Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Induction of Embryogenic Callus and Plant Regeneration by Mature Embryo Culture of Onion (Allium cepa L.)

양파의 성숙배 배양을 통한 체세포배발생 캘러스 유기 및 식물체 재분화

  • Cho Kwang-Soo (Horticulture Research Division, National Institute of Highland Agriculture) ;
  • Hur Eun-Joo (Floriculture Research Division, National Horticulture Research Institute) ;
  • Hong Su-Young (Horticulture Research Division, National Institute of Highland Agriculture) ;
  • Moon Ji-Young (Horticulture Research Division, National Institute of Highland Agriculture)
  • 조광수 (농촌진흥청 고령지농업연구소 원예과) ;
  • 허은주 (농촌진흥청 원예연구소 화훼과) ;
  • 홍수영 (농촌진흥청 고령지농업연구소 원예과) ;
  • 문지영 (농촌진흥청 고령지농업연구소 원예과)
  • Published : 2005.03.01
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Abstract

To obtain regeneration system of onion, we analyzed the effects of 2,4-D and BA concentration on the embryogenic callus induction from mature embryos. The highest embryogenic callus induction ratio was shown on MS medium (Murashie and Skoog 1962) containing $2.5\;\cal{mg/L}\;or\;5\;\cal{mg/L}$ picloram after mature embryos were placed on medium. When induced callus were cultured on half strength of MS medium containing $1\;\cal{mg/L}$ Kinetin, the highest shoot formation ratio was observed on MS medium containing $1\;{mg/L}$ 2,4-D and $1\;{mg/L}$ BA. Embryogenic callus were cultured in MS liquid medium containing $1\;\ccal{mg/L}$ of 2,4-D and $1\;\cal{mg/L}$ BA. The suspension cultured cell clumps could be mass propagated. Embryogenic callus were friable, but non-embryogenic callus included a lot of moisture, hence the identification between embryogenic and non-embryogenic callus as easily achieved. When embryogenic callus as cultured on half strength of MS medium containing $1\;\cal{mg/L}$ Kinetin, shoots were induced. The whole plantlet was obtained on rooting medium containing $0.5\;\cal{mg/}$ of NAA.

양파의 재분화 체계 확립을 위해 성숙된 양파의 종자로부터 성숙배를 적출한 후 2,4-D와 BA의 다양한 농도를 이용하여 체세포배발생 캘러스의 형성율을 조사하였다. 체세포 배발생 캘러스는 쉽게 부서지는 특성을 가지고 있었으며 비체세포배발생 캘러스는 물기를 많이 포함하고 있어 쉽게 구분이 가능하였다. 체세포배발생 캘러스는 picloram $2.5\;\cal{mg/L}$ 또는 $5\;\cal{mg/L}$가 단독으로 포함된 MS배지를 이용하여 $25^{\circ}C$ 암배양 4주 후 가장 높은 유도율을 나타내었다. 그러나 유도된 캘러스를 shoot로 재분화 시킨 결과 2,4-D와 BA $1\;\cal{mg/L}$가 혼합된 처리구에서 가장 높은 shoot 유기율을 나타내었다. 따라서 다른 처리에서 유도된 캘러스를 2,4-D와 BA $1\;\cal{mg/L}$가 포함된 액체배지에 현탁배양을 시도하였다. 현탁배양은 캘러스의 증가뿐만 아니라 캘러스의 cell cycle을 synchronized 함으로서 이후 형질전환이 용이할 것으로 판단되었다. 현탁 배양된 캘러스는 Kinetin $1\;\cal{mg/L}$가 포함된 1/2 MS 배지에서 60일간 광배양 하여 shoot 유기하였으며 유도된 shoot는 NAA $0.5\;\cal{mg/L}$가 포함된 발근배지에서 발근 후 순화하여 완전한 식물체로 재생할 수 있었다.

Keywords

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