Seed dormancy is an indispensable trait for plants. It of barley is gradually decayed by a period of dry storage (after-ripening) or certain environmental condition. Freshly harvested (FH) and after-ripened (AR) grains have been differentially characterized by numerous studies, such as light, temperature and imbibition. Although FH and AR grains are known to change their gene expression during imbibition, phosphosignaling remain unclear. In this study we aim to investigate the effect of after-ripening on phosphosignaling in our cereal model species, barley. Phosphoproteomic analysis was carried out by using following process. Isolated barley seed embryo was used for protein extraction. Trypsin digestion and phosphopeptide enrichment by HAMMOC method were performed, then purified peptide sample was analyzed by TripleTOF5600 (AB-SCIEX). 2019-09-29 Shinnosuke ISHIKAWA Tokyo University of Agriculture and Technology Shinnosuke ISHIKAWA barley, phosphoproteome, seed dormancy, germination, after-ripening 22_3.wiff raw 20959232 17_2.wiff raw 7708672 23_1.wiff raw 25182208 17_3.wiff raw 20099072 23_2.wiff raw 7933952 18_1.wiff raw 25378816 23_3.wiff raw 19378176 18_2.wiff raw 7720960 24_1.wiff raw 26185728 18_3.wiff raw 18751488 24_2.wiff raw 7745536 19_1.wiff raw 28188672 24_3.wiff raw 22798336 19_2.wiff raw 7987200 19_3.wiff raw 19308544 1_1.wiff.scan raw 613556544 20_1.wiff raw 25894912 1_2.wiff.scan raw 426811064 20_2.wiff raw 7778304 1_3.wiff.scan raw 572500940 20_3.wiff raw 20578304 2_1.wiff.scan raw 673745148 21_1.wiff raw 26398720 2_2.wiff.scan raw 409893292 21_2.wiff raw 7647232 2_3.wiff.scan raw 596321320 21_3.wiff raw 19648512 3_1.wiff.scan raw 723400088 22_1.wiff raw 27963392 3_2.wiff.scan raw 417673100 22_2.wiff raw 7712768 3_3.wiff.scan raw 528162884 4_1.wiff.scan raw 628061616 4_2.wiff.scan raw 423518032 4_3.wiff.scan raw 586675292 5_1.wiff.scan raw 632512640 5_2.wiff.scan raw 428581468 5_3.wiff.scan raw 571762284 6_1.wiff.scan raw 613373572 6_2.wiff.scan raw 411046816 6_3.wiff.scan raw 565598496 7_1.wiff.scan raw 699212716 7_2.wiff.scan raw 476531140 7_3.wiff.scan raw 577565076 8_1.wiff.scan raw 828342504 8_2.wiff.scan raw 461784556 8_3.wiff.scan raw 659662360 9_1.wiff.scan raw 769444912 9_2.wiff.scan raw 471093184 9_3.wiff.scan raw 559410144 10_1.wiff.scan raw 715252440 10_2.wiff.scan raw 452339620 10_3.wiff.scan raw 570849848 11_1.wiff.scan raw 655065964 11_2.wiff.scan raw 445924128 11_3.wiff.scan raw 606466376 12_1.wiff.scan raw 748242112 12_2.wiff.scan raw 419577572 12_3.wiff.scan raw 573246464 13_1.wiff.scan raw 967995080 13_2.wiff.scan raw 467924952 13_3.wiff.scan raw 777470160 14_1.wiff.scan raw 825296924 14_2.wiff.scan raw 407567652 14_3.wiff.scan raw 713319188 15_1.wiff.scan raw 874493084 15_2.wiff.scan raw 367315700 15_3.wiff.scan raw 692118648 16_1.wiff.scan raw 778558676 16_2.wiff.scan raw 422203056 16_3.wiff.scan raw 763275740 17_1.wiff.scan raw 872497740 17_2.wiff.scan raw 395984456 17_3.wiff.scan raw 731549272 18_1.wiff.scan raw 855327744 18_2.wiff.scan raw 389918912 18_3.wiff.scan raw 697505420 ._8_1.wiff raw 4096 19_1.wiff.scan raw 1031365948 ._8_2.wiff raw 4096 19_2.wiff.scan raw 423834932 ._8_3.wiff raw 4096 19_3.wiff.scan raw 693983568 ._9_1.wiff raw 4096 20_1.wiff.scan raw 852943244 ._9_2.wiff raw 4096 20_2.wiff.scan raw 419418012 ._9_3.wiff raw 4096 20_3.wiff.scan raw 769972816 ._10_1.wiff raw 4096 21_1.wiff.scan raw 860443824 ._10_2.wiff raw 4096 21_2.wiff.scan raw 391775860 ._10_3.wiff raw 4096 ._11_1.wiff raw 4096 21_3.wiff.scan raw 710559844 ._11_2.wiff raw 4096 22_1.wiff.scan raw 993364344 ._11_3.wiff raw 4096 22_2.wiff.scan raw 412434868 ._12_1.wiff raw 4096 ._12_2.wiff raw 4096 22_3.wiff.scan raw 763387680 ._12_3.wiff raw 4096 23_1.wiff.scan raw 857447676 ._13_1.wiff raw 4096 ._13_2.wiff raw 4096 23_2.wiff.scan raw 380076904 ._13_3.wiff raw 4096 23_3.wiff.scan raw 676375216 ._14_1.wiff raw 4096 24_1.wiff.scan raw 898205256 ._14_2.wiff raw 4096 ._14_3.wiff raw 4096 24_2.wiff.scan raw 358062300 ._15_1.wiff raw 4096 24_3.wiff.scan raw 794410592 ._15_2.wiff raw 4096 ._1_1.wiff raw 4096 ._15_3.wiff raw 4096 ._16_1.wiff raw 4096 ._1_2.wiff raw 4096 ._16_2.wiff raw 4096 ._1_3.wiff raw 4096 ._16_3.wiff raw 4096 ._2_1.wiff raw 4096 ._17_1.wiff raw 4096 ._17_2.wiff raw 4096 ._2_2.wiff raw 4096 ._17_3.wiff raw 4096 ._2_3.wiff raw 4096 ._18_1.wiff raw 4096 ._18_2.wiff raw 4096 ._3_1.wiff raw 4096 ._18_3.wiff raw 4096 ._3_2.wiff raw 4096 ._19_1.wiff raw 4096 ._3_3.wiff raw 4096 ._19_2.wiff raw 4096 ._19_3.wiff raw 4096 ._4_1.wiff raw 4096 ._20_1.wiff raw 4096 ._4_2.wiff raw 4096 ._20_2.wiff raw 4096 ._4_3.wiff raw 4096 ._20_3.wiff raw 4096 ._21_1.wiff raw 4096 ._5_1.wiff raw 4096 ._21_2.wiff raw 4096 ._5_2.wiff raw 4096 ._21_3.wiff raw 4096 ._5_3.wiff raw 4096 ._22_1.wiff raw 4096 ._22_2.wiff raw 4096 ._6_1.wiff raw 4096 ._22_3.wiff raw 4096 ._6_2.wiff raw 4096 ._23_1.wiff raw 4096 ._23_2.wiff raw 4096 ._6_3.wiff raw 4096 ._23_3.wiff raw 4096 ._7_1.wiff raw 4096 ._24_1.wiff raw 4096 ._7_2.wiff raw 4096 ._24_2.wiff raw 4096 ._24_3.wiff raw 4096 ._7_3.wiff raw 4096 1_1.wiff raw 17055744 1_2.wiff raw 7708672 1_3.wiff raw 18919424 2_1.wiff raw 25137152 2_2.wiff raw 7757824 2_3.wiff raw 19820544 3_1.wiff raw 26427392 3_2.wiff raw 7794688 3_3.wiff raw 18333696 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 4_1.wiff raw 23015424 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 4_2.wiff raw 7753728 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 4_3.wiff raw 19603456 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 5_1.wiff raw 22945792 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 5_2.wiff raw 7614464 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 5_3.wiff raw 19316736 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 6_1.wiff raw 22769664 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 6_2.wiff raw 7712768 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 6_3.wiff raw 19021824 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 7_1.wiff raw 24596480 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 7_2.wiff raw 7983104 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 7_3.wiff raw 18632704 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 8_1.wiff raw 26542080 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 8_2.wiff raw 7708672 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 8_3.wiff raw 20643840 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 9_1.wiff raw 23617536 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 9_2.wiff raw 7917568 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 9_3.wiff raw 18554880 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 10_1.wiff raw 23035904 barley embryo Whole TiO2, hydroxy acid-modified metal oxide chromatography (HAMMMOC) Trypsin Carbamidomethyl (C) Phospho (S) Phospho (T) Phospho (Y) Oxidation (M) TripleTOF 5600 Relative quantification - 10_2.wiff raw 7794688 10_3.wiff raw 19046400 11_1.wiff raw 23089152 11_2.wiff raw 7675904 11_3.wiff raw 20185088 12_1.wiff raw 25051136 12_2.wiff raw 7741440 12_3.wiff raw 20234240 13_1.wiff raw 32563200 13_2.wiff raw 10461184 13_3.wiff raw 22503424 14_1.wiff raw 26791936 14_2.wiff raw 7692288 14_3.wiff raw 19574784 15_1.wiff raw 28622848 15_2.wiff raw 7729152 15_3.wiff raw 19677184 16_1.wiff raw 25886720 16_2.wiff raw 7729152 16_3.wiff raw 20164608 17_1.wiff raw 25874432 180710_3.mzid peak 324746896 mzML 180710_4.mzid peak 183692272 mzML 180710_5.mzid peak 255090136 mzML 180710_6.mzid peak 309416683 mzML 180710_7.mzid peak 490365599 mzML 180710_8.mzid peak 279748185 mzML 180710_9.mzid peak 478447197 mzML 180710_10.mzid peak 357923002 mzML 180710_11.mzid peak 316375131 mzML 180710_12.mzid peak 394475911 mzML 180710_13.mzid peak 61647241 mzML 180710_14.mzid peak 321676437 mzML 180710_15.mzid peak 149947740 mzML 180710_16.mzid peak 253529516 mzML 180710_17.mzid peak 203837875 mzML 180710_18.mzid peak 369770077 mzML 180710_19.mzid peak 598379437 mzML 180710_20.mzid peak 389430291 mzML 180710_21.mzid peak 262419645 mzML 180710_22.mzid peak 267995467 mzML 180710_23.mzid peak 353790564 mzML 180710_24.mzid peak 181360622 mzML 180710_1.mzid peak 267167248 mzML 180710_2.mzid peak 152506535 mzML 180710_3.dat result 148256535 other Mascot 180710_4.dat result 92938592 other Mascot 180710_5.dat result 117440678 other Mascot 180710_6.dat result 138457553 other Mascot 180710_7.dat result 202326462 other Mascot 180710_8.dat result 130753396 other Mascot 180710_9.dat result 209141072 other Mascot 180710_10.dat result 159412423 other Mascot 180710_11.dat result 142999896 other Mascot 180710_12.dat result 156296487 other Mascot 180710_13.dat result 37234802 other Mascot 180710_14.dat result 133173486 other Mascot 180710_15.dat result 83063413 other Mascot 180710_16.dat result 113592680 other Mascot 180710_17.dat result 96771263 other Mascot 180710_18.dat result 156043117 other Mascot 180710_19.dat result 202434078 other Mascot 180710_20.dat result 140046793 other Mascot 180710_21.dat result 122360131 other Mascot 180710_22.dat result 116856591 other Mascot 180710_23.dat result 142428131 other Mascot 180710_24.dat result 87212827 other Mascot 180710_1.dat result 131615242 other Mascot 180710_2.dat result 83640111 other Mascot