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目的:研究不同植物生长物质配比对小蔓长春花叶、茎、根愈伤组织诱导的影响,及各生长期无菌植株、愈伤组织中长春胺含量差异。方法:以小蔓长春花无菌苗的叶、茎、根为材料,采用正交设计实验研究2,4-D,6-BA,NAA对小蔓长春花愈伤组织诱导的影响。在无菌植株生长20,40,60 d,愈伤组织形成高峰期30 d,继代培养30 d时,采用HPLC分别对其中的长春胺含量测定,实验数据进行统计分析。结果:培养基中添加的6-BA,NAA对小蔓长春花叶、茎、根愈伤组织的诱导、生长无显著影响,而2,4-D则影响显著。无菌植株生长20,40,60 d时,长春胺分别为(0.015±0.003)%,(0.097±0.001)%,(0.113±0.06)%;诱导培养、继代培养至30 d时,小蔓长春花叶、茎、根愈伤组织长春胺分别为(0.024±0.002 5)%,(0.016±0.001 5)%,(0.010±0.001 5)%,(0.041±0.002)%,(0.019±0.001)%,(0.016±0.002)%。结论:诱导小蔓长春花叶、茎、根愈伤组织形成的最适培养基为MS+2,4-D 1.0 mg.L-1+6-BA 0.5 mg.L-1+NAA 0.5 mg.L-1。不同生长期无菌植株中长春胺含量差异显著,源于不同外植体的愈伤组织中的长春胺含量不同,其中叶诱导的愈伤组织长春胺含量显著高于茎、根产生的愈伤组织。
OBJECTIVE: To study the effects of different plant growth regulators on the callus induction of leaves, stems and roots of Prunus humilis, and the differences of the contents of vincamine in callus from the vegetative plants in each growth stage. Methods: The effects of 2,4-D, 6-BA and NAA on callus induction of Catharanthus roseus were studied by orthogonal design experiments. At 20, 40, and 60 days after germ-free plant growth, the callus formation peaked 30 days, and 30 days after subculture, the content of vincamine and the experimental data were analyzed respectively by HPLC. Results: 6-BA and NAA added in the medium had no significant effect on callus induction, growth of callus, stem and root callus, but 2,4-D had a significant effect. When the aseptic plants were grown for 20, 40 and 60 days, the vincamine concentrations were (0.015 ± 0.003)% and (0.097 ± 0.001)% and (0.113 ± 0.06)%, respectively. (0.024 ± 0.0025)%, (0.016 ± 0.001 5)%, (0.010 ± 0.001 5)%, (0.041 ± 0.002)%, and (0.019 ± 0.001)% respectively in the leaves, %, (0.016 ± 0.002)%. CONCLUSION: The best medium for inducing the formation of callus of stem, root and root of C. villosa was MS + 2,4-D 1.0 mg.L-1 + 6-BA 0.5 mg.L-1 + NAA 0.5 mg. L-1. There was significant difference in the content of vincamine in different vegetative plants in different growth periods. The content of vincamine in callus derived from different explants was different, and the content of vincamine in the callus induced by leaf was significantly higher than that in the stem and root organization.