In the present study, we have used an annealing-control-primer (ACP)-based differentially display RT-PCR method to identify salt-stress-induced differentially expressed genes (DEGs) in barley leaves. Using 120 ACPs, a total of 11 up-regulated genes were identified and sequenced. Temporal expression patterns of some up-regulated DEGs in response to salt stress were further analyzed by Northern blot analysis. The possible roles of these identified genes are discussed within the context of their putative role in response to salt stress. Thus, the identification of some novel genes-such as SnRK1-type protein kinase; 17 kDa, class I, small heat shock protein; and RNase S-like protein precursor genes-may offer a new avenue for better understanding the salt stress response in plants, knowledge which might be helpful for developing future strategies.

In this study, we conducted to select the promising crops for both uses in the bioethanol and forage production in Korea. The result indicated t㏊t Natsukaje (guinea grass), Gwangpyeongok (corn), Jumbo (sorghum×sudangrass hybrid), SS405 (sorghum×sorghum hybrid), Millex32 (pearl millet), Jeju barnyard grass), Alamo (switch grass) and Selection75 (klein grass) showed the production of biomass from the highest to the lowest in order. However, the order of the production of quality forage was, from the highest to the lowest, Natsukaje (guinea grass), Jumbo (sorghum×sudangrass hybrid), SS405 (sorghum×sorghum hybrid), Gwangpyeongok (corn), Millex32 (pearl millet), Selection75 (klein grass), Jeju (barnyard grass), and Alamo (switch grass). We concluded the Natsukaje (guinea grass) was the best bioethanol crop, and also the Natsukaje (guinea grass) was the best for forage production.

To know adaptability and production of silage corn, we studied on paddy field experiment. Heading date of 'Kwangpyongok' and 'P32P75' Hybrids was on 15th July, 14th July respectively. The dry matter yield of 'P32P75' hybrid (20.4ton/㏊) was the highest among 10 hybrids and that of 'Kwangpyongok' hybrid (19.1ton/㏊) showed high in domestic corn hybrids and other characters as like feed vales is almost same in 10 hybrids.

Background : Although ginseng has various bioactive compounds in it, there is lack of study on the variations of bioactive compounds in ginseng according to the cultivation soil and the applied fertilizer types (or amount). Therefore, this study aims to examine the variations of 37 fatty acids (FA) and 8 vitamin E (Vit-E) vitamers in 6-year-old ginseng root cultivated in different soil types with different fertilizers regimes. Methods and Results : The profiling of 37 FAs and 8 Vit-E vitamers in 6-year-old ginseng roots was measured by gas chromatography coupled with a flame ionization detector, and then these results were statistically analyzed with chemometrics. The FA and Vit-E content in ginseng roots varied significantly with respect to soil cultivation conditions due to organic fertilizer types and amounts used. Unsaturated FA in ginseng is approximately 2.7 fold higher than the saturated FA. Linoleic, palmitic, and oleic acids were the most abundant FAs found in the ginseng roots. Also, the major Vit-E vitamer found in ginseng root is α-tocopherol. In particular, the application of rice straw compost or food waste fertilizer was increased to create nutritionally desirable FAs and bioactive Vit-E in ginseng root. In addition, phytonutrient profiling coupled with chemometrics can be used to discriminate the cultivation conditions of ginseng. Conclusion : This study extends our understanding about the variations of FA and Vit-E in ginseng root depending on cultivation conditions. Hence, these results can be useful as basic information for reliable ginseng production containing high amounts of phytonutrients in a paddy-converted field.

Background : The geographical origin of Panax ginseng Meyer, a valuable medicinal plant, is important to both ginseng producers and consumers in the context of economic profit and human health benefits. We therefore aimed to discriminate between the cultivation regions of ginseng using the stable isotope ratios of C, N, O, and S, which are abundant bio-elements in living organisms. Methods and Results : The C, N, O, and S stable isotope ratios were measured by isotope ratio mass spectrometer, and then these isotope ratios profiling was statistically analyzed with chemometrics. The various isotope ratios found in Panax ginseng roots were significantly influenced by region, cultivar, and the interactions between these two factors (P ≤ 0.0002). In particular, δ18O was lower in ginseng roots grown at high altitudes (r = −0.47), while δ34S was higher in ginseng roots grown close to coastal areas (r = −0.48). Chemometric results provided discrimination between the majorities of different cultivation regions. Conclusion : Our case study extends the understanding about the variation of C, N, O, and S stable isotope ratios in ginseng root depending on cultivation region. Hence, the analysis of stable isotope ratios is a suitable tool for discrimination between the regional origins of ginseng samples from Korea, with potential application to other countries.

Background : The natural stable isotope ratio of common bio-elements like carbon (C), nitrogen (N), oxygen (O), or sulfur (S) varies with diverse isotope fractionation processes in nature. Therefore, measuring the variation of these stable isotope ratios in ginseng roots can be a feasible tool to discriminate the geographical origins of ginseng in Korea. Methods and Results : The 3-year-old six Korean ginseng cultivars were cultivated at the five regions in Korea, and then used for measuring the stable isotope ratios of C, N, O, and S by isotope ratio mass spectrometry (IRMS). The mean C, N, O, and S stable isotope ratio values in the ginseng roots significantly differed according to the cultivation regions (p < 0.05). However, these isotope ratios in ginseng roots had relatively weak discriminative power against to the ginseng cultivars at each cultivation region. The interaction of the cultivation region and ginseng cultivar type also significantly affected to the C, N, O, and S stable isotope ratio in ginseng roots (p < 0.0001). The two-dimensional plots associated with the N stable isotope ratio can effectively separate the ginseng roots in Jinan compared to those in the other regions. The partial least squares-discriminant analysis showed more significant separation between ginseng geographical origins compared to the principal component analysis. Conclusion : Our findings improve our understanding of how the isotope composition of ginseng roots varies with respect to cultivation regions and cultivars, and suggest that the analysis of the stable isotope ratios combined with chemometrics can be used as a feasible tool to discriminate geographical origin of ginseng in Korea.

High yield is the most important trait in various agricultural characteristics. Many approaches to improve yield have been tried in conventional agricultural practice and recently biotechnological tools employed for same goal. Genetic transformation of key genes to increase yield is one way to overcome current limitation in the field. We are producing transgenic soybean plants through high efficient transformation method by introducing all gene member with AT-hook binding domain, hoping to obtain manageable delay of senescence. Many transgenic soybean plants are growing in greenhouse and GMO field, and will be evaluated their senescence and any association with yield increase.

Soybean is a crop of importance economically and nutritionally in many parts of the world. Thanks to many new genes brought from genomic research, It is possible to introduce various candidate genes through genetic transformation to see the performance of the genes in field. In our lab, soybean transformations have been tried for last 10 years to probe the possibility of traits improvement by transformation of new gene into soybean. For this purpose, three different genes were transformed into Korean soybean variety, Kwangan. First, the gene that controls early flowering of plant was transformed into Kwangan. Second, a candidate gene for soybean mosaic virus (SMV) resistance was transformed to produce transgenic plants. Third, another candidate gene for drought tolerance was transformed. All the transgenic plants from three genes transformation were produced for their gene insertion and their expression using PCR, qRT-PCR. Further analysis including harvesting seeds is currently undertaken.

Small heat shock proteins (Hsps) are one of most conserved molecular chaperones that protect stress-inducible denaturation of substrates in living organisms. Small Hsps consist of a large subfamily categorized by subcellular localization ranging in size from 12 to 40 kDa. Here, we identified and characterized a small Hsp 16.9 gene (EsHsp16.9) from Siberian wild rye (Elymus sibiricus L.). EsHsp16.9 is a 456-bp cDNA with an open reading frame predicted to encode a 151-amino acid protein. It possesses a conserved ɑ-crystallin domain, which is a unique domain for small Hsps; shares high sequence similarity with cytosolic class I small Hsps among the small Hsp subfamily in Arabidopsis; and is close (96% similarity) to small Hsp in wheat. Northern blot analysis showed that EsHsp16.9 transcripts were enhanced by heat, drought, arsenate, methyl viologen, and H2O2 treatments. Moreover, we expressed and purified recombinant EsHsp16.9 proteins in Escherichia coli to confirm its activity as a molecular chaperone. We found that recombinant EsHsp16.9 exhibits effective molecular chaperone activity, as determined by inhibition of thermal aggregation of malate dehydrogenase (MDH), which is broadly used as a model substrate.

To develop transgenic forage crops with enhanced tolerance to abiotic stress, we introduced an alfalfa Hsp23 gene expression vector construct through Agrobacterium-mediated transformation. Integration and expression of the transgene were confirmed by PCR, northern blot, and western blot analyses. Under normal growth conditions, there was no significant difference in the growth of the transgenic plants and the non-transgenic controls. However, when exposed to various stresses such as salt or arsenic, transgenic plants showed a significantly lower accumulation of hydrogen peroxide and thiobarbituric acid reactive substances than control plants. The reduced accumulation of thiobarbituric acid reactive substances indicates that the transgenic plants possessed a more efficient reactive oxygen species-scavenging system. We speculate that the high levels of MsHsp23 proteins in the transgenic plants protect leaves from oxidative damage through chaperon and antioxidant activities. These results suggest that MsHsp23 confers abiotic stress tolerance in transgenic forage crops and may be useful in developing stress tolerance in other crops.

Abiotic stress is the major limiting factor of forage crops growth and yields. The objective of this work was to study the stress tolerance and regeneration capability of transgenic forage crops carrying a MsHSP23 gene, encoding a alfalfa mitochondrial sHSP protein. The expression of the MsHSP23 gene was confirmed in bacteria, recombinant mHSP23 conferred tolerance to salinity and arsenic stress. Furthermore, mHSP23 was cloned in a plant expressing vector and transformed into forage crops such as alfalfa, tall fescue and bent grass. The transgenic plants exhibited enhanced tolerance to salinity and arsenic stress conditions. In comparison to wild type plants, transgenic plants were exhibited significantly lower electrolyte leakage. Moreover, the transgenic plants had superior germination rates when placed on medium containing arsenic. Taken together, these overexpression results imply that mHSP23 plays an important role in salinity and arsenic stress tolerance in transgenic forage crops. This approach could be useful to develop stress-tolerant plants including forage crops.

The molecular responses to various abiotic stresses were investigated by the approaches with transcriptomic analysis based on an ACP system. Here we identified differentially expressed genes under abiotic stresses in alfalfa seedlings and they were mostly unknown genes and a few common stress-related genes. Among them, mitochondrial small HSP23 was responded by the diverse stress treatment such as heat, salt, As stresses and thus it could be a strong candidate that may confer the abiotic stress tolerance to plants. When expressed in bacteria, recombinant MsHSP23 conferred tolerance to salinity and arsenic stress. Furthermore, MsHSP23 was cloned in a plant expressing vector and transformed into tobacco, a eukaryotic model organism. The transgenic plants exhibited enhanced tolerance to salinity and arsenic stress under ex vitro conditions. In comparison to wild type plants, the transgenic plants exhibited significantly lower electrolyte leakage. Moreover, the transgenic plants had superior germination rates when placed on medium containing arsenic. Taken together, these overexpression results imply that MsHSP23 plays an important role in salinity and arsenic stress tolerance in transgenic tobacco. The results of the present study show that overexpression of alfalfa mitochondrial MsHSP23 in both eukaryotic and prokaryotic model systems confers enhanced tolerance to salt and arsenic stress. This indicates that MsHSP23 could be used potentially for the development of stress tolerant transgenic crops, such as forages.

Tall fescue (Festuca arundinacea Schreb.) is an important cool season forage plant that is not well suited to extreme heat, salts, or heavy metals. To develop transgenic tall fescue plants with enhanced tolerance to abiotic stress, we introduced a MsHsp23 gene expression vector construct through Agrobacterium-mediated transformation. Integration and expression of the transgene were confirmed by PCR, northern blot, and western blot analyses. Under normal growth conditions, there was no significant difference in the growth of the transgenic plants and the non-transgenic controls. However, when exposed to various stresses such as salt or arsenic, transgenic plants showed a significantly lower accumulation of hydrogen peroxide and thiobarbituric acid reactive substances than control plants. We speculate that the high levels of MsHsp23 proteins in the transgenic plants protect leaves from oxidative damage through chaperon and antioxidant activities. These results suggest that MsHsp23 confers abiotic stress tolerance in transgenic tall fescue and may be useful in developing stress tolerance in other crops. Compared with traditional plant breeding, genetic engineering provides a relatively fast and precise means of achieving improved stress tolerance of forage crops. Development of forage crops that are more tolerant to various abiotic stresses could lead to the use of more new lands for cultivation.

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.

Most forage crops growing under field conditions are often being exposed to various environmental stresses such as drought, freezing, high temperature, waterlogging and climate change. A combination of grass breeding approaches will likely be needed to improve significantly the environmental stresses tolerance of forage crops in the field. Attempts have been taken by grass breeders to develop tolerant varieties of different crops for environmental stress. A new tall fescue variety (Festuca arundinacea Schreb.) named ‘Purumi’ was developed by the National Institute of Animal Science, Rural Development Administration from 1999 to 2007. For synthetic seed production of this new variety, 5 superior clones, EFa9108, EFa0010, EFa0020, EFa0108, and EFa0202 were selected and polycrossed. The agronomic growth characteristics and forage production capability of the seeds were studied at Cheonan from 2004 to 2005, and regional trials were conducted in Cheonan, Pyungchang, Jeju, and Jinju from 2008 to 2010. ‘Purumi’ showed enhanced winter hardiness, disease resistance, and regrowth ability as compared to ‘Fawn’. The dry matter yield of ‘Purumi’ was about 5.6% higher as 16,821kg/ha than that of ‘Fawn’. However, the nutritive value of both varieties was similar. When this new variety of tall fescue, Purumi, has been developed and distributed with its most remarkable adaptability for Korean climates and superior value as a livestock feed, it is expected to play an important role for a new restoration of the pasture industry in Korea.

Italian Ryegrass is a high productivity and feed value, and an upright grass that behaves like a biennial or short-lived perennial. It grows vigorously in winter and early spring. Italian ryegrass and a related species, perennial ryegrass, Lolium perenne, are the two common weedy ryegrassses. Italian ryegrass and perennial ryegrass can hybridize, resulting in offspring that are difficult to identify as either species. Ryegrasses are cultivated for turf and forage. Sometimes Italian ryegrass is grown as cover crop. It has the potential to produce high yields and, with proper management, can be high quality with good animal performance. To develop of a high quality, productivity and early variety, 20 varieties examined growth and yield characteristics. The heading date of Green farm with high cold tolerance was on 28 April. Most of Introduced varieties except Grazer have mid and late heading date.