, whereas 421 (37.2%) showed monomorphic patterns. To estimate genetic differentiation of population by various gamma radiation doses, the analysis of molecular variance (AMOVA) was performed using 4 to 5 different radiation doses population of M1 sorghum individuals. This study and marker system will provide valuable information to assist radiation mutation breeding.

Space has many distinguishable characteristics from earth such as strong cosmic radiation, microgravity, supervaccum and weak magnetic field. For this reason, space environments can be used an efficient mutagen for plant breeding nowadays. To identify the affected genes by condition in space with outer space, Brachypodium seeds were placed in the Russia Segment (RS) Biorisk module of International Space Station (ISS). Brachypodium distachyon is a model system for temperature grass, because they represent the characteristics for annual winter grass. Seeds and organs of plants carried by satellite or spacecraft to space can be genetically mutated by exposing space environment. We performed a duplicated RNA sequencing to profile the differentially expressed genes. As a results, about 700 genes were upregulated and 250 genes were downregulated by cosmic environments, respectively. In the molecular function category, protein kinase and transcription activity related genes were upregulated. Among the many transcription factors (TFs), stress related TFs such as ERF, NAC and WRKY were differentially expressed in space exposed samples. In the future, their expression will be identified by using qRT_PCR.

sy-2 (Seychelles-2) is a temperature sensitive natural mutant of Capsicum chinense and native to Seychelles Island in Africa. Previously we showed that sy-2 leaves were irregularly shaped and defective in chlorophyll development at temperatures below 24℃. A segregation test revealed that the sy-2 gene is controlled by a single recessive gene. To identify the sy-2 gene, we performed a map-based cloning approach using a total 600 individual F2 plants derived from crossing sy-2 and the wild type C. chinense ‘No.3341’. Fine-mapping of the locus allowed us to position sy-2 to an approximately 170-kb region flanked by markers IN2-1-1 and SNP-3-7 on chromosome 1. Among the approximately 36 hypothetical genes in this region several candidate genes including: HSP90-like ATPase family proteins, lipid-transfer proteins, calmodulin-domain protein kinases, and zinc finger proteins (ZFPs) were identified. RT-PCR and sequencing of the hypothetical genes are under way to identify sy-2.

UDP-glucose 4-epimerase (UGE; EC 5.1.3.2) is an enzyme that plays an essential role in the interconverts UDP-D-glucose (UDP-Glc) and UDP-Dgalactose (UDP-Gal). Five members of the Chinese cabbage (Brassica rapa) UDP-glucose 4-epimerase gene family, designated BrUGE1 to BrUGE5, have been cloned and characterized. Quantitative PCR shows that the BrUGE1and BrUGE4 mRNA are most abundant among other BrUGE genes, accounting for more than 55% of total BrUGE transcripts in most of the tissues examined. All genes showed organ specific expression pattern, two of which (BrUGE1 and 4) actively responded after Pectobacterium carotovorum subsp. carotovorum infection, while four genes (BrUGE-1, -3, -4 and -5)were shown to respond considerably against salt, drought and abscisic acid (ABA) treatments. To better understand the function of the UGE gene, we constructed a recombinant pART vector carrying the BrUGE1 gene under the control of the CaMV 35S promoter and nos terminator and transformed using Agrobacterium tumefaciens. We then investigated BrUGE1 overexpressing rice lines at the physiological and molecular levels under biotic and abiotic stress conditions. Bioassay of T3 progeny lines of the transgenic plants in Yoshida solution containing 120 mM Nacl for 2 weeks, confirmed that the BrUGE1 enhances salt tolerance to transgenic rice plants. Also T3 progeny lines of the transgenic plants, when exposed to infection caused by Xanthomonas oryzae pv oryzae, showed tolerance to bacterial blight. These results showed that BrUGE1 can be used as potential genetic resource for engineering Brassica with multiple stress resistance.

This study was carried out to determine the amount of lignin, cellulose, and hemicellulose in six kenaf cultivars during different harvesting stages. Three mutant cultivars (Jangdae, Jeokbong and Baekma), two original cultivars (Jinju, C14), and one Chinese cultivar (Auxu) were planted on May 14, 2013. Four harvesting times were made at intervals of 20 days from 15 July to 16 September, 2013. The overall growth characters of mutant cultivar ‘Jeokbong’ such as plant height, stem diameter, flowering time, and dry mass were similar with those of the original variety. The mutant cultivar ‘Baekma’ occurred 10-day late flowering in comparison with the original variety and also displayed higher dry mass than the original variety. Jinju, Auxu and Jangdae, mid-late maturing kenaf cultivars, had high dry weight compared to early maturing cultivars such as Jeokbong, Baekma and C14. In all cultivars, the lignin contents were increased by a late harvest. The Mid-late maturing kenaf cultivars showed high lignin content in comparison with those of the early maturity cultivars. There were no significant differences of cellulose, and hemicellulose content between the cultivars, however cellulose content in stems of these kenaf cultivars were significantly decreased by a late harvest. These results may provide valuable information to assist the parental selection of kenaf breeding.

Kenaf (Hibiscus cannabinus L.) native to Africa can be used as fiber, food, feedstock and bio plastic. This study was carried out to evaluate the mineral, amino acid and vitamin contents of six selected kenaf cultivars which are enable to produce seed under Korean circumstance. The leaves of three mutant cultivars (Jangdae, Jeokbong and Baekma), two original cultivars (Jinju, C14) and one Chinese cultivar (Auxu) were harvested at flowering time. Mineral components of kenaf leaves, such as calcium, potassium, and mineral, did not showed significant differences among the cultivars. As major amino acids including proline and phenylalanine, significant differences were found in these kenaf cultivars. The Auxu cultivar contained the highest amount of essential amino acid (Phenylalanine, Leucine, Isoleucine, Valine, Methionine and Lysine). The amount of vitamin displayed significant differences such as vitamin E and vitamin K among these cultivars. Especially, Jangdae cultivar contained the highest amount of vitamin E and vitamin K. Thus, these data suggested that Jangdae and Auxu is the most desirable cultivar containing high amount of vitamin and amino acid.

Foxglove aphid, Aulacorthum solani (Kaltenbach), is a Hemipteran insect that infected a wide variety of plants worldwide and caused serious yield losses in crops. The objective of this study was to identify the putative QTL for foxglove aphid resistance in wild soybean, PI 366121, (Glycine soja Sieb. and Zucc.). One hundred and forty one F2-derived F8 recombinant inbred lines developed from a cross of susceptible Williams 82 and resistant PI 366121, were used. The phenotyping of antibiosis and antixenosis was done through choice and no-choice assays with total plant damage (TPD) and primary infestation leaf damage (PLD); a genome-wide molecular linkage map was constructed with 504 single nucleotide polymorphism markers utilizing a GoldenGate assay. Using inclusive composite interval mapping analysis for foxglove aphid resistance, one major candidate QTL on chromosome 7 and 3 minor QTL regions on chromosome 3, 6 and 18 were identified. The major QTL on chromosome 7 showed both antixenosis and antibiosis resistance responses. However, the minor QTLs showed only antixenosis resistance response. The major QTL mapped to a different chromosome than the previously identified foxglove aphid resistance QTL, Raso1, from the cultivar Adams. Also, the responses to the Korea biotype foxglove aphid were different for Raso1, and the gene from PI 366121 against the Korea biotype foxglove aphid were different. Thus the foxglove aphid resistance gene from PI 366121 was determined to be an independent gene to Raso1 and designated to Raso2. This result could be useful in breeding for new foxglove aphid resistant soybean cultivars.

Foxglove aphid, Aulacorthum solani (Kaltenbach), is a Hemipteran insect that infected a wide variety of plants worldwide and caused serious yield losses in crops. The foxglove aphid resistance gene, Raso2 was previously mapped from PI 366121 (Glycine soja Sieb. and Zucc.) to a 26cM marker interval on soybean chromosome 7. The development of additional genetic markers, which are mapped closer to Raso2 were required to accurately position the gene to improve the effectiveness of marker assisted selection. The objective of this study was to narrow down the putative QTL region, which is responsible to foxglove aphid resistance in PI366121 using recently developed high-density 180K Axiom SoyaSNP genotyping array. One hundred and forty one F8-derived F12 recombinant inbred lines developed from a cross of susceptible Williams 82 and resistant PI 366121, were used to generate a fine map of Raso2 interval. The phenotyping of antibiosis and antixenosis was done through choice and no-choice assays with total plant damage (TPD) and primary infestation leaf damage (PLD). The composite interval mapping analysis showed that the physical interval between two flanking makers, which was corresponding to Raso2, was narrowed down to 500kb on the Williams 82 genome assembly (Glyma2.0), instead of 4Mb in the previous report using Goldengate assay. In the Raso2 interval, there are about 60 candidate genes, including 4 of NBS-containing putative R genes. This result could be useful in breeding for new foxglove aphid resistant soybean cultivars.