摘要註: |
本試驗以人蔘癒傷組織作為培植體,培養於含不同濃度的化學添加劑、包括ascorabic acid(AA)、activated charcoal(AC)、polyvinylpyrrolidone(PVP)之改良式MS培養基(Murashige and Skoog 1962 medium),進行癒傷組織增殖及試管內形態發生試驗。在癒傷組織增殖試驗方面,培養四週後,0.5 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D)+ 0.1 mg/L 1-fenyl-3-(1,2,3-thiadiazol-5-yl) ureum(TDZ)組別癒傷組織添加100 mg/L AA提升最多增殖率,平均達9.14倍;0.5 mg/L 2,4-D + 0.1 mg/L TDZ 組別癒傷組織則是50mg/L AA提升117%;2 mg/L 2,4-D + 0.5 mg/L TDZ組別添加50mg/L PVP增殖率上升至7.1倍。在試管內形態發生試驗方面,將濃度0.5 mg/L 2,4-D + 0.1 mg/L TDZ 、0.5 mg/L 2,4-D + 0.5 mg/L TDZ和2 mg/L 2,4-D + 0.5 mg/L TDZ的癒傷組織置入含不同濃度cytokinins和auxins之培養基,並於光照下培養。八週後,置於PGRs free的癒傷組織逐漸褐化。然而,2 mg/L 2,4-D + 0.5 mg/L TDZ的癒傷組織置於0.02 mg/L N6-furfuryladenine (kinetin) + 3 mg/Lα-naphthalene acetic acid(NAA)五個月後,有不定根形成,每個癒傷組織平均6.34條根,根長5.11公厘。此外,0.5 mg/L 2,4-D + 0.1 mg/L TDZ和0.5 mg/L 2,4-D + 0.5 mg/L TDZ誘導之癒傷組織在不同濃度kinetin + indole-3-butyric acid(IBA)或kinetin + 2,4-D之處理下雖無根部之發育,但癒傷組織已有轉綠現象。綜合而論,本試驗所採用之癒傷組織具有器官形成之能力,而根器官之形成至少需要五個月左右之培養期間。 Calli of Panax ginseng were used as explants to study the effects of ascorabic acid (AA), activated charcoal (AC) and polyvinylpyrrolidone (PVP) on proliferation rate and in vitro morphogenesis. All the explants were cultured on a modified MS (Murashige and Skoog, 1962) medium in darkness. After four weeks of culture, 100 mg/L AA gave the highest proliferation rate with an average of 9.14 folds on the callus line which was originally induced at 0.5 mg/L 2,4-D + 0.1 mg/L TDZ. Callus cultured on MS medium supplemented with 0.5 mg/L 2,4-D, 0.5 mg/L TDZ and 50 mg/L AA had highest proliferation rate with an average of 13.69 folds. Callus cultured on MS medium supplemented with 2 mg/L 2,4-D, 0.5 mg/L TDZ and 50 mg/L PVP had highest proliferation rate with an average of 7.1 folds.We selected the callus which induced by the concentration of 0.5 mg/L 2,4-D + 0.1 mg/L TDZ, 0.5 mg/L 2,4-D + 0.5 mg/L TDZ and 2 mg/L 2,4-D + 0.5 mg/L TDZ with different ratio of cytokinins and auxins medium for in vitro morphogenesis. Under the condition of the callus which induced by the concentration of 2 mg/L 2,4-D + 0.5 mg/L TDZ after 5 months were cultured on MS medium with 0.02 mg/L Kinetin + 3 mg/L NAA, the treatment could promote root formation and had of 6.34 roots in average. In addition, the callus treated with 0.5 mg/L 2,4-D + 0.1 mg/L TDZ or 0.5 mg/L 2,4-D + 0.5mg/L TDZ were cultured on MS medium with all different ratio of kinetin + IBA or Kinetin + 2,4-D did not possess the ability of root formation, but callus turned green. The result showed that root induction is depends on the culture time while not the high ration of NAA/kinetin. |