생물반응기에서 배양된 마늘 신초와 Microbulb의 형태발달 및 조직관찰
Morphological Development and Histology of Multiple Shoots and Microbulbs of Garlic Cultured in Bioreactors
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초록
마늘 (Allium sativum L.) 다신초를 생물반응기를 이용, sucrose 2%가 포함된 MS 배지에서 3주간 증식시킨 후 microbulb의 형성을 위해 NAA 0.1 mg/L와 sucrose 11%가 포함된 MS 배지에서 9주간 배양하였다. 다신초 증식의 경우, 잎을 제거하지 않았을 때 90% 이상이 과수화되었고 과수화 된 신초의 세포는 크기와 형태가 일정하지 않았으며 microbulb의 형성도 이루어지지 않았다. 신초 배양 3주 후 표피의 부정아 세포에서 분열이 활성화되면서 부정아가 비대되었고 신초의 분열조직에서 수층과 병층으로 분열이 일어나 크게 비대된 후 배양 7주 부터 microbulb를 형성하였다. Ploidy analyzer를 이용한 배수성 검정에서 다신초와 microbulb의 피크 모두, 모주 식물의 상대적인 피크와 동일한 위치에서 나타나 배수성의 변이가 없는 것으로 나타나 배양체가 모주 식물과 유전적으로 동일함을 입증하였다.
Abstract
Multiple shoots of garlic (Allium sativum L.) were propagated in bioreactors containing MS medium supplemented with 2% sucrose for 3 weeks. Microbulbs were induced on MS medium supplemented with 0.1 mg/L NAA and 11% sucrose for 9 weeks. For multiple shoot proliferation, leaves in the shoot must be removed before cultures. When the multiple shoots were cultured without removal of leaves, more than 90% of hyperhydricity and no microbulb formation were observed. Histological observation also indicated irregular size and shape of the cells in hyperhydricity of the shoot. Microbulbs were strarted to form from the shoot after 7 weeks of culture by protuberance of adventitious shoot buds followed by inner periclinal divisions and simultaneous anticlinical division in the epidermis of meristematic bulge. Analysis of ploidy level indicated no phenotypic variations in both multiple shoots and microbulbs induced from the mother plant, suggesting genetic homogeneity among the regenerants.
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