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以朝鲜白头翁幼嫩叶片诱导产生的不定根为材料诱导分生结节。采取Epon 812树脂和石蜡包埋切片技术分别对诱导初期的单个分生结节和培养5~6周的组织进行组织切片研究。结果显示:(1)不定根在诱导培养基上分化形成独立的表面光滑的球形分生结节。(2)单个分生结节顶端为3~4层分生细胞,其表皮和皮层上也观察到分生细胞。(3)分生结节分化后,结构上存在两种类型,一种为由分生细胞、维管束、薄壁细胞和分化形成的小芽原基组成的结构,芽原基基部由排列整齐的表皮细胞组成,不定芽之间结合较松散,与母体结合不紧密,易脱离,为独立的个体;另一种结构由排列规则的表皮细胞环绕而成,其表皮细胞上广泛分布染色较深的球形、椭圆形或心形的体细胞胚状体,整个结构细胞大小一致,无内部结构。结果证实,朝鲜白头翁不定根来源的分生结节上同时发生器官分化和直接体细胞胚分化,分生结节也可分化形成胚性愈伤组织,通过愈伤组织间接再生体细胞胚。
Adventitious roots induced by young leaves of Pulsatilla indica were used as materials to induce meristematic nodules. Epon 812 resin and paraffin embedded sections were used to study the histological sections of the induced single meristematic nodules and the cultured tissues from 5 to 6 weeks respectively. The results showed that: (1) Adventitious roots differentiated into inducing medium to form independent smooth spherical meristematic nodules. (2) The top of a single meristematic nodule is 3-4 meristematic cells, and meristematic cells are also observed on the epidermis and cortex. (3) There are two types of structures after differentiation of meristematic nodules, one is composed of meristematic cells, vascular bundles, parenchyma cells and small budding primordium formed by differentiation. The base of bud primordia consists of neatly arranged Of epidermal cells, adventitious buds are loosely associated with each other, maternal bond is not close, easy to detachment, as an independent individual; the other structure is arranged by the rules of the epidermal cells formed around the epidermal cells are widely distributed deep staining Spherical, oval or heart-shaped somatic embryoid body, the entire structure of the same size cells, no internal structure. The results confirmed that the adventitious root of Pulsatilla North Korea at the same time occurred on the meristems of organ differentiation and direct differentiation of somatic embryos, meristematic nodules can differentiate into embryogenic callus, regenerated indirectly through callus somatic embryos.