사과 갈라 품종의 Agrobacterium이용 형질전환에 영향하는 요인

Factors Affecting the Agrobacterium Mediated Transformation of 'Gala' Apple

  • 발행 : 2001.07.01

초록

사과 갈라 품종의 고효율 형질전환 기술체계를 확립하기 위하여 Agobacterium을 이용한 형질전환에 관여하는 상처유도 방법, 배지 고형물, 엽절편체 기원, acetosyringone의 농도와 MES첨가 등 몇 가지 요인이 재분화 및 형질전환에 미치는 영향에 대하여 연구하였다. 엽절편체의 상처유도는 수술용 핀셋 (non-traumatic forcep)의 사용보다는 단순한 절단 후 균접종과 3일 공동배양을 할 경우 재분화와 형질전환율이 더 증가하는 경향을 나타냈다. Agar (A)와 Gelrit $e^{ }$(G)의 조성에 따른 배지 고형물의 처리에서는 Gelrite의 농도가 높을수록 재분화율이 증가하였으나, 형질전환율에 있어서는 2.5 G, 3.5 A+l.2 G 및 7.0 A의 처리 간에 차이가 없었다. 엽신장배지 또는 증식배지 유래가 재분화 및 형질전환율에 유의한 차이를 보이지는 않았으나, 증식배지 유래가 형질전환율을 더 높이는 경향을 나타냈다. Acetosyringone은 0.1 mM 이상의 농도가 첨가할 경우 높은 재분화 및 형질전환을 나타냈고, 0.15 mM의 농도에서 가장 높은 형질전환율을 나타냈다. MES의 재분화와 형질전환에 미치는 영향은 나타나지 않았다.다.

Some factors of wounding methods, solidifying agents, origin of leaf explants, cone. of acetosyring-one, and MES affecting regeneration and transformation by Agrobacterium tumefaciens were investigated to establish an efficient transformation system of apple. Wounding by cutting the leaves merely showed the tendency of regeneration and transformation with higher efficiency compared with that of wounding by non-traumatic forcep when carrying out co-cultivation for three days after bacterial inoculation. While examining the solidifying agents of medium with the combination of agar (A)+Gelrit $e^{ }$ (G) in g. $L^{-1}$ , the higher concentration of Gelrit $e^{ }$ increased the efficiency of regeneration. However, there was no difference in the efficiency of transformation from the treatments of 2.5 G, 3.5 A+1.2 G, and 7.0 G. The origin of leaf explants showed no difference statistically in the efficiency of regeneration and transformation, but that from the shoots of proliferation medium showed the tendency with higher efficiency. The concentration of above 0.1 mM acetosyringon had an increase in the efficiency of regeneration and transformation and the concentration of 0.15 mM had the highest efficiency of transformation in the treatment of acetosyringone with different concentration. There was no effect of MES on regeneration and transformation.ion.

키워드

참고문헌

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