莴苣原生质体的分离方法

以莴苣无菌苗的幼叶为试材,研究了不同的分离材料、质壁分离时间、苗龄、酶液组合、酶解时间及酶液渗透压等对原生质体分离效果的影响。结果表明:幼叶在含有13%甘露醇的CPW溶液里质壁分离2h,获得的原生质体产量存活率均高于其它质壁分离时间;苗龄为20d的幼叶分离得到的原生质体质量高,原生质体呈圆形、内含物多、活力强;酶液组合为1.0%纤维素酶+0.2%离析酶,25℃黑暗条件下酶解12h可获得大量有较高活力的原生质体;酶解液中添加浓度为9%的甘露醇较适合原生质体分离。

Study on the Method of Isolation Protoplasts of Lactuca sativa L.

Chosen young leaves of asepsis seedling of Lactuca sativa L.as test material,the effect of material resources,plasmolysis time,seedling age,enzyme combination,enzymolysis time and ostimum pressure on protoplast isolation were studied.The results showed that when young leaves were plasmolysed for 2 h in CPW solution containing 13% mannitol,the yield of protoplast and its survival rate were higher than those of other plasmolysis time.20 d of the young leaves were isolated protoplasts high quality,protoplasm takes the shape of circular,many inclusion,strong vitality;The enzyme concent ration was 1.0% cellulase+0.2% macerozyme,isolate ezyme.A large amount of high-active protoplast could be obtained by enzymolysis for 12 h at 25℃ under dark conditions.The minstra of 9% were suitable for protoplast dissociation.