Lim Sun-Hyung, Kim Da-Hye, Cho Myeong-Cheoul, Lee Jong-Yeol
Division of Horticultural Biotechnology, School of Biotechnology, Hankyong National University, Anseong 17579, Korea.
Research Institute of International Technology and Information, Hankyong National University, Anseong 17579, Korea.
Plants (Basel). 2022 Mar 19;11(6):820. doi: 10.3390/plants11060820.
Selecting transformed plants is generally time consuming and laborious. To develop a method for transgenic plant selection without the need for antibiotics or herbicides, we evaluated the suitability of the R2R3 MYB transcription factor gene from purple chili pepper () for use as a visible selection marker. positively regulates anthocyanin biosynthesis. Transient expression assays in tobacco () leaves revealed that actively induced sufficient pigment accumulation for easy detection without the need for a basic helix-loop-helix (bHLH) protein as a cofactor; similar results were obtained for tobacco leaves transiently co-expressing the anthocyanin biosynthesis regulators bHLH B-Peru from maize and R2R3 MYB mPAP1D from Arabidopsis. Tobacco plants harboring were readily selected based on their red color at the shoot regeneration stage due to anthocyanin accumulation without the need to impose selective pressure from herbicides. Transgenic tobacco plants harboring showed strong pigment accumulation throughout the plant body. The ectopic expression of dramatically promoted the transcription of anthocyanin biosynthetic genes as well as regulators of this process. The red coloration of tobacco plants harboring was stably transferred to the next generation. Therefore, anthocyanin accumulation due to expression is a useful visible trait for stable transformation, representing an excellent alternative selection system for transgenic plants.
选择转化植株通常既耗时又费力。为了开发一种无需抗生素或除草剂的转基因植物选择方法，我们评估了来自紫色辣椒（）的R2R3 MYB转录因子基因作为可见选择标记的适用性。正向调节花青素生物合成。烟草（）叶片中的瞬时表达分析表明，可在无需碱性螺旋-环-螺旋（bHLH）蛋白作为辅因子的情况下，积极诱导足够的色素积累以便于检测；对于共瞬时表达来自玉米的花青素生物合成调节因子bHLH B-Peru和来自拟南芥的R2R3 MYB mPAP1D的烟草叶片，也获得了类似结果。由于花青素积累，携带的烟草植株在芽再生阶段很容易根据其红色被筛选出来，而无需施加除草剂的选择压力。携带的转基因烟草植株在整个植株体内都表现出强烈的色素积累。的异位表达显著促进了花青素生物合成基因以及该过程调节因子的转录。携带的烟草植株的红色能够稳定遗传到下一代。因此，由于表达导致的花青素积累是稳定转化的一个有用的可见性状，代表了一种用于转基因植物的优秀替代选择系统。
