Changes in Isozyme Patterns of Peroxidase and Esterase during Regeneration of Pepper (Capsicum annuum L.)

고추 (Capsicum annuum L.)의 재분화에 따른 Peroxidase와 Esterase 동위효소의 양상 변화

The plantlet was regenerated on MS medium containing BAP (2 mg/I) and 1M (1 mg/I) from leaf discs of pepper after 3 weeks of culture. And then, we investigated the activity of peroxidase and esterase and the pattern of their isozymes from leaf, stem and root in order to observe physiological and biochemical changes on the developemental stage, respectively. The peroxidase was expressed with tissue specificity because peroxidase activity according to the developemental stage of the tissue was not only highest in the leaf of the pepper at 10 days after it germinated but also 2 new bands of its isozyme were found in pI 7.2 and pI 5.2. However, a new pI 3.4 band was found in the leaf and root of the pepper after 14 days of germination, and in the stem was found out pI 5.2 band. As regeneration of leaf dises was progressed, its peroxiase activity was increased about 80% more than that of control after 14 days of culture and new pI 3.2 and 6.5 bands of it isozyme were found. The results suggested that peroxidase would be connected with regeneration of pepper. Also, esterase activity was increased about 50% more than that of control after 14 days of culture, the pattern of esterase isozyme was shown to be 3 cathodic bands and 1 anodic band after 7 days of culture.ulture.

고추의 잎 절편을 BAP(2 mg/l)와 IAA(1 mg/l)가 첨가된 MS 배지에서 분화를 유도하여 40일 후에 완전한 식물체를 얻을 수 있었다. 또한 분화 단계에 따라 peroxidase와 esterase의 효소 활성과 isozyme 양상을 조사하였는데, 조직별 peroxidase 활성은 발아 10일째의 잎에서 가장 높았으며, isozyme 양상은 10일째에 잎에서 pI 7.2와 pI 6.9의 새로운 band가 나타났고, 14일째에는 잎과 뿌리에서 새로운 pI 3.4 band가, 줄기에서는 pI 5.2의 새로운 band가 나타나 peroxidase의 조직 특이적 발현을 알 수 있었다. 또한, 잎의 재분화가 진행됨에 따라 peroxidase 효소활성이 증가하기 시작하여 배양 14일째에는 대조구에 비해 80% 정도 높은 효소 활성을 나타냈으며, 분화가 진행됨에 따라 band의 양이 증가하며 배양 14일째에는 pI 3.2와 pI 6.5의 새로운 isozyme이 유도되어 peroxidase가 분화와 발생에 관여하고 있음을 시사하였다. 한편, esterase 활성은 14일째에 대조구보다 2배 이상 높은 활성을 보였으며 esterase isozyme 양상은 배양 7일째에 3개의 cathodic band와 1개의 anodic band가 나타남으로써, 이들 isozyme이 분화시기에 밀접하게 연관되었음을 알 수 있었다.