Plant Regeneration of Bupleurum spp. through Somatic Tissue Culture

자호(紫胡)의 체세포조직배양(體細胞組織培養)에 의한 식물체재분화(植物體再分化)

This study was conducted to determine the optimum conditions of inducing callus, proliferating callus, forming somatic embryos, and regenerating plantlets via somatic embryogenesis, for the purpose of producing artificial seeds and substantially developing plant factory technologies that can be employed to all seasons production of Bupleurum plants. Callus was efficiently induced from leaf tissues at three leaf stage in the MS medium supplemented with 2, 4-D 2mg /1 and thidiazuron(TDZ) 0.lmg /1. Callus induction from leaf tissues at maturity was mostly effective in the mixture of 2,4- D 2mg /1 and TDZ 1.0mg /1 while that from flower bud tissues was fairly good in the MS medium containing 2,4-D 1 or 2mg /1.Callus was formed in 15 to 20 days after culture initiation in the MS media supplemented with 2, 4- D 1-2mg /1 and TDZ 0.l-1.0mg /1. Such hormones as kinetin 3mg /1, GA 1mg /1, and the mixture of GA 1mg /1 and TDZ 1mg /1 effected markedly to proliferate the callus cells.The optimum temperature and light intensity for callus culture were found to be $25^{\circ}C$ and 3000 Lux, respectively. Direct plant regeneration from cultured callus was fairly made on hormone-free MS or half-strength MS medium. Somatic embryogenesis was most frequently observed in hormone-free media:60 somatic embryos per 20ml in MS medium and 28 somatic embryos per 20ml in half -strength MS medium. There were three stages-globular, heart, and torpedo-in development of somatic embryos, among which globular stage was more frequently observed in MS medium rather than in half-strength MS medium. Somatic embryos induced from suspension culture fairly differentiated a number of shoots and roots on hormone-free and half-strength MS solid medium.

1. 캘러스 유기는 잎과 화뢰의 유조직에서 잘 되었으며 유엽(65%)이 화기(44%)보다 양호하였고 엽령별 캘러스 유기율은 5,7엽기에서 각각 46% 와 40%의 캘러스유기율을 나타낸 반면 3엽기에서 74%로 가장 높았다. 2. 성숙엽의 배양에서 참시호는 $2,4-D2mg\;/\;\ell+TDZ\; 1mg\;/\;\ell$ 혼합배지에서 가장 높은 캘러스 유기율(92%)을 나타냈으며 삼도시호는 $2,4-D\;2mg\;/\;\ell+BA\;0.5\;1mg\;/\;\ell$ 에서 캘러스유기 (48%)가 양호하였다. 3. $2,4-D\;1-2mg\;/\;\ell$와 $TDZ\;0.1-lmg\;/\;\ell$가 첨가된 배지에서 치상 후 15-20일에 캘러스유기율이 50%를 넘었으며 캘러스증식은 Kinetin $3mg\;/\;\ell$와 $GA\;1mg\;/\;\ell$ 및 $GA\;1mg\;/\;\ell+TDZ\;1mg\;/\;\ell$ 혼합처리에서 양호하였다. 4. 시호의 체세포조직배양에 적합한 광도는3000Lux이었으며 온도는 $25^{\circ}C$가 적온이었다. 5. 체세포배 형성에 있어서 MS배지가 1 / 2MS배지보다 더 효과적이었으며 식물체의 재분화와 재분화된 식물체의 생장에는 1 / 2X MS배지가 1X MS배지보다 더 효과적이었다.