Steroidal glycoalkaloids which serve the plant defense, are toxic secondary metabolites present in the plants of solanaceae family. The upper safe limit of glycoalkaloids for human consumption is 20mg/KG FW, excess of which may cause severe health disorders. Several factors like drought, high temperature, light exposure, and wounding increase tuber glycoalkaloid content. Among these, drought is an important factor which causes a rapid increase in potato glycoalkaloid content. Glycoalkaloid biosynthetic genes and their expression pattern need to be characterized to regulate the glycoalkaloid accumulation. Three key genes SGT1, SGT2 and SGT3 are demonstrated to be directly participated in the biosynthetic pathway for glycoalkaloid formation. Present study was focused on the study of expression pattern of key genes in GA pathway under drought stress in two different potato cultivars Atlantic and Haryoung which are low and high glycoalkaloid accumulating respectively. Drought stress was imposed by withholding water to the plants grown in pots and control plants kept irrigated. Expression analysis of SGT1, SGT2 and SGT3 was done from the leaf and tuber sample of three time intervals i.e 5, 10 and 20 days after imposing stress. Variation in the expression level of genes was observed in leaf and tuber where the fold increase in expression over control was higher in tuber sample compared to leaf. Expression levels also varied in leaf and tuber among two cultivars. However, expression of SGT1, SGT2 and SGT3 is significant indicating the involvement of these genes in glycoalkaloid accumulation under drought stress.

Chronic gamma irradiation can be used an alternative mutation breeding methods for induction of many useful mutants. Seedlings of purple-colored wheat plants were irradiated with wide range doses of chronic gamma-rays (20, 25, 30, 40, 50, 70, 100, 125, 150, 200, 250, 300 Gy) during 6 weeks at gamma-phytotron in the Korea Atomic Energy Research Institute, respectively. To identify the biological responses purple-colored wheat, we examined the plant height, chlorophyll, carotenoid and total anthocyanin contents in leaf. Plant growth, chlorophyll and carotenoid contents in leaf were decreased when the dose rate increased. Anthocyanin contents were increased with the increase of the radiation dose until 50 Gy treatment. To confirm the real contents of anthocyanin, we also investigated cyanidin-3-glucoside in purple-colored wheat leaf by using UPLC analysis. These results indicate that anthocyanin accumuation was observed under chronic gamma irradiation.

Acanthopanax species is known commonly as Siberian ginseng, touch-me-not, devil’s shrub, prickly eleutherococc, eleutherococc and wild pepper. A diverse group of chemical compounds isolated from Acanthopanax species was named ‘eleutherosides’. Among eleutherosides, eleutherosides B and E were widely known in Acanthopanax species. Acanthopanax species are cultivated and grow wild in a various area of Korea and have a variety of pharmacological effects. But, there are a lot of difficulties on producing excellent Acanthopanax species, according to the cultivated method is different pharmacological ingredients. This study, therefore, analyzed eleutherosides B and E in A. divaricatus and A. koreanum by different fertilizer ratio using HPLC. We will be investigated a high content of eleutherosides B and E by different fertilizer ratio and suggest an efficient fertilizer ratio of A. divaticatus and A. koreanum. All samples of A. divaricatus and A. koreanum were collected at Yeongcheon Agricultural Technology & Extension Center, Yeongcheon, Korea. The sample was prepared by upper and lower parts. The fertilizer ratio are N-P-K(10.5-8.5-8.5: 50 kg/10a), 2N-P-K (21-8.5-8.5: 50 kg/10a), N-2P-K (10.5-17-8.5: 50 kg/10a), N-P-2K (10.5-8.5-17: 50 kg/10a), and 2N-2P-2K (21-17-17: 50 kg/10a), respectively. To analyze eleutherosides B and E, 5 g of A. divaricatus and A. koreanum was extracted with 50% MeOH (3 × 100 ml) by reflux and evaporated in vacuo. The residue was dissolved in 1 ml of MeOH. The resulting solution was used for HPLC analysis. HPLC separation of eleutherosides B and E for qualitative and quantitative analysis was performed using a reverse phase system. A Discovery®C18 (4.6 × 250 mm, 5 μm) column was used with a mobile phase that consisted of water and acetonitrile. A gradient solvent system of water and acetonitrile (90:10 to 70:30 for 20 min) was used for the elution program. UV detection was conducted at 350 nm. The injection volume was 10 μl and the flow rate was 1 ml/min. All injections were performed in triplicate. The different fertilizer ratio yielded total eleutherosides B and E contents of 4.417-6.905 and 3.652-7.227 mg/g in the upper and lower parts of A. divaricatus, respectively. In A. koreanum, the total eleutherosides B and E contents were 4.591-10.108 and 3.834-9.079 mg/g in the upper and lower parts, respectively. The best conditions to increase eleutherosides B and E content in A. divaricatus was determined to be with N-2P-K fertilizer ratio, on the other hand, in A. koreanum was 2N-2P-2K fertilizer ratio.

The change of quality characteristics with storage temperature (room (25°C), low (15°C) and cold (4°C) temperature) and period of unhulled foxtail millet (Setaria italica Beauv. cv. Samdame and Gyeongkwan 1), proso millet (Panicum miliaceum L. cv. Hwanggeum-gijang and Manhongchal) and sorghum (Sorghum bicolor (L.) Moench cv. Hwanggeumchal and Donganme) were evaluated. The 1,000 grain weight, lightness, redness, yellowness and moisture content with storage temperature and period of unhulled foxtail millet, proso millet and sorghum were not showed difference. Germination percent and milling recovery of foxtail millet, proso millet and sorghum decreased with increasing storage temperature and period. Fat acidity of stored foxtail millet, proso millet and sorghum was increased with increasing storage period, and the higher temperature increased more. The results of this study show that the storage of foxtail millet, proso millet and sorghum at the low and cold temperature is better than the room temperature. Especially, in mind of the economics, the storage method is good low temperature than cold temperature.

Soybean mosaic virus (SMV), a member of Potyviridae family, is one of the most typical viral diseases and results in yield and quality loss of cultivated soybean. Due to the depletion of genetic resources for resistance breeding, a trial of genetic transformation to improve disease resistance has been performed by introducing SMV-CP and HC-Pro gene by RNA interference (RNAi) method via Agrobacterium-mediated transformation. Transgenic plants were infected with SMV strain G5 and investigated the viral response. As a result, two lines (3 and 4) of SMV-CP(RNAi) transgenic plants and three lines (2, 5 and 6) of HC-Pro(RNAi) transgenic plants showed viral resistance. In genomic Southern blot analysis, most of lines contained at least one T-DNA insertion in both SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. Subsequent investigation confirmed that no viral CP and HC-Pro gene expression was detected in two SMV-resistant lines of SMV-CP(RNAi) and three lines of HC-Pro(RNAi) transgenic plants, respectively. On the other hand, non-transgenic plants and other lines showed viral RNA expression. Viral symptoms affected seed morphology, and clean seeds were harvested from SMV-resistant line of SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. In addition, strong viral gene expression was detected from seeds of SMV-susceptible non-transgenic plants and SMV-susceptible transgenic lines. When compared the viral resistance between SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants, soybean transgenic plants with the HC-Pro gene using RNAi strategy showed much stronger and higher frequency of viral resistance.

Over the last decades, increasing natural disasters and climate change are considered as the major environmental problems facing the globe. Numerous studies have been indicated it would cause huge losses on agriculture, especially in the grain productivity. Therefore, several alternatives are suggested for boosting up productivity of wheat as one of the main human food crop. One of important strategy is proper management of inflorescence development and DELLA proteins have been elucidated to play pivotal roles in growth of many plant organs. In this study, putative negative regulator of DELLA protein, GAST (Gibberellic acids stimulated transcript) have been isolated to identify their role in the developing spike of wheat. Four genes were isolated from its gene family and designated as TaGAST1, 2, 3, 4. Genomic structure was analyzed to demonstrate chromosomal localization of TaGAST genes and evolutionary relationships were also verified with GAST genes in other plant species. RT-PCR was conducted to detect transcriptional changes of TaGAST genes on external phytohormone. Each of TaGAST genes showed considerable changes in transcription level after GA, ABA, PAC treatment, respectively. Through Yeast two-hybrid assay, one protein for TaGAST1, and four proteins for TaGAST2 was isolated as putative interactive proteins in wheat spikes just before and after emergence.

Glycoalkaloids are a family of toxic secondary metabolites present in the plants of solanaceae family, which serve for plant defense. Two major glycoalkaloids present in plants are a-solanine and a-chaconine. The upper safe limit of glycoalkaloids for human consumption is 20mg/KG FW and its excess may cause severe health disorders. Light is the major factor known to increase the glycoalkaloid content in post harvest potato tuber. Glycoalkaloid pathway is not completely understood. Hence, identification and characterization of SGA biosynthetic genes and the genetic factors that control their expression levels assumes significance. Present investigation was focused on the study of expression pattern of key genes in steroidal glycoalkaloidal pathway under various light qualities in potato (Solanum tuberosum L). Two potato cultivars Atlantic and Haryeong which accumulates low and high glycoalkaloids respectively were used to check the levels of gene expression under various light qualities viz., red, blue, white, green, yellow, purple, UV light and in dark at different time intervals. Expression of three genes viz., SGT1, SGT2 and SGT3 which are directly involved and four other genes, HMG1, SQS1, SMT1 and SMT2 in the pathway envisaged to be indirectly involved in the glycoalkaloid formation was quantified by RT PCR. Varietal variation in the expression among the genes was observed in different light qualities. White, red and green light compared to other light qualities majorly contributed for the increased expression of genes for glycoalkaloid accumulation at different time intervals. Importantly, there is no significant transcript accumulation of these genes in dark condition. However, more efforts would be extended for further understanding of glycoalkaloid accumulation under light.

Olive flounder Paralichthys olivaceus is one of the most widely cultured fish species in Korea. Although olive flounder receive attention from aquaculture and fisheries and extensive research has been conducted eye morphological change in metamorphosis, but little information was known to molecular mechanism and gene expression of eye development- related genes during the early part of eye formation period. For the reason of eyesight is the most important sense in flounder larvae to search prey, the screening and identification of expressed genes in the eye will provide useful insight into the molecular regulation mechanism of eye development in olive flounder. Through the search of an olive flounder DNA database of expressed sequence tags (EST), we found a partial sequence that was similar to crystallin beta A1 and gamma S. Microscopic observation of retinal formation correspond with the time of expression of the crystallin beta A1 and gamma S gene in the developmental stage, these result suggesting that beta A1 and gamma S play a vital role in the remodeling of the retina during eye development. The expression of crystallin beta A1 and gamma S were obviously strong in eye at all tested developing stage, it is also hypothesized that crystallin acts as a molecular chaperone to prevent protein aggregation during maturation and aging in the eye.

Citrus is one of the major fruits produced in Korea. There are about 20 species mainly grown in Jeju Island, Korea. Four representative species, which are quite different in the shape of leaf and the taste of fruit, were selected and were used to profile the transcriptomes. These species are ‘Miyagawa Wase’ (C. unshiu Marcov.) satsuma mandarin, ‘Kiyomi’ (C. unshiu Marcov. × C. sinensis) mandarin hybrid, ‘Dangyuja’ (C. grandis) and ‘Natsudaidai’ (C. natsudaidai). Classification of the up-regulated and down-regulated genes using the Cluster of Orthologous Groups of proteins (COG) database reveals that the number of genes included in each group differed significantly among the four species. Several genes that showed significant differences in expression on the microarray were selected and their expression patterns were examined by reverse transcription- ploymerase chain reaction. Metabolic genes such as tyrosine decarboxylase and β-glucosidase ligase were found to be highly expressed in Miyagawa Wase, relative to other species. On the other hand, the expression level of mannose phosphate isomerase was lower in Miyagawa Wase. An efflux pump gene was found to be up-regulated in Kiyomi, whereas cinnamyl-alcohol dehydrogenase was down-regulated. β-carotene 15,15’-dioxygenase, which is involved in the vitamin metabolism, was up-regulated in Natsudaidai. Interspecific differentiations of gene expression are analyzed in terms of the metabolic pathways and their possible roles in citrus species.

Rye has important genes for biotic and abiotic stress resistance. Introduction of these genes to wheat by breeding wheat-rye translocation have been intensively used in wheat breeding program. Rye chromatin 1RS and/or 2RL show superior performance in unfavorable environments. In order to develop high yielding wheat, we applied various molecular breeding strategies. To develop EST-derived 1RS specific markers, we used comparative genomics with public sequence databases of Poaceae family. Putative rye chromatin specific sequences were used to design 1RS specific markers. To identify genes related to water deficiency, cDNA AFLP analysis was used in PEG treated seedlings of 1RS RILs. For functional analysis of identified genes and markers, we used Brachypodium distachyon, as a new model plant of temperate grasses. B. distachyon were recently applied for transformation and we constructed Agrobacterium-mediated transformation system. Integration of those strategies and conventional breeding method would enhance the usefulness of rye chromatins for wheat improvement.

Bacillus thuringiensis (Bt) crystal protein genes encode insecticidal δ-endotoxins that are widely used for the development of insect-resistant crops. Common soybean is a crop of economic and nutritious importance in many parts of the world. Korea soybean variety Kwangan was transformed with Bacillus thuringiensis (Bt) crystal protein genes. These genes were transformed into Kwangan using highly efficient soybean transformation system. Transgenic plants with Bt crystal protein genes were confirmed for gene introduction and their expression using PCR, real-time PCR and RT-PCR. Currently, the confirmation of stable gene introduction with Bt genes is also performing by southern blot analysis and physiology test and agronomic characters are investigating.

Cry1Ac protein is known as one of toxin crystal proteins synthesized from Bacillus thuringenesis that plays a critical role for the insect resistance. Recently, cry1Ac genes have introduced into many plants in general and soybean as well. However, the gene expression of cry1Ac genes in transgenic plants remains low that need to be improved. Several mutations we reintroduced into the cry1Ac genes in order to enhance the insecticidal effect. In this study, the cry1Ac with mutant #2, #11 and #16 were transformed into Kwangan, a Korean soybean variety by using the “half-seed” method. The plant lets carrying modified cry1Ac genes were primarily selected on media containing Phosphinothricine (PPT), a bar selective agent and Basta leaf painting. Then, the presence of introduced genes in T0 plants and the gene expression were investigated by PCR, RT-PCR and Real-time PCR. PCR and RT-PCR analysis showed expression of bar and cry1Ac genes from tested transgenic soybean plants. The number of copy of bar gene ranged from 1 to 3 by Real-time PCR analysis. These results provided a fundamental back ground for our further experiments: Confirmation of the gene expression by Southern blot and identification of the function of modified cry1Ac by insect bioessays.

Korean soybean variety Kwangan was transformed with ORE7 gene using highly efficient soybean transformation system. The gene is known to exhibit a delayed leaf senescence phenotype in Arabidopsis. To confirm phenotypic characterization of leaf senescence for non-transgenic (NT) and transgenic plants, we transplanted T1 transgenic lines 7, 9, 14 and 15 together with two negative controls (NT and EV) in greenhouse. As a result, line 15 showed dramatic phenotypic characterization of yield increase and senescence delay. In addition, to investigate the agriculture traits for transgenic plants with leaf senescence delaying, T2 transgenic lines and two negative controls were transplanted on GMO fields in Ochang and harvested T3 seeds (2010). Most transgenic lines showed higher total seed weigh than NT. Especially, total seed weight of line 15 was increased by about 180% and 120% compared with the NT and EV, respectively. Therefore, we carried out the second field experiments with T3 transgenic line 15 and NT in Ochang (2011). A total of 117 transgenic plants were divided into two groups, senescence delaying (64 out of 117 plants) and increased yield (53 out of 117 plants), by transcript level of ORE7 gene. Interestingly, among increased yield plants, total seed weight of each 7 plants were increased by more than 200% compared with NT.

Korean soybean variety Kwangan was transformed with coat protein (CP), helper component-proteinase (HC-Pro), and ABRE binding factor 3 (ABF3) genes using highly efficient soybean transformation system. Among these genes, CP and HC-Pro were transformed using RNAi technology. Transgenic plants with CP were confirmed for gene introduction and their expression using PCR, real-time PCR, RT-PCR, Southern blot, and Northern blot. To investigate the response of viral infection with CP, T1 plants were inoculated with SMV-infected leaves and confirmed the existence of mosaic symptom in both leaves and seeds. Two transgenic lines with CP were highly resistant to SMV with clear leaves and seeds while SMV-susceptible lines showed mosaic symptom with seed mottling. The transcript levels of T1 plants with CP were also determined by northern blot, suggesting that SMV-resistant T1 plants did not show viral RNA expression whereas SMV-susceptible T1 plants showed viral RNA expression. Currently, the response of viral infection with HC-Pro is investigating to produce SMV-resistant soybean transgenic plants, and the physiological experiment with ABF3 is also carrying out to produce drought-tolerant soybean transgenic plants.

Wild rice might have previously unidentified genes important for disease resistance and stress tolerance in response to biotic and abiotic stresses. A set of subtractive library was constructed both from leaves of wild rice plants, Oryza grandiglumis (CCDD, 2n=48), treated with fungal elicitor and from wounded leaves. A partial fragment that was homologous to PR10 genes from other plant species was identified via suppression subtractive hybridization and cDNA macroarray. The obtained full-length cDNA sequence (OgPR10) contains an open reading frame of 480 bp nucleotide, encoding 160 amino acids with a predicted molecular mass of 16.944 kDa and an isoelectric point (pI) of 4.91. The multiple alignment analyses showed the higher sequence homology of OgPR10 with PR10 genes identified in rice plants at amino acid level. The OgPR10 mRNA was not expressed by treatment with wounding, jasmonic acid, and salicylic acid, but markedly expressed in leaves treated with protein phosphatase inhibitors cantharidin and endothall, and yeast extract. In addition, the expression of OgPR10 mRNA was induced within 72 h after treatment with probenazole, one of well-known chemical elicitors, and reached the highest level at 144 h. Heterologous expression of OgPR10 caused growth inhibition and seedling lethality in E. coli and Arabidopsis, respectively. Chemically induced OgPR10 expression with glucocorticoid-mediated transcriptional induction system further reconfirmed its lethality on Arabidopsis seedling. In addition, OgPR10-expressing rice plants, Oryzae sativar were resistant against the infection of rice blast fungus, Magnaporthe grisea. These results indicate that OgPR10 is involved in probenazole- and microbe associated molecular patterns-mediated disease resistance responses in plants and is a potential gene for developing disease resistance crop plants.

SOHLH2 is a novel germ cell-specific transcription factor that is crucial for folliculogenesis in the ovary and spermatogenesis in the testis. SOHLH2 represents a candidate gene for infertility with premature ovarian failure. We analyzed whether mutations in the SOHLH2 gene in 98 Korean women with premature ovarian failure. The sequence analysis identified six novel SNPs (c.431-41G>C, c.656A>T, c.1000+27C>T, c.1000+33G>T, c1258-106G>A, and c.2094+ 11T>C) from Korean patients with premature ovarian failure. The c.656A>T found in exon 7 results in change of an amino acid, tyrosine to phenylalanine. Functional mutations in SOHLH2 gene are rare in Korean women with premature ovarian failure.