동백나무 미숙배 배양으로부터 비정상 체세포배 형성과 식물체 재생
Anomalous somatic embryos formation and plant regeneration from the cultures of immature embryos of Camellia japonica L.
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동백의 배발생 캘러스는 1 mg/L 2,4-D가 첨가된 MS배지에서 미숙배 배양을 통해 유도하였으며, 동일배지에 계대 배양하여 4주 이상 증식시켰다. 배발생 캘러스로부터자엽기 체세포배를 생산하기 위하여 2,4-D가 첨가되지않은 MS기본배지 배양하였다. 동백 미숙배로부터 체세포배 발생율은 25.1%였으며, 형성된 대부분의 체세포배중에서 48.3%가 cup모양, 12.6%가 1개 자엽, 9.4%가 3개자엽, 1.9%가 4개 자엽을 갖는 비정상적인 형태의 체세포배였으며, 반면 2개의 자엽을 갖는 정상적인 체세포배는27.5%였다. 정상 및 비정상형태의 체세포배를 MS기본배지, ½MS배지, 0.1 mg/L GA3가 첨가된 ½MS배지 및 0.1mg/L IBA와 GA3가 조합 첨가된 ½MS배지에 치상하였을때 2개의 자엽을 갖는 정상체세포배의 식물체 재생율이11.1%로 0.0 ~ 8.3%를 나타내는 1개 자엽 (0.0 ~ 8.3%), 3개자엽 (0.0 ~ 5.8%), 4개 자엽 (0.0%) 및 cup 모양 (0.3 ~ 4.2%)보다 높았다. 이러한 결과로 볼 때 비정상 자엽을 갖는체세포배 빈도가 높을수록 식물체 재생빈도가 감소될 수있음을 알 수 있었다.
Abstract
Embryogenic callus was induced from the cultures of immature embryos of Camellia japonica L. on Murashige & Skoog’s (MS) solid medium supplemented with 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D), and then the embryogenic callus was proliferated on same medium for 4 weeks over. The embryogenic callus was sub-cultured on MS basal medium without 2,4-D to produce coyledonary stage of somatic embryo. The frequency (%) of somatic embryogenesis was 25.1%, and the majority of somatic embryos formed had a abnormal morphology with cupshaped cotyledon (48.3%), one cotyledon (12.6%), three cotyledons (9.4%), four cotyledons (1.9%), whereas was only normal morphology with two cotyledon (27.5%). When the somatic embryos with normal or abnormal cotyledons transfer to MS basal medium or ½ MS medium with/or without plant growth regulators (GA3, IBA) for regeneration, the frequency (%) of two-cotyledon embryos regenerated into plantlets was higher 11.1% than one cotyledon (0.0~8.3 %), three cotyledons (0.0~5.8%), four cotyledons (0.0%), cup-shaped (0.3~4.2%). These results demonstrated that the anomalous cotyledons of somatic embryos could caused to decrease the rate of plant regeneration.
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