국내 자생 좀개구리밥 (Lemna paucicostata) 및 개구리밥(Spirodela polyrhiza)의 엽상체 증식을 통한 기내 식물체 대량 증식체계 확립
High frequency plant proliferation via direct fronds regeneration of Korean endemic duckweed species
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국내에 자생중인 좀개구리밥 (Lemna paucicostata) 및 개구리밥 (Spirodela polyrhiza)의 엽상체로부터 엽상체의 직접적인 분화 기작 능 통하여 식물체의 대량증식체계를 확립하였다. 좀개구리밥 및 개구리밥의 엽상체는 생장조절제가 첨가되지 않은 1/2MS 기본배 지에서도 증식이 가능하였으며 1~3 mg/L 범위의 BA 처리가 엽상 체 증식을 촉진시킴을 알 수 있었다. 반면에 고농도의 BA 처리는 오히려 엽상체 증식을 억제시켰다. 엽상체 증식에 미치는 배양배 지의 무기염류 농도의 영향을 조사한 결과 1/2MS 배지가 고농도의 무기염류보다 엽상체 증식에 더욱 효과적임을 알 수 있었다. 좀개 구리밥 엽상체로부터 캘러스 형성빈도는 1 mg/L BA 가 첨가된 1/2 MS배지에서 3.3% 로 매우 낮았으며 개구리밥 역시 캘러스 형성빈 도가 매우 낮았다. 백색의 구형 구조물이 발생한 엽상체 조직을 동 일 조성의 배지에 계대배양하여 증식한 결과 다수의 구형 체세포 배 유사구조가 발달하였으며 동시에 캘러스 증식이 가능하였다. 그러나 구형 체세포배 유사구조를 생장조절제가 첨가되지 않은 MS 배양배지로 옮겨 배양하였으나 식물체 재생은 이루어지지 않 았다. 따라서 좀개구리밥(Lemna paucicostata) 및 개구리밥(Spirodela polyrhiza)의 엽상체로부터 직접적인 엽상체의 분화 기작을 통한 식 물체의 증식체계가 .캘러스 형성을 통한 식물체 재생 보다 더욱 효 과적임을 알 수 있었다. 본 연구에서 확립된 좀개구리밥 및 개구리 밥의 식물체 재생체계는 본 식물의 대량증식은 물론 분자 육종을 통한 우량 형질 도입 연구 소재로 직접적인 활용이 가능할 것으로 기대된다.
Abstract
High frequency plant proliferation system via direct frond regeneration of endemic duckweed plants Lemna paucicostata and Spirodela polyrhiza was established. Fronds of L. paucicostata and S. polyrhiza were able to multiply half-strength MS basal medium without plant growth regulators. However, addition of BA at a range of 1 to 3 mg/L was more effective than high concentration of BA treatments for fronds proliferation. Also half-strength MS salts was suitable for the fronds proliferation. Increase of salts concentration had inhibitory effect on fronds proliferation. Also the frequency of callus formation from fronds of L. paucicostata was 3.3%, when they cultured onto 1/2 MS medium supplemented with 1 mg/L of BA. Similarly the frequency of callus formation from S. polyrhiza was very low. After subculture of white globular structures derived from fronds of L. paucicostata, numerous globular somatic embryos and calluses were developed onto the surface of fronds. However these somatic embryos did not fully develop into normal plants when transferred to 1/2 MS basal medium. Therefore direct frond regeneration system was more efficient for mass proliferation of L. paucicostata and S. polyrhiza. The plant regeneration system of L. paucicostata and S. polyrhiza established in this study, might be applied to mass proliferation and genetic transformation for molecular breeding.
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