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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。
The petioles of Pulsatilla koreana were used as materials to induce meristematic nodules. The anatomical characteristics of Pulsatilla koreana in vitro were studied by conventional paraffin section method and resin-embedded section method. The results showed that the induction rate of adventitious nodules was 82.5% after 42 days’ culture on the induction medium of MS + 3 mg zeatin (ZEA) + 0.5 mg L -1 indole acetic acid (IAA) After 56 days of culture, 95% of meristematic nodules are transformed into adventitious buds. The observation of the anatomical structure showed that the anterior meristem, the cambium and the meristematic structure, the apical meristem and the leaf primordia were observed at 7, 14, 28 and 42 days respectively. Adventitious buds of Pulsatilla in North Korea originate from the vascular center within the meristematic nodules. The microdissection of cell clusters similar to the callus at the base of the newly differentiated bud showed that epidermal cells or lower epidermal cells, vascular bundles and meristematic cells are distributed in the basal tissue, which play an important role in the organotypic differentiation of Pulsatilla chinensis effect.