Ruan Yiting, Chen Ke, Su Yangyang, Jiang Suyu, Xu Ping, Murray Jeremy D
CAS-JIC Center of Excellence for Plant and Microbial Science (CEPAMS), Center for Excellence in Molecular Plant Sciences (CEMPS), Shanghai Institute of Plant Physiology and Ecology (SIPPE), Chinese Academy of Sciences, Shanghai 200030, China.
Shanghai Engineering Research Center of Plant Germplasm Resource, College of Life Sciences, Shanghai Normal University, Shanghai 200234, China.
Plants (Basel). 2021 Mar 23;10(3):605. doi: 10.3390/plants10030605.
The hairy root transformation system is widely used in symbiotic studies of model legumes. It typically relies on fluorescent reporters, such as DsRed, for identification of transgenic roots. The MtLAP1 transcription factor has been utilized as a reporter system in based on production of anthocyanin pigment. Here, we describe a version of this reporter driven by a root-cap specific promoter for direct observation of anthocyanin accumulation in root tips, which allows the identification of transgenic hairy roots by the naked eye. Results from our analysis suggest that the reporter had no significant effects on nodulation of . This approach, by virtue of its strong and specific expression in root cap cells, greatly reduces false positives and false negatives, and its use of an easily scored visible pigment should allow greater versatility and efficiency in root biology studies.
毛状根转化系统广泛应用于模式豆科植物的共生研究。它通常依靠荧光报告基因，如DsRed，来鉴定转基因根。基于花青素色素的产生，MtLAP1转录因子已被用作报告系统。在这里，我们描述了一个由根冠特异性启动子驱动的该报告基因版本，用于直接观察根尖中花青素的积累，这使得可以通过肉眼识别转基因毛状根。我们的分析结果表明，该报告基因对[具体植物名称]的结瘤没有显著影响。这种方法由于其在根冠细胞中强烈且特异性的表达，大大减少了假阳性和假阴性，并且其使用易于评分的可见色素应该会在根生物学研究中带来更大的通用性和效率。
