| 摘要註: |
本試驗以人蔘根部作為培植體，培養於含不同濃度的植物生長調節劑：2,4-Dichlorophenoxyacetic acid（2,4-D）及Thidiazuron（TDZ）之改良式Murashige and Skoog medium（MS）進行癒傷組織誘導。培養四週後結果顯示，在PGR–free 的對照組之根培植體逐漸褐化，在不同濃度的2,4-D及TDZ誘導下，根培植體長出不同形態的癒傷組織。癒傷組織增殖率試驗中，以濃度2 mg/L 2,4-D + 0.5 mg/L TDZ 所誘導之癒傷組織增殖率最高，平均達11.5倍。將濃度0.5 mg/L 2,4-D + 0.1 mg/L TDZ 及2 mg/L 2,4-D + 0.5 mg/L TDZ誘導的癒傷組織置入含不同濃度N6-benzyladenine （BA）及α-Naphthaleneacetic acid（NAA）之培養基觀察其形態發生。八週後結果顯示，置於PGR–free的癒傷組織逐漸褐化，此外，NAA濃度越高之誘導下癒傷組織傾向發根，BA濃度越高之誘導下則會使癒傷組織褐化。在濃度0.1 mg/L BA + 10 mg/L NAA之形態發生試驗中，濃度0.5 mg/L 2,4-D + 0.1 mg/L TDZ所誘導之癒傷組織發根率最高，平均發根數為每培植體3.4條根。此外，濃度2 mg/L 2,4-D + 0.5 mg/L TDZ誘導之癒傷組織在不同濃度BA及NAA之處理下皆無發根。在秋水仙素誘導多倍體試驗中，選用三品系癒傷組織，分別是0.5 mg/L 2,4-D + 0.1 mg/L TDZ、2 mg/L 2,4-D + 0.1 mg/L TDZ及2 mg/L 2,4-D + 0.5 mg/L TDZ為增殖率前三高之組別。四週後結果顯示，三品系癒傷組織經由50及100 mg/L秋水仙素處理後，癒傷組織的生長情形明顯受到抑制，其餘秋水仙素濃度在0、0.5、1、2、3、4、5、10 mg/L的處理下，癒傷組織的生長情形則無太大影響。值得注意的是癒傷組織經由秋水仙素處理後，大部分呈現白色鬆軟的外觀。此現象對於未來人蔘癒傷組織誘導多倍體研究可作為參考依據。 Roots of Panax ginseng were used as explants to culture on MS medium supplemented with different concentrations of plant growth regulators. After four weeks, the results showed that callus could be induced by 0.5–2 mg/L of 2,4-D + 0.1–0.5 mg/L of TDZ, but the root explants in PGR–free medium became browning. Callus cultured on MS medium with 2 mg/L 2,4-D + 0.5 mg/L TDZ had highest proliferation rate which had an average of 11.5 folds. We selected the callus which induced by the concentration of 0.5 mg/L 2,4-D + 0.1 mg/L TDZ and 2 mg/L 2,4-D + 0.5 mg/L TDZ with different ratio of BA and NAA medium for in vitro morphogenesis. Under the condition of the callus which induced by the concentration of 0.5 mg/L 2,4-D + 0.1 mg/L TDZ were cultured on MS medium with 0.1 mg/L BA + 10 mg/L NAA, the treatment could promote root formation and had an average of 3.4 roots. In addition, the callus treated with 2 mg/L 2,4-D + 0.5 mg/L TDZ were cultured on MS medium with all different ratio of BA and NAA did not possess the ability of root formation. The result showed that higher NAA/BA ratio favored to promote root induction. 0.5–100 mg/L of colchicines were used to induce polyploidy of callus, the test selected three callus lines comprising 0.5 mg/L 2,4-D + 0.1 mg/L TDZ、2 mg/L 2,4-D + 0.1 mg/L TDZ and 2 mg/L 2,4-D + 0.5 mg/L TDZ﹐respectively. The results showed that at the concentrations of 50 and 100 mg/L colchicine treatment were inhibited callus growth, but other concentrations(0,0.5,1,2,3,4,5 and 10 mg/L)were not affected. It was noteworthy to mention that the callus treated by different concentrations of colchicines became white and soft. |