Alfalfa (Medicago sativa L.) is one of the most important forage crops in the world and it’s has been known as the best feed materials for dairy cows and other high valued animals. The new uses of alfalfa are being explored as bio-energy, food, medical and biochemical uses. R2R3-type MYB transcription factors play important roles in transcriptional regulation of anthocyanin biosynthesis. The R2R3-type IbMYB1 is known to be a key regulator of anthocyanin biosynthesis in the storage roots of sweetpotato. We previously showed that the expression of IbMYB1a led to anthocyanin pigmentation in tobacco and Arabidopsis. In this study, we generated transgenic alfalfa plants expressing the IbMYB1a gene under the control of CaMV 35S promoter. Overexpression of IbMYBa in transgenic alfalfa produced strong anthocyanin pigmentation in seedlings and generated a deep purple color in leaves, stems, roots, and even in seeds. High performance liquid chromatography (HPLC) analysis revealed that IbMYB1a expression led to the production of cyanidin as a major core molecule of anthocyanidins in alfalfa, as occurs in the purple leaves of sweetpotato (cv.Sinzami). We also examined expression of several structural genes in the anthocyanin biosynthetic pathway in alfalfa by RT-PCR analysis. In this presentation, we will further present molecular and biochemical characterization in IbMYB1a-overexpression lines. This result shows that the IbMYB1a transcription factor is sufficient to induce anthocyanin accumulation in the forage legume alfalfa plants.

This paper describes the domestic and international status for melt decontamination, which has been known as the most effective technology for the volume reduction and recycling of the metal wastes generated from nuclear facilities. The recycle or self disposal of metallic wastes can be considered as one of the waste management options under the circumstances of the capacity limitation of a waste disposal in Korea. The limited recycle or self disposal of the metal wastes through an melt decontamination have the merit from the positive view point of the increase in resource recyclability as well as the decrease in the amount of wastes to be disposed resulting the reduction of disposal cost and the enhancement of disposal safety. Among the scenarios for recycle and reuse of the radioactive metallic wastes, the most feasible and reasonable one is limited reuse option, in which the ingot can be recycled as the products such as the waste drums and ISO containers. Prior to recycle and reuse in the nuclear sector, however, the regulatory criteria for the recycle and reuse of metallic wastes should be established in parallel with the development of the recycling technology.

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.

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 (Lolium multiflorum Lam.) is one of important forage crop grass widely cultivated in Korea. Progress in breeding using conventional selection procedure is very slow, since Italian ryegrass is highly self-infertile. Biotechnological approaches, therefore, may contribute to the development of improved cultivars for forage crops. In an effort to optimize tissue culture responses of Italian ryegrass (Lolium multiflorum Lam.) for future genetic manipulations to improve forage characteristics, the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of Korean Italian ryegrass (Lolium multiflorum Lam.) seven cultivars as explant tissues.

Italian ryegrass (Lolium multiflorum Lam.) is one of important forage crop grass widely cultivated in Korea. The genetic manipulation of Italian ryegrass (Lolium multiflorum Lam.) necessitates a reliable and efficient, genotype-independent method of transformation. We are interested in developing molecular breeding methods to improve its nutritional quality and abiotic stress resistance. Development of a rapid and efficient transformation system is the basis for genetic manipulation of Italian ryegrass. In order to establish an efficient Agrobacterium-mediated transformation system was applied to transfer genes into seven genotypes of Italian ryegress, namely cv. 'Kogreen', 'Kopeed', 'Kowinearly', 'Kowinmaster', 'Hwasan 101', 'Hwasan104' and 'Kowinner.' The transformation system developed in this study would be useful for Agrobacterium-mediated genetic transformation of Italian ryegrass plants with genes of agronomic importance.