title: Comparative Study of Early Cold-Regulated Proteins by Two-Dimensional Difference Gel Electrophoresis Reveals a Key Role for Phospholipase Dα1 in Mediating Cold Acclimation Signaling Pathway in Rice.
abstract: To understand the early signaling steps that regulate cold responses in rice, two-dimensional difference gel electrophoresis (2-D DIGE)(1)was used to study early cold-regulated proteins in rice seedlings. Using mass spectrometry, 32 spots, which represent 26 unique proteins that showed an altered expression level within 5 min of cold treatment were identified. Among these proteins, Western blot analyses confirmed that the cellular phospholipase D α1 (OsPLDα1) protein level was increased as early as 1 min after cold treatment. Genetic studies showed that reducing the expression ofOsPLDα1makes rice plants more sensitive to chilling stress as well as cold acclimation increased freezing tolerance. Correspondingly, cold-regulated proteomic changes and the expression of the cold-responsive C repeat/dehydration-responsive element binding 1 (OsDREB1) family of transcription factors were inhibited in thepldα1mutant. We also found that the expression ofOsPLDα1is directly regulated by OsDREB1A. This transcriptional regulation ofOsPLDα1could provide positive feedback regulation of the cold signal transduction pathway in rice. OsPLDα1 hydrolyzes phosphatidylcholine to produce the signal molecule phosphatidic acid (PA). By lipid-overlay assay, we demonstrated that the rice cold signaling proteins, MAP kinase 6 (OsMPK6) and OsSIZ1, bind directly to PA. Taken together, our results suggest that OsPLDα1 plays a key role in transducing cold signaling in rice by producing PA and regulatingOsDREB1s' expression by OsMPK6, OsSIZ1, and possibly other PA-binding proteins.
authors.affiliation: From the ‡Hebei Collaboration Innovation Center for Cell Signaling; Key Laboratory of Molecular and Cellular Biology of Ministry of Education; Hebei Key Laboratory of Molecular and Cellular Biology; College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei 050024, China; ‖College of Biology Science & Engineering, Hebei University of Economics & Business, Shijiazhuang, Hebei 050061, China.
authors.forename: Chenmin
authors.initials: C
authors.lastname: Huo
authors.affiliation: From the ‡Hebei Collaboration Innovation Center for Cell Signaling; Key Laboratory of Molecular and Cellular Biology of Ministry of Education; Hebei Key Laboratory of Molecular and Cellular Biology; College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei 050024, China;
authors.forename: Baowen
authors.initials: B
authors.lastname: Zhang
authors.affiliation: From the ‡Hebei Collaboration Innovation Center for Cell Signaling; Key Laboratory of Molecular and Cellular Biology of Ministry of Education; Hebei Key Laboratory of Molecular and Cellular Biology; College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei 050024, China;
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.affiliation: §State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China;
authors.forename: Fawei
authors.initials: F
authors.lastname: Wang
authors.affiliation: From the ‡Hebei Collaboration Innovation Center for Cell Signaling; Key Laboratory of Molecular and Cellular Biology of Ministry of Education; Hebei Key Laboratory of Molecular and Cellular Biology; College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei 050024, China;
authors.forename: Meng
authors.initials: M
authors.lastname: Liu
authors.affiliation: From the ‡Hebei Collaboration Innovation Center for Cell Signaling; Key Laboratory of Molecular and Cellular Biology of Ministry of Education; Hebei Key Laboratory of Molecular and Cellular Biology; College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei 050024, China;
authors.forename: Yingjie
authors.initials: Y
authors.lastname: Gao
authors.affiliation: §State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China;
authors.forename: Wenhua
authors.initials: W
authors.lastname: Zhang
authors.affiliation: ¶State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Deng
authors.affiliation: From the ‡Hebei Collaboration Innovation Center for Cell Signaling; Key Laboratory of Molecular and Cellular Biology of Ministry of Education; Hebei Key Laboratory of Molecular and Cellular Biology; College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei 050024, China;
authors.forename: Daye
authors.initials: D
authors.lastname: Sun
authors.affiliation: From the ‡Hebei Collaboration Innovation Center for Cell Signaling; Key Laboratory of Molecular and Cellular Biology of Ministry of Education; Hebei Key Laboratory of Molecular and Cellular Biology; College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei 050024, China; tangwq@mail.hebtu.edu.cn.
authors.forename: Wenqiang
authors.initials: W
authors.lastname: Tang
mesh_terms: D000064:Acclimatization; D003080:Cold Temperature; D018506:Gene Expression Regulation, Plant; D012275:Oryza / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D010712:Phosphatidic Acids / Q000378:metabolism; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D015398:Signal Transduction; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods*
chemical_list: D010712:Phosphatidic Acids; D010940:Plant Proteins; D010739:Phospholipase D