title: A novel Glycine soja cysteine proteinase inhibitor GsCPI14, interacting with the calcium/calmodulin-binding receptor-like kinase GsCBRLK, regulated plant tolerance to alkali stress.
abstract: It has been well demonstrated that cystatins regulated plant stress tolerance through inhibiting the cysteine proteinase activity under environmental stress. However, there was limited information about the role of cystatins in plant alkali stress response, especially in wild soybean. Here, in this study, we focused on the biological characterization of a novel Glycine soja cystatin protein GsCPI14, which interacted with the calcium/calmodulin-binding receptor-like kinase GsCBRLK and positively regulated plant alkali stress tolerance. The protein-protein interaction between GsCBRLK and GsCPI14 was confirmed by using split-ubiquitin based membrane yeast two-hybrid analysis and bimolecular fluorescence complementation assay. Expression of GsCPI14 was greatly induced by salt, ABA and alkali stress in G. soja, and GsCBRLK overexpression (OX) in Glycine max promoted the stress induction of GmCPI14 expression under stress conditions. Furthermore, we found that GsCPI14-eGFP fusion protein localized in the entire Arabidopsis protoplast and onion epidermal cell, and GsCPI14 showed ubiquitous expression in different tissues of G. soja. In addition, we gave evidence that the GST-GsCPI14 fusion protein inhibited the proteolytic activity of papain in vitro. At last, we demonstrated that OX of GsCPI14 in Arabidopsis promoted the seed germination under alkali stress, as evidenced by higher germination rates. GsCPI14 transgenic Arabidopsis seedlings also displayed better growth performance and physiological index under alkali stress. Taken together, results presented in this study demonstrated that the G. soja cysteine proteinase inhibitor GsCPI14 interacted with the calcium/calmodulin-binding receptor-like kinase GsCBRLK and regulated plant tolerance to alkali stress.
authors.affiliation: Key Laboratory of Agricultural Biological Functional Genes, Northeast Agricultural University, Harbin, 150030, People's Republic of China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Sun
authors.forename: Shanshan
authors.initials: S
authors.lastname: Yang
authors.forename: Mingzhe
authors.initials: M
authors.lastname: Sun
authors.forename: Sunting
authors.initials: S
authors.lastname: Wang
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Ding
authors.forename: Dan
authors.initials: D
authors.lastname: Zhu
authors.forename: Wei
authors.initials: W
authors.lastname: Ji
authors.forename: Hua
authors.initials: H
authors.lastname: Cai
authors.forename: Chaoyue
authors.initials: C
authors.lastname: Zhao
authors.forename: Xuedong
authors.initials: X
authors.lastname: Wang
authors.forename: Yanming
authors.initials: Y
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D017931:DNA Primers; D008969:Molecular Sequence Data; D010802:Phylogeny; D030821:Plants, Genetically Modified / Q000235:genetics; D011480:Protease Inhibitors / Q000737:chemistry*; D060888:Real-Time Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000737:chemistry*
chemical_list: D017931:DNA Primers; D011480:Protease Inhibitors