National Key Laboratory for Tropical Crop Breeding, School of Life and Health Sciences, Hainan University, Haikou 570228, China.
National Key Laboratory for Tropical Crop Breeding, Key Laboratory of Biology and Genetic Resources of Tropical Crops, Sanya Research Institute, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China.
Int J Mol Sci. 2024 Nov 3;25(21):11808. doi: 10.3390/ijms252111808.
Cassava ( Crantz) is a staple crop in tropical and subtropical regions, valued for its high starch content in roots. Effective genetic transformation and genome editing of cassava require efficient screening methods for transgenic and edited plants. In this study, a visual selection marker system using an R2R3-MYB transcription factor anthocyanin 1 gene ( LOC110667474) from a rubber tree ( Müll. .) has been developed to facilitate the identification of transgenic cassava plants. Transgenic cassava lines expressing accumulated anthocyanins in their leaves, allowing for easy visual identification without the need for destructive assays or specialized equipment. Importantly, the accumulation of anthocyanins did not affect the regeneration or transformation efficiency of cassava. Additionally, the AR-CRISPR/Cas9-gRNA system with the gene as a marker produced gene-edited cassava mutants with purple leaves, demonstrating successful editing. This anthocyanin-based visual reporter (AR) system will provide an effective tool for genetic transformation and genome editing in cassava.
木薯（Crantz）是热带和亚热带地区的主食作物，其根茎富含淀粉。木薯的有效遗传转化和基因组编辑需要高效的转基因和编辑植物筛选方法。本研究开发了一种利用橡胶树（Müll..）的 R2R3-MYB 转录因子花青素 1 基因（LOC110667474）的可视化选择标记系统，以促进转基因木薯植物的鉴定。表达 的转基因木薯系在叶片中积累花青素，无需破坏性测定或专用设备即可轻松进行直观鉴定。重要的是，花青素的积累不影响木薯的再生或转化效率。此外，以 基因为标记的 AR-CRISPR/Cas9-gRNA 系统产生了叶片呈紫色的 基因编辑木薯突变体，表明编辑成功。这种基于花青素的可视化报告基因（AR）系统将为木薯的遗传转化和基因组编辑提供有效工具。
