露地菊品种‘紫秋裳’遗传转化受体再生体系的建立

Establishment of Regeneration System for Genetic Transformation for Chrysanthemum morifolium ‘Ziqiushang'

以露地菊品种‘紫秋裳’的无菌苗叶片为试验材料,在附加不同质量浓度的6-BA和NAA的MS培养基上进行培养,通过器官发生途径获得再生植株。结果表明:当6-BA和NAA的质量浓度均为0.5 mg·L-1时,获得的再生率高达92.4%;腋芽在附加6-BA和NAA的质量浓度分别为0.5、0.1 mg·L-1的培养基上丛生芽增殖效果最好,增殖倍数达到4.20;茎段的最适生根培养基为MS+NAA0.05 mg·L-1,生根率为100%。在筛选结果中发现,叶片对卡那霉素比较敏感。卡那霉素筛选叶片再生的最适质量浓度为30 mg·L-1;卡那霉素筛选茎段生根的最适质量浓度为40 mg·L-1。头孢霉素质量浓度为200 mg·L-1时,能完全抑制农杆菌菌株EHA105的生长,而且菊花叶片再生率受头孢霉素的影响较小,所以选定200 mg·L-1的头孢霉素为最适抑菌抗生素。

An experiment was conducted to obtain the regeneration plantlets from young leaf explants of the Ground-grow Den- dranthema induced on MS medium supplemented with different concentrations of 6-BA and NAA by organogenesis. The re- generation rate of plantlets is 92.37% on MS medium supplemented with 0.5 mg·L^-1 6-BA and 0.5 mg·L^-1 NAA. The regeneration buds are well developed on MS medium supplemented with 0.5 mg·L^-1 6-BA and 0.1 mg·L^-1 NAA, and the multiplication coefficient is 4.20. The optimal culture medium for the stems rooting of Chrysanthemum morifolium is MS supplemented with 0.05 mg·L^-1 NAA and a rooting rate of 100%. In the screening process, the explants are sensitive to kanamycin sulfate（Kin）. The optimal content of Km screening leaf regeneration is 30 mg ·L^-1. The optimal content of Km screening stem rooting is 40 mg·L^-1. Cefotaxime sodium salt （Cef） with the content of 200 mg ·L^-1 can inhibit the growth of living agrobacterium strains EHA105, however, the Chrysanthemum morifolium leaf regeneration rate is affected less. Therefore, 200 mg ·L^-1 of Cef is considered to be an appropriate antimicrobial antibiotic.