title: Populus euphratica APYRASE2 Enhances Cold Tolerance by Modulating Vesicular Trafficking and Extracellular ATP in Arabidopsis Plants.
abstract: Apyrase and extracellular ATP play crucial roles in mediating plant growth and defense responses. In the cold-tolerant poplar, Populus euphratica, low temperatures up-regulate APYRASE2 (PeAPY2) expression in callus cells. We investigated the biochemical characteristics of PeAPY2 and its role in cold tolerance. We found that PeAPY2 predominantly localized to the plasma membrane, but punctate signals also appeared in the endoplasmic reticulum and Golgi apparatus. PeAPY2 exhibited broad substrate specificity, but it most efficiently hydrolyzed purine nucleotides, particularly ATP. PeAPY2 preferred Mg(2+) as a cofactor, and it was insensitive to various, specific ATPase inhibitors. When PeAPY2 was ectopically expressed in Arabidopsis (Arabidopsis thaliana), cold tolerance was enhanced, based on root growth measurements and survival rates. Moreover, under cold stress, PeAPY2-transgenic plants maintained plasma membrane integrity and showed reduced cold-elicited electrolyte leakage compared with wild-type plants. These responses probably resulted from efficient plasma membrane repair via vesicular trafficking. Indeed, transgenic plants showed accelerated endocytosis and exocytosis during cold stress and recovery. We found that low doses of extracellular ATP accelerated vesicular trafficking, but high extracellular ATP inhibited trafficking and reduced cell viability. Cold stress caused significant increases in root medium extracellular ATP. However, under these conditions, PeAPY2-transgenic lines showed greater control of extracellular ATP levels than wild-type plants. We conclude that Arabidopsis plants that overexpressed PeAPY2 could increase membrane repair by accelerating vesicular trafficking and hydrolyzing extracellular ATP to avoid excessive, cold-elicited ATP accumulation in the root medium and, thus, reduced ATP-induced inhibition of vesicular trafficking.
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Shurong
authors.initials: S
authors.lastname: Deng
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Jian
authors.initials: J
authors.lastname: Sun
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Rui
authors.initials: R
authors.lastname: Zhao
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Mingquan
authors.identifier: http://orcid.org/0000-0002-0835-6963
authors.initials: M
authors.lastname: Ding
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Yinan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Yuanling
authors.initials: Y
authors.lastname: Sun
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Yeqing
authors.initials: Y
authors.lastname: Tan
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Dandan
authors.initials: D
authors.lastname: Liu
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Xujun
authors.initials: X
authors.lastname: Ma
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Peichen
authors.initials: P
authors.lastname: Hou
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Meijuan
authors.initials: M
authors.lastname: Wang
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Cunfu
authors.initials: C
authors.lastname: Lu
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.).
authors.forename: Xin
authors.initials: X
authors.lastname: Shen
authors.affiliation: College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, People's Republic of China (S.D., R.Z., Y.Z., Y.S., W.W., Y.T., D.L., X.M., M.W., C.L., X.S., S.C.);College of Life Science, Jiangsu Normal University, Xuzhou 221116, People's Republic of China (J.S.);College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Hangzhou 311300, People's Republic of China (M.D.); andNational Engineering Research Center for Information Technology in Agriculture, Beijing 100097, People's Republic of China (P.H.) lschen@bjfu.edu.cn.
authors.forename: Shaoliang
authors.identifier: http://orcid.org/0000-0001-7511-3067
authors.initials: S
authors.lastname: Chen
mesh_terms: D000222:Adaptation, Physiological / Q000187:drug effects; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors; D000255:Adenosine Triphosphate / Q000378:metabolism*; D001081:Apyrase / Q000037:antagonists & inhibitors / Q000378:metabolism; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D002413:Cations, Divalent / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D003080:Cold Temperature; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005110:Extracellular Space / Q000737:chemistry; D006868:Hydrolysis; D010940:Plant Proteins / Q000378:metabolism*; D018517:Plant Roots / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D030821:Plants, Genetically Modified; D032107:Populus / Q000201:enzymology*; D021381:Protein Transport / Q000187:drug effects; D011994:Recombinant Proteins / Q000378:metabolism; D013312:Stress, Physiological / Q000187:drug effects; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D013379:Substrate Specificity / Q000187:drug effects
chemical_list: D002413:Cations, Divalent; D004791:Enzyme Inhibitors; D010940:Plant Proteins; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D001081:Apyrase