Establishment of regeneration and transformation system of ground-cover chrysanthemum Yuhuaxunzhang | 地被菊雨花勋章再生和遗传转化体系的建立

chinois. 以匍匐型地被菊雨花勋章叶片为外植体，建立了其高频再生体系及根癌农杆菌介导的稳定遗传转化体系。结果表明，不同激素配比对叶片离体再生影响很大，以MS＋1．0mg・L. 16-BA＋1．0mg・L. 1NAA的诱导率最高，达91．1％；分化前期一定时间的暗培养可防止愈伤组织褐变，提高再生率。预培养3d，侵染10min，共培养3d，延迟培养3d为最优转化体系；乙酰丁香酮导致外植体褐化加速，不利于转化。叶片对潮霉素十分敏感，叶盘再生筛选以6―8mg・L. 1为宜，生根筛选以7mg・L. 1为宜。羧苄青霉素抑菌质量浓度以250～350mg・L. 1为宜。获得的抗性芽部分株系经PCR检测初步证实外源基因已转入植物基因组DNA中。[著者文摘]

anglais. Using leaf disks of ground-cover chrysanthemum Yuhuaxunzhang as explants, an efficient and reliable regeneration and transformation system was established. The results showed that the content and proportion of different plant regulators affected greatly the regeneration of leaves. The optimum medium for the highest differentiation rate amounting to 91.1% was MS medium with supplement of 1.0 mg ・ L. 1 naphthalene-acetic acid （NAA） and 1.0 mg ・ L. 1 6-benzylamino purine （ 6-BA ）. A period of culture in dark could effectively prevent callus browning and promote shoot regeneration rate. The transformation was performed, with the optimized pre-cuhure duration 3 d, the suitable agroinfection duration 10 min, and the durations for appropriate co-cuhure and postponed antibiotics selecting 3 d. Browning of callus was accelerated when acetosytingone （AS） was used in the transforma- tion experiments. The leaf was very sensitive to hygromycin （Hyg） , and the optimal concentration of it for callus selecting was 6 -8 mg ・ L. 1 , while 7 mg ・ L. 1 of Hyg is appropriate for rooting selecting. For the control of agrobacterium growth, the optimal concentration of carbenicillin （Carb） was 250 - 350 mg・ L. 1. Several putative transformants were obtained, and PCR analysis indicated that the exogenous gene had been integrated into the genome of transformed chrysanthemum.