珠眉海棠根原生质体分离条件研究

Isolation method of root protoplasts of Malu zumi

以苹果属珠眉海棠(Maluzumi Mati)根为供体进行原生质体分离条件的研究,结果表明,酶解过程中酶液浓度、酶液渗透压、酶解时间均对原生质体分离效果产生重要影响。对于珠眉海棠,一步酶解法,两步酶解法皆可以分离根原生质体,但两步酶解法优于一步酶解法。两步酶解法的最佳分离条件为:Cellulysin 28 mg/mL+纤维素酶(Cellulase RS)15 mg/mL+果胶酶(PectolaseY-23)2 mg/mL。

Factors that influence root protoplasts isolation were studied with apple species of M.zumi as the experimental material.The results showed that factors of osmoticum,enzyme concentration of enzymesolution and incubation period could all have influence on isolation of the root protoplasts.Although either one-step or two-step method could isolate the root protoplasts,the most suitable method for protoplast isolation was the two-step method and the condition of the method was as follows： Cellulysin 28 mg/mL ＋ Cellulase RS 15 mg/mL ＋ PectolaseY-23 2 mg/mL.