Supplementary Material Table S1. BLAST database with the downloaded EST sequences involved in the wood production process. In parentheses are the additional keywords used during the search, leading to the resulted sequence collection, presented in the right part of the table No. of the Name of the encoded EST sequences with accession number in NCBI downloaded protein EST sequences GO660523.1, DR749492.1, CO905428.1, CO904299.1, BI975131.1, AW278473.1, BM269700.1, CA936889.1 CA935704.1, BU761220.1, AW428831.1, AW428803.1, AW428739.1, AW428711.1, AJ388815.1, AW163987.1, AI484155.1, AA660372.1, FE969312.1, 14-3-3 like protein FE969243.1, FE969340.1, CN748203.1, CN747244.1, (eudicots) CN746811.1, CN745511.1, CN743519.1, 64 CN747598.1, CN747325.1, CN745584.1, CN745563.1, CN743598.1, CN748506.1, CN748440.1, CN747023.1, GW452329.1, GT662944.1, GW458864.1, GT674095.1, GT674094.1, GT674093.1, GT674092.1, GT712481.1, GT691439.1, GT715099.1, GT704346.1, GT704345.1, GT695499.1, GT695498.1, GT732071.1 GW445847.1, GW450736.1, GW491421.1, GT656115.1, GW431117.1, GW475407.1, GT651591.1, GT651590.1, GT651589.1, GT651588.1, GT660655.1, GW457915.1, GW455608.1, GW457695.1, GW457678.1, GT660154.1, GW430551.1, GW444839.1, GW463204.1, GW462962.1, GW447573.1, GW442110.1, GW441905.1, GW482541.1, GW482491.1, ADP ribosylation factor GT662664.1, GT662618.1, GT668217.1, (eudicots) GW460053.1, GW462758., GW474884.1, 129 GW462640.1, GW462638.1, GW474267.1, GW451892.1, GW441327.1, GW468462.1, GW468212.1, GW451445.1, GW482171.1, GW481960.1, GT664729.1, GW459165.1, GW484383.1, GW473985.1, GW468075.1, GW461895.1, GT663973.1, GW440652.1, GW475895.1,, GW455988.1, GW437599.1, GT666617.1, GT682454.1, GT682162.1, GT718282.1, GT718281.1, GT712414.1, GT728977.1, GT728976.1, GT728975.1, GT728974.1, GT673913.1, GT673912.1, GT673911.1, GT673910.1, GT673909.1, GT692480.1, GT679122.1, GT710914.1, GT710913.1, GT710912.1, GT710911.1, GT710910.1, GT695364.1, GT682952.1, GT682875.1, GT731110.1, GT678817.1, GT681668.1, GT681591.1, GT681497.1, GT678340.1, GT728973.1, GT728972.1, GT682327.1, GT709991.1, GT715014.1, GT715013.1, GT681071.1, GT673412.1, GT686454.1, GT686353.1, GT683696.1, GT683695.1, GT685749.1, GT671675.1, GT706769.1, GT706768.1, GT706767.1, GT706766.1, GT701553.1, GT701552.1, GT701551.1, GT701550.1, GT672765.1, GT716290.1, GT716289.1, GT683400.1, GT683321.1, GT682805.1, GT685666.1, GT685226.1, GT686499.1, GT686292.1, GT686220.1, GT704237.1, GT722589.1, GT722588.1, GT722587.1, GT722586.1, GT722585.1, GT721198.1, GT721197.1, GT669342.1, GT713591.1, GT727190.1, GT727189.1, GT727188.1 blue copper binding XM_002308941 1 protein (Populus) caffeic acid/5- hydroxyferulic acid O- methyltransferase U13171.1, KX348778.1, KX348777.1, X62096.1 4 (Populus)
EB710382.1, GO499735.1, ES441913.1, GR989713.1 , GO550006.1 , GO504476.1 , DT043844.1, CO653968.1 , FP050575.1, FP049308.1, FP042726.1, FP042487.1, FP054109.1, FP074248.1, FP053483.1, FP036938.1, FP038948.1, FP038536.1, FP034112.1, FP034092.1, FP033773.1, FP033699.1, FP040592.1, FP040274.1, FP066936.1, FP024707.1, FP036236.1, FP033517.1, FP033516.1, FP040044.1, FP033320.1, CCoAOMT2 (eudicots) 60 FP066490.1, FP029494.1, FP029493.1, FP029386.1, FP029338.1, FP029250.1, FP072009.1, FP044955.1, FP041025.1, FP037193.1, FP034356.1, FP035308.1, FP035307.1, FP038131.1, FP038126.1, FP026858.1, FP027113.1, FP035518.1, FP035517.1, FP034949.1, FP034885.1, FP041257.1, FP034520.1, FP025164.1, GO529797.1 , DY650719.1, DY649828.1, DY648571.1, BM497443.1 KY584513.1, KJ138996.1, KJ138982.1, KJ138971.1, KJ138954.1, KJ138951.1, KJ138946.1, KJ138934.1, KJ138908.1, KJ138894.1, KJ138893.1, KJ138883.1, KJ138878.1, KJ138877.1, KJ138874.1, KJ138843.1, KJ138835.1, KJ138806.1, KJ138804.1, KJ138784.1, GT47 (eudicots) 39 KJ138760.1, KJ138748.1, KJ138742.1, KJ138726.1, KJ138717.1, KJ138707.1, KJ138697.1, KJ138684.1, KJ138679.1, KJ138677.1, KJ138671.1, KJ138667.1, KJ138665.1, KJ138662.1, KJ138661.1, KJ138652.1, KJ138648.1, KJ138631.1, KJ138630.1 K transporter (Populus) XM_006371877.1, XM_002308277.1 2
GO564158.1, GO555283.1, GO555432.1, laccase (eudicots) GO550501.1, GO497739.1, CA760339.1, 8
GT661358.1, GT679927.1 potassium channel 2 (Populus) AJ271447, XM_006385150 2
S-adenosylmethionine synthase 1 (eudicots) DR990403.1, AI727497.1 2
potassium channel XM_006372459.1, XM_002317669.2, SKOR (Populus) XM_002305858.2, XM_011045605.1, 5 NM_001304315.1 GH10 (eudicots) BI977446.1, HX871788.1, CB074467.1 3
GH16 (eudicots) FP054031.1, FP055271.1, FP055257.1, HX871793.1 4
GR717395.1, AW201079.1, BE210142.1, GT2 (eudicots) AW596787.1, BE607585.1, BE555293.1, 9 BE555145.1, BE331246.1, AI900211.1 GT8 (eudicots) XM_006352620 1 XM_018954768.1, XM_018954767.1, GT14 (eudicots) 4 XM_018954766.1, XM_018953813.1 KR052154.1, AY216900.1, AB696958.1, KP762452.1, NW_010363192.1, XM_010103620.1, KJ139035.1, KJ139030.1, KJ139021.1, KJ139014.1, GT31 (eudicots) KJ138995.1, KJ138985.1, KJ138984.1, KJ138975.1, 35 KJ138974.1, KJ138973.1, KJ138964.1, KJ138933.1, KJ138928.1, KJ138899.1, KJ138897.1, KJ138882.1, KJ138861.1, KJ138838.1, KJ138829.1, KJ138825.1, KJ138762.1, KJ138758.1, KJ138741.1, KJ138729.1, KJ138651.1, KJ138642.1, KJ138635.1, KJ138629.1, KE345037.1, GT43 (eudicots) NM_126123.5, NM_102525.3, NM_202193.1, 7 KJ138981.1, KJ138863.1, KJ138720.1, KJ138625.1 KY584513.1, KJ138996.1, KJ138982.1, KJ138971.1, KJ138954.1, KJ138951.1, KJ138946.1, KJ138934.1, KJ138908.1, KJ138894.1, KJ138893.1, KJ138883.1, KJ138878.1, KJ138877.1, KJ138874.1, KJ138843.1, KJ138835.1, KJ138806.1, KJ138804.1, KJ138784.1, GT47 (eudicots) 39 KJ138760.1, KJ138748.1, KJ138742.1, KJ138726.1, KJ138717.1, KJ138707.1, KJ138697.1, KJ138684.1, KJ138679.1, KJ138677.1, KJ138671.1, KJ138667.1, KJ138665.1, KJ138662.1, KJ138661.1, KJ138652.1, KJ138648.1, KJ138631.1, KJ138630.1 AY216902.1, KJ138921.1, KJ138920.1, KJ138856.1, GT61 (eudicots) KJ138832.1, KJ138821.1, KJ138820.1, KJ138785.1, 9 KJ138637.1 XM_006372459.1, XM_002317669.2, SKOR (Populus) XM_002305858.2, XM_011045605.1, 5 NM_001304315.1
Table S2. List of the designed 55 primers (with each pairs’ nucleotide succession, melting temperature, and length of the amplified sequence) N DNA sequence named after the Fwd, T Sequence Primers r. coded protein Rev m size 54, f CGTTCACTTTTGGCAGATCC 3 1. ADP ribosylation 1 617 GCAAACAGTGTGGACAAGG 55, r A 7 55, f GATGAACTTCGGGATGCTGT 0 2. ADP ribosylation 2 147 58, r CTGCCTGAGGGAGTGCAG 2 55, f GCCATCGTCTATGCCTTGTT 4 3. ADP ribosylation 3 431 55, r TGAATGCTGCAGAGATCACC 1 56, f GATAAGCTTGGGCTGCACTC 2 4. ADP ribosylation 4 606 54, r GCCAATCAAGACCCTCGTAA 8 GGCAGTAGATCATGAGGTTG 55, f G 2 5. blue copper binding protein 1 115 54, r TCACCAAGAGCGAAGGTTTT 6 56, f TTCACTACAGGTGCACACGA 3 6. blue copper binding protein 2 137 55, r TTGTTGCTGGTCCAGTTGTC 7 51, f CGGGACAAAAATTAGCCATT 3 7. blue copper binding protein 3 145 CATCAAAGCAAAGGTAGCA 54, r ACA 0 55, f GATGCGCCTGTCATGTTAGA 0 8. caffeic acid 5-hydroxiferulic 1 456 56, r GAGGACCTTGTCCTGGTTCA 5 57, f ATCGGCACCACAGACTTCTC 0 9. caffeic acid 5-hydroxiferulic 2 457 54, r ACTTTAGGGGATGCCCATTT 2 55, f ACCTTCGACGAAAGCATCAC 10 4 caffeic acid 5-hydroxiferulic 3 340 . TGAGTAAAGAAAAAGCAAT 51, r GCAA 7 55, f CCATGTCCAAGCTCCAAGTT 11 3 caffeic acid 5-hydroxiferulic 4 513 . 56, r TTTGAGGGAGGCAGTGAAGT 3 CGGAGAGGAACAGCAGACT 57, f 12 C 2 Caffeoyl-CoA O-methyltransferase 5 207 . 53, r CAAAACTCGCACCAAGATCA 4 CCAGTGTGTACCCAAGAGAG 57, f 13 C 5 Caffeoyl-CoA O-methyltransferase 1 269 . 54, r ACACCACGCCATAAACATCA 7 55, f ATCCCTGAGGATGGAAAGGT 14 5 Caffeoyl-CoA O-methyltransferase 2 233 . 54, r TTTCTGAATCACCGGGAGAC 4 54, f TTTGTGGATGCTGACAAGGA 15 4 Caffeoyl-CoA O-methyltransferase 3 263 . 54, r CAGCTGCAAGTGCCTTATTG 6 54, f CATGCTTCCTGTTGGTGATG 16 1 Caffeoyl-CoA O-methyltransferase 4 265 . 54, r AATGGAGTTTCTCGGCCATT 6 GGAAGAGCATTAGACGGAA 52, f 17 AA 6 Cyclophilin 2 109 . 54, r GATTGGTTCGTTGGAGAGGA 4 55, f GGTTCGTTTCCCTCCTCTTC 18 2 elongation factor 1 300 . 56, r CACATCACCTCCGCAGACTA 5 55, f GGTTCGTTTCCCTCCTCTTC 19 2 elongation factor 2 362 . 56, r CACATCACCTCCGCAGACTA 5 54, f TCCGTTTGACTTCCTTGACC 20 9 GH10 1 265 . 57, r AGTCCTGACTGCTTCCTCCA 9 54, f TCCGTTTGACTTCCTTGACC 21 9 GH10 2 265 . 57, r AGTCCTGACTGCTTCCTCCA 9 55, f TGAAGCCTCATTGTGGTGTG 22 5 GH10 3 249 . CCTAAAAGTACAGTGATGTT 53, r TTCTTGA 6 53, f TTGCAGGTGCAATTCATACC 23 6 GH10 4 300 . 53, r GGGTGCTCTTTTGCCATATT 4 55, f CCACGCCTTCTTTCACTCTC 24 7 GH16 1 323 . 56, r GCGACCATCCTGGAGTGTAT 6 57, f CAGCATAGCGGACCTCTCTC 25 1 GH16 2 318 . 55, r GGTTTGCTCAAAGTCGTGGT 9 58, f TGATAGGAGTCGCTGCTGGT 26 1 GH16 3 244 . 55, r CCGCTCATGTCTACTTCACG 4 54, f ATATTTCGCCGACTCTCACG 27 7 GH16 4 255 . 55, r GAATTACTAACGGCGGAGCA 0 55, f TCCCACTCCTCTTTGTGTGA 28 6 GT2 1 234 . GAAAGACGAAGTTTGTGGTT 53, r GA 5 GAAGAAGGAGAAGGAGACA 54, f 29 ATGA 0 GT2 2 242 . 50, r TTTTCTTCAACCCATTGTGAA 6 54, f CGGAAACGGAGACTTGCTTA 30 9 GT8 1 341 . AAGGGAGAAATTGCGAAGG 54, r T 5 TTTTGAGTATGTGCGTGTTAT 53, f 31 CAA 1 GT8 2 398 . 53, r CGCACGGACAAGTATTTCAA 4 54, f CCGATGAAAGCCGAGAAGT 32 9 GT14 1 210 . 53, r TCCCTTGTTTTCCTCCATCA 7 54, f ATTGATGGTCGGAAACTGCT 33 5 GT14 2 291 . 53, r ATTCTTTAAACGCGGTGCAG 9 54, f ATTTGGCCACCAGAACTTCA 34 8 GT31 1 336 . 54, r CACGCTGAACTTTCCATGTTT 0 55, f TGGCGTCTATGTCGCTATCA 35 7 GT31 2 392 . 53, r TTTGCTTGCCACATCTCAAA 2 52, f TCCATTCCAATCGTGTTTCA 36 1 GT43 1 269 . 52, r TTTCACAGCTCCCAAATCAA 4 54, f CGAGAATGGAGGTTCACACA 37 6 GT43 2 253 . 55, r GTAAAACGGTCGCCTCCAT 2 54, f CCGTTACTCCGGTGATTGAT 38 5 GT47 1 392 . 53, r CCCATTGACCTTTCAAGCAT 3 54, f AGCGGATTTGTTCTTTGTGC 39 2 GT47 2 455 . 53, r CGGAAATACGGCATTTGACT 2 GGATTCTTAAAGGTTGATTT 52, f 40 GCAT 1 GT61 1 188 . CCTTGAGAGAAACTTAACGA 53, r ACG 7 GCAAAACCCAAAAGTGATG 52, f 41 G 8 GT61 2 255 . 55, r TTTCTCGACAAGTGGTGCTG 3 56, f GCCCACTCTCCATCTGTGAT 42 7 K-transporter 1 774 . 54, r CTGTAATGGTTTGGGGGAGA 3 55, f TGACGAACTTGGTTGTGCAT 43 1 K-transporter 2 874 . 58, r AGGAGGCAGCTTGTTCCTCT 5 55, f AGTTGTTGAAACGGGGACTG 44 4 K-transporter 3 725 . 56, r ACTTCCTTTGGACGCTGCTA 5 55, f CACAGATGGGTTCAAGCTCA 45 0 laccase 4 323 . 54, r TCGGTCTTAAATGGCTTGGT 2 54, f GGCACAATCAACAACGTGAC 46 9 laccase 1 340 . 53, r TTGAAGGTTTTTGGGACCAC 4 54, f CCATCGAGAGGAACACCATT 47 5 laccase 2 358 . 55, r CCACAAAGGCCATCTTGAGT 3 55, f TGTTAGGGCTGCCTAAATGC 48 4 potassium channel 2 1 913 . 57, r AGTGCTCTATACGGCGAGGA 5 57, f CACTGTGCTGGTTGCCTCTA 49 3 potassium channel 2 2 738 . 53, r CATGGAGTTTTGAGCATGGA 0 54, f TGACAGATACCCGCACAAAG 50 9 potassium channel 2 3 738 . 53, r CATGGAGTTTTGAGCATGGA 0 54, f CCCCGAGTATATGCCTTTGA 51 1 S-adenosyl-L-methionine synthase 1 576 . 50, r CGGCAATTTCATCATTTGTG 6 55, f TCTTGCTCGTAGGTGCATTG 52 2 S-adenosyl-L-methionine synthase 2 502 . 55, r CCGTATGCAGCTGTCTTCAA 2 54, f 53 CTCCTCAGGTTTCGTTATCCA 0 potassium channel SKOR 1 336 . CAAGAGGCTAACCCAAAGC 55, r A 3 55, f AACTCTATTGCACGCACACG 54 9 potassium channel SKOR 2 389 . 56, r TGTTGTGTACCGCTTCCAGA 3 55, f GCAGGAGCTGATCCCAATAG 55 2 potassium channel SKOR 3 280 . 56, r TGTGCCAGCATTAGAGGTGT 6
Table S3. List of the poplar samples used in the study Clone name or Parents Place of DNA code identifier/sample Origin (female × male) breeding/selection type Lébény NYF 480 Populus nigra L. Populus nigra L. Hungary 211/wood Populus
NYF 481 S 307-24/wood deltoides Populus deltoides MARSH. Belgium
MARSH.
Populus Populus deltoides MARSH. A
NYF 482 S-1-54/wood deltoides 142 (Iowa) × Populus deltoides Belgium
MARSH. MARSH. A 148 (Missouri) Lassicsárda NYF 483 Populus nigra L. Populus nigra L. Hungary 7/wood Populus
NYF 484 S 332-1/wood deltoides Populus deltoides MARSH. United States
MARSH. NYF 485 Győr 203/wood Populus nigra L. Populus nigra L. Hungary
Populus × Populus deltoides MARSH. NYF 486 Sv-656/wood euramericana Choctown S 55 × Populus nigra Hungary
(DODE) GUINIER L. Győr 203
Populus Populus deltoides MARSH. V-18
NYF 487 Sv-761/wood deltoides × Populus deltoides MARSH. S Hungary
MARSH. 332-1 Populus × Populus deltoides MARSH. S 9-2 NYF 488 Sv-874/wood euramericana Hungary × Populus nigra L. Lébény 211 (DODE) GUINIER
Populus × Populus deltoides MARSH. S NYF 489 Sv-890/wood euramericana 307-24 × Populus nigra L. Hungary
(DODE) GUINIER Lébény 211 Populus × euramericana
Populus × (DODE) GUINIER ‘Canadese NYF 25 I-214/ leaf euramericana bianco’ × Populus deltoides Italy
(DODE) GUINIER MARSCH. ‘Caroliniano prodigioso’
Populus × Populus deltoides MARSCH. S- NYF 27 Pannónia/ leaf euramericana 1-54 × Populus nigra L. Lébény Hungary
(DODE) GUINIER 211
Populus × Populus deltoides MARSCH. S- NYF 35 Koltay/ leaf euramericana 1-526 [Populus deltoides Hungary
(DODE) GUINIER MARSCH. V-5 (Iowa) × Populus deltoides MARSCH. V-9 (Missouri)] × Populus nigra L. Lassi 7 Populus alba L. ‘No. 2 Instituto NYF 41 Villafranca/ leaf Populus alba L. Pignatelli’× Populus alba L. Italy ‘No. 2 Lucca’ Populus Populus maximowiczii maximowiczii A.HENRY × Populus × A.HENRY × NYF 7 Kornik-21/ leaf berolinensis DIPPEL (Populus Poland Populus × laurifolia LEDEB. × Populus berolinensis nigra L. cv. Italica) DIPPEL
Populus trichocarpa TORR. ET
GRAY cv. Fritzi Pauley × Populus × Populus deltoides MARSCH. NYF 8 Raspalje/ leaf interamericana Belgium (Populus deltoides MARSCH. BROCKH. Iowa × Populus deltoides
MARSCH. Missouri)
Populus trichocarpa TORR. ET
GRAY cv. Fritzi Pauley × Populus × Populus deltoides MARSCH. NYF 26 Beaupré/ leaf interamericana Belgium (Populus deltoides MARSCH. BROCKH. Iowa × Populus deltoides
MARSCH. Missouri)
Populus deltoides MARSCH. S- 4-231 (Populus deltoides
MARSCH. A 142 Iowa ×
Populus Populus deltoides MARSCH. A NYF 11 Durvakérgű/ leaf deltoides 12 Ontario, Canada) × Populus Belgium
MARSH. deltoides MARSCH. var. deltoides [syn.: Populus deltoides var. missouriensis (A.
HENRY) A. HENRY] (tetraploid)
Populus × Populus deltoides MARSCH. S- NYF 3 Kopecky/ leaf euramericana 1-54 × Populus nigra L. Lébény Hungary
(DODE) GUINIER 211
Populus Populus trichocarpa TORR. ET Muhle Larsen/ NYF 105 trichocarpa GRAY × Populus trichocarpa Belgium leaf TORR. ET GRAY TORR. ET GRAY Populus NYF 40 227/ leaf Populus grandidentata MICHX. Canada grandidentata MICHX. Populus× BA 65 65/ leaf Populus × canescens SM.* Barcs, Hungary canescens SM. BA 105 105/ leaf P. tremula L. P. tremula L. Darány, Hungary Note. * Natural hybrid; the actual origin (crossing direction between Populus alba L. and Populus tremula L. parents) has not been identified. 1 Table S4. BLASTN search results against the NCBI database of the seven candidate genes. As results are 2 presented only the best BLAST hits’ accession numbers and description of the sequences in the NCBI 3 database Max Total Query E- Accession nr. in Chromosomal Query sequence Abbrev. Ident Description
score score cover value NCBI location Populus trichocarpa 2e- chromosome K transporter 1 Kt 1 292 292 100% 99% XM_006371877.2 potassium channel 75 18 AKT1 Populus kitakamiensis (P. sieboldii X P. grandidentata) caffeate O- 8e- homt1 gene for chromosome methyltransferase COMT 3 121 121 70% 87% D49710.1 24 caffeic acid O- 12 3 methyltransferase, complete cds (exon1-4)
Populus fremontii caffeate O- isolate COMT2_15- 7e- chromosome methyltransferase COMT 4 200 200 98% 99% JX552711.1 E_10 caffeate O- 48 12 4 methyltransferase (COMT2) gene Caffeoyl-CoA O- CCoAOMT 3e- Populus trichocarpa chromosome methyltransferase 215 215 99% 92% AJ223620.1 4 52 CCoAOMT2 gene 1 4 Populus tremula x potassium 1e- Populus tremuloides chromosome ptk 2/2 243 243 97% 99% AJ271447.1 channel 2 2 60 potassium channel 3 2 (ptk2 gene) Populus trichocarpa S-adenosyl-L- 7e- S- chromosome methionine SAMS 1 307 307 99% 98% XM_006388164.2 80 adenosylmethionine 2 synthase 1 synthase 1-like Populus trichocarpa potassium 2e- chromosome SKOR 3 108 108 83% 97% XM_024600186.1 potassium channel channel SKOR 3 20 4 SKOR 4 Table S5. Number of significant pairwise comparisons by Fisher's exact test and the sites between the test was significant Number of significant pairwise Number of No. of No. comparisons by Significant polymorphic Abbrev. sequences of Fisher's exact between sites used sites test sites analyzed (* 0.01 Figure S1. The presence/absence of seq. 1-18 on gel electrophoresis photos, amplified with primers designed on Populus trichocarpa (taxid 3694) genome. For sequence abbreviation see Table S2. L- ladder; 1- sample I-214; 2- sample Koltay; 3- sample Pannónia; 4- H2O (control). Exposure time: 3.000. Photo: ERTI AI0120 Figure S2. The presence/absence of seq. 19-36 on gel electrophoresis photos, amplified with primers designed on Populus trichocarpa (taxid 3694) genome. For sequence abbreviation see Table S2. L- ladder; 1- sample I-214; 2- sample Koltay; 3- sample Pannónia; 4- H2O (control). Exposure time: 3.000. Photo: ERTI AI0120 Figure S3. The presence/absence of seq. 37-55 on gel electrophoresis photos, amplified with primers designed on Populus trichocarpa (taxid 3694) genome. For sequence abbreviation see Table S2. L- ladder; 1- sample I-214; 2- sample Koltay; 3- sample Pannónia; 4- H2O (control). Exposure time: 3.000. Photo: ERTI AI0120