Establishment of a Homologous Silencing System with Intact-Plant Infiltration and Minimized Operation for Studying Gene Function in Herbaceous Peonies

oleh: Kaijing Zhang, Xiaobin Wang, Xiaoxuan Chen, Runlong Zhang, Junhong Guo, Qiyao Wang, Danqing Li, Lingmei Shao, Xiaohua Shi, Jingtong Han, Zhiyang Liu, Yiping Xia, Jiaping Zhang

Format: Article
Diterbitkan: MDPI AG 2024-04-01

Deskripsi

Gene function verification is a crucial step in studying the molecular mechanisms regulating various plant life activities. However, a stable and efficient homologous genetic transgenic system for herbaceous peonies has not been established. In this study, using virus-induced gene silencing technology (VIGS), a highly efficient homologous transient verification system with distinctive advantages was proposed, which not only achieves true “intact-plant” infiltration but also minimizes the operation. One-year-old roots of the representative species, <i>Paeonia lactiflora</i> Pall., were used as the materials; prechilling (4 °C) treatment for 3–5 weeks was applied as a critical precondition for <i>P. lactiflora</i> to acquire a certain chilling accumulation. A dormancy-related gene named <i>HOMEOBOX PROTEIN 31</i> (<i>PlHB31</i>), believed to negatively regulate bud endodormancy release (BER), was chosen as the target gene in this study. GFP fluorescence was detected in directly infiltrated and newly developed roots and buds; the transgenic plantlets exhibited remarkably earlier budbreak, and <i>PlHB31</i> was significantly downregulated in silenced plantlets. This study established a homologous transient silencing system featuring intact-plant infiltration and minimized manipulation for gene function research, and also offers technical support and serves as a theoretical basis for gene function discovery in numerous other geophytes.