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Method|26 Nov 2023|OPEN
Tracking organelle activities through efficient and stable root genetic transformation system in woody plants 
Jinli Gong1,2 , Yishan Chen1,2 , Yanna Xu1,2 , Miaofeng Gu1,2 and Haijie Ma1,2 , Xiaoli Hu1,2 , Xiaolong Li1,2 , Chen Jiao3 , Xuepeng Sun,1,2 ,
1Collaborative 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
2Key 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
3Institute of Biotechnology, Zhejiang University, Hangzhou 310058, Zhejiang, China
*Corresponding author. E-mail: xs57@zafu.edu.cn

Horticulture Research 11,
Article number: uhad262 (2024)
doi: https://doi.org/10.1093/hr/uhad262
Views: 21

Received: 10 Sep 2023
Revised: 13 Dec 2023
Published online: 26 Nov 2023

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.