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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (33): 1-5.

所属专题： 生物技术； 水稻

• 农学 农业基础科学 •    下一篇

提高农杆菌介导的籼稻成熟胚愈伤组织再生频率研究

周杰 王春台 刘学群 谭艳平   

  • 收稿日期:2011-08-03 修回日期:2011-10-27 出版日期:2011-12-25 发布日期:2011-12-25

A Study on Improving Regeneration Rate of Agrobacterium-Mediated Callus of Mature Embryo for Indica Rice

  • Received:2011-08-03 Revised:2011-10-27 Online:2011-12-25 Published:2011-12-25

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摘要：

为了提高籼稻红莲型保持系粤泰B(YTB)成熟胚愈伤组织再生频率和遗传转化效率，利用培养基种类、菌液浓度、侵染方法、共培养方法、愈伤组织干燥处理方法、筛选剂浓度等6个方面设计农杆菌不同的培养条件进行试验。结果表明：LB和YEB培养基都可以作为农杆菌的培养方法；真空负压处理可以明显提高转化率；共培养时在共培养基上加滤纸有利于后续的除菌工作，能够提高转化效率；愈伤组织经过适当的干燥处理可以明显提高转化效率；适宜的潮霉素浓度在40~50 mg/L之间；采用V型筛选的方法，可以使转基因植株的阳性率有所提高。总之，通过对农杆菌培养条件的优化能有效地提高农杆菌介导的籼稻成熟胚愈伤组织再生频率。

关键词: 肉制品, 肉制品, 细菌素, 乳酸菌, 分离, 鉴定

Abstract:

In order to improve the regeneration frequency and the rate of genetic transformation of agrobacterium-mediated callus of mature embryos for maintainer line (YTB) of HL-CMS, indica rice, a series of experimental program were carried out, such as, composition of culture medium, concentration of bacilli, infection method, co-culture methods, callus drying method, concentration of selection agent. The results revealed that both of LB and YEB medium can culture agrobacterium; vacuum treatment can significantly improve the rate of conversion; adding filter paper on co-culture medium is in favor of subsequent sterilization and can improve the conversion efficiency; callus dried properly can significantly improve the conversion efficiency; the appropriate concentration of hygromycin is between 40-50mg.L-1-1; the use of V-type screening method can increase the positive rate of transgenic plants. So, the systematic optimization of culture methods for agrobacterium can improve regeneration frequency of callus of mature embryos for indica rice.

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