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Warning Text @8^ĉ 2XTableStyleMedium9PivotStyleLight16`FSupporting materialsJTable S1Table S2Table S37Table S4Table S5VV\# _Hlk296016657:ACCCGCGUUAUUg TGATTGAGCCGTGCCAATATA Hvu-miR171mTTGAGCCGCGTCAATATCTCT Hvu-miR171nTGATTGAGCCGCGCCAATATA Hvu-miR172aAGAATCTTGATGATGCTGCAT Hvu-miR172bGAATCTTGATGATGCTGCAT Hvu-miR172fAGAATCCTGATGATGCTGCAT Hvu-miR395bTGAAGTATTTGGGGGAACTC Hvu-miR395nTGAAGTGTTTGGGGGAACTC Hvu-miR396dCTCCACAGGCTTTCTTGA Hvu-miR396eTTCCACAGCTTTCTTGAACTTHvu-miR396e-3pGTTCAAGAAAGCCCATGGAAA Hvu-miR396fTCCACAGGCTTTCTTGAACTGHvu-miR396f-3pGTTCAAGAAAGTCCTTGGAAA Hvu-miR396hTTCCACAGCTTTATTGAACTTHvu-miR396i-3pTTCAATAAAGCTGTGGGATA Hvu-miR397bTTGAGTGCAGCGTTGATGAAC Hvu-miR399jTGCCAAAGGAGAGTTGCCC Hvu-miR444eTGCAGTTGCTGCCTCAAGCTT Hvu-miR530-5p Hvu-miR530bAGCTGCATTTGCACCTGCATCcsi Hvu-miR1520nACACTCAGAACATGACACGTGACG Hvu-miR1520qACAAGTACCAATCAGAACATGACA Hvu-miR1863bACGCTCTGATACCATGTTAAGCAT Hvu-miR2275dTTTGTTTTTCTCCAATATCTCA Hvu-miR5175aACTTAAATTTAGGAACGGAGGGA Hvu-miR5205bCTTATAATTAGGAACGGAGGGAGT Hvu-miR5205dTTATAATTTGGGACGGAGGTAGtae: T. aestivum; osa: O. sativa; bdi: B.Distachyon; far: F.arundinaceea; zma: Z.mays; sbi: S.bicolor; sof: S.officinarum; ssp: S.ssp;vvi: V.vinifera; smo: S.moellendorffii; rgl: R.glutinosu; sly: S.lycopersium; ptc: P.taeda; ppt: P.patens; peu: P.euphratica; mtr: M.truncatula; gma: G.max; csi: C.sinensis; cre: C.clementine; ath:A. thaliana; aly: A.lyrata** completely ortholog* 1or 2bp mismatch miRNA namemature sequencelocationMFE(kj/mol) Hvu-miRn001aAGTATTTAGGTACAGAGGGAGHB28N03r:335:432:- Hvu-miRn001bgnl|UG|Hv#S12497639:306:390:+ Hvu-miRn001cgnl|UG|Hv#S12621510:52:137:- Hvu-miRn001dgnl|UG|Hv#S12642695:392:476:- Hvu-miRn001egnl|UG|Hv#S12776522:254:338:- Hvu-miRn001fgnl|UG|Hv#S12851783:48:133:- Hvu-miRn001ggnl|UG|Hv#S12852157:45:130:- Hvu-miRn001hgnl|UG|Hv#S15209280:206:290:- Hvu-miRn001ignl|UG|Hv#S41545272:34:119:- Hvu-miRn001jgnl|UG|Hv#S41554889:64:149:- Hvu-miRn001kgnl|UG|Hv#S41554890:351:436:+ Hvu-miRn001lgnl|UG|Hv#S41560827:354:439:+ Hvu-miRn001mgnl|UG|Hv#S41560832:69:154:- Hvu-miRn001ngnl|UG|Hv#S41567714:351:435:+ Hvu-miRn001ognl|UG|Hv#S42547467:461:545:+ Hvu-miRn002TCTGTAACTTAATATAAGACGHD07I19r:262:368:+ Hvu-miRn003GAAGCTGTAGTGCAGCCGGCGTTHT11L02r:406:539:+ Hvu-miRn004TGTTGGCTCGACTCACTCAGAHU13M08r:329:422:+ Hvu-miRn005aTACCATGGACAGAAGGCACTTAHW04N16u:103:378:- Hvu-miRn005bHW09H09u:103:378:- Hvu-miRn005cgnl|UG|Hv#S12497598:47:322:- Hvu-miRn005dgnl|UG|Hv#S12519926:248:523:+ Hvu-miRn005egnl|UG|Hv#S12541825:124:399:- Hvu-miRn005fgnl|UG|Hv#S12543211:124:399:- Hvu-miRn005ggnl|UG|Hv#S12711263:382:657:+ Hvu-miRn005hgnl|UG|Hv#S12841392:264:539:+ Hvu-miRn005ignl|UG|Hv#S12843614:57:332:- Hvu-miRn005jgnl|UG|Hv#S12843719:1:269:- Hvu-miRn005kgnl|UG|Hv#S15196148:69:344:- Hvu-miRn005lgnl|UG|Hv#S32304408:89:364:- Hvu-miRn005mgnl|UG|Hv#S42547372:50:325:- Hvu-miRn006aCACGGAACTGGGCGGTTGCACTgnl|UG|Hv#S12495391:331:428:+ Hvu-miRn006bgnl|UG|Hv#S12839456:207:304:+ Hvu-miRn007AACCAGTCGAGGCCAGGGGGTTgnl|UG|Hv#S12495627:135:227:+ Hvu-miRn008aAGACAATTATTTTGGGACGGAGGgnl|UG|Hv#S12499374:390:497:+ Hvu-miRn008cAGACAATTATGTTGGGACGGAgnl|UG|Hv#S41550800:63:165:- Hvu-miRn008bgnl|UG|Hv#S48843262:228:335:- Hvu-miRn008dgnl|UG|Hv#S41550801:499:601:+ Hvu-miRn008egnl|UG|Hv#S41560602:85:187:- Hvu-miRn008fgnl|UG|Hv#S41562929:38:140:- Hvu-miRn009AGGGTGGAAGAAAGAGGGCGgnl|UG|Hv#S12511307:1:98:+ Hvu-miRn010TGCTCTTCATGGACTGCGGCGCCgnl|UG|Hv#S12523587:539:614:+ Hvu-miRn011aTGAGTGTGTGATGGATGGCTTTgnl|UG|Hv#S12532968:125:327:+ Hvu-miRn011bgnl|UG|Hv#S12561290:182:384:+ Hvu-miRn011cgnl|UG|Hv#S12612923:165:367:+ Hvu-miRn011dgnl|UG|Hv#S12687849:198:400:+ Hvu-miRn011egnl|UG|Hv#S12687887:198:400:+ Hvu-miRn011fgnl|UG|Hv#S12688943:231:433:+ Hvu-miRn011ggnl|UG|Hv#S12798737:96:298:+ Hvu-miRn011hgnl|UG|Hv#S12803729:96:298:+ Hvu-miRn011ignl|UG|Hv#S15195305:410:601:+ Hvu-miRn012aTGAGCGAGTTGGCTGCAAGTTCgnl|UG|Hv#S12538067:181:256:+ Hvu-miRn012bgnl|UG|Hv#S12778801:531:606:+ Hvu-miRn013aCGGCGTCGGATCTGGCCGGCCTgnl|UG|Hv#S12539028:58:154:+ Hvu-miRn013bgnl|UG|Hv#S39523603:687:783:+ Hvu-miRn013cgnl|UG|Hv#S48841089:58:154:+ Hvu-miRn014aTCTGGCAGCGACGGGAACATAgnl|UG|Hv#S12565538:73:259:+ Hvu-miRn014bgnl|UG|Hv#S12576685:90:276:+ Hvu-miRn014cgnl|UG|Hv#S23836902:214:400:+ Hvu-miRn014dgnl|UG|Hv#S23839248:75:261:+ Hvu-miRn014egnl|UG|Hv#S23879320:119:305:- Hvu-miRn015aCGAGGAAGGCGCTGAGAGAGAgnl|UG|Hv#S12569610:36:190:+ Hvu-miRn015bgnl|UG|Hv#S12763471:54:208:+ Hvu-miRn016aTGAACAGGTTCGGCGACCTCAgnl|UG|Hv#S12569698:319:399:+ Hvu-miRn016bgnl|UG|Hv#S12709533:343:423:+8 GUU UUUAAACUGGGACUCUGUGAGUAUGGCGAU GA UCGG UAU A8C AGU U C^ UAAG GCC 8 100 90 80 70 60 Initial dG = -60.30 < 10 20 30 40 50 60<UG| UUCU C A A C U UA < AAAAGU ACUCCC CCGAUCCA AUUAAUUGAC CUGC UUAGU CAACUUU G< UUUUCA UGAGGG GGCUAGGU UAAUUAACUG GACG AAUCA GUUGAAA U<UG^ ---- A A C C U CA < 110 100 90 80 70 Initial dG = -23.70 7 10 20 30 40 7CU ------- UG - CUG--- -| UC 7 AUGUGA UG AAGAUUUUAAGCAUUG AAC AUC GGC \7 UACACU GC UUCUAAGAUUUGUAAU UUG UAG UUG G7CU UUUNGGG CU G UCUUCA U^ UA 7 90 80 70 60 50 Initial dG = -59.10 R 10 20 30 40 50 60 RUG AUA------------ G CCU A CC C .-UUC U--| CCCAAAA R UGCCGA GAC AUUAAGAAGA GCA GG ACAU CAG CAGU GCAAUA \R GCGGCU CUG UAGUUCUUCU UGU CC UGUA GUC GUCG CGUUGU GRUG CCUUCCUCUUUUUUC - CG- A C- A \ --- CUU^ CUUAUAA R270 260 250 240 230 220 80 70 t 90 100 110 120 t AUCUAA-------------- .-UUUUUAUCAGAUCAAACUCUUU A t CACAUUGCC UCC Ut GUGUGAUGG AGG GTC202631)Squamosa promoter-binding-like protein 16TC222091Teosinte glume architecture 1CA032492)Squamosa promoter-binding-like protein 17BF258419Promoter binding proteinEX598089TC203114(Squamosa promoter-binding-like protein 3BF630636SBP transcription factorBU990948SH3 domain-containing proteinTC208671$Triosephosphate isomerase, cytosolic Hvu-miR159d 7TC225094GZinc finger AN1 and C2H2 domain-containing stress-associated protein 16 7TC208432NAC domain protein NAC1TC205144TC213127*Mitogen-activated protein kinase homolog 1TC199406%Peptidyl-prolyl isomerase PASTICCINO1TC2347580Glutathione gamma-glutamylcysteinyltransferase 1BQ462194BARE-1 polyproteinDN156119Sperm protamine-P1CX627597$Peptidoglycan-recognition protein-LDTC226007Growth-regulating factor 2TC237714Glutathione transferase F4 Hvu-miR167TC223859Auxin response factor 17TC232622NP315934TC215572G-box-binding factorTC205164SRPK4BG414720TC219866BQ461013 Scl1 proteinTC231973TC196893APETALA2-like proteinTC223285Sporulation related precursorAL499992 Succinyl-CoA ligase beta subunitAL501298-Succinyl CoA ligase beta subunit-like proteinTC2056323Protein kinase domain containing protein, expressedTC199627TC219499< ATP sulfurylaseBG344292TC200840CB860915Growth-regulating factor 1TC236309Growth-regulating factor 3TC227397Growth-regulating factor 3 7TC228863TC210608GH213244Growth-regulating factor 7TC217627Growth-regulating factor 8TC223723Kinesin heavy chainAV836786UPF0344 protein lin2366BI960433Laccase-10 precursorBY839519Laccase-12/13 precursorBI960140TC230973BI957301Laccase LAC5-6TC220141TC204108CPD-photolyaseBM816251Superoxide dismutase [Cu-Zn] 4AGH228935-MIKC-type MADS-box transcription factor WM32ATC221988 Cyclin-B1-5CB864936Src homology-3 domain protein 3BQ759855Phytocyanin homologTC209865Superoxide dismutase [Cu-Zn]TC201898Expansin EXPB2TC216069RegA/RlsA-like proteinTC234153Protein rough sheath 2TC204555TC226505#Serine/arginine repetitive matrix 1BJ486485-Polyubiquitin containing 7 ubiquitin monomersBF625481&Transcriptional regulator, LysR familyBM816541+Serine/arginine repetitive matrix protein 2BI949150Ubiquitin carrier proteinTC195365TC201949CB866855Proteasome subunit alpha typeTC206699 MAS1 proteinTC196192BU986355Calmodulin-like proteinTC220355Tobacco fibrillarin homologTC208940 Pyruvate decarboxylase isozyme 2BI955549/Guanine nucleotide-binding protein beta subunitExpressed proteinTC220056!C2 domain-containing protein-likeBE411757 E3 ubiquitin-protein ligase PRT1TC228269-Protein phosphatase 2C-like protein precursorAV835198 Periplasmic iron-binding proteinTC209002DMT family transporter NP1677122NPR1-like 1 proteinTC226861Protein kinaseTC208489BG310342chloroplast rRNA-operonTC214142TC2092112Kinesin motor domain containing protein, expressedTC223827*GDSL-motif lipase/hydrolase family proteinTC2103756Pentatricopeptide (PPR) repeat-containing protein-likeTC200241CAAX peptidaseCB862207'RRNA intron-encoded homing endonucleaseBE421835 Rurm1 proteinTC203938TC222438&4Fe-4S ferredoxin, iron-sulfur bindingTC237943PHomolog of Brachydanio rerio "Muscle-specific beta 1 integrin binding protein 2.TC215432Transmembrane protein kinaseBY864876TC236919LMW-m1 glutenin subunitTC199906BU999651EComponent of a complex containing the Tor2p kinase and other proteinsBU998718Phospholipase A2 precursorTC231632FERONIA receptor-like kinaseBJ483286%Retinitis pigmentosa GTPase regulatorTC220835Transcriptional regulatorTC231259!Peroxisomal membrane protein-likeTC226208TC237091DDual specificity mitogen-activated protein kinase kinase hemipterousTC236255`Glycine betaine/L-proline ABC transporter, periplasmic glycine betaine/L-proline-binding proteinTC225370Ocs-element binding factor 1TC220919Cdc2 protein kinases-likeTC207007TC229614BY855949Tetraspanin family proteinTC214946>Phospholipid hydroperoxide glutathione peroxidase-like proteinTC224622 Glutathione S-transferase GST 34TC238394CONSTANS-like protein CO5TC196281+Efflux transporter, RND family, MFP subunitTC209974Cinnamyl alcohol dehydrogenaseTC228335TC236673TC225137Kinase binding proteinTC215165)ABA induced plasma membrane protein PM 194 CGUU A4 - UUG Initial dG = -51.90 1 10 20 30 40 1- U U U A C - A-| G GA 1 G CACUA GAUGUUGGC CG CUCA UCAG ACC CGCCG AGG G1 C GUGAU CUAUAACCG GC GAGU AGUC UGG GCGGC UCU C1U - U U C U U CG^ G AC 1 90 80 70 60 50 Initial dG = -77.90 : 10 20 30 40 :C UAUA-------- - .-AAGAAGAAAA| GAG : UCGUUCA CCAUGGACA GAAGGCACUU UG A: GGUAAGU GGUACCUGU CUUUCGUGAA AC U:C UGAUUAGAGAUG A \ ----------^ GGA : 270 260 250 240 50 n 60 70 80 90 100 110 n AGUACAGAUGAAA U A - ACA-- .-AAG U AGGA UG n CC GGACC CA CCUA UUUUCC UUUUGUGGC UGUA GG Gn GG UCUGG GU GGAU AAAAGG GAAACAUCG GUAU CC An CGGG--------- U - C AAGAC \ --- C GUG- CU n 230 220 210 130 120 h 140 150 h ACG--- .-AU G h ACGAG GCC \h UGUUC CGG Ah AAAAAA \ -- A w 160 170 w ---- G A- A U w CAU AA AGUG UGA Aw GUA UU UUAC ACU Gw AAAA G GA - U w 190 180 Initial dG = -79.00 G 10 20 30 40 50 60GC UAUA-------- - .-AAGAAGAAAA| A CAAG A G UCGUUCA CCAUGGACA GAAGGCACUU UGG UAUAGG UUC GG GGUAAGU GGUACCUGU CUUUCGUGAA ACC AUGUCC AAG AGC UGAUUAGAGAUG A \ ----------^ - A--- U G 270 260 250 240 70 X 80 90 100 110 X CA- .-CA .-AAG U AGGA UG X CCUAA UUUUCC UUUUGUGGC UGUA GG GX GGGUU AAAAGG GAAACAUCG GUAU CC AX CGG \ -- \ --- C GUG- CU X 230 210 130 120 R 140 150 R ACG--- .-AU G R ACGAG GCC \R UGUUC CGG AR AGAAAA \ -- A a 160 170 a ---- G - A U a CAU AA AAGUG UGA Aa GUA UU UUUAC ACU Ga AAAA G G - U a 190 180 > < > ---- AA > CAG U> GUC A> CUGG GG Initial dG = -65.80 D 10 20 30 40 50 60 D C CA A - - .-AAGAAGAAAAUGGAGAUAGGCAAGUA| AUG DCU GUU UAU CCAUGG ACA GAAGGCACUU CAG \DGA UAG AUG GGUACC UGU CUUUCGUGAA GUC AD U AG - A A \ --------------------------^ CAA D 260 250 240 70 ? 80 90 ? .-GN A- A UUUU AGU miR-n026a 7miR-n029 7 350 340 330 320 310 300 290 280 270 260 210 130 140 150 A-------------- .-GA AAAAA AA UCCAU CUAA GC A AGGUA GGUU UG A AAAAACAAUUUGAAA \ -- AAUAA AA 200 190 160 GUG--- U UUGUG U AACAC C GAAAGA U 220 230 AA--------- GA UUUG GAU AUGC \ CUA UACG A AAUAGUAAUAC AG UAUU 250 240 Initial dG = -42.40 , 10 20 30 ,.-AUAUUAAUUUGU GCA GUU--| CUUG , GGAAAGUGG GUUGG GAG \, CCUUUUACU CAACC CUC A,\ ------------ AA- ACUGU^ UAAU , 60 50 F 70 80 90 100 110 120 F UCUCAUU UUCCUA -- --- - GCAUAUCAUUU C UG U F GUCC UGUGA GCU CGU UUG GGU UGGAAUUCA GAU \F CGGG ACACU CGG GCG AAC UCA GCCUUAAGU UUA UF ------- UUUAG- GU UUU C ----------- A UU C F 170 160 150 140 Initial dG = -39.00 .-GC| U CGCUG A GUGAU C \ --^ A 9 20 30 40 50 60 9 UUUC-------- GGA G U G G GU UUC 9 GUCUCU UCUCCCU UCCU GUGG GA UCCUU UG \9 UAGGGA GGGGGGA AGGA UACU UU AGGAG AC U9 GGGAUUUUGUUC A-- G - G G GU UGA 9 110 100 90 80 70 Initial dG = -67.40 = 10 20 30 40 50=U UGUU U - AC ------- .-AC -| ACU = AGGGG GUUGC CUG CC CGGUCCAU GACGAC CUGA GCG C= UCCCU CGAUG GAC GG GUUAGGUA CUGUUG GACU CGU C=C UU-- - U A- ACGGAAG \ -- A^ CAC = 190 180 170 160 60 G A---- - .-AA AG G GCU UCGG GCAG UG CGA AGCC CGUC UG CCUUU G \ -- UU G 150 80^ 90 100 110 ^ CAUCCCUCCAA UC- G UU ^ UACA UCUGC GUGUC U^ GUGU AGACG CGUAG U^ GC--------- UCA A UG ^ 140 130 120 Initial dG = -46.90 G 10 20 30 40 50 60 GA A A GU UG G - -| .-ACACCAACAUUACCUGCAA AA G UGA GCA CAACUGCA ACUUG G GAAAG CACAAGUA GG GC \G ACU CGU GUUGACGU UGAAC U CUUUC GUGUUCAU CC CG GGA C C GC GU - G A^ \ ------------------- CA G 130 120 110 100 90 70 = AAAAUUAAUA AA = G \= C C= A--------- UA = Initial dG = -21.60 . 10 20 30 40 .CG- CA |GG CAG AACG UGCC AU . CC CUUU GUUU AGAUC CCCAUAUA GC G. GG GAGG CAGA UCUAG GGGUAUGU CG U.GAA AA ^AG CAG ---- CCUC AC . 80 70 60 50 Initial dG = -57.60 8 10 20 30 40 508- CG A U AGU UGAG-- .-AACG -| ACC 8 GCGC UGC UGAG ACGU AGGGA GAUCGU CCU GC G8 UGUG GCG GCUC UGCG UCCCU CUGGCG GGA CG G8C CU - C GC- CUGGAA \ ---- C^ ACC 8 140 130 120 110 60 D 70 80 miR-n035ACATTATGGGACGGAGGGAGTmiR-399bmiR-156dmiR-396d miRNA_Acc. Target_Acc. ExpectationUPE InhibitionTargetTC223235CleavageGAMYB-binding proteinCV060003Starch branching enzyme IIaDN187880Catalase isozyme 2CK566281HV1LRR1TC219375Serine/threonine protein kinaseTC195122,Serine/threonine kinase-like protein ABC1036DN155589Myb-related protein Hv33TC203666 Translation)Carbonic anhydrase, chloroplast precursorTC228991 Ribophorin ITC19647560S ribosomal protein L17-2BJ450193&Pathogenesis-related protein precursorTC1976813Chloroplast Mg-chelatase subunit XANTHA-G precursor< TC195197TC232544Alcohol dehydrogenaseTC196726NAC transcription factorCV060414Cystatin Hv-CPI8TC195133/ CCU CC A/ A A CCAACAA / 70 60 * 10 20 30 *AUC AGAU--- -| CU GC CG * GGAC GCUCUUC AUGGA GCG GCCGG \* CCUG UGGGGAG UACCU UGU CGGCC C*GCU GCCGGAG C^ -- A- AC * 70 60 50 40 Initial dG = -55.40 + 10 20 30 +.-U CAC U GA .-A| A GU + GCGAG AUGAGUG GU UGG UGGCUUUG GC \+ CGUUU UACUUAC CA ACC ACUGGAAC CG U+\ - CC- U AA \ -^ C UU + 160 150 50 40 O 60 70 80 90 O CUU C---- CAACA CU G G- AAA O GAUCAC CUCUUCUCGG AACUA GU GUAC GC \O UUGGUG GGGAAGGGUC UUGAU UA CGUG CG CO CUG UUCUA ----- UC A GA CAA O 140 130 120 110 100  170 180  CACCUGCCAUUACAU U-- G  CUCC GA A GAGG CU C CAC------------ CCC A  200 190Initial dG = -27.80 ' 10 20 30 '- AAUUAU-| G AGU UGAA ' GACA UGAGCGAGUUGGCU CA UC G' CUGU GCUUGCUCGACUGA GU AG U'A CGUUACU^ G AAU UAAU ' 70 60 50 40 Initial dG = -52.40 5 10 20 30 40 5AAG CA----- A-| U G C U GUU 5 ACGGU CGGCGUCGG UC GGCCG CCUGGG GGCC CG \5 UGCCG GCCGCGGCC AG UCGGC GGGCUC CUGG GC U5GGG CGGGACC CG^ C - - C GAC 5 90 80 70 60 50 Initial dG = -55.30 : 10 20 30 40 50 :CG AAG GCA G A .-CG AG .-CGA| U A : GGGUU UCUG GC ACGGG ACAUA CA CAGUG CGUC UC A: CUUAG AGAC CG UGUCU UGUGU GU GUCGC GCAG AG C:UG --- A-- G A \ -- CU \ ---^ U U : 180 170 110 60 I 70 80 I AGAGGGAUCGAGAAAU U A I GCA AUUCC \I CGU UAGGG CI AU-------------- - C @ 120 130 140 @ AAGUCUGAACUA GC GU A @ GUGUAC GCAUGC C \@ CACAUG UGUACG G A@ AAAG-------- -- UG A @ 160 150 Initial dG = -63.50 S 10 20 30 40 50 60 70 SC G C G UUCUU-- U- U U- CC--| UC U - AAAC S UCUUCUCU UUCUCUCUCGGC UCU CCUC CUC CUCG CC CUG UCU UUGAUU GCG GUA \S AGAAGAGA GAGAGAGAGUCG GGA GGAG GAG GAGU GG GAC AGG AACUGG CGC CGU US- G C A CACCGUC CC - UU AUAU^ UC - A CCGA S 150 140 130 120 110 100 90 80 Initial dG = -63.00 S AGAGGAGA GAGAGAGAGUCG GGA GGAG GAG GAGU GG GAC AGG AACUGG CGC CGU UInitial dG = -30.30 % 10 20 30 %ACC .-CA| AG U AACC % GUCGG UGAAC GU CGGCGACCUC \% CGGCC ACUUG CG GCUGCUGGAG C%CUU \ --^ A- C CGAC %80 50 40  CAG CC  GUU \ CAA A A-- CA Initial dG = -40.50 7 10 20 30 40 50 7G|UG AG G GGC U U GUUU UA CAUC 7 U GUG UU GCUGGGUU UGUCGGU CC CC GCAU CCUC U7 G CGC GA CGGCCCAA GUAGCUA GG GG UGUG GGGG U7G^GU AG G --- - U AC-- GC ACGC 7 100 90 80 70 60 Initial dG = -61.90 5GC G CU U U -| AU 5 AG UA GUUCCAGUUUCAAGGU UCAAAGCUGAAGGCU CCCAG AG C5 UC AU CAGGGUCAAAGUUCCA GGUUUCGAUUUCUGG GGGUC UC C5UU G UU C U G^ GU 5 100 90 80 70 60 Initial dG = -45.40 0 10 20 30 40 0AUUCUA| A A C A GA 0 CUUCCUCUGU CC AAAUAAUUG AGUUGG GAGAAAUA C0 GAGGGAGGCA GG UUUAUUAAC UCAACC CUCUUUAU U0UUUUAA^ A A A C GA 0 90 80 70 60 50 Initial dG = -52.50 C 10 20 30 40 50 CGGGUUGAAAA GA U - .-UACUU U---- - --| CU C AGGU UGC GU GGUGAUC GCCGAUGC CAUG GAGC UGC \C UCCG ACG CA UCACUAG UGGUUACG GUAC UUCG ACG AC 150 140 80 70 60 @ 90 100 110 @ GCUGUUCA UC GU -- UU @ CUCUG GU UUGC UGGC C@ GAGAC CG AACG GCCG U@ -------- C- AU UA UC @ 130 120 Initial dG = -36.70 = 10 20 30 40 50 60=A GUUUC -| A GAUUUUGG UC GA CCU = UUAUU GAGGAA AGGAAGAAGCC UG UCUC GGC CGGCGG A= GGUAA CUUCUU UUUUUUUUCGG AC AGGG CCG GUCGUU A=G ACUGU C^ A AAGUA--- U- AG CUC = 110 100 90 80 70 * 10 20 30 *- C A U --| U * AUUCUUGU UUAG UUUGUCUAGA AUGAA UAUCAAG \* UAAGAACA AAUC AAAUAGAUCU UACUU AUAGUUU A*U U C U AC^ U *. 70 60 50 40 Initial dG = -94.40 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 - AC GGU GGUG U U --- U U U C U - U A -- CCCA- UC UGACUAAAAA .-AAAAAGUAAUAA| UGU UCUA AAUAUAUUUGCA UUUAGGUCUAAGU AAUA CGA CUCU GAA AUA UGA AACCUU ACAAA UUGC UGAUUAUUU GUAG GCU AUUAUUGU CAUGCAUA CA AAGCA UGGGG U GGAU UUAUAUGAACGU AGAUCCAGAUUUA UUAU GUU GAGA CUU UAU AUU UUGGAA UGUUU AAUG ACUAGUAAG UAUC UGA UAGUAAUA GUACGUAU GU UUCGU ACUCU GA CU AC- AUUG - - ACG C U U - - A U G AA CCUAA UA ---------- \ ------------^ UGU 340 330 320 310 300 290 280 270 260 250 240 230 220 160 170 180 190 CAAAGAAAGAUGGAA AA < AAGUUU \ UUCAAA C AAGUAGAAGGAAGUG AA 210 200 AAGUUU \Initial dG = -97.20 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 - C GGU GGUG U U --- U U U C U - U A -- CCCA- UC UGACUAAAAA .-AAAAAGUAAUA| GGGUG U UCUAG AAUAUAUUUGCA UUUAGGUCUAAGU AAUA CGA CUCU GAA AUA UGA AACCUU ACAAA UUGC UGAUUAUUU GUAG GCU AUUAUUGU CAUGCAUA CA AAGCA AUU UUGUG U GGAUC UUAUAUGAACGU AGAUCCAGAUUUA UUAU GUU GAGA CUU UAU AUU UUGGAA UGUUU AAUG ACUAGUAAG UAUC UGA UAGUAAUA GUACGUAU GU UUCGU UAG AACAC CA U AC- AUUG - - ACG C U U - - A U G AA CCUAA UA ---------- \ -----------^ AAAGA U 340 330 320 310 300 290 280 270 260 250 240 230 220 170 180 190 GGAA----------- AA Initial dG = -103.70 10 20 30 40 50 60 70 80 90 100 110 120 - C GGU GGUG U U --- U U U C U - UU UG .-AA| AU UC UCUAG AAUAUAUUUGCA UUUAGGUCUAAGU AAUA CGA CUCU GAA AUA UGA AACCUU ACAAA UUGC UGAUUAUUU GUAGCGCU--CUUUCA AG AUG UG \ GGAUC UUAUAUGAACGU AGAUCCAGAUUUA UUAU GUU GAGA CUU UAU AUU UUGGAA UGUUU AAUG ACUAGUAAG UAUCGUGA GGAAGU UC UAC AC CA U AC- AUUG - - ACG C U U - - A U- \ GU \ --^ GU AC s Table S1. Conserved miRNAs from barley and its evolutionary conservation compared with miRNA of other plant species  7. Table S2.The putative novel miRNAs from barley 73 GCA GCCGC GUC AGCAGG UGCC G A3 AUGA -- GG C -------- - G 3 120 110 100 90Initial dG = -37.50 . 10 20 30 40 .AU AG GGC U .-GAA| A GA . AC GUGCAGA GCU GUUUUGGUGUAC GU CGA \. UG CACGUCU CGG UAGAACUACGUG CA GCU C.GU CU ACU C \ ---^ - GA . 100 90 80 50 / AGUUGA - A / GG CG \/ CC GC C/ AAAGA- A A Initial dG = -20.20 * 10 20 30 *- C-| AGGU - AGA- C CUCGU * GCC CAA GAGAUCA GAA UGUG GGG C* CGG GUU CUCUGGU CUU ACAU CCC A*A UA^ ---- A GGCC - AAUUA * 70 60 50 40 Initial dG = -37.40 & 10 20 30 &C .-CAA| C U ACU & CCGUGC ACUC UUGGUUA CAACUUCGA G& GGCGCG UGGG AACCAGU GUUGGAGCU C&C \ ---^ C - CGA & 60 50 40  AUCUGAAG G  GCU U CGA C CCA----- C : 10 20 30 40 50 miFAMNameSeqlengthtaeosabdi farzmasbisofsspMIR156** Hvu-miR158aTCCCAAATGTAGACAAAGCAMIR158alyMIR159MIR160MIR164 Hvu-miR164bTGGAGAAGCAGGGCACTTGCA Hvu-miR165bTCGGACCAGGCTTCATCCMIR166TCAAGCTGCCAGCATGATATAMIR168 Hvu-miR169aCAGCCAAGGATGACTTGCCGACTAGCCAAGAATGACTTGCATCMIR169gma Hvu-miR169qTAGCCAGGATGACTTGCCGAvvi Hvu-miR171fTGAGCCGAACCAATATCACTCMIR172 Hvu-miR390bAAGCTCAGGAGGGATAGCGCCMIR390 Hvu-miR393bTTCCAAAGGGATCGCATTGATMIR393 Hvu-miR395aGTGAAGTGTTTGGGGGAACTCMIR395MIR396 Hvu-miR396iTCCACAGGCTTTCTTGAACGAGAMIR397 Hvu-miR398TGTGTTCTCAGGTCGCCCCCMIR398 Hvu-miR399bTGCCAAAGGAGAATTGCCCMIR399*MIR444 Hvu-miR482TTGCCAATTCCACCCATTCCTAMIR482sly Hvu-miR528TGGAAGGGGCATGCAGAGGAGMIR528CTGCATTTGCACCTGCACCTAMIR530 Hvu-miR535TGACAACGAGAGAGAGCACMIR535 Hvu-miR821eGAAGTCATCAAAATAAAAGMIR821 Hvu-miR845bAAGCTCTGATACCAATTGATAGCA Hvu-miR845cGAGGCTCTGATACCAATTGTTGGCMIR845 Hvu-miR845eTGGCTCTGATACCAATTGAAAGGA Hvu-miR894TTTCACGTCGGGTTCACCAMIR894ppt Hvu-miR916AGAGGTCATCGGTTCGATCCCGTTMIR916cre Hvu-miR1083TAGCCTGGAACGAAGCACGTMIR1083smo Hvu-miR1117TCTAGTACCGGTTCGTGGCACGAMIR1117 Hvu-miR1432TCAGGAGAGATGACACCGACAMIR1432 Hvu-miR1511AAACCAGGCTCTGATACCATGTTAMIR1511 Hvu-miR1520kTCAGAACATGACACATGACGACATMIR1520 Hvu-miR1863GCTAGCTCTGATACCATGTTAGGCAMIR1863 Hvu-miR2118qTTCCCGATGCCTCCTATTCCTAMIR2118 Hvu-miR2120AGGATCTTTAGTCCCGGTTGGTAAMIR2120Hvu-miR2275c-3pTTCAGTTTCCTCTAATATCTCAMIR2275 Hvu-miR2911GCCGGCCGGGGGACGGGCTGGGMIR2911peu Hvu-miR2916AGTTGGGGACTCGAAGACGATCAMIR2916 Hvu-miR4362AAGGACAGACGTCATGTAGATCTMIR4362 Hvu-miR4992ATAAGATGGTTTTTGTTAGAGTAAMIR4992 Hvu-miR4995TAGGCAGTGGCTTGGTTAAGGMIR4995 Hvu-miR5021CGACGGAGAAGAAGAAGAAGAAACMIR5021athTATTTGCAGGTTTTAGGTCTAATTTGGCACCTTGAAACTGGGAMIR5067 Hvu-miR5139AACCTGGCTCTGATACCAMIR5139rgl Hvu-miR5141AGACCCGACGCGACTGACGGATMIR5141Hvu-miR5176-3pGATGATGTGGCATAGAATTAAMIR5176 Hvu-miR5200TAGATACTCTCTAAGGCTTGGMIR5200 Hvu-miR5205aAGACAATTTGGGACGGAGGGAGTMIR5205mtr Hvu-miR5210AAATGTGTTGGAATTAAGMIR5210 Hvu-miR5339CAGATAGAGAATCTTCTCAGAMIR5339 Hvu-miR5368AGGGACAGTCTCAGGTAGACAGTMIR5368 Hvu-miR5486AGGGGCTTGCATATTCTACCAMIR5486 Hvu-miR5497CAGAATATCTGGGACGAGCATMIR5497 Hvu-miR5503TTCGGATCTTTCTAGAGGCATTMIR5503 Hvu-miR5522GAACAATAGGAATGGGAGGCAMIR5522 Hvu-miR5532ATGGAATATATGACAAAGGTGGMIR5532 Hvu-miR5538CTACTGAACTCAATCACTTGCTGCMIR5538Supporting materials:1Table S5. Target gene prediction of barley miRNAs  Conservativecountdicot Hvu-miR156aTGACAGAAGAGAGAGAGCACA Hvu-miR156bTGACAGAAGAGAGAGAGCAC Hvu-miR156cTGTCAGAAGAGAGTGAGCAC Hvu-miR156dTGACAGAAGAGAGTGAGC Hvu-miR156eTGACAGAAGAGAGCGAGCAC Hvu-miR156gTTGACAGAAGATAGAGGGCAC Hvu-miR156hTGACAGAAGAAAGAGAGCAC Hvu-miR156jTGACAGAAGAGGGTGAGCAC Hvu-miR156lTGACAGAAGAGAGTGAGCATA Hvu-miR156oTTGACAGAAGAGAGTGAGAAC Hvu-miR159dTTTGGATTGAAGGGAGCTCCG Hvu-miR159eTTTGGATTGAAAGGAGCTHvu-miR159e-3pTTTGGATTGAAGGGAGCTCATT Hvu-miR319MIR319TGGACTGAAGGGAGCTCCCTCHvu-miR319a-3pACTGGATGACGCGGGAGCTAA Hvu-miR319bTTGGACTGAAGGGTGCTCCCT Hvu-miR160bTGGCTCCCTGAATGCCATCC Hvu-miR160dTGCCTGGCTCCCTGTATGCCA Hvu-miR160eTGCCTGGCTCCCTGAATGCCA Hvu-miR160hTGCCTGGCTCCCTGCATGCCAptc Hvu-miR164aTGGAGAAGTAG< GGCACGTGCT Hvu-miR164cTGGAGAAGCAGGGTACGTGCA Hvu-miR164eTGGAGAAGCAGGGCACGTGCA Hvu-miR164fTGGAGAAGAAGGGCACATGCA Hvu-miR164hTGGAGAAGCAGGGCACGTGTA Hvu-miR166aTCGGACCAGGCTTCATTTCCCC Hvu-miR166eCTCGGACCAGGCTTCATTCCC Hvu-miR166fTCTCGGACCAGGCTTCATTCC Hvu-miR166iTCGGATCAGGCTTCATTCCCC Hvu-miR166kTCGGACCAGGCTTCATTCCTT Hvu-miR166lCCGGACCAGGCTTCATTC Hvu-miR166mTCGGACCAGGCTTCATTCCCT Hvu-miR166nTCGGACCAGGCTTCAATCCCT Hvu-miR166pTCGGACCAGGCTCCATTCCCC Hvu-miR167aMIR167 Hvu-miR167bTGAAGCTGCCAGCATGATCTA Hvu-miR167cTGAAGCTGCCAGCATGATCTT Hvu-miR167dTGAAGCTGCCAGCATGATCTGA Hvu-miR167hTGAAGCTGCCAACATGATCTA Hvu-miR168aTCGCTTGGTGCAGATCGGGAC Hvu-miR168bTCGCTTGGTGCAGCTCGGGAC Hvu-miR169dTAGCCAAGAATGACTTGCCT Hvu-miR169eTAGCCAAGGATGACTTGCCTG Hvu-miR169fCAGCCAAGGATGACTTGCCGG Hvu-miR169hTAGCCAAGGATGACTTGCCTATHvu-miR169j-5p Hvu-miR169kTAGCCAAGGATGATTTGCCT Hvu-miR169lCTGCCAAGAATGACTTGCCGC Hvu-miR171bMIR171TTGAGCCGTGCCAATATCAC Hvu-miR171dD GAUUUGC U AG AG D CG GGUCAGGG CUC CD GC CCAGUCCC GGG AD CA----- U AG CU D 100 90 Hvu-miRn29bInitial dG = -58.60 H 10 20 30 40 50 60 HG| C CGCA - GGU G CCAU GG-- --- A - C H CG UGC CUCG CCCUACU ACG ACUCA CGG GCCU CCUCGCA CGGG UGCC \H GC AUG GAGU GGGAUGA UGC UGAGU GCC CGGG GGGGCGU GCUC ACGG AH-^ A CUAG A --- A ACGU GCGA AAC C G C H 130 120 110 100 90 80 70 Initial dG = -61.40 1 10 20 30 40 1C ----- AC AGA .-AA| CA 1 GAGCGA UGGAGG GAGG UGGAGAGG GGUGCAG G1 CUCGCU GCCUCU CUCC ACCUCUCC CCACGUC U1U CUGCA CC ACC \ --^ CA 1 130 120 110 50 A 60 70 A GCAAG A C CU A GA UCG A GG UU CGC C CC GC \A CC AA GCG G GG CG CA G---- - C AG A A- UCG A 100 90 Initial dG = -42.90 / 10 20 30 40 /UU| AC CC C N NNCAA / GUAC CCUCUGUU UAAAUGUAAGA GU UUUGGCAG \/ CAUG GGAGACAA AUUUACAUUCU CA AAACCGUC U/AU^ AU AN A - AAGNU / 90 80 70 60 50 Initial dG = -32.80 0 10 20 30 40 0---|UU AUAUUGAC G U AUG G 0 G CUCACCGA GGCUCU UCU GG GUCAUUCU U0 U GGGUGGUU UCGAGG AGA CC UAGUGAGA U0AGU^GG ACUUCAAC - - AA- G 0. 80 70 60 50 Initial dG = -30.40 0GU-- -| AUAUUGAC G U AUG G 0 UCUCC CCGA GGCUCU UCU GG GUCAUUCU U0 GGGGG GGUU UCGAGG AGA CC UAGUGAGA U0AGUU U^ ACUUCAAC - - AA- G Initial dG = -52.20 @ 10 20 30 40 50 @AAG A- C- - .-GCCUCCUC - - -| AG @ CUUGC GCCACAGAAUU UCACAG UGAUGG UUG CC CCGU GUC U@ GGAUG UGGUGUCUUAA AGUGUC ACUACC AAC GG GGUA CGG C@AUA AC AC A \ -------- U U C^ AA @ 140 130 120 70 60 4 .-AGCC UUU 4 AUCC C4 UAGG U4 \ ---- UAU 2 CA CUU 2 ACAG \2 UGUC C2 G- CUA Initial dG = -51.80 1 10 20 30 40 1A| U C C A AG 1 UCUAU UACUUCCUC GU CCUAAAUAAUUGUAGUU GAGAGAAU U1 GGAUG AUGAGGGAG CA GGAUUUAUUAACAUCAA CUUUCUUG U1A^ U A U C AU 1 90 80 70 60 50Initial dG = -33.60 * 10 20 30 40 *UAGAAAU| AU GUG GG * UACUCCCUCUGU ACCUAAAUAA UAGUU G* AUGAGGGAGACA UGGAUUUAUU AUCAA A*UCAUUAC^ -- A-- GA *. 70 60 50 Initial dG = -56.00 S 10 20 30 40 50 60 70 SUUG UU-- U-- C - - UCUU .-AAUGGGAGAA G .-UU| CAU G S GGC UGU UGG CAA CU AGAUCUAUGAU GC GUGCUU GUU GGGUU ACC US CUG ACA ACC GUU GA UUUAGAUACUA CG CACGGA CGA UCCAG UGG GSCUA UACU UCU U A C CUUU \ ---------- A \ --^ --- C S . 180 170 160 150 110 80 N UU AGUG N CCC AN GGG CN CG AAGA A 120 130 A ACAU G AU A CAU UUGUUGCC \A GUA AACAAUGG CA G--- G GA Initial dG = -57.80 8 10 20 30 40 50 8- CU- C -| UGGA UUU 8 CAA GAGUUUGACCCUGAGGCACUCAUACCGCUA CU AGUU AUG \* 10 20 30 *- CCAUC UC - .-UUC| AU * AGCAG AACCAG GAG GCCAGGGGG GA \* UCGUC UUGGUC CUC UGGUUUCCC CU U*C ----- -- G \ ---^ AG * 90 80 70 40 2 UUGCUU U C 2 CAUC CAA C2 GUAG GUU U2 ------ U U Initial dG = -39.90 < 10 20 30 40 <GUUUC C A ------------| CC G < UACUUCC CUGUUCCAAAAUAA UGUCU CAA UUGUACUA C< AUGAAGG GGCAGGGUUUUAUU ACAGA GUU AAUAUGAU U<UUAUU A A UCUCCGAAUCAU^ AA C < 100 90 80 70 60 50 Initial dG = -39.20 ' 10 20 30 'GUUUC C A | A ' UACUUCC CUGUUCCAAAAUAA UGUCU--CA C' AUGAAGG GGCAGGGUUUUAUU ACAGA GU C'UUAUU A A \ ^ U ' 100 90 80 40 5 UACUA U A 5 GCUU AGUAUA A5 CGAA UCAUGU A5 UCUC- - U 5 70 60Initial dG = -48.20 5 10 20 30 40 50 5AAG| G C G A - U 5 UACUCCUUCCGUCUCAAUAUAA UGUCU AA UUU GUAU AAUUUUA \5 AUGAGGGAGGCAGGGUUGUAUU ACAGA UU AAA CAUG UUGAAAU A5UAG^ A A G - A U 5 100 90 80 70 60 Initial dG = < -34.20 ( 10 20 30 (- ACU GGU AA A .-CA| GG ( GGAA GAG GGAAG AG GGGCGG GCC G( CCUU CUC UCUUC UC CCCGCC CGG C(C CC- AU- C- G \ --^ GU ( 90 80 40 / A - AAGAAAG / GGA GG \t CUUGUACUAUAACUACUUAC \ ---------------------- U t 210 200 130 y 140 150 160 y CAACUU A UUCCAAAG AU y GAGC ACUGUAAU GU Uy UUUG UGACGUUA CA Uy ------ A CCCUGUA- GU y 190 180 170 Initial dG = -42.60 & 10 20 30 &U - .-G|U AGUUG UGCUC & GGGUC CA G CGGUGUGGGU AGC \& CUCAG GU C GUCGCACCCG UCG C&U U \ -^- GUGG- UCUUU &. 50 40 . 60 70 80 90 . AGUACUGC GA- U GAAGU . CGCAGU AGUU UUCUUCU A. GUGUCG UUAA AAGAGGA A. AA------ AGA U AAGGU . 120 110 100  Initial dG = -41.70 ' 10 20 30 'U - .-G U ----| U CUG ' GGGUC CA G CGGUGUGGGU AGU GAA C' CUCAG GU C GUCGCACCCG UCG CUU U'U U \ - - GUGG^ U UCC '. 50 40 Initial dG = -43.70 &U - .-G|U AGUUA UGCUC  Initial dG = -154.70 .-C -| AC  GAGUGC GUCG \ CUCGCG CGGU A\ - U^ CC 0 30 40 50 60 0 .-CAA C C CCGUC C C--- .-A C 0 CU CU CUGC CUGC GC GCCCC UCGUC \0 GA GA GACG GAUG CG UGGGG AGCAG U0 \ --- A U AU--- C UUCU \ - C 0 200 190 180 70 c 80 90 100 110 120 c CC - CAA - AA -- GACCAUCCU CGU c CCGCG CCGCC GCGG UGCCGC GU GCGAGGA GGAGGC \c GGCGC GGCGG CGCC ACGGCG CG UGCUCCU CCUCCG Cc -- C CGG G GC UC --------- CUA c 170 160 150 140 130 : 210 220 230 240 : A C .-CCCCUG - -- GU UGU- UU : AG GUCAUGU GCGG GUGGGU CCAG CGG GGGUAG \: UC CAGUGUA CGUC CACCCG GGUC GUC CCCGUC G: - U \ ------ G GU -- UUUC GA : 350 280 270 260 4 290 300 310 4 AGUACUGC GA- U GAAGU 4 CGCAGU AGUU UUCUUCU A4 GUGUCG UUAA AAGAGGA A4 A------- AGA U AAGGU 4 340 330 320 Initial dG = -19.20 + 10 20 30 40 +GC| GCAU UUUCA CC-- A + GUUA AUUG UGUUCAAACGUCC UUUGC \+ UAGU UGAU AUAGGUUUGUAGG AAACG U+AU^ ACUU UUUGC AUUC C + 80 70 60 50 Initial dG = -26.40 0 10 20 30 0CG ----- GU U C- ACUCA---| U 0 AGG CUGUG UCUGCA CUC GUGCC CUCU A0 UCC GAUAC GGACGU GGG CACGG GAGG U0UG CUUAA -- C CA AUCGUUCG^ U 0 80 70 60 50 40Initial dG = -59.20  10 20 C .-GUG| A G AAG  ACGAGG GGCC CAUGGGC UG G UGCUCC CCGG GUACCUG AC U- \ ---^ - A CGG  40 303 50 60 70 80 3 ACCA GA AU C AACAAGAA G C 3 CGU CGGUG CAG UCGUCC GCGG C GE Table S3.The secondary structure of the newly identified Barley miRNA 7" Table S4. Primers used for qRT-PCR 7TC208562Glutathione S-transferase TSI-1TC221252SOUL heme-binding protein-likeTC216584&Multidrug resistance protein 1 homologBU9704431Phospholipid-hydroperoxide glutathione peroxidaseTC223207TC217005TC210538FTTC223667Actin-6DN156857 Glucan endo-1,3-beta-glucosidase 7BG309224 otopetrin 1TC224113&Bacterial-induced peroxidase precursorBU987855TC232080TC223241$Chloroplast 30S ribosomal protein S8TC223711$Chloroplast 30S ribosomal protein S3CK5683390Protoporphyrin IX Mg-chelatase subunit precursorCX632312KI domain interacting kinase 1BM443289Early nodulin 75-like proteinTC205541 Two-component response regulatorCA009933CChromosome undetermined scaffold_104 whole genome shotgun sequenceBG418690Cytochrome reductaseTC209134Probable transmembrane proteinCK566959 VitellogeninTC196924Sec61 alpha subunitTC213187BU974512)Meiotically up-regulated gene 154 proteinTC201826'Short-chain dehydrogenase/reductase SDRTC233191Metallo-beta-lactamase-likeBG299802NirSTC216185.Transcriptional regulator, LysR family proteinBJ480073!coiled-coil domain containing 136DN156749LProbable histone-lysine N-methyltransferase NSD2 (EC 2.1.1.43), partial (1%)TC222488Beta-defensin 123 precursorBY858005!Winged-helix transcription factorTC237450TC227440%Phosphoethanolamine methyltransferaseTC227983TC200155TC197523Transcription factor MYB39TC205883#Uncharacterized protein At1g18620.2TC229003Aquaporin NIP1-3TC2188951SET-domain transcriptional regulator-like proteinBI956664Malate dehydrogenaseTC220447.Emsy N terminus domain-containing protein-likeTC2064319Branched-chain-amino-acid aminotransferase-like protein 1TC211932CX626387 TSBP proteinBF064522GUDP-glucoronosyl and UDP-glucosyl transferase family protein, expressedTC221409TC217173Cyanate hydrataseBI947058&Uncharacterized protein yadL precursorTC221159Cytochrome c oxidase subunit ITC214518Arm repeat proteinTC204557P450TC218364TC195124,Serine/threonine kinase-like protein ABC1040TC210261Cyclin-dependent kinase F-3TC223315TC229461C-mycGH213613DNA binding protein-likeBQ754039J-domain proteinTC229758TC214914Hin1-like proteinTC205531SMeltrins, fertilins and related Zn-dependent metalloproteinases of the ADAMs familyBE4214380Vacuolar ATP synthase 16 kDa proteolipid subunitTC199748Phenylalanine ammonia-lyaseTC231325TC230590GTP-binding protein SAR1ATC196742TC236970+Chlorophyll synthase, chloroplast precursorTC201556 RNA helicase Hvu-miRn028 7TC231355)Zinc finger (C3HC4-type RING finger)-likeTC234130 FibrillarinBG300599Calmodulin-binding protein-likeBY861926 HSP20 proteinBJ480160Beta adaptin-like proteinTC196598;Cyclic nucleotide-gated calmodulin-binding ion channel-likeTC196614Proteasome sub< unit alpha type-5AJ476175BQ468979(Pollen coat oleosin-glycine rich proteinTC207713Pseudouridylate synthaseGH228307Phospholipase D alphaTC204396TC202472TC223782Quinone reductase 2TC224164"Early transcribed membrane proteinFD5224242Type II and III secretion system protein precursorTC198932Surface antigen protein 2TC224987Homeodomain proteinTC230450%Stress-induced receptor-like kinase 2TC231123TC197323TC207657 Fibroin 1TC228739Expansin EXPA6TC224940TC224133TC230161Glutathione transferaseTC202587+Cinnamyl-alcohol dehydrogenase-like proteinTC228542LigABI947651%One repeat myb transcriptional factorTC208702TC229055%Hepatocyte erythrocyte protein 17 kDaTC218605Expansin EXPB4TC202593BM098091/DNA-directed RNA polymerase iii largest subunitBM816796F-box family proteinTC2354437180-200 kDa membrane protein scavenger receptor homologTC215360Ribonuclease BNBE437939HHomolog of Homo sapiens 'Splice Isoform 2 of Tripartite motif protein 39TC237488TC201998SR protein factorBF256221 Wali2 proteinTC231374PE-PGRS family proteinTC203730ERD4-related membrane proteinTC204205&Ubiquitin-conjugating enzyme E2-23 kDaTC195833Pollen caleosinTC220359TC230772@Magnesium-protoporphyrin IX monomethyl ester [oxidative] cyclase 7Transcription factor GAMYB 7 flame chlorosis virus-like agent 7;CCAAT-binding transcription factor subunit B family protein 7'6,7-dimethyl-8-ribityllumazine synthase 7GATA zinc finger family protein 74Ribulose bisphosphate carboxylase small chain PWS4.3 7)Hydrolase, alpha/beta fold family protein 7)Hydrolase, alpha/beta fold family protein 7KH domain containing protein 7KH domain containing protein 7(Protein kinase domain containing protein 7Triosephosphate isomerase 7VHS domain containing protein 730S ribosomal protein S17 730S ribosomal protein S17 7@ATP-dependent Clp protease ATP-binding subunit clpA homolog CD4B 7+Harpin-induced protein 1 containing protein 7AIG1 family protein 7Conserved serine-rich protein 7.Chlorophyll a/b-binding protein WCAB precursor 7)Mitogen-activated protein kinase kinase 3 7Phenylalanine ammonia-lyase 7? CCAC CCU ACA CCA \? GGUG GGA UGU GGU N? \ -- GA A UC-- GUU ? 110 100 P 120 130 140 150 P A- U G - .-NCAAAGA .-GANAU G P UCCCG GUAU C GCU CGAC GCC \P GGGGU CGUG G CGG GUUG CGG AP CG U G U \ ------- \ ----- A P 230 220 V 160 170 V CAUGAAAA UGA V GUGA \V CACU UV AUUA---- UGA K 190 200 K AUGAAAA----- GU K CUU AK GAA AK AUAAGACAAAAG AA Initial dG = -25.00 , 10 20 30 40 ,CUUUCAAAGA UG C C .-GAA| UU , CACGGAAC GG GGUUG ACU GACA \, GUGCUUUG CC CCAGU UGA CUGU A,UAUUCGAGAA CA - - \ ---^ UA , 90 80 70 . AUACA UGC . UCG \. AGC C. AG--- UUC . Initial dG = -30.70 :- .-GUCAA A C .-CA| CCACAU CAA UC : CCUCC CAG UCAAGA GCUCCGGCA AGUAC GC GU \: GGAGG GUC GGUUUU CGGGGCCGU UCAUG CG CA A:C \ ----- A A \ --^ AGC--- C-- UU : 110 100 60 6 A----------- ACC 6 CAA C6 GUU U6 AUAGAUUCUUAC CCU 6 90 80  120 130  AAUA G CCUCC  GCA CACAAC \ CGU GUGUUG U A--- G UCCAC  150 140 Initial dG = -19.90 0 10 20 30 40 0CUA A C A - CAUC UUA----| U 0 AUAC GUUUA AAUGC C GAUG GCCUCAG UG G0 UAUG CAGAU UUGUG G CUGC CGGGGUU AC U0ACA A C A C AA-- UUAAAAG^ A 0 80 70 60 50 Initial dG = -48.10 ` 10 20 30 40 50 60 70`---------- U A A ----- G U --- .-GAAA .-A UCUG- C---| U ` GAUUAGUC UG GG UGAUC GUUU G CAUG CUGG GACAG AACUU CUC GGAAAAGAAU U` CUGGUCAG AC UC GUUAG CAAA C GUAU GACC CUGUC UUGAG GAG UCUUUUUUUA U`CAAUUAUCGU - A - ACAUA G - UGA \ ---- \ - UUUUA AAUA^ U ` 210 200 190 180 170 140 90 80 \ 100 110 120 \ GAUUCUUUUUG C -- CU \ CUG AGA UUG \\ GAC UCU AAC A\ A---------- U UA AC F AUAAUU A F UUUCAGA \F GAAGUUU UF AU---- U Initial dG = -70.50 I 10 20 30 40 50 60 IACGUUUAC -- A A A C UC U .-CUUCA .-A -| AG I CACGAC GACG CG GC CGA AAG GG GGGCGG UGCUC GG UCGGCGC AI GUGCUG CUGC GC CG GCU UUU CU UCCGCU ACGGG CC AGCUGCG CICUAGCCUA UA - G C - GA U \ ----- \ - G^ CC I 190 180 170 160 70 M 80 90 M GG - CA M CGGCG GC--GGC \M GCCGU CG CCG AM < -- A \ AC R A----- UU R GG \R CC UR UACUAA AA @ A------ AAGA @ UUCAC A@ AGGUG A@ UGCUUAA ACCU @ 150 140analysis by mfold version 3.5*M. Zuker, Rensselaer Polytechnic Institute Primer nameSequence (5' to 3')18s rRNA forwardGTGACGGGTGACGGAGAATT18s rRNA reverseGACACTAATGCGCCCGGTAT Hvu-miRn016cgnl|UG|Hv#S46888350:343:423:+ Hvu-miRn017GGTGGATCGATGAACCCGGCGAgnl|UG|Hv#S12577759:17:120:+ Hvu-miRn018agnl|UG|Hv#S12603856:235:339:- Hvu-miRn018bgnl|UG|Hv#S12645774:280:384:+ Hvu-miRn018cgnl|UG|Hv#S15199467:161:265:- Hvu-miRn018dgnl|UG|Hv#S15202936:334:438:+ Hvu-miRn019aTACAATTATTTAGGAACGGAGgnl|UG|Hv#S12610725:126:221:- Hvu-miRn019bgnl|UG|Hv#S12611217:148:243:- Hvu-miRn019cgnl|UG|Hv#S12851315:151:246:- Hvu-miRn019dgnl|UG|Hv#S12851593:111:206:- Hvu-miRn020AGGTGATGCTGTGGTGATCTgnl|UG|Hv#S12614112:174:331:+ Hvu-miRn021aCGAGGAAAGGAAGAAGCCATGgnl|UG|Hv#S12617378:78:194:- Hvu-miRn021bgnl|UG|Hv#S12618143:158:274:- Hvu-miRn021cgnl|UG|Hv#S12618144:145:261:- Hvu-miRn021dgnl|UG|Hv#S12780980:152:268:- Hvu-miRn021egnl|UG|Hv#S12783667:152:268:- Hvu-miRn022aTAGATTTGTCTAGATATGAAgnl|UG|Hv#S12617700:39:118:- Hvu-miRn022bgnl|UG|Hv#S12818417:40:119:- Hvu-miRn023agnl|UG|Hv#S12626198:196:539:+ Hvu-miRn023bgnl|UG|Hv#S12684203:187:530:+ Hvu-miRn023cgnl|UG|Hv#S12808034:191:547:+ Hvu-miRn023dgnl|UG|Hv#S39523879:212:555:+ Hvu-miRn023egnl|UG|Hv#S48849876:196:539:+ Hvu-miRn023fgnl|UG|Hv#S12684445:187:530:+ Hvu-miRn024GTGGAAAGTGGGCAGTTGGGTTgnl|UG|Hv#S12682787:90:267:+ Hvu-miRn025TAGGAGAGGGGGGAAGGGATCTgnl|UG|Hv#S12696085:174:287:- Hvu-miRn026aCTCTGCCACCGGTCCATGACGACgnl|UG|Hv#S12698195:219:412:- Hvu-miRn026bgnl|UG|Hv#S15197490:229:422:- Hvu-miRn027AACTGCAGTACTTGTGGGGAAgnl|UG|Hv#S12700874:31:163:+ Hvu-miRn028TATGGGGATCTGACAGACGAGgnl|UG|Hv#S12703572:305:391:- Hvu-miRn029aTGAGTACGTAGTAGGGATGAGgnl|UG|Hv#S12712711:26:170:+ Hvu-miRn029bgnl|UG|Hv#S32760078:171:302:+ Hvu-miRn030AGGACGAGGAGATGGAGAGGAgnl|UG|Hv#S12746049:57:192:+ Hvu-miRn031TCTGTTCCTAAATGTAAGACGgnl|UG|Hv#S12777146:235:326:+ Hvu-miRn032aGCTCTGTCTTGGATGGTCATTCgnl|UG|Hv#S12791026:455:544:+ Hvu-miRn032bgnl|UG|Hv#S12797319:455:544:+ Hvu-miRn032cgnl|UG|Hv#S42550728:66:155:- Hvu-miRn033aCCACAGAATTCTCACAGTGATGGgnl|UG|Hv#S12810862:176:323:+ Hvu-miRn033bgnl|UG|Hv#S12811004:299:446:+ Hvu-miRn034aAATTATTTAGGTACAGAGGGAgnl|UG|Hv#S15195843:200:297:- Hvu-miRn034bgnl|UG|Hv#S15209220:172:251:+ Hvu-miRn034cgnl|UG|Hv#S41556714:467:546:+ Hvu-miRn034dgnl|UG|Hv#S41556718:88:167:- Hvu-miRn035TTTGGCCAACTAGATCTATGAgnl|UG|Hv#S15197625:297:487:+ Hvu-miRn036TGACCCTGAGGCACTCATACCGgnl|UG|Hv#S15199857:241:348:+ Hvu-miRn037AATTAATATGGATCGGAGGGAgnl|UG|Hv#S15204353:407:522:+ Hvu-miRn038TGAAGATTTTAAGCATTGAAgnl|UG|Hv#S15206357:3:99:- Hvu-miRn039AGACGATTAAGAAGACCTGCAAgnl|UG|Hv#S15345790:143:414:- Hvu-miRn040aAGGAGAATAATTAGAGCTGTGAgnl|UG|Hv#S21599653:162:291:- Hvu-miRn040bgnl|UG|Hv#S21600472:162:291:- Hvu-miRn040cgnl|UG|Hv#S21602081:162:291:- Hvu-miRn040dgnl|UG|Hv#S21605165:162:291:- Hvu-miRn040egnl|UG|Hv#S21606378:162:291:- Hvu-miRn040fgnl|UG|Hv#S21607363:162:291:- Hvu-miRn040ggnl|UG|Hv#S21608905:162:291:- Hvu-miRn040hgnl|UG|Hv#S39524333:663:1016:+ Hvu-miRn041AGGATGTTTGGATACGTTTTAGTHK06D16r:117:199:- Hvu-miRn042GGCACACGGGCTGCAGGCATAGgnl|UG|Hv#S23835075:4:88:+ Hvu-miRn043aTGGACGACCTGGGCGCCGACGgnl|UG|Hv#S23837460:2:127:+ Hvu-miRn043bgnl|UG|Hv#S23879357:483:608:- Hvu-miRn044aGTGCATCAAGATCGGCTCATCTgnl|UG|Hv#S32301119:132:237:- Hvu-miRn044bgnl|UG|Hv#S32743122:83:188:- Hvu-miRn045TGAGATCAGAAAGATGTGCGgnl|UG|Hv#S32302937:186:260:- Hvu-miRn046ACTCCTTGGTTATCAACTTCGAgnl|UG|Hv#S39523167:797:885:+ Hvu-miRn047CAGATCAAGACGCTCCGGCAgnl|UG|Hv#S39523871:814:970:+ Hvu-miRn048ATACAGTTTACAATGCACGATgnl|UG|Hv#S41566329:1:89:+ Hvu-miRn049ACAGATTGCTACAGACTGGTCgnl|UG|Hv#S42548914:1:218:+ Hvu-miRn050CGACGACGACGAGCACGACAgnl|UG|Hv#S48832926:275:473:- Hvu-miRn023*aAGACCTAGACATGCAAGTATA Hvu-miRn023*b Hvu-miRn023*c Hvu-miRn023*d Hvu-miRn023*e Hvu-miRn023*f Hvu-miRn024*CACCAACAATCATTTTCCTCTC Hvu-miRn026*aGGCAATGGATTGAGGTCAGGTA Hvu-miRn026*b Hvu-miRn027*TCTTGCAAGTCGTGCAGTTGC Hvu-miRn032*aGAGTGATAACCAGAGGAGCTCA Hvu-miRn032*b Hvu-miRn032*c Hvu-miRn036*GTATGAGTGTCTCAGGGTCAAA Hvu-miRn045*ACACCGGTTCATGGTCTCTTGInitial dG = -45.30 2 10 20 30 40 502AUCACUAU| G AC UA U UG 2 UACUCCCU UGUAUC AAUACUUG GU GGGGGCACU U2 AUGAGGGA ACAUGG UUAUGAAC CA CCCUUGUGA A2CUUCAANG^ G AU -- C UU 2 90 80 70 60 Initial dG = -33.90 + 10 20 30 40 +GU| ACA A GGUC CUCC + CGC UACUCCCU UGUACCUAAAUA UAGUU \+ GCG AUGAGGGA ACAUGGAUUUAU AUCAA C+AU^ ACA G GAAC UCAC + 80 70 60 50 Initial dG = -24.20 / 10 20 30 40 /U - C - GA U-- - -| UAAAA / GCCC UGU AC CU UCUGUACCU UAC UAUA UUG U/ UGGG ACA UG GG AGACAUGGA AUG AUAU AAC U/C U - A G- UUU A C^ CCCUC / 80 70 60 50 Initial dG = -48.70 6 10 20 30 40 50 6N| C A G GCUAA UUUCUG 6 UAUACUCC UCUGU ACUUAAUAUAAGAC UUU ACACUGUC \6 GUAUGAGG AGGCA UGAAUUAUAUUCUG AAA UGUGACAG A6G^ A C G AAAAA CAUCAC 6 100 90 80 70 60 Initial dG = -79.30 A 10 20 30 40 50 60 A- C C C - .-C| C GA A A GGG GGGGG GAAGCUGUAGUGCAG CGGCGUU GUGCCG GCAUCGA GGCG GCUGC \A CUC CUCCC UUUCGACGUUGCGUC GUUGCAG CACGGC CGUAGUU CCGC CGACG GAG - U - U \ -^ A -- U A 130 120 110 100 70 4 U UGG 4 GCAA \u Table S1. Conserved miRNAs from barley and its evolutionary conservation compared with miRNAs of other plant species.  7/ Table S2.The putative novel miRNAs from barley.  7$ Table S4. Primers used for qRT-PCR.  7G Table S3. The secondary structure of the newly identified Barley miRNA.  73 Table S5. Target gene prediction of barley miRNAs.  7 $ 9%9n&n'*l+l,#.# / 1 24t5t6J:J>_@_BB@D!iEJFlGwHaJBK&M Np XP9 7RV]bl&go tv|{tt1 j^zf] Xږt$̥B 8εD.s[A<:?JB9r}NlN~ L:meA c>Cqg#X0O}` jB L hr#'Z)@*G,--7/ 0 &2 35~6d79V;<L>2ccB   GJ  dMbP?_*+%&?'?(?)?" dXX??&U} @ gN;N;N,N;N ;N N N N LO [2 \\\\\\\\\\\\\O [3 ]]]]PPPPPPPPPO MX PPPPPPPPPPPPPO ^Y __PPPPPPPPPPP O ^ `````PPPPPPPP<<<<<>@<`b " 7ggD  $KaxҢ  dMbP?_*+%&?'?(?)?"??&U} @} @!;C;;;;;;; ; ; ; ; ;;;;;;;;;;;;;;;;;;; ; e fffffffffffff DDDDD ggggggggg EV EU EW EX E EY EZ E[ E\  E]  E^  E_  E`  E B Ba AA5@C@LA ~ A h@A AbA Ab  Ab  Ab  AAA BB AA4@~ A@A Ab Ab Ab  Ab  Ab  AAA BB AA4@~ A@U@A Ab AbA  Ab  Ab  AAA BB AA2@~ AX@A Ab AbA  Ab  Ab  AAA BB AA4@ ~ AȄ@A Ab AA  Ab  Ab  AAA B B A A5@ ~ A@ A Ab AA Ab Ab AAA B B A  A4@ ~ A@ A Ab AA Ab Ab AAA B! B A" A4@ ~ A]@ A Ab AA Ab AAAA B# B A$ A5@ ~ AW@ A Ab AA Ab AAAA B% B A& A5@~ Ab@ AAAAAAAA Au Bc Be AdA4@~ A@AAAAAAAA  Af B' Bg A(A5@~ A?A Ab AA  Ab A  Ab  AA B)B A*A2@~ A@A AbAAAA  A  AA B+B A,A6@~ A@AAAAAAAA  Au B- B. A/A5@~ A@ AbAAAA  Ab  AAA B0B A1A5@~ A@P@A AbAAAAAAA B2B A3A5@~ A@@A Ab AbA  Ab  Ab  AAA B4 Bh A5A4@~ A?A Ab AbA  Ab  Ab  AAA B6B A7A5@~ AA@A Ab AbA  Ab  Ab  AAA B8B A9A5@~ A&@A Ab AbA  Ab  Ab  AAA B:B A;A5@~ A?AAAAAAAA  A< B= Bi A>A5@~ A8@A Ab Ab Ab  Ab AAAA BjB AkA5@~ A"@A Ab Ab Ab  Ab  Ab  AAA B?B A@A5@~ A$@A Ab AbA  Ab  Ab  AAA BAB ABA5@~ AP@A Ab AbA  Ab  Ab  AAA BCB ADA5@~ A"@A Ab AbA  Ab AAAA BEB AFA5@~ A@AAAA  Ab AAAA Bl Bn AmA2@ ~ A@AAAAAAAA  Af BG B AH A6@!~ A? A Ab Ab A Ab Ab AAA Dl2<{{{!;";#;$;%;&;';(;);*;+;,;-;.;/;0;1;2;3;4;5;6;7;8;9;:;;;<;=;>;?;@; !BI!B !AJ!A5@"~ !AB@!A !Ab !Ab!A ! Ab ! Ab ! AAA "BK"B "AL"A5@#~ "A?"A "Ab "Ab"A " Ab " Ab " AAA #BM#B #AN#A5@$~ #AA@#A #Ab #AA # Ab # Ab # AAA $BO$B $AP$A5@%~ $AX@$A $Ab $AA $ Ab $ Ab $ AAA %BQ%B %AR%A2@&~ %A?%A %Ab %AA % Ab% AAAA &BS&B &AT&A5@'~ &A@&A &Ab &AA & Ab& AAAA 'BU'B 'AV'A5@(~ 'A@'A 'Ab'AAAAAAA (BW(B (AX(A5@)~ (A5@(AAAAAAAA ( A< )BY )BZ )Ao)A5@*~ )A@ )Ab)AAAAAAAA *B[*B *A\*A5@+~ *A@ *Ab *Ab *Ab*A * Ab * Ab * Ab * Ab* A +B]+B +A^+A5@,~ +AX@+A +Ab +Ab+A + Ab + Ab + AAA ,B_,B ,A`,A6@-~ ,AP$A,A ,Ab ,Ab,A , Ab , Ab , AAA -Ba-B -Ab-A5@.~ -A:@-A -Ab -AA - Ab - Ab - AAA .Bc .Bp .Ad.A5@/~ .A0$@.A .Ab .AA . Ab. A . Ab . Ab. 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