荔枝转基因悬浮细胞再生松散性胚性愈伤组织的原生质体分离与初步培养

Isolation and preliminary culture of protoplasts from the friable embryogenic calli derived from transgenic suspension cells in litchi

以荔枝优良品种下番枝转gus和hpt基因悬浮细胞再生的松散性胚性愈伤组织(TSFEC)为试材,进行原生质体分离研究及初步培养。结果表明:潮霉素会延长荔枝TSFEC原生质体分离的酶解时间,比一般的酶解时间多2 h,但对原生质体产量和存活率的影响不大;TSFEC培养时间对原生质体分离的影响很大,其中以培养15-20 d时的原生质体产量最大,为(10.1-10.5)×107个.g-1,此时原生质体的活力最高,存活率可以达到93.1%-93.4%;酶解方式对原生质体产量和活力影响都不大,常规的酶解方式酶解的最适宜时间为14 h。

In this experiment, the isolation of protoplasts from the friable embryogenie calli derived from transgenic suspension ceils （TSFEC） with hpt and gus （ uid A） was studied in Litchi chinensis Sonn. cv. Xiafanzhi. The results showed that the time of the protoplast isolation was prolonged for 2 hours when TSFEC had been cultured in media with Hygromycin, but the protoplast output and livability were little affected. The protoplast isolation was remarkably affected by the cultural time of TSFEC, the maximal output （10.1 -10.5 ）×10^7 ·g^-1 and activity （93.1% -93.4% ） of protoplast was achieved in the calli culturing for 15 -20 days. The protoplast output and livability were little affected by enzyme decomposing modes. The feasible time of enzyme decomposing was 14 hours by the general enzyme decomposing mode.