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Article|01 Oct 2021|OPEN
Development of potent promoters that drive the efficient expression of genes in apple protoplasts
Xianpu Wang1, Lili Xu1, Xiuxia Liu1, Li Xin1, Xuesen Chen1, & Shujing Wu1
1College of Horticultural Science and Engineering, State Key Laboratory of Crop Biology, Shandong Agricultural University, Tai’an, Shandong, PR China

Horticulture Research 8,
Article number: 211 (2021)
doi: 10.1038/hortres.2021.211
Views: 51

Received: 01 Feb 2021
Revised: 29 Jun 2021
Accepted: 05 Jul 2021
Published online: 01 Oct 2021

Abstract

Protoplast transient expression is a powerful strategy for gene functional characterization, especially in biochemical mechanism studies. We herein developed a highly efficient transient expression system for apple protoplasts. The abilities of the Arabidopsis thaliana and Malus domestica ubiquitin-10 (AtUBQ10 and MdUBQ10) promoters to drive the expression of multiple genes were compared with that of the CaMV 35S promoter, and the results revealed that the AtUBQ10 and MdUBQ10 promoters were more efficient in apple protoplasts. With this system, we demonstrated that active AtMKK7ac could activate MAPK6/3/4 signaling cascades, which further regulated MdWRKY33 phosphorylation and stability in apple. Furthermore, the ligand-induced interaction between the immune receptor AtFLS2 and the coreceptor AtBAK1 was reconstituted in apple protoplasts. We also found that the stability of the bacterial effector AvrRpt2 was regulated by feedback involving auxin and the immune regulator RIN4. The system established herein will serve as a useful tool for the molecular and biochemical analyses of apple genes.