The wheat E3 ligase TaPUB26 is a negative regulator in response to salt stress in transgenic Brachypodium distachyon

Abstract

Various abiotic stresses, including high salinity, affect the growth and yield of crop plants. We isolated a gene, TaPUB26, from wheat that encodes a protein containing a U-box domain and armadillo (ARM) repeats. The TaPUB26 transcript levels were upregulated by high salinity, temperature, drought and phytohormones, suggesting the involvement of TaPUB26 in abiotic stress responses. An in vitro ubiquitination assay revealed that TaPUB26 is an E3 ubiquitin ligase. We overexpressed TaPUB26 in Brachypodium distachyon to evaluate TaPUB26 regulation of salt stress tolerance. Compared with the wild type (WT) line, the overexpression lines showed higher salt stress sensitivity under salt stress conditions, but lower chlorophyll (Chl) content, lower photosynthetic levels and overall reduced salt stress tolerance. Additionally, the transgenic plants showed more severe membrane damage, lower antioxidant enzyme activity and more reactive oxygen species (ROS) accumulation than WT plants under salt stress, which might be related to the changes in the expression levels of some antioxidant genes. In addition, the transgenic plants also had higher Na⁺ and lower K⁺ contents, thus maintaining a higher cytosolic Na⁺/K⁺ ratio in leaves and roots than that in WT plants. Further analysis of the molecular mechanisms showed that TaPUB26 interacted with TaRPT2a, an ATPase subunit of the 26S proteasome complex in wheat. We speculated that TaPUB26 negatively regulates salt stress tolerance by interacting with other proteins, such as TaRPT2a, and that this mechanism involves altered antioxidant competition and cytosolic Na⁺/K⁺ equilibrium.

Keywords: Antioxidant compete; Brachypodium distachyon; Na(+)/K(+) equilibrium; Protein interaction; Salt stress; U-box protein.