目的：优化王枣子愈伤组织的诱导和探究疏松愈伤组织的继代条件，为王枣子悬浮细胞培养奠定初步基础。方法：以王枣子无菌苗幼叶为外植体，采用单因素试验研究不同培养基类型对愈伤组织诱导的影响；采用L9 ( 32 ) 正交试验设计，研究2，4-D、KT不同浓度配比对王枣子疏松愈伤组织的增殖培养的影响。结果：愈伤组织的诱导率在MS、1/2MS和B5培养基上呈上升趋势。MS的出愈时间最晚，1/2MS的出愈时间次之；在B5培养基上出愈时间最早，且愈伤组织的质地和生长状态较好。正交试验的结果显示，愈伤组织的在B5培养基上添加2，4-D 2.5 mg·L-1 + KT 1.0 mg·L-1增殖倍数最小为19.78倍，添加2，4-D 1.5 mg·L-1 + KT 0.2 mg·L-1 达到最大增殖倍数27.34倍；王枣子疏松愈伤组织增殖试验中2，4-D ( P < 0.01 ) 的影响效应比KT ( P = 0.091 ) 大。结论：B5培养基是最佳的王枣子愈伤组织诱导培养基，B5培养基中添加2，4-D 1.5 mg·L-1 + KT 0.5 mg·L-1 为疏松愈伤组织增殖的最佳配比。
Objective: To optimize the systems for callus induction and loose callus subculture of I. amethystoides and to lay the initial foundation for its suspension cell culture. Methods:Using the young leaves of Isodon amethystoides aseptic explants as explants and the effects of different medium types (MS,1/2MS and B5) on callus induction were studied by single factor experiment; the effect of different concentration of 2,4-D and KT on the proliferation culture of loose callus was studied by L9(32) orthogonal design. Results: The induction rate of callus showed an increased trend in the three basic media of MS, 1/2 MS and B5. The callus production time of MS was the last, and that of 1/2MS was the second, while that of callus on B5 medium was the earliest, and the texture and growth state of callus were better than the other two media. The results of orthogonal experiment showed that the proliferation-fold of callus was 19.78 times on medium supplemented with 2,4 D 2.5mg ·L-1 and KT 1.0mg ·L-1 on B5 medium, while it reached the maximum multiplication rate of 27.34 times with the addition of 2, 4 D 1.5mg ·L-1 and KT 0.2mg ·L-1. The effect of 2,4-D (P<0.01) in the proliferation of loose callus was greater than that of KT (P=0.091). Conclusion: B5 medium is the best medium for callus induction of I. amethystoides and the B5 medium supplemented with 2,4-D 1.5mg·L-1 and KT 0.5mg·L-1 is the best proportion for the proliferation of loose callus.