Expanding agroinfiltration host range with broad-spectrum effectors monitored by autoluminescent reporter
Tiange Wang1 , Jieyu Ge1,2 , Chengyi Qu1 , Zhenkai Wang1 , Hongyu Chen1 , Jie Li3 , Shun Wang1,4 , Xueying Guan1 , Tao Chen5 , Zhanfeng Si1 , Keming Hu6 , Xinhua Ding7 , Li Liu8 , Jian Zhang9 and Alexander S. Mishin10 , Ilia V. Yampolsky10 , Hao Du,1,2,
1College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China 2ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou 311215, China 3Hebei International Research Centre of Vegetable Functional Genomics, College of Horticulture, Hebei Agricultural University, Baoding 071000, China 4Yazhouwan National Laboratory, Sanya 572024, China 5College of Life and Environmental Science, Hangzhou Normal University, Hangzhou 311121, China 6Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China 7State Key Laboratory of Crop Biology, Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests, College of Plant Protection, Shandong Agricultural University, Taian 271018, China 8State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, School of Life Sciences, Hubei University, Wuhan 430062, China 9State Key Laboratory of Rice Biology and Breeding, China National Rice Research Institute, Hangzhou 311400, China 10Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117997, Russia *Corresponding author. E-mail: du_hao@zju.edu.cn
Received: 20 Oct 2025 Accepted: 27 Mar 2026 Published online: 06 Apr 2026
Abstract
Agrobacterium-mediated transient transformation is a fundamental tool for plant research and molecular pharming, but its application is often limited by host immune responses, post-transcriptional gene silencing, and the lack of a robust, quantitative reporter for species beyond Nicotiana benthamiana. To address these challenges, we developed a sensitive reporter system based on the fungal bioluminescence pathway (FBP). As an autonomous, substrate-free metabolic pathway, the FBP enables high-throughput, quantitative screening without external manipulations. Using this system, we systematically evaluated factors affecting transformation efficiency across more than 20 plant families. This cross-lineage analysis identified that co-expressing NahG and P19, termed the NaP19 module, not only mitigates P19-induced cytotoxicity but also synergistically enhances transient expression in diverse species, including agroinfiltration-recalcitrant crops and horticultural plants. We demonstrate the platform's in vivo utility in its native genetic background for applications, such as protein localization, interaction assays, and transcriptional regulation studies. By integrating a quantitative, substrate-free reporter with a broadly effective NaP19 enhancer, this work establishes a robust and versatile platform for advancing functional genomics and biotechnology throughout the land plant lineages.
