Tracking organelle activities through efficient and stable root genetic transformation system in woody plants

Jinli Gong; Yishan Chen; Yanna Xu; Miaofeng Gu; Haijie Ma; Xiaoli Hu; Xiaolong Li; Chen Jiao; Xuepeng Sun

doi:10.1093/hr/uhad262

Tracking organelle activities through efficient and stable root genetic transformation system in woody plants

Hortic Res. 2023 Nov 26;11(1):uhad262. doi: 10.1093/hr/uhad262. eCollection 2024 Jan.

Authors

Jinli Gong^{1

2}, Yishan Chen^{1

2}, Yanna Xu^{1

2}, Miaofeng Gu^{1

2}, Haijie Ma^{1

2}, Xiaoli Hu^{1

2}, Xiaolong Li^{1

2}, Chen Jiao³, Xuepeng Sun^{1

2}

Affiliations

¹ Collaborative Innovation Center for Efficient and Green Production of Agriculture in Mountainous Areas of Zhejiang Province, College of Horticulture Science, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China.
² Key Laboratory of Quality and Safety Control for Subtropical Fruit and Vegetable, Ministry of Agriculture and Rural Affairs, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China.
³ Institute of Biotechnology, Zhejiang University, Hangzhou 310058, Zhejiang, China.

Abstract

Due to the protracted transgenic timeline and low efficiency in stable genetic transformation of woody plants, there has been limited exploration of real-time organelle imaging within stable transgenic woody plant cells. Here, we established an efficient in vivo genetic transformation system for woody plants using an Agrobacterium rhizogenes-mediated approach. This system was successfully validated in multiple perennial woody species. Using citrus as a model, we introduced organelle-targeted fluorescent reporters via genetic transformation and investigated their subcellular localization and dynamics using advanced imaging techniques, such as confocal microscopy and live-cell imaging. Moreover, we subjected transgenic MT-GFP-labeled mitochondria in root cells to stress conditions simulating agricultural adversities faced by fruit crops. The stress-induced experiments revealed notable alterations in mitochondrial morphology. Our study contributes novel insights into membrane trafficking processes, protein localization dynamics, and cellular physiology in woody plants, while also providing stable and efficient genetic transformation methods for perennial woody species.