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Method|10 Mar 2021|OPEN
An effector–reporter system to study cellular signal transduction in strawberry fruit (Fragaria ananassa) 
Baozhen Zeng1 ,† , Tianyu Li1 ,† , Wei Wang1 ,† , Zhengrong Dai1 ,† , Jie Li1 , Zhiyuan Xi1 , Kenan Jia2 , Yu Xing3 , Bingbing Li1 , Jiaqi Yan1 and Wensuo Jia,1 ,
1College of Horticulture, China Agricultural University, Beijing 100193, China
2College of International Education, Beijing University of Chemical Technology, Beijing 100029, China
3College of Plant Science and Technology, Beijing University of Agriculture, Beijing 102206, China
*Corresponding author. E-mail: jiaws@cau.edu.cn
†Baozhen Zeng,Tianyu Li,Wei Wang and Zhengrong Dai contributed equally to the study.

Horticulture Research 8,
Article number: 60 (2021)
doi: https://doi.org/10.1038/s41438-021-00493-3
Views: 241

Received: 06 Jun 2020
Revised: 24 Dec 2020
Accepted: 29 Dec 2020
Published online: 10 Mar 2021

Abstract

An effector–reporter system is a powerful tool used to study cellular signal transduction, but this technique has been traditionally used in protoplasts. A similar system to study cellular signal transduction in fruits has not yet been established. In this study, we aimed to establish an effector–reporter system for strawberry fruit, a model nonclimacteric fruit. We first investigated the characteristics of transient gene expression in strawberry fruits and found marked variation in gene expression levels among individual fruits, and this variation has complicated the establishment of a technical system. To overcome this difficulty, we investigated a sampling strategy based on a statistical analysis of the activity pattern of four different reporters (GUS, GFP, FLuc, and RLuc) among individual fruits and combinations of pairs of reporters (GUS/GFP and RLuc/FLuc). Based on an optimized sampling strategy, we finally established a step-by step protocol for the effector/reporter assay. Using FaMYB10 and FaWRKY71 as the effectors and GUS driven by the FaCHS promoter as the reporter, we demonstrated that this effector/reporter system was practical and reliable. This effector/reporter technique will contribute to an in-depth exploration of the signaling mechanism for the regulation of strawberry fruit ripening.