Pan Zhao-Jun, Hung Yu-Ling, Chen Tsun-Ying, Shih Yu-An, Lin Ying-Chung Jimmy, Wang Chun-Neng
Department of Life Science National Taiwan University Taipei Taiwan.
Institute of Plant Biology National Taiwan University Taipei Taiwan.
Appl Plant Sci. 2022 Jun 15;10(3):e11476. doi: 10.1002/aps3.11476. eCollection 2022 May-Jun.
Transient gene expression systems are powerful tools for studying gene interactions in plant species without available or stable genetic transformation protocols. We optimized a petal protoplast transformation protocol for , a model plant, to study the development of floral symmetry.
A high yield of petal protoplasts was obtained using a 6-h enzyme digestion in a solution of 1.5% cellulase and 0.4% macerozyme. Modest transfection efficiency (average 41.4%) was achieved. The viability of the transfected protoplasts remained at more than 90%. A fusion of green fluorescent protein and CYCLOIDEA (SsCYC), the Teosinte branched 1CincinnataProliferating cell factor transcription factor responsible for floral symmetry, was subcellularly localized inside the nuclei of the protoplasts. Transiently overexpressing indicates the success of this system, which resulted in the predicted increased (but nonsignificant) expression of its known target (), consistent with gene network expectations.
The transient transfection system presented herein can be effectively used to study gene-regulatory interactions in Gesneriaceae species.
对于没有可用的或稳定的遗传转化方案的植物物种，瞬时基因表达系统是研究基因相互作用的有力工具。我们优化了一种用于模式植物金鱼草的花瓣原生质体转化方案，以研究花对称性的发育。
在含有1.5%纤维素酶和0.4%果胶酶的溶液中进行6小时酶解，获得了高产的花瓣原生质体。实现了适度的转染效率（平均41.4%）。转染后的原生质体活力保持在90%以上。绿色荧光蛋白与CYCLOIDEA（SsCYC）融合，CYCLOIDEA是一种负责花对称性的玉米分枝1-辛辛那提增殖细胞因子转录因子，在原生质体细胞核内进行亚细胞定位。瞬时过表达表明该系统成功，导致其已知靶标（）的表达预期增加（但不显著），这与基因网络预期一致。
本文介绍的瞬时转染系统可有效地用于研究苦苣苔科物种中的基因调控相互作用。
