《Table 3 Effect of different types and concentrations of auxins combined with 5 mg/L zeatin on shoot

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《卡琪花蒂玛茎段增殖及生物反应器培养（英文）》

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Z means seperation within columns by Duncan’s multiple range test at 5%level.NS:nonsignificant；**:significant at P≤0.01；***:significant at P≤0.001.

After nodal explants were cultured on the MS medium supplemented with different concentrations of cytokinins for more than 12 weeks，the multiple shoot induction was significantly influenced by the cytokinin type and concentrations（Table 1，Fig.1）.Multiple shoots developed profusely in the medium supplemented with the TDZ and zeatin，whereas just one shoot or two shoots were observed in the cytokinin-free medium（the control）and in the medium supplemented with kinetin and BA.Therefore，the TDZ and zeatin were more effective for shoot propagation.As shown in Table 1，the TDZ showed the highest cytokinin-like activity and provided the maximum number of shoots（21.5 shoots per explant）at the TDZ concentration of 0.1 mg/L.However，the shoot elongation（length of shoots<0.50cm）and leaf development（No.leaves<3 and Leaf area≤0.3 cm2）were not observed（Table 1，Fig.1）.The inhibition of shoot elongation by the TDZ may be related with its high cytokinin activity，as previously reported[14].But the zeatin in the medium significantly promoted the shoot elongation（Fig.1）.The medium supplemented with 5.0 mg/L zeatin grew 6.8 relatively long shoots per explant among the zeatin treatments（Table 1）.Zeatin was superior compared with the BA and kinetin in terms of shoot proliferation and shoots length.In addition，the zeatin treatment also produced large leaves as compared with other cytokinins（Fig.1，Table 1）.These findings were similar to earlier reports regarding the positive effective of zeatin on promoting shoot proliferation and shoot length in the culture of Bixa orellana[15].HARTINIE M and JUALANG G A[3]reported the substantial role of zeatin（1μM）on shoot elongation but not on shoot proliferation of L.pumila.This study also showed that the lower-concentration zeatin（0.5 or 1.0 mg/L）just promoted shoot elongation，while the higherconcentration zeatin（3.0，5.0，or 7.0 mg/L）led to both shoot proliferation and shoot elongation.Therefore，cytokinins increased to a certain concentration could stimulate shoot multiplication but inhibit its elongation.In general，the BA was more effective than the other cytokinins，which was considerably affected by the plant species and explant types.The variation in the activity of different cytokinins could be explained by their different uptake rate in different genomes reported[16-17]，different translocation rates to meristematic regions and different metabolic processes，in which the cytokinin may be degraded or conjugated with sugars or amino acid to form biologically insert compounds as reported by TRAN THANH VAN K and TRINH T H and KAMíNEK M[18-19].