Gene manipulation of Siberian wildrye grass through transgenic technology can modify grass feature for further improvement in the forage production. In order to optimize transformation conditions of Siberian wildrye grass, the effects of transformation efficiency was investigated with GUS gene. Seed-derived calli were infected and co-cultured with Agrobacterium tumefaciens carrying the binary vector pCAMBIA3301 encoding the GUS gene in the T-DNA region. The effects of several factors on transformation and the expression of the GUS gene were investigated. The highest transformation efficiency was obtained when embryogenic calli were inoculated with Agrobacterium strain EHA101 in the presence of 200 μM acetosyringone and coculture for 5 days. The present study was framed and materialized to standardize an efficient, high frequency and reproducible genetic transformation protocol for aromatic Siberian wildrye grass using Agrobacterium tumefaciens.

우라늄 오염토양을 동전기제염 시 많은 양의 동전기 침출액이 발생한다. 발생된 우라늄 침출액을 재이 용하기 위한 처리기술이 개발되었다. 동전기제염 시 발생된 우라늄침출액 내의 우라늄농도는 180 ppm이 었고, Mg(II), K(I), Fe(II), Al(III) 농도는 20 ppm∼1,210 ppm이었다. 우라늄침출액의 최적 처리공정은 혼합, 응집, 침전, 농축, 그리고 여과로 구성된다. 침전액의 pH를 11로 맞추기 위해, calcium hydroxide 는 3.0g/100ml 그리고 sodium hydroxide는 2.7g/100ml이 필요했다. 여러 침전실험 결과 NaOH+0.2g alum+0.15g magnetite가 여과를 위한 최적 침전혼합제로 선정되었다. NaOH+0.2g alum+0.15g magnetite 투입 시 침전입자의 평균크기는 600 ㎛이었다. pH=9에서 침전 후 상등액에 총 금속농도가 가 장 낮았기 때문에, 최적 침전을 위하여 먼저 0.2g/100 ml alum와 0.15g/100ml magnetite 투입한 후pH=9일 때까지 sodium hydroxide을 투입하여야 한다

Antibiotics marker-free with herbicide-resistant tall fescue plants were produced through Agrobacterium: mediated transformation system. Embryogenic calli derived calli were infected with Agrobacterium tumefaciens strain EHA105 harboring the pCAMBIA3300 vector containing the bar and the CP4-EPSPS genes. The PPT-resistant calli and plants were selected with 10 ㎎/L PPT, respectively. Soil-grown plants were obtained about 14-16 weeks after Agrobacterium-mediated transformation. Genetic transformation of the regenerated plants growing under selection was demonstrated by PCR, and Southern blot analysis revealed that one copies of the transgene were integrated into the plant genome of each transgenic plant. Expression of the bar and CP4-EPSPS genes in transgenic plants was confirmed by Northern blot analysis and CP4-EPSPS genes in transgenic plant ELISA uses antibody protein by ELISA Assays. Transgenic plants sprayed of two herbicides with glufosinate ammonium or glyphosate remained green and healthy. We therefore report here a successful and reliable Agrobacterium-mediated transformation of two herbicide-resistances and this method may be useful for routine transformation and has the potential to develop new varieties of tall fescue with several important genes.

In the present study, we have developed a high-frequency plant regeneration system for Italian ryegrass via callus culture using mature seeds as explants. Optimal embryogenic callus induction was found to occur in MS medium containing 5 ㎎ 1?¹ 2,4-D, 0.5 ㎎ 1?¹ BA, 500 ㎎ 1?¹ L-proline, 1 g 1?¹ casein hydrolysate, 30 g 1?¹ sucrose, 7 ㎎ 1?¹ AgNO₃, 2 ㎎ 1?¹ CuSO₄ and solidified with 3 g 1?¹ Gelrite. The highest regeneration rate was obtained in MS medium containing 1 ㎎ 1?¹ 2,4-D, 5 ㎎ 1?¹ BA, 500 ㎎ 1?¹ L-proline, 1 g 1?¹ casein hydrolysate, 1 ㎎ 1?¹ thiamine-HCl, 30 g 1?¹ sucrose, 7 ㎎ 1?¹ AgNO₃, 2 ㎎ 1?¹ CuSO₄ and solidified with 3 g 1?¹ Gelrite. By using the most effective treatment determined for each parameter, the highest rates of embryogenic callus formation (48.9%) and regeneration (47.6%) were obtained with the Hwasan 101 cultivar. The overall plant regeneration rates of the examined cultivars ranged from 7.5% to 23.2%. Thus, optimization of regeneration frequency using mature seeds as explant material may offer a simple and efficient protocol for Italian ryegrass that may improve molecular breeding of this species.

In the present study, genotypic variation of Agrobacterium-mediated transformation of Korean Italian ryegrass has been evaluated. Mature seed-derived calli of a total of seven cultivars were infected and co-cultured with Agrobacterium tumefaciens carrying the binary vector pCAMBIA1301, which contains a reporter gene (gus) and a plant selectable marker gene conferring resistance to hygromycin (hpt) in the T-DNA region. The effects of several factors such as callus type and callus age on transformation frequency and the expression of the GUS gene were investigated. The highest transformation frequency (6.7%) was obtained with the Hwasan 101 cultivar when 9-week-old calli (type-I) were inoculated with Agrobacterium. The overall transformation rates of the examined cultivars ranged from 0.4% to 6.7%. GUS histochemical assays, PCR, and southern analysis of transgenic plants demonstrated that transgenes were successfully integrated into the genome of Italian ryegrass. Thus, optimization of transformation frequency and selection of a suitable cultivar of Italian ryegrass may improve molecular breeding of this species.

Environmental stress is a growing problem for the productivity of forage crops. Reactive oxygen species (ROS) formation due to several abiotic stresses is a fundamental process that interrupts several physiological processes and causes a significant reduction on growth and yield of many forage crops. Molecular breeding such as genetic transformation has become a popular biotechnological tool for improving forage quality as well as improves tolerance to various abiotic stresses. As a first step of genetic transformation in tall fescue, we established an efficient Agrobacterium-mediated genetic transformation protocol using mature seed-derived embryogenic callus. After optimization of the transformation system, several genes of interest have been used to generate abiotic stress tolerant forage grasses. We generated transgenic tall fescue plants expressing NDPK genes under the control of the oxidative stress-inducible SWPA2 promoter. Transgenic plants showed enhanced tolerance to several abiotic stresses. Results in the current study, suggest that NDPK mediated multiple stress tolerance by increasing the expression of genes involved in antioxidant and protective functions, possibly through activation of an MAPK cascade.

The experimental work was conducted to investigate the effects of the application level of composted cattle manure (CCM) on forage productivity using whole crop barely (WCB)-whole crop rice (WCR) double cropping system for 3 years. Main plot was consisted of application level of CCM such as 150%, 200% and control plot. The total DMY of forages under WCB and WCR in treatment of CF significantly decreased as compared with that of treatment of CCM 150%. However, NDF, ADF and CP content of WCB and WCR were hardly influenced by CCM application.

The experimental work was conducted to investigate the effects of the application level of composted cattle manure (CCM) on forage productivity using Italian ryegrass-WCR double cropping system for 3 years (2006~2008). Main plot was consisted of application level of CCM such as 150%, 200% and control plot. The total day matter yield (DMY) of Italian ryegrass and WCR in treatment of CCM 150% and CCM 200% significantly decreased as compared with that of treatment of chemical fertilizer (CF). However, NDF, AND and CP content of Italian rye grass and WCR were hardly influenced by CCM application.

The experimental work was conducted to investigate the effects of the application level of composted cattle manure (CCM) on forage productivity using rye-whole crop rice (WCR) double cropping system for 3 years. Main plot was consisted of application level of CCM such as 150%, 200% and control plot. The total DMY of of rye-WCR in treatment of CCM 150% significantly decreased as compared with that of treatment of CF and CCM 200%. However, NDF, ADF and CP content of rye and WCR were hardly influenced by CCM application.

Managing the fatty acid composition of grazing ruminant diets could lead to meat and milk products t㏊t have higher conjugated linoleic acid (CLA) concentrations, but forage fatty acid dynamics must be more fully understood for a range of forages before grazing systems can be specified. The fatty acid profiles of 23 different forages, including grasses, legumes and wild forage plants were determined. Linolenic(C18:3, 41.1~58.6% of fatty acid), linoleic(C18:2, 10.1~27.4% of fatty acid), and palmitic(C16:0, 9.8~18.0% of fatty acid) acids were the most abundant fatty acids in all species, together representing ~96% of the fatty acids present. Concentrations of fatty acids declined as plants developed, but the fraction contribution of each fatty acid to total fatty acids remained relatively stable over time.

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.