H+-pyrophosphatases enhance low nitrogen stress tolerance in transgenic Arabidopsis and wheat by interacting with a receptor-like protein kinase

Huijuan Zhang; Ming Chen; Chengjie Xu; Rongbang Liu; Wensi Tang; Kai Chen; Yongbin Zhou; Zhaoshi Xu; Jun Chen; Youzhi Ma; Weiguo Chen; Daizhen Sun; Hua Fan

doi:10.3389/fpls.2023.1096091

H⁺-pyrophosphatases enhance low nitrogen stress tolerance in transgenic Arabidopsis and wheat by interacting with a receptor-like protein kinase

Front Plant Sci. 2023 Jan 27:14:1096091. doi: 10.3389/fpls.2023.1096091. eCollection 2023.

Authors

Huijuan Zhang^{1

2}, Ming Chen², Chengjie Xu^{1

2}, Rongbang Liu², Wensi Tang², Kai Chen², Yongbin Zhou², Zhaoshi Xu², Jun Chen², Youzhi Ma², Weiguo Chen¹, Daizhen Sun¹, Hua Fan¹

Affiliations

¹ College of Agriculture, Shanxi Agricultural University, Shanxi, China.
² Institute of Crop Sciences, Chinese Academy of Agricultural Sciences (CAAS)/National Key Facility for Crop Gene Resources and Genetic Improvement, Key Laboratory of Biology and Genetic Improvement of Triticeae Crops, Ministry of Agriculture, Beijing, China.

Abstract

Introduction: Nitrogen is a major abiotic stress that affects plant productivity. Previous studies have shown that plant H+-pyrophosphatases (H+-PPases) enhance plant resistance to low nitrogen stress. However, the molecular mechanism underlying H+-PPase-mediated regulation of plant responses to low nitrogen stress is still unknown. In this study, we aimed to investigate the regulatory mechanism of AtAVP1 in response to low nitrogen stress.

Methods and results: AtAVP1 in Arabidopsis thaliana and EdVP1 in Elymus dahuricus belong to the H+-PPase gene family. In this study, we found that AtAVP1 overexpression was more tolerant to low nitrogen stress than was wild type (WT), whereas the avp1-1 mutant was less tolerant to low nitrogen stress than WT. Plant height, root length, aboveground fresh and dry weights, and underground fresh and dry weights of EdVP1 overexpression wheat were considerably higher than those of SHI366 under low nitrogen treatment during the seedling stage. Two consecutive years of low nitrogen tolerance experiments in the field showed that grain yield and number of grains per spike of EdVP1 overexpression wheat were increased compared to those in SHI366, which indicated that EdVP1 conferred low nitrogen stress tolerance in the field. Furthermore, we screened interaction proteins in Arabidopsis; subcellular localization analysis demonstrated that AtAVP1 and Arabidopsis thaliana receptor-like protein kinase (AtRLK) were located on the plasma membrane. Yeast two-hybrid and luciferase complementary imaging assays showed that the AtRLK interacted with AtAVP1. Under low nitrogen stress, the Arabidopsis mutants rlk and avp1-1 had the same phenotypes.

Discussion: These results indicate that AtAVP1 regulates low nitrogen stress responses by interacting with AtRLK, which provides a novel insight into the regulatory pathway related to H+-pyrophosphatase function in plants.

Keywords: H+-pyrophosphatase; arabidopsis; low nitrogen stress; receptor-like protein kinase; transgenic wheat.

Grants and funding

This research was sponsored by the National Science and Technology Major Project of the Ministry of Science and Technology of China (SQ2022YFD1200002), Science and Technology Innovation Foundation of Shanxi Agricultural University (2020BQ42), and Agricultural Science and Technology Innovation Program of CAAS and National Natural Science Foundation of China (32272099, 31900256, and 32101719).