Large-scale production of enhancer trapping lines for rice functional genomics
Large-scale production of enhancer trapping lines for rice functional genomics
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DOI：
10.1016/j.plantsci.2004.03.026
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发表时间：
2004-08
期刊：
影响因子：
5.2
通讯作者：
Yongzhi Yang;Hao Peng;Hongmei Huang;Jinxia Wu;Shi-Ring Jia;Dafang Huang;Tiegang Lu
中科院分区：
文献类型：
--
作者：
Yongzhi Yang;Hao Peng;Hongmei Huang;Jinxia Wu;Shi-Ring Jia;Dafang Huang;Tiegang Lu
使用快速高效的根癌农杆菌介导的水稻转化方案，我们获得了约 100,000 个粳稻品种日本晴的独立增强子捕获系。 T1 种子已从 60,000 个品系中获得。转基因植株的再生从愈伤组织诱导开始仅需3个月，从而将体细胞无性系变异的发生降至最低水平。潮霉素抗性愈伤组织诱导率和植株再生率分别超过90%和80%。 PCR和Southern印迹分析表明，大约95%的潮霉素抗性植物是转基因的。 T1种子的Southern印迹和分离分析表明，约40%的转基因植株含有单拷贝插入，平均拷贝数为1.7。 T0植物的组织化学分析揭示了报告基因β-葡萄糖醛酸酶（GUS）在根、叶、胚胎和胚乳中的表达模式。这种大规模的增强子捕获池为水稻基因组的功能分析提供了有价值的工具。
Using a rapid and efficient Agrobacterium tumefaciens mediated rice transformation protocol, we had got about 100,000 independent enhancer trapping lines of japonica rice variety Nipponbare. T1seeds had already been obtained from 60,000 lines. Regeneration of transgenic plants could be as short as 3 months starting from callus induction, thus the occurrence of somaclonal variation was reduced to a minimum level. The frequencies of hygromycin-resistant callus induction and plant regeneration were over 90 and 80%, respectively. PCR and Southern blot analysis showed that about 95% hygromycin resistant plants were transgenic. Southern blot and segregation analysis of T1seeds showed that about 40% transgenic plants contained single copy insertion, and the average copy number was 1.7. Histochemical assay of T0plants revealed expression patterns in root, leaf, embryo and endosperm of the reporter gene β-glucurodinase (GUS). This large-scale enhancer trapping pool provides a valuable tool for functional analysis of rice genome.