Supplementary Material
Theoretical and Experimental Plant Physiology
DIMBOA levels in hexaploid Brazilian wheat are not associated with antibiosis against the cereal aphids Rhopalosiphum padi and Sitobion avenae
Jorge F. Pereira • André L. F. Sarria • Stephen J. Powers • Gudbjorg I. Aradottir • John C.
Caulfield • Janet Martin • Lesley E. Smart • John A. Pickett • Michael A. Birkett • Paulo R. V.
S. Pereira
J. F. Pereira ( ) • P. R. V. S. Pereira
Embrapa Trigo, Rodovia BR 285 km 294, Passo Fundo, RS, 99001-970, Brazil
E-mail: [email protected] Phone: +55(32)3311-7400 Fax: +55(32)3311-7401
Present address:
J. F. Pereira
Embrapa Gado de Leite, Rua Eugênio do Nascimento 610, Juiz de Fora, MG, 36038-330,
Brazil
A. L. F. Sarria • G. I. Aradottir • J. C. Caulfield • J. Martin • L. E. Smart • J. A. Pickett • M. A.
Birkett
Biological Chemistry and Crop Protection Department, Rothamsted Research, West
Common, Harpenden, Hertfordshire, AL5 2JQ, UK
S. J. Powers
Computational and Systems Biology Department, Rothamsted Research, West Common,
Harpenden, Hertfordshire, AL5 2JQ, UK
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0 50 100 150 200 2500
500000
1000000
1500000
2000000
2500000
3000000
3500000
f(x) = 13135.9555434026 x − 24179.9126147631R² = 0.999076115972712
DIMBOA concentration (µgmL-1)
Area
(mAU
)
Fig. S1 Calibration curve obtained with DIMBOA standard solutions (n = 27)
Fig. S2 1H NMR (500 MHz, CD3OD) spectrum of the 2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA).
2
N
O O
O
H3CO
OH
R2
5
78
8a
4a346
1
DIMBOA (R = H)
Fig. S3 13C NMR (125 MHz, CD3OD) spectrum of the 2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA).
Fig. S4 1H-1H COSY (500 MHz, CD3OD) spectrum of the 2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA).
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Fig. S5 HSQC (500 MHz, CD3OD) spectrum of the 2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA).
Fig. S6 HMBC (500 MHz, CD3OD) spectrum of the 2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA).
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Fig. S7 1H NMR (500 MHz, CD3OD) spectrum of the 2-O-β-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA-Glc).
Fig. S8 13C NMR (125 MHz, CD3OD) spectrum of the 2-O-β-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA-Glc).
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N
O O
O
H3CO
OH
R2
5
78
8a
4a346
1
DIMBOA-Glc (R = Glc)
Fig. S9 Projection of the 13C NMR from HSQC (125 MHz, CD3OD) spectrum of the 2-O-β-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA-Glc).
Fig. S10 Projection of the 13C NMR from HMBC (125 MHz, CD3OD) spectrum of the 2-O-β-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA-Glc).
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Fig. S11 1H-1H COSY (500 MHz, CD3OD) spectrum of the 2-O-β-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA-Glc).
Fig. S12 HSQC (500 MHz, CD3OD) spectrum of the 2-O-β-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA-Glc).
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Fig. S13 HMBC (500 MHz, CD3OD) spectrum of the 2-O-β-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA-Glc).
m/z100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 300
%
0
100 164
152
149113113101 131119
119127 136 143
155
158
234208
165
184180166
169173
194192
189198
206
232220
210 220
213222
224
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241290262248 260
257 276264 274 286278 300293
m/z100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
%
0
100 152
123
122
113
149
131
164
454372
192
165
178
356235
194 234
220
248 354
341312274 302
404
440432
430
476
767
652536492
516
552630552 574 618 653
722700 732
789
790
849791821 850 970910887
924950 975 996
Fig. S14 Mass spectrum of DIMBOA (above) and DIMBOA-Glc (below) both in negative mode [M-H]-.
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Table S1 Mean concentration of DIMBOA in nine wheat cultivars at three time points. Data are presented on the natural log scale for statistical comparison of means using the maximum (assuming n = 3) standard error of the difference 0.1456 on 71 degrees of freedom invoking the least significant difference (LSD) value at the 5% level of significance: 0.2309.
Genotype 7 days 9 days 14 daysBR 32 2.392 0.295 0.007BRS 327 2.659 2.568 1.719BRS 328 2.716 1.809 1.373BRS Guabiju 1.667 0.808 -0.206BRS Guamirim 3.303 2.768 1.947BRS Parrudo 1.745 2.055 0.685PF869120 2.623 2.208 1.804Solstice 2.575 2.193 1.503BRS Timbaúva 3.418 2.452 1.836
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