오차드그래스의 종자유래 캘러스배양 및 재분화에 미치는 배지첨가물질의 영향

Effects of Medium Supplements on Seed-Derived Callus Culture and Regeneration of Orchardgrass

  • 이상훈 (경상대학교 응용생명과학부) ;
  • 이동기 (경상대학교 응용생명과학) ;
  • 이병현 (경상대학교 응용생명과학부)
  • 발행 : 2004.06.01

초록

오차드그래스의 최적 조직배양조건을 확립하기 위하여 'Roughrider' 품종의 성숙종자로부터 최적 배발생 캘러스 유도 및 식물체 재분화에 미치는 배지첨가물질의 영향을 조사하였다. 성숙종자로부터 배발생 캘러스 유도시 첨가되는 생장조절 물질로는 3 mg/L 2,4-D가 가장 효율적이었으며, 식물체 재분화에는 1 mg/L 2,4-D와 3 mg/L BA가 첨가된 배지에 캘러스를 배양했을 때 36.3%의 재분화율을 나타내었다. 캘러스 유도배지와 재분화배지에 1 g/L의 casein hydrolysate와 300 mg/L의 proline을 동시에 첨가해주었을 때 캘러스 유도율과 재분화율이 각각 57.3%와 60.7%로 증가되었다. 또한 항산화물질로서 10 mg/L의 $\textrm{AgNO}_3$ 와 40 mg/L의 cysteine을 첨가해준 과 켈러스 유도율 재분화율 각각 68.7%와 71.7%까지 증가되었다. 본 연구를 통하여 확립된 성숙종자로부터 효율적인 배발생 캘러스의 유도 및 식물체 재분화 체계는 유전자 형질전환을 통한 신품종 오차드그래스 개발에 유용하게 응용되어질 수 있을 것이다.

In order to optimize tissue culture conditions for genetic transformation of orchardgrass (Dactylis glomerata L.), the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of a cultivar, 'Roughrider', as explant tissues. The optimal concentration of 2,4-D for the induction of embryogenic callus from mature seeds was 3 mg/L. Plant regeneration frequency was 36.3% when embryogenic calli were cultured on the regeneration medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Addition of 1 g/L casein hydrolysate and 300 mg/L L-proline improved frequencies of embryogenic callus induction and plant regeneration up to 57.3 and 60.7%, respectively. Supplementation of the media with 10 mga $\textrm{AgNO}_3$ and 40 mg/L cysteine enhanced frequencies of callus induction and plant regeneration. Efficient regeneration system established in this study will be useful for molecular breeding of orchardgrass through genetic transformation.

키워드

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