中国水稻科学

中国水稻科学

• 研究报告 • 上一篇    下一篇

提高稻瘟病菌转化频率的研究

黄大年; 王金霞; 杨炜; 倪云美   

  1. 中国水稻研究所，杭州310006
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1995-01-10 发布日期:1995-01-10

Improving Transformation Efficiency of Pyricularia oryzae

Huang Danian; Wang Jinxia; Yang Wei; Ni Yunmei   

  • Received:1900-01-01 Revised:1900-01-01 Online:1995-01-10 Published:1995-01-10

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被引次数

8

摘要： 为提高pAN7-1质粒导入稻瘟病菌建立的遗传转化体系的转化率，进行了对转化效率影响因子的研究。稻瘟病菌转化溶液中使用50 mmol/L CaCl2浓度，PEG4000 40%或PEG80000 40%多聚物，添加30 μg鲱鱼精DNA，供体质粒DNA呈直线状都有利转化频率的提高。其中鲱鱼精DNA作用尤为显著，由于添加30 μg鲱鱼精DNA，转化频率高达71.6转化子/μg DNA。42℃热冲击对稻瘟病菌转化并非必需，转化溶液中添加受体菌2539w DNA后，反而引起转化频率的明显下降。

关键词: 稻瘟病菌, 转化频率, 鲱鱼精DNA, 聚乙二醇

Abstract: The transformation system of rice blast fungus(Pyricularia oryzae) with hygromycin B as selective marker had been established in our laboratory. In order to improve transformation efficiency the effective conditions were studied. The transformation efficiency of using 40% PEG4000(or 40% PEG8000) were greater than that of 20% PEG4000(or 20% PEG8000). The best suitable CaCl2 concentration was 50-100 mmol/L in transformation. Adding 5-30 μg herring sperm DNA can improve transformation efficiency obviously and the herring sperm DNA may be one of indispendable condition of the transformation. Our result indicated that receipt strain DNA had negative influence on transformation, and 42℃ heat shock for 2 or 8 minutes had not active effect. Transformation with linear DNA was better than circular DNA, which conformed to the results of literature.

Key words: herring sperm DNA, PEG, Pyricularia oryzae, transformation frequence