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.

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.

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.

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.

Capsinoids, low-pungent compounds, have the same biological effects as capsaicinoids such as anticancer and anti-obesity. A precursor of capsinoids, vanillyl alcohol, is known to be produced by mutations in the putative-aminotransferase (pAMT) gene. In the previous study, ‘SNU11-001’ (Capsicum chinense) containing high levels of capsinoids was identified in germplasm collections of Capsicum. This collection has a unique mutation in the pAMT gene that can cause dysfunction of this gene. In order to develop pepper varieties containing high capsinoids contents, marker-assisted foreground and background selections were performed during backcross breeding. Compared to the conventional backcrossing, marker-assisted backcrossing (MABC) is extremely useful for recovery of a recurrent parent’s genetic background. For foreground selection, plants carrying the pAMT/pamt genotype were selected from a BC1F1 and BC2F1 populations using SCAR markers derived from the unique pAMT mutation of ‘SNU11-001’. To obtain background selection markers, a total of 412 single nucleotide polymorphism (SNP) markers was screened on ‘Shinghong’ parental lines and ‘SNU11-001’ to obtain polymorphic SNP markers. Of the 412 SNP markers, 144 and 204 polymorphic SNP markers evenly distributed in pepper genome were finally selected. BC1F1 and BC2F1 plants carrying the pAMT/pamt genotype were subjected to background selection using the selected marker sets. Multiple genotype analysis was done using a high-throughput genotyping system (EP1TM, Fluidigm®, USA). As a result, one BC1F1 plant 84% similar to the recurrent parent and several BC2F1 plants more than 96% recovery rate of the recurrent parent were selected. Genetic backgrounds of the selected BC2F1 plants were evaluated by the genotype-by-sequencing (GBS) method in order to confirm the background selection results using the SNP marker set. GBS results showed that recovery rate and positions of introgressed segments were well matched between two methods demonstrating MABC can be successfully done with a couple hundred SNP markers.

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.

Root-knot nematode, Meloidogyne incognita is a virulent pest of solanaceaous crops worldwide. The M. incognita resistance gene Me7 derived from Capsicum annuum CM334, is located on chromosome 9. In the present study, an F2 population derived from a cross between ECW03R and CM334 was used to locate the Me7 gene. An F2 population was inoculated using approximately 1,000 second-stage juveniles per individual plant. Phenotype screening was done 45 days after inoculation by using gall index system. The phenotype study of 503 F2 individual showed 391 resistant and 112 susceptible plants. The 3:1 phenotypic ratio confirmed that resistance phenotype is controlled by a single dominant gene. Previously reported two markers were tested to reveal the linkage of markers to phenotype. Two markers, CAPS_F4R4 and SCAR_PM6a were located at 4.3 and 2.7 cM from the resistance gene, respectively. Additional SNP markers were developed using CM334 reference genome information to narrow down the position of the gene, but no closer markers could be developed due to errors of DNA sequence assembly. The closest marker was positioned on telomere of the chromosome 9 long arm, where tens of other NB-LRR genes are clustered. NB-LRR genes are being used as candidates to identify the Me7 gene.

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.

Phytophthora capsici an Oomycete pathogen is a major challenge to the pepper (Capsicum spp.) production around the world. Control measures are proved ineffective, so breeding resistant cultivars are the most promising strategy against the pathogen. Resistance against P. capsici is governed by quantitative trait loci (QTL). According to previous studies on QTL detection, the QTL on pepper chromosome 5 is a major contributor to resistance. In this study, to exploit the involvement of this QTL and identify its contributing genes, the F2 population derived from a cross between ECW30R and CM334 was inoculated with a medium virulence P. capsici strain JHAI1-7 zoospores at the 6-8 leaf stage. Composite interval mapping revealed two major QTLs; QTL5-1 from 7 days post inoculation (dpi) and QTL5-2 from 16 dpi on chromosome 5. To characterize and detect interactions of the two QTLs, near isogenic lines (NIL) were constructed by crossing Tean and recombinant inbred line (RIL) derived from a cross between YCM334 and Tean. RILs were screened with P. capsici strain MY-1 and resistant lines were selected. Among the resistance RILs most closely related to Tean were selected using AFLP and SSR genotyping data. These RILs were named as YT39-2 and YT143-2. To develop more advanced NILs, two rounds of marker-assisted backcrossing were done using a high-throughput SNP genotyping system (EPI Fluidigm, USA). Among the NILs derived from YT39-2, YT39-2-64 contains only QTL5-1 whereas YT39-2-61 and YT39-2-69 were identified to have both QTLs. On the other hand, YT143-2-55-7 with the highest Tean genetic background contains QTL5-1 only. In the next step, the 3 different NILs having QTL5-1, QTL5-2 individually and both QTLs will be identified. Furthermore, phenotyping and fine mapping will be done for the analysis of individual and interaction effects of QTLs.

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.

Rose (Rosa Hybrida Hort.) are of a high symbolic value and a great cultural importance in different societies. They are widely used as garden ornamental plants and as cut flowers. For the induction of mutation, gamma-rays are widely used as a mutagen. This study was carried out to establish a system for mutation breeding by irradiation of gamma-ray in rose. The rooted cuttings of five cultivar roses (Lovelydia, Vital, Aqua, Yellowbabe and Haetsal) are grown by in a greenhouse. They were two difference treatment (Before rooting gamma-ray irradiation, After rooting gamma-ray irradiation) were exposed to dose of 70 Gy using a 60Co gamma-irradiator (150 TBq of capacity ; ACEL, Canada) at the Korea Atomic Energy Research Institute. The irradiated plants were planted in a greenhouse, and investigated survival rate, mutation rate, flower buds number, and shoot length were planted after 80days. The two treatments of and growth characters was significantly reduced to 20% to 40% compared with the control. In addition, survival rate and mutation rate were ‘after rooting γ-ray irradiation (37.4~67.3% and 0.5~5.6%)’ higher than ‘before rooting γ-ray irradiation (18.3~50.8% and 0.3~3.4%)’. Mutation types were solid type, chimeric and mosaic petal mutants with various colors were induced from five rose. These results indicate that efficiency of mutation induction in rose by gamma-ray irradiation on petal colors and petal shapes in two difference treatment with rooted cutting system.

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.

The depletion of stratospheric ozone has resulted in increased amount of ultraviolet-B radiation (UV-B: 280-320 nm) reaching the Earth’s surface and could cause significant biological effect in plants. In this study, putative quantitative trait loci (QTL), which is responsible to UV-B resistance in soybean, was identified using recently developed high-density 180K Axiom SoyaSNP genotyping array. A population of 115 recombinant inbred lines (RILs) derived from a cross between susceptible Keunolkong and resistant Iksan 10 was analyzed. A total 8,970 polymorphic SNP markers were used to construct linkage map. The both parents and RILs were grown with supplemental UV-B radiation in a greenhouse condition. Three categories of UV-B induced morphological damage, degree of leaf chlorosis, leaf shape change, and total plant damage were evaluated. Using composite interval mapping analysis, one major QTL associated with all of the phenotypic traits was detected on 7.7cM of soybean chromosome 7 with 22 of LOD score accounting for about 60% of phenotypic variance. Also, the allele from Iksan 10 were responsible for the UV-B resistance. Thus, the UV-B resistance QTL on chromosome 7 from Iksan 10 was designated to qUVBR1, corresponding to 30kb on the Williams 82 genome assembly (Glyma2.0) including 7 candidate genes. This result could be useful in breeding for new foxglove aphid resistant soybean cultivars. In addition, these results provided useful information not only for marker-assisted selection for UV-B resistance soybean, but also for the future identification of putative candidate genes, responsible for UV-B resistance in soybean.

This study analyzes the buckling safety in the area of circumferentially edge-stiffened door openings without any additional longitudinal stiffeners of offshore tubular steel towers. The tubular steel tower is subjected to six (6) different load situations which are deemed to be normal and abnormal operating cases for the ultimate limit state. Analytical method using parametric equations based on Eurocode 3 - Design of Steel Structures and numerical method of finite element are used to analyze the critical meridional buckling stress. ABAQUS, a finite element program, is used for the numerical method analysis. Buckling safety analysis in the localized area near the opening is studied, and points of interest are defined for comparison between the two aforementioned analyses. Findings are tabulated and shown in illustrative charts, and conclusions are made.

This parametric study investigates the effect of significant parameters on the elastic global lateral torsional buckling capacity of horizontally curved twin I-girder systems. Included are the effect of curvature and the effect of girder spacing. Twin I-girder systems analyzed in this study are those which are interconnected by intermediate cross frames and are subjected to uniform moment. The finite element analysis software, ABAQUS, is used to model the horizontally curved twin I-girder system and conduct the buckling analysis. To see the significant effect of curvature and girder spacing, the results from the finite element analysis of horizontally curved twin I-girder systems are compared to the results of a closed form solution for straight twin I-girder systems. The findings of this parametric study will be shown using illustrative figures and tabulated data and conclusions are made.