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农杆菌介导的玉米自交系愈伤组织转化条件的优化
引用本文:权瑞党,尚梅,张举仁.农杆菌介导的玉米自交系愈伤组织转化条件的优化[J].植物生理与分子生物学学报,2003,29(3):245-250.
作者姓名:权瑞党  尚梅  张举仁
作者单位:山东大学生命科学学院,济南,250100
基金项目:国家高技术发展计划项目 (No .2 0 0 2AA2 12 0 71)资助
摘    要:采用农杆菌介导法,将外源基因导入玉米骨干自交系掖515和掖502胚性愈伤组织并再生玉米植株。在农杆菌浸染玉米胚性愈伤组织过程中,采取真空渗入、部分酶解和超声波处理,可以显著提高转化率。农杆菌浸染玉米胚性愈伤组织时,在50kPa下真空渗入5min,掖515和掖502胚性愈伤组织的转化率分别从6.3%和4.5%提高到8.7%和7.8%。愈伤组织在农杆菌浸染前,以0.2%离析酶酶解10min,掖515和掖502胚性愈伤组织的转化率分别提高到8.3%和8.9%。在农杆菌浸染时,以功率100W超声波处理90-120s,掖515和掖502胚性愈伤组织的转化率最高可达9.1%和9.4%。

关 键 词:农杆菌  玉米  愈伤组织  转化  真空渗入  酶解  超声波
修稿时间:2002年12月11

Optimization of Transformation Conditions in Maize Inbred Line Calli via Agrobacterium tumefaciens
Abstract:Embryogenic calli of elite maize inbred lines Ye515 and Ye502 were transformed by mediation with Agrobacterium tumefaciens , and fertile plantlets were regenerated from the transformed calli. In the process of Agrobacterium infection, vacuum infiltration, partial enzymolysis and sonication could improve transformation efficiency significantly. During Agrobacterium infection, by vacuum infiltration at 50 kPa for 5 min, transformation efficiency of embryogenic calli of Ye515 and Ye502 increased from 6.3% and 4.5% to 8.7% and 7.8%, respectively (Fig.3). Partial enzymolysis of calli with 0.2% Macerozyme R 10 for 10 min before infection improved transformation efficiency of Ye515 and Ye502 to 8.3% and 8.9% (Fig.4). After sonication for 90-120 s at 100 W during infection, transformation efficiency of embryogenic calli was up to 9.1% and 9.4% for Ye515 and Ye502, respectively (Fig.5).
Keywords:Agrobacterium  maize  calli  transformation  vacuum infiltration  enzymolysis  sonication  
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