PARMS在水稻基因编辑后代基因分型中的应用研究

Application of PARMS Technology in Genotyping of Rice Gene Edited Progeny

基因编辑技术为植物基因功能研究和作物遗传改良提供了有效途径。目前基因编辑植株基因分型多采用PCR/RE法、PAGE检测法、Sanger测序法等手段,但这些方法存在操作复杂、耗时长或成本高等问题。PARMS技术(penta-primer amplification refractory mutation system,五引物扩增受阻突变体系)是最新开发出的基于荧光检测的SNP基因分型方法,具有操作简便、耗时短、成本低的优势。本研究利用PARMS技术对水稻CRISPR/Cas9基因编辑植株进行了基因分型,鉴定结果与PAGE检测和Sanger测序法一致,证明PARMS技术可用于水稻基因编辑植株后代基因分型,也为其它作物基因编辑后代基因分型技术选择提供了参考。

Gene editing technology provides an effective method for plant gene function research and crop genetic improvement.At present,genotyping of gene edited plants often adopts PCR/RE,PAGE or Sanger sequencing methods,which have disadvantages such as complicated operation,long time consuming or high cost.PARMS(Penta-primer amplification refractory mutation system)technology is a newly developed SNP genotyping method based on fluorescence detection,which has the advantages of simple operation,short time consuming and low cost.In this study,the rice CRISPR/Cas9 gene edited plants were genotyped by PARMS and the results were consistent with that detected by PAGE and Sanger sequencing methods,which proved that PARMS technology could be used for genotyping of rice gene edited progeny.This study also provided a reference for the selection of genotyping techniques of other crops.