《植物生理学报》 2010, 46(3): 217-222

药用野生稻TAC 克隆转化籼稻的体系初探

汪暖¹, 陈志雄^1,*, 刘蕊¹, 江奕君³, 潘小芬², 刘向东^1,**, 刘耀光^2,**

华南农业大学1 农学院; 2 生命科学学院, 广州510642; 3 广东省农业科学院水稻研究所, 广州510640

通信作者：陈志雄；E-mail: xdliu@scau.edu.cn, ygliu@scau.edu.cn；Tel: 020-85285752;020-85280187

摘　要：

采用已构建的药用野生稻TAC文库, 比较研究4 个籼稻品种在不同培养基的诱导率及愈伤组织对潮霉素抗性。通过 2 种不同诱导方法产生的愈伤组织, 将携带药用野生稻大片段DNA的TAC 克隆转化到籼稻品种中的结果表明, 除了‘ 粤香 占’ 以外, 其它3 个品种在N6 诱导培养基中添加微量B5 后, 其愈伤诱导率最高。不同品种的潮霉素筛选适合浓度存在差异, 其中 ‘ 华粳籼 74’ 的适合筛选浓度50 mg•L-1, ‘ 粤香占 ’ 为 40 mg•L-1。用预培养 5 d 的愈伤组织进行遗传转化, 在潮霉素筛 选之前先以头孢拉定抑菌处理的, 容易获得转基因植株。4 个品种中以‘ 粤香占’ 的抗性愈伤组织筛选率和分化率最高, 分别为 22.58% 和 14.86%。分子检测 11 株转基因水稻的结果表明, 其中 8 株含有抗性标记基因。据此认为, 药用野生稻 TAC 文库在水稻创新育种中可能有一定的利用前景。

关键词：药用野生稻; TAC 克隆; 遗传转化

收稿：2009-11-24   修定：2010-02-24

资助：国家自然科学基金(30771328)和广东省自然科学基金(7006659)。

Preliminary Study of Transformation System of indica Rice with TAC Clones Containing Large Genomic Sequences of Oryza officinalis Wall.

WANG Nuan¹, CHEN Zhi-Xiong^1,*, LIU Rui¹, JIANG Yi-Jun³, PAN Xiao-Fen², LIU Xiang-Dong^1,**, LIU Yao-Guang^2,**

¹College of Agriculture, ²College of Life Science, South China Agriculture University, Guangzhou 510642, China; ³Rice Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

Corresponding author: CHEN Zhi-Xiong; E-mail: xdliu@scau.edu.cn, ygliu@scau.edu.cn; Tel: 020-85285752;020-85280187

Abstract:

By using TAC library of Oryza officinalis, induction rates of four indica rice varieties in different media and resistance to hygromycin of calli were compared. TAC clones with large genomic sequences of Oryza officinalis were transferred into indica rice varieties by using calli generated by two different induction methods. The results showed that except ‘Yuexianzhan’, the callus induction rates of three rice varieties were highest in N6 induction media plus B5. The appropriate concentrations of hygromycin for four indica rice varieties were detected, 50 mg·L^-1 hygromycin was appropriate for ‘Huajinxian 74’ and 40 mg·L^-1 for‘Yuexianzhan’. In the genetic transformation of 5-day precultured calli, it was more easily to get transgenic plants with anti-bacterium treatment of Cefotaxime before the screening by hygromycin. The transformation frequency and the regeneration frequency of ‘Yuexiangzhan’ was 22.58% and 14.86% respectively, highest among four indica rice varieties. Molecular analysis of 11 transgenic plants showed that, 8 plants carried the transformed selective marker gene. The conclusion suggested that TAC library of Oryza officinalis might have the prospects of utilization in the innovation of rice breeding.

Key words: Oryza officinalis; TAC clones; genetic transformation