摘要
从角叉菜(Chondrus ocellatus Holm)的孢子体和雌配子体中,分离了大量的成活原生质体。分离的最适条件为:纤维素酶3％,粉红鲍鱼酶8％,山梨醇1.2mol/dm^3,pH6.0,温度为15℃。细胞壁的完全解离,已用荧光增白剂的染色不着色和原生质体在低渗溶液中的易碎性证实。培养后再生的细胞,经均等分裂,随后发育成实心球状体;或出芽后不等分裂,发育成丝状体和愈伤组织状细胞团。在PES培养基中,经30～40d培养,部分球状体和愈伤组织状细胞团分化出完整植株,但丝状体未能分化。低浓度的IAA、6—FA和C—751对球状体和愈伤组织状细胞团的生长和分化都有促进作用。由原生质体培养获得完整的再生植株,这在真红藻纲(Florideophyceae)中为首例报道。
Large numbers of viable protoplasts were isolated enzymacically from sporophytes and female gametophytes of Chondrus ocellatus Holm The optimum isolating conditions were: purple abalone enzyme 8% cellulase 3%,sorbitol 1.2mol/dm3,pH6.0,temperature 15℃.The lack of cell wall was confirmed by the absence of Calcoflour White staining and the osmotical fragility of the protoplasts.3-4 days after cell wall regeneration,the cells divided symmetrically and developed into spherical structures,or divided asymmetrically after a budding process,then developed into filaments or brown colored callus-like cell masses.The spherical structuresand callus-like cell masses,but none of the filaments,differentiated whole plants after 30-40 days of culture in PES medium.Low concentrations of IAA 6-FA and C-751 could acceleratetheir growth and differentiation The complete thallus regeneration from protoplasts was reported for the first time in the Florideophyceae.
关键词
角叉菜
原生质体
分离
培养
再生
Chondrus ocellatus Holm
protoplasts
Regeneration
Differentiation
