• Journal of The Korean Society of Grassland and Forage Science
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• v.20 no.2
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• pp.103-108
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• 2000

To produce of transgenic orchardgrass, the seed-derived calli of orchardgrass (Dactylis glomerata L.) co-cultivated with Agrobacterium turnefaciens EHAlOl harboring binary vector pIG121-Hm were selected with hygromycin and then transferred onto N6 regeneration medium containing 1 rngl l of NAA, 5 rngl l of kinetin, 250 rngl l of carbenicillin and 50 mg/ l of hygromycin. The efficiency of transformation was differed on cultivars, that is, 'Potomac' appeared 12% of transformation efficiency while 'Amba' did 5.5%. The addition of acetosyringone during co-cultivation was a key to successhl transformation of orchardgrass. Transgene fragments were identified by PCR analysis and the constitutive expression of GUS gene was confirmed by Northern blot analysis. (Key words : Acetosyringone, Agrobacterium tumefaciens, Orchardgrass (Dactylis glomerata L.), Transformation)

• Korean Journal of Plant Tissue Culture
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• v.28 no.2
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• pp.75-79
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• 2001

An experiment was carried out to introduce OsHSP17.9, a low molecular HSP gene isolated from rice plant to orchardgrass (Dactylis glomerata L.) using Agrobacterium. Mature seed-derived calli of orchardgrass were co-cultured with Agrobacterium tumefaciens EHA101 harboring the plasmid pIG-HSP17.9 for transformation. Calli selected by hygromycin were transferred to N$_{6}$ medium containing 1 mg/L NAA, 5 mg/L kinetin, 250 mg/L cefotaxime and 50 mg/L hygromycin and several hygromycin resistant plants were obtained. Stable incorporation of the introduced OsHSP17.9 to the genome of the hygromycin resistant plants was confirmed by PCR and Southern blot analysis. Transformation efficiency was variable between cultivars in which it was 16.5% in Potomac and 8.0% in Frontier. Constitutive expression of the transgene in the transformed orchardgrass tissues was identified by Northern blot analysis but transcript levels were different among individual plants.s.

• Journal of The Korean Society of Grassland and Forage Science
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• v.20 no.1
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• pp.7-12
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• 2000

This study was carried out to improve the ability of embryo formation and the efficiency of plant regeneration from suspension cultured cells of seed derived calli of orchardgrass (Dactylis glomerata L.). The frequency of formation of round cell and cell colonies was highest at 50 days after suspension culture in $N_6$ medium supplemented with $4\;g/{\ell}$ casein hydrolysate (CH), $20\;g/{\ell}$ sucrose and $30\;g/{\ell}$ sorbitol. The highest frequency of plant regeneration and somatic embryo formation was obtained from suspension cultured cells of 60 days. Addition of CH ($4\;g/{\ell}$) in suspension culture medium gave the highest frequency of embryo formation (39.6%) and plant regeneration (73.0%).

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.341-346
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• 2003

In an effort to optimize tissue culture conditions for genetic transformation of orchardgrass (Dactylis glomerata L.), an efficient and high-frequency plant regeneration system from seed-derived calli was established. Embryogenic calli induced on MS medium containing 3mg/L 2,4-D and 0.1mg/L BA had significantly improved regeneration ability. Plant regeneration rate was 92％ when embryogenic calli were cultured on N6 medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Among three kinds of medium, MS and N6 medium were optimal for embryogenic callus induction and plant regeneration, respectively. Ho difference in callus induction frequency was observed among four cultivars of orchardgrass, however, "Roughrider" cultivar showed higher regenerability with the frequency of 61％. Addition of maltose to the regeneration medium as a carbon source dramatically increased regeneration frequency up to 69％. A short tissue culture period and high-frequency regeneration system would be beneficial for molecular breeding of orchardgrass through genetic transformation.

• Journal of The Korean Society of Grassland and Forage Science
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• v.21 no.1
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• pp.21-26
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• 2001

To develop transgenic orchardgrass resistant to reactive oxygen species produced from environmental stresses, a vector with the cytosolic glutathione reductase cDNA (BcGRl) from Chinese cabbage was constructed under the control of the cauliflower mosaic virus 35S promoter and was introduced into orchardgrass using Agrobacterium tumefaciens EHA101. Transgenic plants from hygromycin-selected calli of orchardgrass did not show any morphological difference from wild-type plants. The results of PCR amplification and genomic Southern blot analysis confirmed the integration of foreign gene into the chromosome of transgenic orchardgrass. Northern blot analysis with total RNA from leaves also confirmed the constitutive expression of BcGR1 in transgenic orchardgrass.

• Journal of The Korean Society of Grassland and Forage Science
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• v.24 no.1
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• pp.25-28
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• 2004

To determine lethal temperature of orchardgrass (Dactylis glomerata L. cv. Janbeol 102) developed in Korea at heat-stressed conditions, seedlings grown in a amall pots for 4 weeks were treated at $45^{\circ}C$, $50^{\circ}C$ or $55^{\circ}C$ for 1 h. Heat treatments at $60^{\circ}C$ and $65^{\circ}C$ for 1 h, several plants were withered and showed damage symptom on their leaves. When the plants were exposed to $70^{\circ}C$ for 1 h, most of leaves were severely withered, but it was not lethal conditions for the whole plants. By contrast, most of plants were died within one day after heat treatment at $80^{\circ}C$ for 1h. Furthermore, plants exposed to $80^{\circ}C$ for 55 min were also died within 7 days. It was found that new shoots were regenerated from the plants that had been treated at $80^{\circ}C$ within 50 min. These results indicate that heat treatment at $80^{\circ}C$ for 55 min is an optimum condition to distinguish the lethality of orchardgrass plants. Simple viability assay system established in this study will be useful for selection and characterization of heat-tolerant transgenic orchardgrass plants.

• Journal of The Korean Society of Grassland and Forage Science
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• v.10 no.3
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• pp.141-146
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• 1990

Changes in dry matter yield, crude components, nutrients uptake($P_2O_5$, $K_2O$, CaO, MgO) and sulphur containing amino acid(cysteine, methionine) of orchardgrass(Dacty1is glomerata. L) and alfalfa(A4edicago sativa. L) by gypsum application(as sulphur source, 0, 2. 5, 10, 20kg SIlOa) were investigated to understand the effect of sulphur on herbage production in pasture, which was established in 1987 as means of hand broadcasting. The effect of gypsum on dry matter yield at different cutting times during growing seasons has not been found both in orchardgrass and in alfalfa, but in respect to annual total dry matter yield there were increment in herbage yield (P<0.05) of alfalfa at 5, 10, 20kg SIlOa in 1989 and the amounts of sulphur taken up in herbage slightly increased according to the rates of gypsum application. Maximum apparent recovery of sulphur was 7.55% at 2kg SIlOa in orchardgrass and was 17.8% at 5kg S/lOa in alfalfa. There were no any great differences in the content of crude components of both species and this trend was similar with the mineral contents of orchardgrass. But in alfalfa, the amounts of $K_2O$, CaO, and $P_2O_5$ taken up were increased by gypsum application and the increment in the amounts of minerals taken up in herbage at 20kg SIlOa were 14.9 of $K_2O$, 9.1 of CaO, and 2.5kgIlOa of $P_2O_5$ as compared to those of at untreated plot. Cysteine and methionine were not influenced by gypsum applicaton not only in orchardgrass but also cysteine in alfalfa, however, the content of methionine in alfalfa was slightly increased at 2, 5, lOkg SIlOa and at 20kg SIlOa was reverse.

• Journal of The Korean Society of Grassland and Forage Science
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• v.13 no.3
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• pp.184-189
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• 1993

Orchardgrass(Dactylis glomerata L.) hays, which had been harvested in different seasons and grown with different levels of potassium (K)fertilizer, were fed to foats in metabolic cages in order to know the effect forage mineral content on mineral balance of goats. The K contents of the feed were 3.4, 4.9 and 5.8% and magnesium (Mg) contents were 0.26, 0.21 and 0.21% on a det matter(DM) basis in K-low, K0medium and K-high treatments, respectively. Urinary K excretion was significantly more on K-high than on K-low treatment. Faecal and urinary Mg excrerions on K-low were higher than those on K-medium and K-high feeds. And there was a tendency of positive relation between urinary Mg excretion and serum Mg concentrarion. Goats on K-high treatment seemed to drink more water and excrete more urine than those on the other feeds.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.1
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• pp.83-87
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• 2003

The study evaluates the enzymatic dry matter (DM) degradability and water holding capacity of leaf and stem fractions of orchardgrass (Dactylis glomerata L.) at different growth stages with or without the presence of surfactant Tween 80. While Tween 80 significantly (p<0.05) increased water and enzyme holding capacities in the leaf blades fraction, less was observed in the fraction of leaf sheath and stem of orchardgrass. The enzyme holding capacity in the leaves was also altered more than that for water holding capacity. This resulted in the increased rate and extent of enzymatic hydrolysis of the leaf blade fractions at two growth stages, whereas little was with leaf sheath and stem fractions. It was also observed that at 0.005% concentrations of Tween 80 the enzymatic DM degradability of young leaf blades was higher (p<0.05) by 20-30% compared to that of the control, as well as for water and enzyme holding capacity. For matured leaf blades the DM degradability were increased with over 0.01% concentrations of the surfactant, but the increase was less than leaf blades of young orchardgrass. This result suggests the possibility of using the surfactant Tween 80 to improve forage digestibility in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.5
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• pp.657-662
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• 2008

To develop transgenic orchardgrass (Dactylis glomerata L.) resistant to high temperature, the recombinant DgHSP17.2 gene was introduced into orchardgrass plants using the Agrobacterium-mediated transformation method and expressed constitutively under the control of the CaMV 35S promoter. The results of genomic DNA PCR and Southern analysis showed a DNA band and hybridization signal on agarose gel and X-ray film in transgenic orchardgrass plants harboring the recombinant DgHSP17.2 gene, but a DNA band and hybridization signal were not observed in the wild type and empty vector control plants. The same result was also obtained in RT-PCR and Southern blot analysis, and these transgenic orchardgrass plants did not show any morphological aberration both in the culture bottle and soil mixture. When leaf discs cut from transgenic orchardgrass plants with recombinant DgHsp17.2 gene were exposed to lethal temperature (heat treatment at $60^{\circ}C$ for 50 min), 60-80% of the leaf discs showed only damage symptoms, but non-transgenic leaf discs showed a lethal condition. These results indicate that the DgHsp17.2 gene may act as a protector from heat stress in plants.