Dong Chongmei, Beetham Peter, Vincent Kate, Sharp Peter
Plant Breeding Institute, University of Sydney, PMB 11, Camden, NSW, 2570, Australia.
Plant Cell Rep. 2006 May;25(5):457-65. doi: 10.1007/s00299-005-0098-x. Epub 2006 Jan 11.
Oligonucleotide-directed gene repair is a potential technique for agricultural trait modification in economically important crops. However, large variation in the repair frequencies among the scientific reports indicates that there are many factors influencing the repair process. We report here a transient assay system using GFP as a reporter for testing the efficiency of plasmid DNA repair in cultured wheat cells. This assay showed that osmotic medium supplemented with 2,4-D increased the oligo-targeting frequency, and that the repair of a point mutation was more efficient than repair of a single base deletion mutation in cultured scutellum cells of immature wheat embryos. This study provides the first evidence that oligonucleotide-directed mutagenesis is applicable to regenerable cultured wheat scutellum cells.
寡核苷酸定向基因修复是一种用于重要经济作物农业性状改良的潜在技术。然而，科学报告中修复频率的巨大差异表明，有许多因素影响修复过程。我们在此报告一种使用绿色荧光蛋白（GFP）作为报告基因的瞬时检测系统，用于测试培养的小麦细胞中质粒DNA修复的效率。该检测表明，添加2,4-D的渗透培养基提高了寡核苷酸靶向频率，并且在未成熟小麦胚的培养盾片中，点突变的修复比单碱基缺失突变的修复更有效。本研究首次证明寡核苷酸定向诱变适用于可再生的培养小麦盾片细胞。
