Table S2. Environmental Differences in Dhurrin Accumulation

Figure S1. Schematic diagram of the revised dhurrin cyanogenic glucoside pathway adapted from Gleadow and Møller, 2014. The branches of the pathway are highlighted in different colors, where the blue region refers to biosynthetic branch, the orange region refers to known catabolic branch involving HCN production and the purple region refers to the new and developing turnover/catabolic branch of the pathway which does not invoke HCN production.

List of Supplemental Tables

Table S1. Canonical genes for dhurrin biosynthesis and catabolism. Positions reported are based on the Sorghum bicolor assembly version 2.1.

Gene / Chr / Start / End / Locus Name / Function
CYP71E1 / 1 / 1,047,338 / 1,050,876 / Sobic.001G012000 / Dhurrin Biosynthesis
CYP79A1 / 1 / 1,138,289 / 1,141,020 / Sobic.001G012300 / Dhurrin Biosynthesis
UGT85B1 / 1 / 1,146,448 / 1,149,561 / Sobic.001G012400 / Dhurrin Biosynthesis
Dhurrinase 1 / 8 / 13,288,579 / 13,293,251 / Sobic.008G079800 / Dhurrin Catabolism
Dhurrinase 2 / 8 / 13,680,347 / 13,683,471 / Sobic.008G080400 / Dhurrin Catabolism
Environment / Dhurrin† / Nitrogen Application
2011 USDA / 45.6 / 30-0-0
2013 USDA / 53.1 / 30-0-0
2013 TAES / 36.4 / None
L.S.D. / 7.6

Table S2. Environmental differences in dhurrin accumulation.

† L.S. Mean values for leaf dhurrin measured as µg/cm2

Table S3. Average leaf dhurrin concentrations for the 700 SC lines used in this study.

Data is combined means of three environments (2011 USDA, 2013 USDA, 2013 TAES).

Pedigree / Working Group / Dhurrin† / S.E.
SC309 / Bicolor / 103.3 / 16.3
SC610 / Bicolor / 98.2 / 27.1
SC1338 / Bicolor / 76.0 / 11.7
SC609 / Bicolor / 70.7 / 14.2
SC987 / Bicolor / 70.7 / 23.2
SC311 / Bicolor / 68.5 / 5.5
SC1333C / Bicolor / 65.9 / 23.6
SC1339C / Bicolor / 60.9 / 18.5
SC66 / Bicolor / 60.4 / 5.1
SC1341C / Bicolor / 59.3 / 18.6
SC307 / Bicolor / 57.0 / 20.4
SC616-11E / Bicolor / 55.5 / 9.5
SC306 / Bicolor / 54.3 / 22.7
SC147 / Bicolor / 49.7 / 4.8
SC1107-9 / Bicolor / 48.9 / 3.2
SC156 / Bicolor / 48.5 / 6.6
SC17 / Bicolor / 47.1 / 6.5
SC67 / Bicolor / 46.3 / 8.9
SC16 / Bicolor / 44.8 / 9.2
SC224 / Bicolor / 42.5 / 10.6
SC19 / Bicolor / 41.3 / 9.8
SC1337C / Bicolor / 40.1 / 15.6
SC679 / Bicolor / 38.6 / 21.2
SC1332C / Bicolor / 37.2 / 9.3
SC614 / Bicolor / 36.2 / 7.7
SC950 / Bicolor / 35.6 / 6.9
SC1426 / Bicolor / 35.1 / 8.4
SC1342C / Bicolor / 32.9 / 6.9
SC20 / Bicolor / 31.9 / 8.9
SC44 / Bicolor / 31.4 / 14.1
SC621 / Bicolor / 31.3 / 17.0
SC145 / Bicolor / 29.5 / 1.9
SC301 / Bicolor / 28.8 / 6.2
SC941 / Bicolor / 27.6 / 6.6
SC214 / Bicolor / 27.1 / 8.2
SC263-11E / Bicolor / 26.9 / 1.4
SC1429 / Bicolor / 23.6 / 7.3
SC618-6 / Bicolor / 23.1 / 8.4
SC1203 / Bicolor / 21.8 / 7.2
SC1177C / Bicolor / 21.7 / 2.0
SC213 / Bicolor / 18.6 / 1.0
SC12 / Bicolor / 17.1 / 3.8
SC620 / Bicolor / 13.6 / 5.1
SC942 / Bicolor / 11.8 / 2.4
SC937 / Bicolor / 9.7 / 1.7
SC728 / Caudatum / 121.8 / 12.9
SC373 / Caudatum / 119.2 / 38.8
SC738 / Caudatum / 118.2 / 1.1
SC239 / Caudatum / 117.7 / 33.7
SC60 / Caudatum / 112.1 / 8.3
SC54 / Caudatum / 107.9 / 34.9
SC685 / Caudatum / 107.4 / 35.0
SC686 / Caudatum / 106.3 / 41.0
SC713 / Caudatum / 105.9 / 23.7
SC1120 / Caudatum / 100.8 / 4.4
SC73 / Caudatum / 100.6 / 24.3
SC50 / Caudatum / 100.4 / 36.7
SC1463 / Caudatum / 100.1 / 16.2
SC338 / Caudatum / 98.5 / 28.1
SC724 / Caudatum / 97.2 / 40.3
SC347 / Caudatum / 96.8 / 16.5
SC1351C / Caudatum / 94.9 / 23.7
SC702 / Caudatum / 94.9 / 28.4
SC574 / Caudatum / 94.6 / 4.3
SC692 / Caudatum / 94.4 / 41.6
SC237 / Caudatum / 93.8 / 23.0
SC557 / Caudatum / 93.6 / 28.1
SC335 / Caudatum / 92.8 / 25.7
SC701 / Caudatum / 91.7 / 39.6
SC396 / Caudatum / 88.9 / 11.8
SC228 / Caudatum / 87.4 / 27.1
SC964 / Caudatum / 87.1 / 22.0
SC323 / Caudatum / 86.6 / 14.8
SC1451 / Caudatum / 86.5 / 7.2
SC146 / Caudatum / 86.2 / 23.3
SC330 / Caudatum / 84.2 / 33.1
SC114 / Caudatum / 84.1 / 15.9
SC730 / Caudatum / 84.0 / 30.3
SC423 / Caudatum / 83.7 / 15.9
SC1116 / Caudatum / 83.4 / 24.9
SC569 / Caudatum / 83.0 / 32.7
SC339 / Caudatum / 82.5 / 14.5
SC55 / Caudatum / 82.5 / 12.1
SC972 / Caudatum / 82.1 / 11.2
SC328 / Caudatum / 81.0 / 25.9
SC567 / Caudatum / 80.7 / 11.8
SC970 / Caudatum / 79.9 / 18.7
SC704 / Caudatum / 79.8 / 35.3
SC1345C / Caudatum / 79.1 / 7.0
SC731 / Caudatum / 78.0 / 28.1
SC1117 / Caudatum / 77.9 / 23.0
SC715 / Caudatum / 77.8 / 18.0
SC63 / Caudatum / 77.2 / 10.7
SC680 / Caudatum / 76.6 / 14.1
SC1079 / Caudatum / 76.6 / 17.5
SC51 / Caudatum / 75.0 / 10.6
SC422 / Caudatum / 74.6 / 23.8
SC803 / Caudatum / 74.5 / 11.1
SC687 / Caudatum / 74.3 / 8.8
SC118 / Caudatum / 73.4 / 30.5
SC397 / Caudatum / 73.4 / 17.7
SC1019 / Caudatum / 73.4 / 3.8
SC1328C / Caudatum / 72.7 / 23.0
SC805 / Caudatum / 71.7 / 28.7
SC708 / Caudatum / 71.4 / 14.3
SC368 / Caudatum / 70.5 / 0.4
SC322 / Caudatum / 70.1 / 15.5
SC979 / Caudatum / 69.7 / 7.0
SC695-9 / Caudatum / 69.6 / 21.7
SC906 / Caudatum / 69.4 / 13.9
SC737 / Caudatum / 69.3 / 27.4
SC52 / Caudatum / 68.1 / 16.1
SC112 / Caudatum / 68.0 / 13.2
SC75-11E / Caudatum / 67.4 / 10.1
SC732 / Caudatum / 66.4 / 10.3
SC343-9 / Caudatum / 66.1 / 17.8
SC599 / Caudatum / 66.1 / 12.3
SC1111 / Caudatum / 65.8 / 7.2
SC562 / Caudatum / 65.6 / 31.1
SC165 / Caudatum / 65.5 / 17.0
SC352 / Caudatum / 65.0 / 13.5
SC719 / Caudatum / 64.7 / 22.2
SC394 / Caudatum / 64.6 / 5.9
SC58 / Caudatum / 64.3 / 15.9
SC176 / Caudatum / 64.1 / 8.1
SC707 / Caudatum / 62.9 / 12.9
SC324 / Caudatum / 62.8 / 24.7
SC1317C / Caudatum / 62.8 / 6.7
SC716 / Caudatum / 62.7 / 17.0
SC185 / Caudatum / 62.7 / 17.6
SC747 / Caudatum / 62.6 / 18.5
SC723 / Caudatum / 62.3 / 23.2
SC1057C / Caudatum / 62.3 / 9.5
SC345 / Caudatum / 61.9 / 32.7
SC720 / Caudatum / 61.6 / 29.9
SC380 / Caudatum / 61.4 / 1.0
SC705 / Caudatum / 61.3 / 16.4
SC798 / Caudatum / 61.2 / 15.9
SC689 / Caudatum / 61.1 / 14.3
SC344 / Caudatum / 61.1 / 13.7
SC1056 / Caudatum / 60.9 / 20.3
SC1263 / Caudatum / 60.6 / 14.1
SC57 / Caudatum / 60.4 / 14.7
SC334 / Caudatum / 60.4 / 1.2
SC398 / Caudatum / 60.0 / 8.6
SC1212C / Caudatum / 59.9 / 12.9
SC347 / Caudatum / 59.8 / 14.3
SC683 / Caudatum / 59.5 / 11.8
SC329 / Caudatum / 58.4 / 14.8
SC784 / Caudatum / 58.3 / 7.8
SC1222 / Caudatum / 58.2 / 11.5
SC564 / Caudatum / 57.8 / 30.1
SC93 / Caudatum / 57.6 / 7.2
SC694 / Caudatum / 57.5 / 17.6
SC726 / Caudatum / 56.9 / 12.8
SC333 / Caudatum / 56.7 / 20.6
SC1271 / Caudatum / 56.1 / 9.2
SC558 / Caudatum / 55.8 / 10.6
SC1262 / Caudatum / 55.3 / 18.1
SC120 / Caudatum / 54.8 / 22.0
SC712 / Caudatum / 54.7 / 9.7
SC790-6 / Caudatum / 54.3 / 18.4
SC387 / Caudatum / 54.2 / 19.8
SC727 / Caudatum / 53.5 / 25.9
SC103 / Caudatum / 53.4 / 19.2
SC336 / Caudatum / 53.3 / 4.4
SC121 / Caudatum / 53.0 / 10.7
SC401 / Caudatum / 52.8 / 17.8
SC175 / Caudatum / 52.6 / 6.6
SC1307C / Caudatum / 52.4 / 8.5
SC370 / Caudatum / 52.3 / 9.1
SC398 / Caudatum / 52.1 / 4.0
SC1302 / Caudatum / 51.9 / 11.1
SC797 / Caudatum / 51.8 / 6.4
SC1055C / Caudatum / 51.8 / 20.6
SC1251 / Caudatum / 51.4 / 20.4
SC367 / Caudatum / 51.2 / 22.1
SC172 / Caudatum / 51.2 / 12.0
SC690 / Caudatum / 50.7 / 10.9
SC173 / Caudatum / 50.5 / 8.4
SC725 / Caudatum / 49.9 / 15.5
SC369 / Caudatum / 49.0 / 13.4
SC77 / Caudatum / 49.0 / 7.7
SC693 / Caudatum / 48.5 / 2.9
SC353 / Caudatum / 48.2 / 10.6
SC804 / Caudatum / 48.2 / 13.2
SC68 / Caudatum / 48.2 / 3.3
SC96 / Caudatum / 48.0 / 11.1
SC563 / Caudatum / 47.5 / 13.4
SC1261 / Caudatum / 46.8 / 10.7
SC733 / Caudatum / 46.7 / 18.9
SC349 / Caudatum / 46.4 / 7.4
SC109 / Caudatum / 46.3 / 11.7
SC62 / Caudatum / 46.3 / 1.4
SC1133-6 / Caudatum / 46.2 / 22.2
SC42 / Caudatum / 46.1 / 7.0
SC56 / Caudatum / 45.9 / 7.5
SC706 / Caudatum / 45.8 / 10.6
SC1305C / Caudatum / 45.0 / 5.0
SC111 / Caudatum / 45.0 / 6.4
SC229 / Caudatum / 44.8 / 13.4
SC700 / Caudatum / 44.6 / 9.1
SC48 / Caudatum / 44.5 / 8.9
SC709 / Caudatum / 44.3 / 8.3
SC782 / Caudatum / 43.6 / 13.1
SC393 / Caudatum / 42.8 / 8.9
SC358 / Caudatum / 42.8 / 7.3
SC800 / Caudatum / 42.8 / 5.9
SC110 / Caudatum / 42.6 / 17.3
SC1300C / Caudatum / 42.5 / 12.8
SC1314C / Caudatum / 41.9 / 10.6
SC119 / Caudatum / 41.2 / 19.7
SC337 / Caudatum / 40.5 / 20.1
SC1441 / Caudatum / 40.3 / 10.3
SC108 / Caudatum / 40.1 / 16.1
SC982 / Caudatum / 39.7 / 3.6
SC504 / Caudatum / 39.6 / 10.3
SC1287C / Caudatum / 39.6 / 6.0
SC736 / Caudatum / 39.3 / 11.8
SC1316C / Caudatum / 39.2 / 6.9
SC842 / Caudatum / 39.0 / 5.1
SC170 / Caudatum / 38.9 / 12.1
SC101 / Caudatum / 38.6 / 7.9
SC308 / Caudatum / 38.5 / 5.1
SC340 / Caudatum / 37.9 / 13.0
SC1356C / Caudatum / 37.8 / 4.5
SC1313C / Caudatum / 37.7 / 3.5
SC963 / Caudatum / 37.2 / 16.5
SC399 / Caudatum / 37.0 / 3.6
SC1114-6 / Caudatum / 36.9 / 8.9
SC348 / Caudatum / 36.0 / 14.0
SC69 / Caudatum / 35.6 / 13.7
SC691 / Caudatum / 35.3 / 3.2
SC356 / Caudatum / 35.0 / 11.0
SC350 / Caudatum / 34.7 / 4.4
SC559 / Caudatum / 34.6 / 7.8
SC1205C / Caudatum / 34.5 / 6.5
SC721 / Caudatum / 34.2 / 10.0
SC424 / Caudatum / 33.7 / 11.4
SC400-6 / Caudatum / 33.4 / 5.1
SC1065 / Caudatum / 33.1 / 11.8
SC113 / Caudatum / 32.7 / 7.0
SC568 / Caudatum / 32.0 / 10.4
SC1103 / Caudatum / 31.7 / 12.3
SC403 / Caudatum / 31.6 / 5.3
SC402 / Caudatum / 30.9 / 8.5
SC1077 / Caudatum / 30.8 / 9.6
SC984 / Caudatum / 30.8 / 10.6
SC276 / Caudatum / 30.4 / 10.3
SC681 / Caudatum / 30.4 / 8.3
SC566 / Caudatum / 30.3 / 9.9
SC386 / Caudatum / 30.1 / 11.7
SC682 / Caudatum / 29.2 / 8.3
SC325 / Caudatum / 27.5 / 11.3
SC351 / Caudatum / 27.4 / 2.1
SC171 / Caudatum / 26.9 / 3.3
SC384 / Caudatum / 26.5 / 7.1
SC382 / Caudatum / 25.5 / 6.3
SC1424 / Caudatum / 25.4 / 9.1
SC374 / Caudatum / 24.7 / 2.8
SC734 / Caudatum / 23.9 / 2.6
SC371 / Caudatum / 23.2 / 7.0
SC354 / Caudatum / 22.7 / 3.6
SC362 / Caudatum / 22.1 / 4.4
SC391 / Caudatum / 20.9 / 3.9
SC1293C / Caudatum / 20.8 / 4.1
SC392 / Caudatum / 20.6 / 5.5
SC342 / Caudatum / 19.8 / 7.3
SC389 / Caudatum / 19.8 / 3.7
SC377 / Caudatum / 19.7 / 0.8
SC388 / Caudatum / 19.3 / 2.3
SC366 / Caudatum / 19.1 / 2.6
SC372 / Caudatum / 16.7 / 4.4
SC249 / Caudatum / 14.8 / 0.8
SC565 / Caudatum / 10.2 / 2.0
SC319 / Caudatum-Bicolor / 88.1 / 21.2
SC412 / Caudatum-Bicolor / 75.0 / 16.4
SC1330C / Caudatum-Bicolor / 64.8 / 24.8
SC85-6 / Caudatum-Bicolor / 62.0 / 17.4
SC949 / Caudatum-Bicolor / 51.4 / 13.0
SC411 / Caudatum-Bicolor / 44.4 / 9.6
SC74-3 / Caudatum-Bicolor / 42.1 / 10.0
SC115 / Caudatum-Bicolor / 40.0 / 1.8
SC59-12E / Caudatum-Bicolor / 36.5 / 6.2
SC1172 / Caudatum-Bicolor / 30.3 / 12.0
SC1076 / Caudatum-Bicolor / 29.4 / 10.2
SC749 / Caudatum-Bicolor / 26.9 / 8.9
SC991 / Caudatum-Bicolor / 26.6 / 6.3
SC413 / Caudatum-Bicolor / 19.6 / 3.0
SC144 / Caudatum-Durra / 125.2 / 24.2
SC142 / Caudatum-Durra / 97.1 / 44.6
SC808 / Caudatum-Durra / 88.3 / 12.0
SC428 / Caudatum-Durra / 82.4 / 28.5
SC1329C / Caudatum-Durra / 74.0 / 15.5
SC427 / Caudatum-Durra / 65.8 / 13.0
SC425 / Caudatum-Durra / 60.7 / 11.6
SC1325C / Caudatum-Durra / 59.3 / 17.9
SC188 / Caudatum-Durra / 58.7 / 28.4
SC186 / Caudatum-Durra / 55.5 / 4.0
SC187 / Caudatum-Durra / 55.3 / 21.2
SC575 / Caudatum-Durra / 53.2 / 10.0
SC577 / Caudatum-Durra / 52.1 / 18.8
SC951 / Caudatum-Durra / 43.9 / 14.2
SC810 / Caudatum-Durra / 41.4 / 13.7
SC1058 / Caudatum-Durra / 38.4 / 8.7
SC501 / Caudatum-Durra / 36.2 / 10.5
SC426 / Caudatum-Durra / 35.6 / 12.7
SC807 / Caudatum-Durra / 27.6 / 8.5
SC159 / Caudatum-Durra / 25.6 / 8.7
SC578 / Caudatum-Durra / 10.5 / 4.2
SC1494 / Caudatum-Guinea / 82.3 / 31.4
SC409 / Caudatum-Guinea / 75.7 / 28.7
SC1067 / Caudatum-Guinea / 73.9 / 10.3
SC1322C / Caudatum-Guinea / 69.5 / 28.2
SC1218 / Caudatum-Guinea / 68.4 / 13.7
SC406 / Caudatum-Guinea / 63.4 / 0.7
SC1321C / Caudatum-Guinea / 61.6 / 33.3
SC407 / Caudatum-Guinea / 56.9 / 8.6
SC405 / Caudatum-Guinea / 54.9 / 19.2
SC408 / Caudatum-Guinea / 52.6 / 20.6
SC49 / Caudatum-Guinea / 39.1 / 10.8
SC748 / Caudatum-Guinea / 36.0 / 12.3
SC1201C / Caudatum-Guinea / 31.1 / 12.1
SC1214C / Caudatum-Guinea / 31.0 / 3.3
SC913 / Caudatum-Guinea / 12.3 / 1.0
SC756 / Caudatum-Kafir / 102.3 / 34.5
SC1237 / Caudatum-Kafir / 87.6 / 37.0
SC751 / Caudatum-Kafir / 85.3 / 26.9
SC773 / Caudatum-Kafir / 75.3 / 40.8
SC780 / Caudatum-Kafir / 72.7 / 20.1
SC1082-6 / Caudatum-Kafir / 72.6 / 49.2
SC417 / Caudatum-Kafir / 63.6 / 23.6
SC777-6 / Caudatum-Kafir / 62.3 / 28.6
SC78 / Caudatum-Kafir / 61.9 / 4.9
SC79 / Caudatum-Kafir / 54.0 / 20.1
SC757 / Caudatum-Kafir / 51.6 / 22.3
SC762 / Caudatum-Kafir / 51.6 / 20.1
SC1104 / Caudatum-Kafir / 51.4 / 15.5
SC652-9 / Caudatum-Kafir / 50.8 / 19.1
SC223 / Caudatum-Kafir / 48.8 / 19.3
SC753-3 / Caudatum-Kafir / 47.8 / 19.6
SC420 / Caudatum-Kafir / 47.6 / 19.2
SC418 / Caudatum-Kafir / 46.6 / 8.5
SC672 / Caudatum-Kafir / 46.0 / 17.6
SC770 / Caudatum-Kafir / 43.7 / 17.0
SC764 / Caudatum-Kafir / 43.3 / 22.5
SC640-6 / Caudatum-Kafir / 42.4 / 12.5
SC760 / Caudatum-Kafir / 42.4 / 20.1
SC1179-6 / Caudatum-Kafir / 42.3 / 16.1
SC64 / Caudatum-Kafir / 40.0 / 13.9
SC637 / Caudatum-Kafir / 39.9 / 14.3
SC759-6 / Caudatum-Kafir / 39.8 / 14.9
SC642 / Caudatum-Kafir / 38.2 / 5.1
SC1246 / Caudatum-Kafir / 38.0 / 15.9