Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
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Efficient Plant Regeneration from Alfalfa Callus by Osmotic Stress Treatment

알팔파 캘러스로부터 삼투압 스트레스 처리에 의한 효율적인 식물체 재분화

  • Kim, J.S. (Division of Applied Life Science, Gyeongsang National University) ;
  • Lee, D.G. (Division of Applied Life Science, Gyeongsang National University) ;
  • Lee, S.H. (Division of Applied Life Science, Gyeongsang National University) ;
  • Woo, H.S. (Division of Applied Life Science, Gyeongsang National University) ;
  • Lee, B.H. (Division of Applied Life Science, Gyeongsang National University)
  • 김진수 (경상대학교 응용생명과학부, 낙농학전공) ;
  • 이동기 (경상대학교 응용생명과학부, 낙농학전공) ;
  • 이상훈 (경상대학교 응용생명과학부, 낙농학전공) ;
  • 우현숙 (경상대학교 응용생명과학부, 낙농학전공) ;
  • 이병현 (경상대학교 응용생명과학부, 낙농학전공)
  • Published : 2004.10.31
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Abstract

Effects of culture mediwn supplements and osmotic stress treatment on embryogenic callus induction and somatic embryogenesis were investigated in order to optimize tissue culture conditions of alfalfa(Medicago sativa L.). SH mediwn containing 5mgIL 2,4-D and 0.2mgIL kinetin was optimal for embryogenic callus induction from cotyledon tissue of alfalfa. Somatic embryos were formed when the embryogenic callus was cultured on SH mediwn supplemented with ImgIL 2,4-D and 2mgIL BA. Supplementation of 5mM L-proline and IgIL casein hydrolysate into the regeneration mediwn further increased plant regeneration frequency. Osmotic stress treatment of callus appeared to improve the frequency of somatic embryo formation, but the frequency of somatic embryo formation differed by the osmotic stress treatment using different osmotic stressors. The highest plant regeneration frequency of 30.7% was observed when embryogenic callus was treated with 0.7M sucrose for 18h. Efficient regeneration system established in this study will be useful for molecular breeding of alfalfa through genetic transformation.

알팔파의 최적 조직배양 조건을 확립하기 위하여 자엽으로부터 배발생 캘러스 유도 및 캘러스로부터 식물체 재분화에 미치는 배지첨가물질과 삼투압 스트레스 처리의 영향을 조사하였다. 배발생 캘러스 유도시에 첨가되는 식물생장조절물질로는 5mg/L 2,4-D와 0.2mg/L kinetin이 혼용 첨가된 SH 배지에서 배발생 캘러스가 가장 높은 빈도로 유도되었다. 배발생 캘러스를 1mg/L 2,4-D와 2mg/L BA가 첨가된 SH 배지에서 배양했을 때 체세포 배가 형성되었다. 재분화 배지에 첨가되는 질소원으로 5mM L-proline과 1g/L casein hydrolysate를 동시에 첨가해주었을 때, 식물체 재분화율이 증가되었다. 배발생 캘러스를 0.7M 농도의 삼투압 스트레스 조건에서 12 ${\sim}$ 18시간 처리한 후, 재분화 배지에서 배양한 결과 재분화율이 현저히 증가되었다. 가장 높은 식물체 재분화율은 0.7M sucrose가 첨가된 배지에서 18시간 처리한 후, 재분화 배지에서 배양했을 때 30.7%의 효율을 나타내었다. 본 연구를 통하여 확립된 알팔파의 효율적인 배발생 캘러스의 유도 및 식물체 재분화체계는 분자육종을 통한 신품종 알팔파의 개발에 유용하게 이용되어질 수 있을 것이다.

Keywords

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