遗传 ›› 2007, Vol. 29 ›› Issue (7): 844-850.doi: 10.1360/yc-007-0844

• 研究报告 • 上一篇    下一篇

提高水稻插入突变体库利用效率的尝试

张亚芳1; 潘存红1; 李爱宏1,2; 汤雯1; 武茹1; 陈宗祥1; 许爱霞1; 潘学彪1   

  1. 1.扬州大学作物遗传生理江苏省重点实验室, 扬州大学植物功能基因组学教育部重点实验室, 扬州 225009; 2.江苏里下河地区农业科学研究所, 扬州 225007
  • 收稿日期:2006-10-18 修回日期:2006-12-18 出版日期:2007-07-10 发布日期:2007-07-10
  • 通讯作者: 潘学彪

Attempting to enhance the efficiency of T-DNA insertional mutant application in rice

ZHANG Ya-Fang1;PAN Cun-Hong1; LI Ai-Hong1, 2; TANG Wen1; WU Ru1; CHEN Zong-Xiang1; XU Ai-Xia1; PAN Xue-Biao1   

  1. 1. Key Laboratory of Crop Functional Genomics of Jiangsu Province; Key Laboratory of Plant Functional Genomics of Ministry of Educa-tion, Yangzhou University, Yangzhou 225009, China;
    2. Lixiahe Agricultural Research Institute of Jiangsu Province, Yangzhou 225007, China
  • Received:2006-10-18 Revised:2006-12-18 Online:2007-07-10 Published:2007-07-10
  • Contact: PAN Xue-Biao

摘要: T-DNA标签法是一种以农杆菌介导的遗传转化为基础来创造插入突变体库, 从而高通量地分离和克隆植物功能基因的方法。但由于种种原因, 水稻插入突变体库的利用效率较低。为了提高水稻插入突变体库的利用效率, 结合水稻一个双拷贝T-DNA插入突变体的发现和鉴定研究, 通过特异PCR检测、侧翼序列与目标性状的共分离分析, 在1个双插入位点均为杂合的植株的后代株系中分拆了2个插入事件, 分离出目标性状存在遗传分离且只带有1个插入事件的后代株系, 为后续的共分离检测和基因克隆研究打下了重要的基础。由此产生了对插入突变体库中的非串联多拷贝插入标签系进行研究的一些思路和方法, 提出来与同行商榷。

关键词: 水稻, T-DNA标签系, 双拷贝T-DNA插入

Abstract: T-DNA tagging method is a high throughput system for identifying and cloning novel genes from T-DNA-inserted mutant population created via genetic transformation by Agrobacterium tumefaciens. However, the efficiency of using T-DNA-inserted mutant population to clone genes in rice was much lower than in Arabidopsis. In this study, a rice tagged line with two copies of T-DNA segments inserted independently to each other was screened out via a series of verification tests, including the co-segregated analysis between the mutated character and the sequence of T-DNA or the genomic sequence flanking inserted T-DNA. From this tagged line, two inserted incidents were separated from the progeny population of a plant heterozygous in two tagged sites, and some plants with the target trait and one of the inserted incidents were obtained, which were important basic materials for the subsequently co-segregated analysis between the mutated character and the sequence of inserted T-DNA, and for cloning the mutant gene in future. Based on this study, we have some thoughts about the gene cloning from the T-DNA tagged lines with more than one inserted sequence independently and put forward to discuss with colleagues.

Key words: rice, T-DNA tagged line, a line with T-DNA copies inserted independently