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

Abstract

IntroductionNitrogen 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 ResultsAtAVP1 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.DiscussionThese 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.

Genetic Dissection of Heat Stress Tolerance in Soybean through Genome-Wide Association Studies and the Use of Genomic Prediction to Enhance Breeding Applications

“…The second SNP, ss715610072 , is near the candidate gene Glyma.11G206700 . This gene is a cysteine-rich receptor-like kinase (CRK), which is a family of receptor-like kinases (RLKS) that are important in plant immunity, response to abiotic stress, and regulating growth and development (Zhang et al, 2023). CRKs are highly evolutionarily conserved across vascular plants and have been found to modulate the tradeoff between growth and stress response in many species (Zhang et al, 2023).…”

“…This gene is a cysteine-rich receptor-like kinase (CRK), which is a family of receptor-like kinases (RLKS) that are important in plant immunity, response to abiotic stress, and regulating growth and development (Zhang et al, 2023). CRKs are highly evolutionarily conserved across vascular plants and have been found to modulate the tradeoff between growth and stress response in many species (Zhang et al, 2023). The SNP on chromosome 13 associated with DRB in heat has two promising candidate genes nearby, Glyma.13G223000 and Glyma.13G223100 .…”

The inorganic pyrophosphatases of microorganisms: a structural and functional review

“…In this review, we propose that the use of nanotechnology in the future may further enhance the direct measurements of PPi. For instance, Zhang et al ( 2023a ; 2023b ) developed a carbon quantum dot system in which highly versatile carbon quantum dots are coupled to nanotubes synthesized using metal–organic frameworks. The detection method worked via a “turn-off” mechanism, where Cu 2+ ions quench the fluorescence from the quantum dots, and the detection is based on the decomposition of the metal–organic framework containing the Cu 2+ ions, which are released by the coordination of the Cu 2+ ions with PPi, leading to fluorescence recovery.…”

“…Concomitantly, the authors also investigated whether m-PPase could interact with other proteins and found that it interacts with the A. thaliana receptor-like protein kinase (AtRLK) protein via a yeast two-hybrid system and colocalization experiments. The most striking finding is that the AtRLK mutant and AtAVP1 mutant exhibit the same phenotype under low-nitrogen conditions ( Zhang et al, 2023b ). Overall, there are still aspects to uncover for m-PPases in plants; perhaps their roles in subcellular localization and protein interactions in plants may lead to future experiments that will help engineer plants tolerant to the increasing stress conditions in the soil, either as a direct effect of temperature or the secondary effects due to the reduction of rainfall and availability of nitrogen.…”

Mutations in type II Golgi-localized proton pyrophosphatase AVP2;1/VHP2;1 affect pectic polysaccharide rhamnogalacturonan-II and alter root growth under low boron condition in Arabidopsis thaliana

“…The functional characterization of AVP1 in A. thaliana has been established by studies in mutants and overexpression lines. Some of its known roles are regulation of auxin-mediated organ development and involvement in various abiotic stress tolerance ( Li et al., 2005 ; Kim et al., 2014 ; Nepal et al., 2020 ; Zhang et al., 2023 ). It has also been shown that rather than proton pumping, PPi hydrolysis activity is the major contribution of AVP1 in organ development as demonstrated using fugu5 mutants ( Ferjani et al., 2011 ; Asaoka et al., 2016 ).…”