Background : Glycyrrhiza uralensis Fischer is one of most important medicinal (Glycyrrhizae radix) herbs in traditional oriental medication and they are also important commercial products used worldwide in sweetening and flavoring. They contain the similar compounds, such as the triterpenoid saponins and flavonoids. Therefore, it is necessary to develop more efficient and sustainable methods for the production of G. uralensis Fischer and its medicinal constituents. This study was accomplished to investigate the effect of medium composition on in vitro propagation and plantlet regeneration from nodal explants of G. uralensis Fischer, and to establish a reliable protocol for micropropagation.
Methods and Results : Young and actively growing stem segments were excised from adult plants of new cultivar ‘manju’. They were cut into a 1cm nodal segments with single node after sterilization, and cultivated in the different medium supplemented with various plant growth regulators for two weeks. For shoot multiplication, one-node stem segments, approximately 1 ㎝ in length, were taken from in vitro derived shoots and subcultured. After four weeks of culture, the efficacy of each medium on shoot proliferation and growth was determined, and these shoots were transferred onto medium with different auxin hormones for rooting.
Conclusion : After seven to ten days of culture, shoots began to emerge from axillary buds. They showed a vigorous growth and elongation in multiplication medium. During culture period, in vitro cultured plantlets showed significantly different responses to the respective medium with different plant growth regulators. Our experiments confirmed that in vitro growth and multiplication of palntlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.
Buckwheat sprout is used as vegetable, and also flour for making noodles, and so on. Currently, information about tissue culture in buckwheat is limited and restricted to micro-propagation. We carried out somatic embryogenesis and plant regeneration using hypocotyl segments as explant of the cultivated buckwheat species, Fagopyrum esculentum which differs from existing studies in the growth regulator combinations used. Maximum callus regeneration was induced on MS medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) 2.0 mg · L-1, benzyladenine (BA) 1.0 mg · L-1 and 3% sucrose. Friable callus was transferred to solidified MS media containing BA (1.0 mg · L-1) with various concentrations of 2,4-dichlorophenoxyacetic acid for the induction of embryogenesis. The optimum concentrations of growth regulators (for regeneration of plantlet) were indole-3-acetic acid (2.0 mg · L-1), Kinetin (1.0 mg · L-1), BA (1.0 mg · L-1). Only 2,4-D did not show any significant effect on callus induction or embryogenesis. Regeneration of embryonic callus varied from 5% to 20%. Whole plants were obtained at high frequencies when the embryogenic calli with somatic embryos and organized shoot primordia were transferred to MS media with 3% sucrose. The main objective of this research was to develop an efficient protocol for plant regeneration for common buckwheat, and to apply in future for genetic transformation.
반하의 조직배양을 통한 대량증식방법 확립의 일환으로 실시한 기내배양에서의 소괴경형성 및 식물체 분화에 영향을 미치는 배지 NAA와 Thidiazuron의 최적조건을 구명하고져 실시한 실험의 결과는 다음과 같다. 1, 식물생장조절물질로 NAA와 TDZ을 단독처리 할 때 TDZ 0.5 μM에서 shoot수가 45개로 가장 양호하였으며 root분화는 NAA 2.0 mg/l 에서 가장 양호하였다. 2. NAA 0.1 mg/l +TDZ 2.0 μM 조합처리에서 shoot분화가 가장 양호하였으며 NAA 2.0 mg/l +TDZ 2.0 μM 처리시에 가장 저조하였다. 3. 반하의 소괴경 형성은 MS배지에서는 TDZ 5.0 μM 단독처리와 NAA 0.1 mg/l +TDZ 2.0 μM 처리에서 소괴경 형성이 가장 양호하였다. B5배지에서는 TDZ 1.0 μM 단독처리와 NAA 1.0 mg/l +TDZ 5.0 μM 처리에서 소괴경 형성이 가장 양호하였으나 생체중은 NAA 0.1 mg/l 와 TDZ 5.0 μM의 단독처리에서 생체중이 가장 무거웠다. 4. MS, MG, B5배지조성에 따른 소괴경형성은 MG배지에서 30일배양후 가장 양호하였으며 생체중도 좋았다. 5. 분화된 식물체 뿌리분화에는 IAA보다 NAA가, MS 보다 1/2MS가 더 효과적인 결과를 보여 1/2MS배지에 NAA 2 mg/l 를 처리하였을 때 23.3개의 뿌리가 유도되어 가장 높은 결과를 보였다. 완전분화된 식물체를 vermiculite가 담긴 포트에 이식하여 온실에서 순화시킨 결과 80%정도의 생존율을 보였다.