凹叶厚朴愈伤组织的诱导及其有效成分含量的比较

目的:研究凹叶厚朴愈伤组织的诱导、生长情况并比较愈伤组织内有效成分的含量。方法:采用单因素比较、正交设计试验,研究影响凹叶厚朴愈伤组织诱导和生长的各种因素;采用高效液相色谱法测定不同来源愈伤组织内有效成分的含量。结果:外植体脱分化形成愈伤组织的能力,以顶芽及茎段最好,雌蕊次之;愈伤组织诱导的最适培养基为B5+NAA0.1mg.L-1+6-BA1.0mg.L-1,而B5+NAA0.5mg.L-1+6-BA4.0mg.L-1对愈伤组织生长促进作用明显。以WPM为基本培养基诱导愈伤组织,继代后厚朴酚与和厚朴酚含量的总和可达0.1679%~0.2344%,但愈伤组织诱导率低,为33.08%~48.65%;培养条件为MS+NAA0.5mg.L-1+6-BA1.0mg.L-1时,愈伤组织诱导率为62.12%,继代后2酚总量为0.1686%。结论:本试验结果对凹叶厚朴愈伤组织的诱导及高产细胞系的筛选具有一定的参考价值。

Induction of Magnolia officinalis Rehd.et Wils.var.biloba Rehd.et Wils.Callus and Contents of Active Components in Callus from Different Sources

OBJECTIVE： To study the induction and growth of Magnolia officinalis Rehd.et Wils.var.biloba Rehd.et Wils. callus and to compare the content of active components in callus from different sources.METHODS： The factors influencing the induction and growth of Magnolia officinalis Rehd .et Wils .var. biloba Rehd .et Wils. callus were studied by single factor analysis and orthogonal design. The contents of active components in callus from different sources were determined by HPLC. RESULTS： In the various explants, the apical buds and stem fragments exhibited the best ability of dedifferentiation to form callus, followed by pistils. The optimunl culture medium of the callus induction was B5 ＋ NAA0.1 mg ·L^-1＋ 6- BA1.0 mg ·L^-1, while the culture medium B5＋ NAA0.5 mg ·L^-1＋ 6- BA4.0 mg ·L^-1 markedly improved the growth of the callus; after secondary culture of the callus induced by the WPM basic medium, the total content of Magnolol plus Honokiol ranged from 0.167 9% to 0.234 4%, but the induction rate of the callus was low at 33.08% -48.65%. Under the culture condition of MS ＋ NAA0.5 mg ·L^-1＋ 6-BA1.0 mg ·L^-1, the rate of callus induction was 62.12% and the total content of Magnolol plus Honokiol after secondary culture was 0.168 6%. CONCLUSION： The results serve as reference in the tissue culture of Magnolia officinalis Rehd.et Wils.var.biloba Rehd.et Wils and the screening of the high production cell line.