• 한국초지조사료학회지
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• 제20권2호
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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)

• 식물조직배양학회지
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• 제28권2호
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• pp.75-79
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• 2001

벼에서 분리한 저분자량 HSP 유전자 (OsHSP17.9)를 오차드그래스 (orchardgrass)에 도입하기 위하여 Agrobacterium을 이용한 형질전환을 실시하여 다음의 결과를 얻었다. 오차드그래스의 종자유래의 캘러스를 OsHSP17.9 유전자가 도입된 Agrobacterium EHA101과 공동배양한 다음, hygromycin 선발된 캘러스로부터 hygromycin 저항성 식물체를 얻었다. PCR 및 Southern blot 분석 결과, 벼의 저분자량 HSP 유전자가 재분화된 식물체에 안정적으로 도입되었음을 확인하였으며, 품종 간의 형질전환 효율은 '포토맥'의 경우 16.5%, '프론티어' 의 경우 8.0%를 나타내었다. 또한 Northern blot 분석 결과, 도입된 유전자가 형질전환체에서 정상적으로 발현된다는 것을 확인하였으며, 형질전환체의 계통 간에 발현량의 차이를 나타내었다.

• 한국초지조사료학회지
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• 제20권1호
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• pp.7-12
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• 2000

오차드그래스의 종자배양에서 형성된 캘러스를 현탁배양하여 배양기간별 부정배 형성정도와 식물체 재분화율 등에 대한 몇 가지 실험을 수행하여 얻은 결과를 요약하면 다음과 같다. 종자유래의 캘러스를 현탁배양하였을 때, 세포 모양이 둥근것과 그들의 세포괴는 배양 50일 후에 최대치를 나타내었고 그 이후는 감소하였다. 현탁배양 기간에 따른 갤러스의 부정배 형성과 식물체 분화율은 배양 60일째 가장 높았으며, 그 이후는 감소하는 경향이었다. 현탁배양에서 배지 내에 첨가되는 casein hydrolysate (CH)의 적정농도는 $4\;g/{\ell}$인 것으로 나타났다.

• Journal of Plant Biotechnology
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• 제30권4호
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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.

• 한국초지조사료학회지
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• 제21권1호
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• pp.21-26
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• 2001

환경 스트레스에 의해 야기되는 활성 산소종에 의한 피해에 내성을 가지는 목초의 개발을 위하여 오차드그래스의 배반 조직 유래의 캘러스에 배추유래의 cytosolic glutathione reductase 유전자(BcGRl)를 Agrobucterium tumefaciens EHA101을 매개로 형질전환시켰다. Hygromcin으로 선발된 캘러스로부터 재분화된 식물체는 야생형과 비교하여 형태적으로 차이를 나타내지 않았다. PCR 및 Southern blot 분석을 통하여 형질전환 식물체의 염색체 내에 BcGRl 유전자가 integration 되었음을 확인하였다. 오차드그래스의 잎으로부터 total RNA를 분리하여 Northern blot 분석을 실시한 결과, 도입된 유전자가 형짙전환 식물체 내에서 지속적으로 발현된다는 것을 확인하였다.

• 한국초지조사료학회지
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• 제24권1호
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• pp.25-28
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• 2004

오차드그라스(Dactylis glomerata L.)의 치사온도를 결정하기 위하여, 국내 육성품종인 "장벌 102호"(Jangbeol 102) 품종을 시험재료로 하여 종자를 petri dish에서 발아시켜 작은 화분에 10 개체씩 이식, 생장실에서 4주간 재배하였다. $45^{\circ}C$, $50^{\circ}C$ 및 $55^{\circ}C$에서 처리한 경우에는 60분간 처리했을 때에도 거의 식물체 손상이 없었다. $60^{\circ}C$에서 30분간 이상 처리했을 때에는 잎 끝이 마르고 약간 시들었으나 식물체의 손상은 약한 편이었으며, $65^{\circ}C$에서 처리한 결과에서도 $60^{\circ}C$에서 처리했을 때보다는 좀 더 시들었지만 역시 심한 정도는 아니었다. $70^{\circ}C$에서 30분간 처리했을 때에 오차드그라스는 잎의 중간 정도까지 말랐으며, 60분간 처리했을 때에는 지상부의 잎이 2/3 이상 마른 상태였다. $80^{\circ}C$에서 처리한 결과에서는 처리 후 1일 이내에 50분 이상 처리에서 거의 죽어가는 현상이 나타났다. 처리 후 7일에는 55분 이상의 처리에서만 모두 죽었으며, 50분 이하의 처리에서는 새로운 shoot가 재생됨을 확인하였다. 결론적으로 오차드그라스의 치사온도는 $80^{\circ}C$에서 55분 처리하는 것으로 결정되었다.

• 한국초지조사료학회지
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• 제10권3호
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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.

• 한국초지조사료학회지
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• 제13권3호
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• pp.184-189
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• 1993

수확시기와 칼륨비료의 시용수준이 각각 다른 오차드그라스(Dactylis glomerata L.)를 대사시험용 케이지 안의 염소(산양)에게 급여하여 목초중 무기질함량이 염소의 무기물질 출납에 어떤 영향을 주는가를 알아보고자 본 실험을 실시하였다. 저칼륨급여구의 오차드그라스 건초중 칼륨과 마그네슘함량은 건물량 기준으로 각각 3.4%와0.26%이었고 중칼륨구의 함량은 4.9%와0.21%이었으며 고 칼륨구의 함량은 5.8%와0.21% 이었다. 고칼륨급여구에서 뇨중칼륨배설량은 저칼륨구보다 유의하게 많았고, 저칼륨구의 분중및 뇨중 마그네슘배설량은 중칼륨구 및 고 칼륨구보다 많았다. 뇨중마그네슘배설량은 혈장의 마그네슘농도 정상관의 경향이 있었으며, 고칼륨급여구의 염소(산양)는 다름 급여구에 비해 음수량과 배뇨량이 많은 경향이었다.

• Asian-Australasian Journal of Animal Sciences
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• 제16권1호
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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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• 제21권5호
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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.