Metabolomics of Potato Genotypes, Varying in Quantitative Resistance in Leaf to Late Blight

Metabolomics of potato genotypes, varying in quantitative resistance in leaf to late blight, discovers candidate genes in phenylpropanoid pathway

Kalenahalli N. Yogendra, Doddaraju Pushpa, Kareem A. Mosa, Ajjamada C. Kushalappa, Agnes Murphy, and Teresa Mosquera

Table S1 Primer sequences used for quantifying RR gene expression in potato genotypes.

Genbank name / Gene name / Forward primer (5'-3') / Reverse primer (5'-3')
A.  qRT-PCR primers
AF150686.1 / St4CL / ATCCTGAGGCCACAGCTAGA / TTACGGGACAACAGCAGCAT
JX896425.1 / StTHT / TGTTGTTGTCAGCTGTGGCT / TTTGCAAAGGCTTGAAGGGC
PGSC0003DMT400038501a / SpTyDC / AGTGGTGGTGGAGGTGTACT / AGGTCCCAGATAGGGTCGTT
AB061263.1 / StEf1-α / ATTGGAAACGGATATGCTCCA / TCCTTACCTGAACGCCTGTCA
B.  Primers for sequencing
AB061243.1 / StTHT / TATCTCATGGCTCCTGCTCCT / ATACTAACATGTCTCTTCCTCAATTTTC
AF150686.1 / St4-CL / AAGATGCCGATGGATACCG / CCATCAATTTGAAATACCAGCA
Sotub03g010370.1.1a / StTyDc / ATGGGTACCCTCAACATCAACC / TTAAACATAGCCTTGATTTAGCAG

a Solanaceae Genomics Resource, Michigan State University, USA.

5

Table S2 Putatively identified resistance related metabolites in potato genotypes, as compared to susceptible genotype, following P. infestans or mock inoculation

Observed mass (Da) / Exact mass (Da) / AME / RT
(min) / Putative Name / Chemical groups / Observed fragmentation / Database fragmentation / P value / Fold change / Database ID
164.0483 / 164.0485 / 1.29 / 0.96 / coumarinate / Phenyl proponoid / 148.25, 145.16, 135.20, 119.14, 115.23, 91.04 / 162.800, 134.700, 119.800, 119.000, 116.900, 92.800 / 0.018 / 1.52*
(RRI) / CPD-7418
165.0797 / 165.0790 / 4.37 / 1.60 / L-phenylalanine / Amino acid / 163.90,149.05,147.12, 136.15, 120.22, 103.08, 90.97, 72.05 / 163.9, 148.8, 147.2, 146.5, 120.1, 118.4, 118.1, 115.7, 103,95, 92.3, 91.2, 77.1, 71.9, 70, 64.8, 45.1, 42 / 0.040 / 1.04*
(RRI) / PHE
172.1472 / 172.1463 / 4.79 / 21.18 / Caprate / Fatty acid / 171.18, 152.82, 141.29, 127.21, 125.20 / 171.13, 153.12, 151.11, 76.95, 68.99, 59.01 / 0.028 / 1.42*
(RRC) / CPD-3617
174.0536 / 174.0528 / 4.64 / 21.07 / shikimate / Carboxylic Acids / 129.16, 145.15, 111.04, 101.22, 85.05 / 128.900, 111.400, 101.300, 85.000, 83.300, 82.200 / 0.027 / 2.44*
(RRC) / SHIKIMATE
192.0430 / 192.0423 / 3.66 / 9.61 / 5,7-dihydroxy-2-methylchromone / Polyketide / 0.026 / 8.57*
(PRr) / CPD-0
192.0641 / 192.0634 / 3.52 / 3.70 / L-quinate / Carboxylic Acids / 173.14, 171.17, 155.07, 127.04, 111.14, 109.15,
93.01, 87.06, 85.11 / 173.400, 170.800, 154.800, 127.000, 111.400, 109.300, 93.000, 87.300, 85.100 / 0.050 / 2.13*
(RRI) / QUINATE
224.0690 / 224.0685 / 2.51 / 9.59 / Sinapic acid / amide / 164.16, 179.16, 208.19, 149.03, / 207.900, 193.100, 164.900, 149.100, 146.100 / 0.008 / 5.71**
(RRC) / 45738
250.1325 / 250.1317 / 3.06 / 8.28 / N-Caffeoylputrescine / amide / 135.03, 207.24, 249.25, 234.33 / 0.026 / 1.54*
(RRC) / C03002
264.1479 / 264.1474 / 2.02 / 10.61 / Feruloylputrescine / amide / 248.22, 247.21, 219.38, 205.23, 201.22 / 0.009
0.00 / 4.78**
(RRC)
2.04**
(RRI) / C10497, In silico, (Muroi et al., 2009; Gunnaiah et al. 2012)
276.1593 / 276.1586 / 2.51 / 11.49 / 4-coumaroylagmatine / amide / 119.07, 233.33, 258.30, / 119.04, 233.129, 258.14 / 0.016 / 7.91*
(PRr) / C00028060, In silico , (Muroi et al. 2009; Gunnaiah et al. 2012)
278.2255 / 278.2246 / 3.16 / 27.51 / γ-linolenate / fatty acid / 277.44, 259.33, 257.30, 247.16, 241.35, 233.38, 194.97, 141.02, 83.24, 79.98 / 277. 2168, 233.2223, 164.9045 / 0.035 / 1.29*
(RRI) / CPD-8117
280.2411 / 280.2402 / 3.16 / 28.41 / Stearolic acid / Fatty Acids and Conjugates / 279.42, 262.70, 261.38, 251.43, 235.19, 234.21, 217.16, 197.25, 142.35, 135.19, 95.20, 82.01 / 0.014 / 1.30*
(RRI) / LMFA01030455
280.2411 / 280.2402 / 3.08 / 28.41 / Linolenic acid / fatty acid / 279.42, 262.70, 261.38, 251.43, 235.19, 234.21, 217.16, 197.25, 142.35, 135.19, 95.20, 82.01 / 279.400, 261.100, 259.300, 243.500,233.000, 219.500, 96.900, 83.10 / 0.029
0.022 / 1.43*
(RRI)
1.47*
(PRr) / 191, C01595, KOX00402, LMFA01030120, McGill MD
280.2411 / 280.2402 / 3.09 / 28.41 / 5Z,11Z-octadecadienoic acid / fatty acid / 279.42, 262.70, 261.38, 251.43, 235.19, 234.21, 217.16, 197.25, 142.35, 135.19, 95.20, 82.02 / 0.014 / 1.55*
(PRr) / LMFA01030322
281.1058 / 281.1052 / 2.16 / 13.86 / Annolobine / Alkaloid / 160.05,145.18,119.06,237.25,253.21 / 0.023 / 29.62*
(PRr) / C09338
283.1218 / 283.1208 / 3.29 / 16.33 / p-coumaroyltyramine / amide / 162.15, 145.15, 136.16, 134.07, 131.96, 119.10, 117.12 / 0.008 / 1.28**
(RRI) / CPD-8943
283.1217 / 283.1208 / 3.12 / 16.36 / N-Phenylacetyl phenylalanine / Amino Acids and Derivatives / 162.15, 145.15, 136.16, 134.07, 131.96, 119.10, 117.12 / 164.071, 147.0443, 103.0525, 72.0991 / 0.047 / 14.86*
(PRr) / 58126
283.1213 / 283.1208 / 1.44 / 16.32 / morphinone / alkaloid / 119.10, 145.15, 162.15, 134.07, 136.16, 131.96 / 0.002 / 15.41**
(PRr) / MORPHINONE
283.1218 / 283.1208 / 3.24 / 16.33 / p-coumaroyltyramine / amide / 162.15, 145.15, 136.16, 134.07,131.96,119.10,117.12 / 0.008 / 17.99**
(PRr) / CPD-8943
286.0936 / 286.0927 / 3.22 / 3.48 / N-glucosylnicotinate / pyridines / 225.28, 203.25, 267.16, 194.16 / 0.035 / 1.65*
(RRC) / CPD-446
291.1466 / 291.1470 / 1.34 / 12.24 / Convolvine / Alkaloids / 272.21, 247.27, 273.24, 248.25, 218.15, 190.17 / 0.007 / 6.30**
(PRr) / C10856
292.1542 / 292.1535 / 2.10 / 10.45 / 4-coumaroyl-3-hydroxyagmatine / amide / 274.24, 273.20, 260.80, 249.21, 243.00, 209.30, 135.06, 111.51 / 0.028
0.01 / 4.14*
(RRI)
3.01*
(RRC) / CPD-12237
295.1067 / 295.1056 / 3.76 / 7.46 / indican / glucoside / 264.01, 251.14, 186.32, 161.12, 146.45,131.99 / 0.017 / 2.47*
(RRC) / INDICAN
297.1007 / 297.1001 / 2.15 / 12.83 / Longifolonine / Alkaloids / 176.05, 145.22, 186.15, 253.19, 119.20 / 0.018 / 68.36*
(PRr) / C09569
299.1166 / 299.1157 / 2.93 / 14.22 / 2,3,9,10-Tetrahydroxyberbine / alkaloid / 135.11, 145.18, 161.17, 178.15, 177.12, 256.29, 152.20 / 0.008
0.014 / 3.91**
(RRC)
23.39*
(PRr) / C12330
299.1165 / 299.1157 / 2.45 / 14.22 / Benzyloxyphenyliso serine Methylester / 135.11, 145.18, 178.15, 161.17, 177.12, 152.20 / 238.08, 210.09, 220.07, 160.03, 135.04, 104.04 / 0.037 / 2.98*
(PRr) / 73564
300.0844 / 300.0845 / 0.38 / 1.27 / salicylate β-D-glucose ester / Phenol / 280.16, 255.14, 239.33, 217.05, 201.30, 161.33, 137.11, 93.08 / 179.26, 137.15 / 0.049 / 3.52*
(RRI) / PMN, J355.695F, In silico, (Torras-Claveria et al. 2011; Gunnaiah et al. 2012)
304.2413 / 304.2402 / 3.63 / 29.97 / Eicosatetraenoate / Fatty acid / 0.039 / 1.36*
(RRC) / CPD-8121
306.1700 / 306.1692 / 2.55 / 12.21 / feruloylagmatine / amide / 288.23, 263.25, 248.28, 149.12, 134.15 / 289.14, 263.13, 177.05, 149.06 / 0.006
0.00 / 29.04**
(RRC)
1.32**
(RRI) / PMN, J817.816J, In silico , (Muroi et al., 2009; Gunnaiah et al. 2012)
311.1162 / 311.1157 / 1.33 / 14.49 / Actinodaphnine / Alkaloids / 295.23, 161.10, 135.17, 252.27, 267.24, 283.30, 160.12 / 0.016 / 35.19*
(PRr) / C09322
313.0956 / 313.0950 / 1.87 / 15.39 / Melicopine / Alkaloids / 166.15, 227.26, 145.08, 192.11, 206.12 / 0.033 / 8.65*
(PRr) / C10724
313.1320 / 313.1314 / 1.86 / 16.78 / N-Feruloyltyramine / amide / 297.34, 296.66, 270.33, 253.18, 191.15, 178.13, 177.12, 166.15, 148.24, 135.01 / 178.0,122,137 / 0.009
0.008 / 4.32**
(RRI)
7.87**
(PRr) / C02717
315.1113 / 315.1107 / 1.92 / 12.89 / Hippeastrine / Alkaloids / 296.29, 180.43, 161.18, 204.18 / 0.009 / 55.72**
(PRr) / C08528
327.1118 / 327.1107 / 3.33 / 13.50 / Terrestriamide / amide / 311.21, 283.31, 267.41, 243.42, 216.27, 202.05, 190.11, 176.17, 175.02, 161.05, 149.18, 135.10, / 0.02 / 5.84*
(RRI) / C17839
329.1269 / 329.1263 / 1.91 / 15.57 / Melicopicine / Acridone alkaloids / 313.23, 257.32, 192.06, 190.96, 178.19, 175.15, 161.04, 149.20, 135.09, 134.07 / 0.011 / 1.43**
(RRI) / C10722
329.1267 / 329.1263 / 1.27 / 14.79 / 3-Epimacronine / Alkaloid / 313.19, 178.12, 175.14, 135.15, 149.18, 161.11, 192.10, 152.13 / 0.041 / 5.10*
(RRC) / C12178
329.1269 / 329.1263 / 1.90 / 14.80 / 3-O-Acetylhamayne / Alkaloid / 313.19, 178.12, 175.14, 135.15, 149.18, 161.11, 192.10, 152.14 / 0.007 / 5.64**
(RRC) / C12165
329.1272 / 329.1263 / 2.64 / 13.12 / 1-O-Acetyllycorine / Alkaloid / 313.23, 257.32, 192.06, 190.96, 178.19, 175.15, 161.04, 149.20, 135.09, 134.07 / 0.009 / 14.83**
(RRC) / C12166
338.1007 / 338.1002 / 1.63 / 8.48 / p-Coumaroyl quinic acid / Polyphenol / 293.37, 277.46, 269.31, 247.41, 232.19, 193.23, 173.22, 163.25 / 293.10, 191.09, 277.01 / 0.016 / 2.03*
(RRI) / PMN, , In silico, Torras-Claveria et al. (2011)
338.1004 / 338.1002 / 0.59 / 4.86 / 4-methylumbelliferyl glucoside / Phenol / 293.37, 277.46, 269.31, 247.41, 232.19, 193.23, 173.22, 163.25 / 0.02
0.000 / 1.27*
(RRI)
3.33**
(PRr) / CPD-11641
338.1006 / 338.1002 / 1.37 / 4.95 / 4-p-Coumaroylquinic acid / Phenol / 293.37, 277.46, 269.31, 247.41, 232.19, 193.23, 173.22, 163.25 / 0.002 / 2.53*
(PRr) / C10441
342.0957 / 342.0951 / 1.89 / 1.29 / Caffeic acid 3-glucoside / Phenolic compound / 323.25, 296.37, 279.29, 266.50, 243.40, 231.44, 179.08, 161.27, 135.24, 134.04 / 0.049 / 1.07*
(RRI) / C10431
342.1317 / 342.1315 / 0.72 / 8.41 / coniferin / glucoside / 179.16, 161.17, 135.14, 251.09, 203.16, 281.15 / 0.030 / 3.31*
(RRC) / CPD-1777
345.1219 / 345.1212 / 2.03 / 13.50 / Candimine / Alkaloids / 343.84, 329.23, 326.32, 314.43, 306.23, 234.22, 202.08, 135.13 / 0.014 / 4.71*
(RRI) / C08520
352.1412 / 352.1423 / 3.07 / 1.58 / Feruloylserotonin / amide / 307.17, 240.40, 215.29. 195.23, 177.34, 173.04, 133.02 / 135.04, 175.08, 177.05, 336.11 / 0.050 / 1.04*
(RRI) / CPD-8935, PMN, J1.973.598B, In silico (Gunnaiah et al. 2012)
354.0958 / 354.0951 / 2.06 / 5.72 / scopolin / polyphenol / 271.00, 191.22, 179.19, 173.10, 135.03 / 0.044 / 1.64*
(RRI) / SCOPOLIN
356.1113 / 356.1107 / 1.55 / 10.28 / 1-O-Feruloyl-beta-D-glucose / Phenylpropanoids / 193.15, 217.17, 175.30, 134.13, 295.16, 235.28 / 0.020 / 2.67*
(PRr) / C17759
356.1114 / 356.1107 / 1.81 / 10.32 / Gentiopicrin / Alkaloid / 193.15, 217.17, 175.30, 134.13, 295.16, 235.29 / 0.027 / 2.65*
(PRr) / C09782
360.1422 / 360.1420 / 0.51 / 8.49 / 7-deoxyloganate / Terpenoid / 299.23, 277.13, 242.44, 217.02, 197.13, 179.19, 153.43 / 0.033 / 1.79*
(RRI) / CPD-9981
368.1114 / 368.1107 / 1.93 / 9.95 / 5-O-Feruloylquinic acid / Phenol / 235.26, 161.21, 160.39, 142.77, 125.26 / 0.029 / 4.80*
(RRI) / C02572
380.2930 / 380.2926 / 0.88 / 28.15 / Persin / Fatty acid / 325.24, 297.02, 349.01, 361.42, 152.38 / 0.011 / 2.07*
(PRr) / 36469
381.2601 / 381.2600 / 0.20 / 23.53 / Sphinganine-1-phosphate / ceramides / 366.30, 348.14, 336.71, 320.67, 298.89, 298.17, 264.09, 142.17 / 0.035 / 2.38*
(RRI)
386.1220 / 386.1213 / 1.91 / 10.93 / 4-O-beta-D-Glucosyl-sinapate / phenylpropanoids / 247.23, 223.04, 205.19, 161.12, 153.14 / 247.0612, 223.0609, 205.0494, 164.0474, 149.0241, 147.0079, 119.0138, 91.0194 / 0.031 / 1.19*
(RRI) / C02919, In silico (Gunnaiah et al. 2012)
386.1221 / 386.1213 / 2.16 / 10.94 / 1-O-Sinapoyl-beta-D-glucose / Phenylpropanoids / 247.23, 223.04, 205.19, 161.12, 153.14 / 247.0612, 223.0609, 205.0494, 164.0474, 149.0241, 147.0079, 119.0138, 91.0194 / 0.050 / 1.21*
(RRI) / C01175
388.1727 / 388.1733 / 1.68 / 12.16 / tuberonic acid glucoside / Jasmonate and its glucoside / 305.35, 284.54, 365.80, 320.37, 342.32 / <.0001 / 5.63**
(PRr) / CPD-11510
402.1525 / 402.1526 / 0.34 / 8.48 / Benzyl alcohol beta-D-xylopyranosyl (1->6)-beta-D-glucopyranoside / 341.12, 343.20, 281.36, 240.28, 359.14 / 0.023 / 2.94*
(RRC) / 130622-31-0
408.1806 / 408.1784 / 5.43 / 3.51 / Vernoflexuoside / Sesquiterpene / 203.21, 319.47, 349.77, 341.27, 226.15 / 0.026 / 2.03*
(RRC) / C09579
414.1163 / 414.1162 / 0.33 / 8.16 / Asperuloside / iridoid glycoside / 0.041 / 4.36*
(PRr) / C09769
414.1174 / 414.1162 / 2.87 / 8.24 / Futalosine / terpenoid / 0.018 / 4.22*
(PRr) / C16999
428.2057 / 428.2046 / 2.47 / 13.32 / Cuminoside B / sesquiterpenoid / 0.003 / 8.27**
(RRC) / 597565-59-8
430.1623 / 430.1628 / 0.95 / 3.56 / Sesartemin / Phenylpropanoids / 225.22, 203.28, 369.16, 130.07 / 0.019 / 2.60*
(RRC) / C10884
430.1638 / 430.1628 / 2.52 / 3.67 / Aliarin 4'-methyl ether / Flavonoids / 225.20, 203.27, 369.03, 159.26, 130.21 / 0.028 / 2.88*
(RRC) / 51268
430.2211 / 430.2203 / 1.86 / 15.20 / Dendroside F / sesquiterpene / 369.05, 239.01, 179.00, 161.07, 143.18 / 0.037 / 1.43*
(RRI) / 503443-15-0
432.1634 / 432.1631 / 0.54 / 9.93 / Zizybeoside I / Lipid / 395.19, 387.42, 385.08, 354.13, 300.15, 299.15, 269.15, 259.22, 191.17 / 0.047 / 4.35*
(RRI) / C17564
442.2576 / 442.2567 / 2.21 / 18.58 / Dendronpholide K / Diterpenes / 422.52, 411.18, 380.99, 359.18, 277.46, 237.36, 161.06 / 0.003 / 8.83**
(RRC) / 1041055-14-4
446.2161 / 446.2152 / 2.00 / 12.13 / Dendromoniliside D / Stilbenoids / 366.86, 384.97, 423.90, 437.01, 348.11, 341.27, 205.28, 161.15 / 0.005 / 2.32**
(RRC) / 603995-11-5
446.2163 / 446.2152 / 2.38 / 14.62 / 7,7-O-Dihydroebuloside / 385.14, 415.47, 179.01, 161.03, 277.15, 233.57 / 0.017 / 3.91*
(RRC)
448.2467 / 448.2468 / 0.23 / 19.75 / Glutathionylaminopropylcadaverine / Amino acid / 315.24,293.19,233.01,191.05,161.13,149.03, 143.09 / 0.047 / 1.13*
(RRI) / C16566
461.1696 / 461.1686 / 2.32 / 14.36 / Morphine 3-glucuronide / Alkaloid / 298.20, 298.84, 295.78, / 460.16, 286.14, 268.13, 229.08 / 0.017 / 10.79*