以朝鲜白头翁(Pulsatilla koreana)的叶柄为材料诱导分生结节, 利用常规石蜡切片法和树脂包埋切片法研究了朝鲜白头翁离体形态发生的解剖学特点。结果表明: 叶柄在MS+3 mg⋅L–1玉米素(zeatin, ZEA)+0.5 mg⋅L–1吲哚乙酸(IAA)的诱导培养基上培养42天后, 分生结节诱导率为82.5%; 培养56天后, 95%的分生结节转化成不定芽。组织解剖结构观察结果显示, 培养7、14、28和42天时分别观察到前分生结节结构、形成层和拟分生组织结构、顶端分生组织、叶原基结构。朝鲜白头翁的不定芽起源于分生结节内部的维管中心。新分化芽基部类似愈伤组织的细胞团的显微切片结果显示, 基部组织上分布着表皮细胞或下表皮细胞、维管束形成层和拟分生组织细胞, 它们在朝鲜白头翁器官分化中发挥重要作用。
Paraffin sections of Pulsatilla koreana petiole explants were studied to illustrate the microstructure of the meristematic nodule and plant regeneration pathway. Meristematic nodules could be induced from petiole explants on Murashige and Skoog medium containing 3 mg⋅L–1 zeatin with 0.5 mg⋅L–1 indole acetic acid. After 42 days of culture the frequency of nodule induction was 82.5%, and 95% of nodules converted to adventitious shoots after 56 days culture. Globular structures were observed after 14 days’ induction. Prenodular structures of cambial origin appeared from the calluses and gave rise to nodules; shoot buds formed after 28 days’ of culture. Nodules kept giving rise to sub-nodules, from which arose an increasing number of shoot buds. Most apical meristem structures with leaf primordia were observed after 42 days’ culture. Meristemoid regions and cells were much smaller in callus-like tissue than in surrounding cells. Some parts of the sections showed epidermal or sub-epidermal cells, vascular cambium, and meristemoid cells, which were regularly or irregularly distributed on the callus-like tissue. These structures provide evidence that callus-like tissue plays an important role in Pulsatilla species for shoot or root differentiation.
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