STDs� +GLU� +INV�Fructose�

Fructose Glucose�

Sucrose�

Gentiobiose�

Gentianose�

Gentianose�

Supplemental Figure 1. TLC analysis of sugars hydrolyzed from

gentianose.

Gentianose (G2F) were hydrolyzed by β-glucosidase obtained from

Ustilago esculenta (GLU; Nakajima et al., 2012) and purified

recombinant protein of INV. After hydrolysis, sugars were separated by

silica gel TLC with ethyl acetate-acetic acid-water (3:2:1) as the mobile

phase and stained thymol-sufuric acid reagent. Fructose was stained pink

color, but glucose was stained violet color.

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631�

0

0.05

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0

100

200

300

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Feb.

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.

nmol

mg-1

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cv. SpB�cv. Maciry�

0

50

100

150

200

Oct

. N

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Dec

. Ja

n.

Feb.

M

ar.

Apr

.

nmol

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G2F�

G2�

0

50

100

150

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. N

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Dec

. Ja

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Feb.

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.

nmol

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0

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INV� INV�

G2F�

0

100

200

300

Oct

. N

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Dec

. Ja

n.

Feb.

M

ar.

Apr

.

nmo

mg-1

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Supplemental Figure 2. Variety comparison of changes in gentio-

oligosaccharide concentrations and INV expression in OWBs.

The concentration of gentiobiose (G2) and gentianose (G2F), as well as

the expression level of INV in OWBs of cv. Maciry and cv. SpB were

compared. Each value represents the mean ± SD calculated from three

independent experiments.

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631�

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631�

Supplemental Figure 3. Characterization of recombinant INV protein.

(A) SDS-PAGE and western-blot analysis of recombinant INV. Purified INV protein

produced in P. pastoris was detected by Coomassie brilliant blue (CBB) staining or western-

blotting using the anti-Myc antibody.

(B) The effect of pH and temperature on gentianose (G2F) hydrolytic activity of INV. Each

value represents the mean ± SD calculated from triplicate measurements. �

19�

26�

37�

49�

64�

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15�

[kDa]�

15�

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CBB� anti-Myc antibody�

G2F

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ity (%

)�

A� B�

0 20 40 60 80

100 120

3 4 5 6 7 pH

0 20 40 60 80

100 120

10 20 30 40 50 60 70 80 Temperature (oC)

GFP� BF� Merge�IN

V�

At-G

RP5�

Con

trol�

Supplemental Figure 4. Subcellular localization of INV-sGFP.

Transiently expressed GFP (control, upper-row) and GFP-fusion protein

(INV, middle row) in Nicotiana benthamiana leaves. GFP channel (left),

bright field (BF, center) and merged image (right) are shown. Bars

indicate 50 µm. The subcellular localization of the GFP fusion protein

was examined 3 days after incubation under constitutive dark conditions.

The At-GFP5 protein was used as a vacuolar protein marker (Mangeon et

al, 2009).�

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631�

Supplemental Figure 5. Uptake of G2-AP into IOWBs.

IOWBs were cultured on solid MS medium containing 0.5% G2-AP at

22oC with a 16/8 h light/dark photoperiod. After 24 h, IOWBs (n=3) were

divided in half, and metabolites in the top (T) and bottom (B) halves of

IOWBs were extracted as described in METHODS. G2-AP was separated

by TLC and the fluorescence of G2-AP was detected.

G2-AP (0.5 mg/ml)�

T� B� T� B� T� B�IOWB 1� IOWB 2� IOWB 3�

T�

B�

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631�

0

1

2

3

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Mar

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Supplemental Figure 6. Changes in PDH expression and the sum

concentrations of gentiobiose and gentianose in field grown OWBs.

The expression level of PDH and the sum of gentiobiose (G2) and

gentianose (G2F) concentrations in OWBs of cv. SpB was measured.

Each value represents the mean ± SD calculated from three independent

experiments.

0

100

200

300

400

Oct

. N

ov.

Dec

. Ja

n.

Feb.

M

ar.

Apr

.

nmol

mg-1

DW�

PDH� G2+G2F�

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631�

Supplemental Figure 7. Changes in gentiobiose-affected gene

expression in field-grown OWBs before and during budbreak.

Expression levels of genes that gentiobiose induced in IOWBs were

compared in field grown OWBs. Each value represents the mean ± SD

calculated from three independent experiments.

0

10

20

Jan

Feb

Mar

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pr

0.0 0.2 0.4 0.6 0.8

Jan

Feb

Mar

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100

200

300

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Feb

Mar

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Jan

Feb

Mar

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Jan

Feb

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50

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Feb

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Feb

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1.5

Jan

Feb

Mar

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0.5

1

1.5

Jan

Feb

Mar

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pr 0

0.5

1

1.5

Jan

Feb

Mar

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0.2

Jan

Feb

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0.1

0.2

Jan

Feb

Mar

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pr 0

10 20 30 40

Jan

Feb

Mar

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2

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SiR� OASL1� OASL2� GCL�

GSHS� EGS� CBL� MeS1� MeS2�

SAMS1� SAMS2� SAHH1�

SAHH2�

SAMDC�

DHAR1� DHAR2�

0

5

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Jan

Feb

Mar

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0

5

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Jan

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0 0.01 0.02 0.03 0.04

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ACO�

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631�

Supplemental Figure 8. Effect of treatment of ethylene precursors on

budbreak of gentian OWBs.

Field-grown endodormant OWBs (G. triflora cv. SpB) harvested in

September were treated with water (Control), 140 µM ethephon, or 5 mM

ACC. After incubation at 22oC for 3 weeks, budbreak of OWBs was

measured. Scale bars indicate 2 cm.

Control 0 week�

Control 3 weeks�

140 µM ethephon 0 week�

140 µM ethephon 3 weeks�

5 mM ACC 0 week�

5 mM ACC 3 weeks�

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631�

Supplemental Figure 9. Effect of gentiobiose treatment on

germination of Arabidopsis seeds.

Arabidopsis seeds were placed on sugar-free (SF) or 1% gentiobiose

(G2)-containing MS agar medium and incubated at 4oC for 4 days before

transferring to a growth chamber at 22oC under continuous light (60 µmol

m-2 s-1). Metabolites in the seeds were extracted and quantified as

described in METHODS. Each value represents the mean ± SD

calculated from three independent experiments.

SF (1 day)�

SF (2 days)�

SF (3 days)�

1% G2 (1 day)�

1% G2 (2 days)�

1% G2 (3 days)�

0

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0 12 24 36

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(nm

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Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631�

PC1 PC2Gly 144.02 ± 66.19 80.49 ± 24.52 205.39 ± 17.62 0.79 -0.31Ala 1042.80 ± 457.39 890.32 ± 206.37 1034.70 ± 237.13 0.24 -0.14Asn 23095.54 ± 6697.22 9524.78 ± 3250.30 71582.78 ± 7704.58 0.96 -0.07Asp 7161.58 ± 1077.12 4590.30 ± 698.22 8162.24 ± 808.54 0.69 -0.57Lys 311.53 ± 56.98 524.20 ± 123.59 1344.66 ± 310.59 0.87 0.30Gln 5678.61 ± 1409.99 1048.14 ± 306.89 21238.19 ± 4531.39 0.98 -0.06Glu 10529.49 ± 1993.13 8662.74 ± 1984.99 13275.41 ± 939.12 0.77 -0.27His 570.93 ± 242.94 546.21 ± 158.34 5578.94 ± 1547.26 0.95 0.14Arg 5332.56 ± 1674.87 2083.70 ± 892.34 11081.49 ± 4991.90 0.77 -0.18Ser 1322.28 ± 251.97 1247.35 ± 166.34 2488.25 ± 489.95 0.94 0.13Pro 430.74 ± 119.04 170.97 ± 67.55 1430.42 ± 257.78 0.96 -0.04Val 574.36 ± 78.58 732.44 ± 71.79 1352.32 ± 131.75 0.91 0.30Thr 495.20 ± 112.48 264.63 ± 91.88 985.34 ± 136.57 0.93 -0.16Leu/Ile 411.14 ± 95.73 1016.40 ± 360.70 1679.97 ± 472.02 0.70 0.53Met 121.34 ± 47.92 87.06 ± 27.49 421.03 ± 126.43 0.96 0.07Phe 216.07 ± 58.07 618.38 ± 271.21 460.99 ± 122.20 -0.01 0.79Tyr 111.10 ± 20.93 260.85 ± 138.72 1150.68 ± 749.45 0.77 0.22Trp 137.94 ± 24.07 983.53 ± 219.59 2788.21 ± 497.14 0.87 0.44Putrescine 35.87 ± 13.97 156.75 ± 43.51 190.50 ± 80.42 0.50 0.74GABA 664.89 ± 250.97 427.83 ± 173.20 1188.38 ± 1040.69 0.57 -0.01Spermidine 78.69 ± 30.78 486.44 ± 140.86 297.03 ± 57.14 -0.10 0.89Citrulline 126.17 ± 20.25 123.23 ± 14.16 234.18 ± 17.53 0.96 0.11Spermine 9.68 ± 2.49 24.87 ± 7.70 48.35 ± 15.53 0.68 0.39Arg-suc 4.31 ± 0.47 4.00 ± 1.01 10.38 ± 2.72 0.92 0.08SAM 25.44 ± 6.11 35.08 ± 5.07 42.52 ± 14.72 0.57 0.51Glucose 51832.01 ± 9506.34 25482.80 ± 9691.22 86292.99 ± 37891.61 0.74 -0.25Sucrose 95711.42 ± 6408.32 105375.75 ± 11629.42 56825.65 ± 24731.40 -0.82 0.03Gentiobiose 36034.78 ± 11962.49 3116.67 ± 607.29 64196.07 ± 34612.87 0.64 -0.42Gentianose 23790.61 ± 20678.04 72913.03 ± 15621.24 10752.35 ± 4160.33 -0.65 0.55Arabinose 228.00 ± 76.62 178.00 ± 54.95 312.00 ± 66.86 0.66 -0.09Galactose 1216.67 ± 294.22 501.67 ± 85.07 755.00 ± 150.65 -0.01 -0.87AMP 24.75 ± 8.76 47.78 ± 23.64 35.82 ± 17.50 0.02 0.66ADP 79.41 ± 22.78 140.73 ± 15.72 144.36 ± 20.91 0.41 0.81ATP 183.24 ± 30.85 90.38 ± 6.36 287.13 ± 105.07 0.71 -0.37GMP 8.91 ± 2.87 12.35 ± 1.64 10.41 ± 2.29 -0.02 0.59GDP 13.11 ± 4.04 30.44 ± 13.44 25.00 ± 9.88 0.14 0.78GTP 36.17 ± 11.41 29.46 ± 6.41 62.82 ± 24.61 0.67 -0.06UDPG 957.72 ± 140.33 512.14 ± 50.18 1319.17 ± 236.97 0.79 -0.44NAD 88.78 ± 9.81 153.04 ± 24.64 154.37 ± 51.48 0.37 0.78NADP 115.35 ± 35.74 111.18 ± 17.84 141.07 ± 32.66 0.54 0.04NADPH 56.44 ± 19.59 6.35 ± 2.04 77.19 ± 33.73 0.63 -0.51Pyruvate 166.39 ± 31.18 191.44 ± 45.82 185.04 ± 47.52 0.00 0.32Fumarate 2379.81 ± 649.44 4363.92 ± 734.53 4470.54 ± 1014.06 0.24 0.64Succinate 251.78 ± 59.12 67.91 ± 22.79 212.40 ± 83.38 0.38 -0.772OG 99.46 ± 11.13 42.95 ± 19.48 95.99 ± 38.06 0.48 -0.56PEP 10.55 ± 2.59 8.71 ± 2.96 44.67 ± 7.51 0.97 0.10DHAP 31.74 ± 8.07 25.04 ± 5.18 32.22 ± 8.80 0.31 -0.31Aconitate 62.66 ± 27.87 21.91 ± 4.37 62.40 ± 34.82 0.45 -0.42Shikimate 112.92 ± 27.68 89.15 ± 26.48 93.74 ± 12.94 -0.07 -0.483PGA 69.92 ± 9.45 75.30 ± 18.85 231.40 ± 56.52 0.93 0.17Ru5P/R5P 366.12 ± 292.02 107.53 ± 35.36 205.13 ± 138.70 0.03 -0.54G6P 1339.84 ± 400.14 1003.30 ± 155.11 1521.59 ± 172.54 0.61 -0.256PG 122.23 ± 62.40 64.39 ± 22.54 79.23 ± 55.44 -0.09 -0.54RuBP 3.33 ± 0.81 5.28 ± 2.17 9.09 ± 2.21 0.75 0.35FBP 40.07 ± 16.21 25.28 ± 4.62 60.65 ± 29.15 0.72 -0.12Isocitrate 757.52 ± 153.04 1543.56 ± 199.20 1186.67 ± 468.25 -0.14 0.67Citrate 32948.76 ± 14720.26 55351.82 ± 23177.19 41762.71 ± 9831.45 -0.20 0.43Malate 25881.88 ± 6914.06 60272.95 ± 11358.89 57208.58 ± 10034.76 0.23 0.79

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631

PC loadings

Supplemental Table 1. Metabolite profiles in OWBs (G. triflora x G. scabra (05-543)) harvested in October, January, andMarch

Data presented are means ± SD of five independent experiments. DW, dry weight; GABA, γ-amino butyrate; SAM, S-adenosyl methionine; UDPG, UDP glucose; 3PGA, 3-phosphoglycerate; PEP, phosphoenolpyruvate; DHAP,dihydroxyacetone phosphate; Ru5P, ribulose 5-phosphate; R5P, Rib 5-phosphate; G6P, Glc 6-phosphate; 6PG, 6-phosphogluconate; RuBP, ribulose 1,5-bisphosphate; FBP, Fru 1,6-bisphosphate.

Concentrations (nmol g-1DW)October January March

Metabolites Ratio F-test T-testSF 1% G2 SF 1% G2 G2/SF

Alanine 0.368 0.345 0.078 0.052 0.939 0.526 0.649Arginine 9.313 6.729 0.224 1.111 0.723 0.026 0.017Asparagine 3.486 3.373 0.192 0.464 0.968 0.184 0.724Aspartate 1.294 1.288 0.08 0.407 0.995 0.024 0.975Citrulline 0.195 0.044 0.061 0.029 0.224 0.263 0.004Cysteine 0.005 0.015 0 0.001 3.186 0.111 1.27E-04GABA 0.297 0.274 0.148 0.06 0.923 0.175 0.787Glu-Cys 0.003 0.008 0.001 0.001 2.814 0.777 0.001Glutamate 5.611 7.427 0.579 1.082 1.324 0.332 0.035Glutamine 10.562 9.702 0.345 1.894 0.919 0.019 0.434Glycine 0.02 0.021 0.007 0.004 1.048 0.459 0.825Histidine 0.479 0.344 0.189 0.021 0.719 0.005 0.206Homocysteine 0.014 0.027 0.007 0.005 1.979 0.69 0.02Isoleucine 2.068 1.705 0.646 1.016 0.824 0.476 0.572Leucine 1.204 1.4 0.439 0.313 1.163 0.59 0.496Lysine 2.864 1.887 0.722 0.492 0.659 0.545 0.073Methionine 0.228 0.469 0.064 0.037 2.061 0.408 0.001Ornithine 0.267 0.114 0.035 0.011 0.426 0.088 0.002Phenylalanine 2.145 1.974 0.881 0.804 0.92 0.884 0.784Proline 0.462 0.445 0.158 0.239 0.962 0.518 0.906SAM 0.022 0.035 0.004 0.003 1.599 0.941 0.002Serine 0.344 0.414 0.079 0.074 1.203 0.919 0.244Spermidine 0.217 0.188 0.125 0.045 0.864 0.124 0.682Spermine 0.032 0.005 0.014 0.002 0.167 0.016 0.008Threonine 0.564 0.669 0.134 0.117 1.186 0.828 0.285Tryptophan 0.703 0.776 0.587 0.324 1.104 0.354 0.837Tyrosine 0.779 0.746 0.818 0.201 0.957 0.045 0.942Valine 4.572 3.682 3.065 3.365 0.805 0.881 0.709ACC 3.095 2.907 0.457 0.443 0.939 0.961 0.577GSH 0.127 2.597 0.08 0.381 20.47 0.029 1.46E-05GSSG 0.054 0.119 0.027 0.041 2.212 0.496 0.0442OG 0.431 0.426 0.062 0.086 0.99 0.596 0.9363PGA 0.219 0.195 0.13 0.117 0.89 0.776 0.7756PG 0.79 0.717 0.683 0.185 0.908 0.06 0.849Aconitate 0.144 0.102 0.078 0.019 0.708 0.042 0.335Ascorbate 0.141 3.628 0.103 0.506 25.695 0.01 1.24E-06DHA 2.161 2.788 1.102 1.009 1.29 0.889 0.434Citrate 8.904 5.527 4.901 0.792 0.621 0.014 0.223DHAP 1.249 0.92 0.554 0.106 0.736 0.022 0.322Fumarate 6.376 5.222 1.891 1.356 0.819 0.599 0.363G6P 0.974 1.42 0.139 0.082 1.458 0.407 0.003Malate 12.334 11.59 4.691 3.163 0.94 0.534 0.803NAD 0.04 0.035 0.016 0.009 0.878 0.36 0.614NADH 0.017 0.016 0.003 0.001 0.945 0.68 0.806NADP 0.039 0.052 0.011 0.004 1.311 0.113 0.114NADPH 0.008 0.008 0.003 0.001 0.931 0.163 0.774PEP 0.23 0.137 0.106 0.05 0.596 0.252 0.183Shikimate 0.068 0.065 0.023 0.008 0.963 0.113 0.846Succinate 0.089 0.045 0.062 0.008 0.504 0.008 0.207

AVR (nmol mg-1DW) SD

Supplemental Table 2. Metabolite concentrations in IOWBs (G. triflora (cv. Ihatovo)) cultured withsugar free (SF) or with 1% gentiobiose containing MS medium (1% G2).

Data presented are calculated from four independent experiments. DW, dry weight; GABA, γ-aminobutyrate; Glu-Cys, γ-glutamyl cysteine; SAM, S-adenosyl methionine; ACC, 1-Aminocyclopropane-1-carboxylic acid; 2OG, 2-oxoglutarate; 3PGA, 3-phosphoglycerate; 6PG, 6-phosphogluconate; DHAP,dihydroxyacetone phosphate; G6P, Glc 6-phosphate; PEP, phosphoenolpyruvate.

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631

Supplementary Table 3. List of primers used in this studyForward (5´→ 3´) Reverse (5´→ 3´)

CloningINV ATGCCATCCAAAAATTCAAAATCT ATAGGATCTAATGTCGGCGGAGTTHXK ATGGGGAAGGTAGCTGTGACGGCGG AGACTCCTCTAAACCAATGTACTGPGM ATGACATTCTGTACC TGTTATCACTGTTGGCTTCTCUGP ATGGAAACTCCTGCAA TATGTCTTGAGGACTGCTAACSPS ATGGCGGGAAATGATTGGATAAACA GAGAACTTGTAGCTTCTCTAACGATGSUS ATGGCGCCTAAGTTAGAGAAAATA ATGCTCATCATCCACAGCAAGSiR ATGGCGACGTCGTATGGAGCAGCAGC CTTCTCAGCAAAACGGGAAGAGTGAGOASL1 ATGGCACAGGAAAAGATTGGTATT GGGATCAAAGGTCATGTTCTCTGCOASL2 ATGGCATCATCTTCTTCTTCTTC CAATCAGGGCACTGGTACCATCTTOASL3 ATGGCTGCTGCTGCTTTAGCAAATCT ATCAACTGACACAGGCTGCATATTCECS ATGGCACTTATGTCACAAGCGGGATC GTATAGTAGCTCTTCGAAAACAGGTGSHS ATGGGTTCTGCATATTCTTCTTCTGA CCAACCGTAACAGCTGCATTTGCACCBL ATGGCGTCTTCCTTGTTCCGTCAA AGCCGGTGCAGTTCTTAGTGCGTAATCGS ATGGCCGTCTCCAGCTGCGCTAGGGT ATGGTCTCCAAAGCCTGCAGGATAMeS1 ATGGCATCACACATCGTTGGATACCC TTTGGCACTTGAGAGCTGTGTGCGGMeS2 ATGGCATCCCACATCGTTGGATATC TTTAGCACTGGCAAGCTGTGTGCGGSAMS1 ATGGACACTTTTCTATTCACCTCA AGCTTTGGGCTTGAGGATCTTGACASAMS2 ATGGATACCTTCTTGTTCACTTCTG TGGCTTGTCGCTTATGAGGGGCTTGSAMDC ATGTCGGAAATGCCGGTCTCTGCGA CTCCTTTTCTTCATCTTCATCCTCCTACO ATGGCATTCCCAATAGTAAACATGGAGAAG TCAAACGGGGGCAATGGGTCCCAAATTAATGSAHH1 ATGTCTCTCTCCGTCGAGAAGACCA ATACCTATAGGCCAAAGGCTTGTATSAHH2 ATGTCTTTGCTCGTGGACAAAACCA ATACCTATAGTGAAGAGGCTTATAADHAR1 ATGTCGACCGCCAAGTTAACATCA ACCACCCATCACCTTCGGCCGCCADHAR2 ATGGCTCCGCCCGTTGAAGTAGCT TGCCTCAACCTTTGGCTTCCATCCGR1 ATGTCGGGTTTGGGAAGGAAGATG AAGGTTTGTTTTCGGTTTAACCGCGR2 ATGGCGACGTCTCTTGCTACGCCG GACCCCAGCAGTGGCTTTGATCTCAtGRP5 ATGGCTTCCAAGTCACTCTTTCTTGT ATGATGTCCACCACCGAAACCACCT

Real-time PCRINV GTGGGTCTCGGGTTGAGATA AATCCAGCCCCATAAAATCCHXK TGTGATATTGGGAACGGGAACTA ATCTTGCCACTTGGGAATCGPGM TTGGGTGGTGATGGTCGAT ACCATTGCCTGCGGAAATTUGP TGAGTTGGGACCCGAGTTTAAG CACGATACTGGGAATGGATTTGASPS CCTTTGGAGAATGTCGACCTTT CTCATCGATATTATCCCGGTTACCSUS CTGGAGTTTATGGCTTCTGGAAAT GAACATCTCAAGGTAACGCCTTGTSiR CCACTTTGTCCTTTGGCAATC CCCTCTTAAGCACATCCGGTATOASTL1 AAAACCTGGCCCGCATAAA CAAAACACCAGGAATGAATCCAOASTL2 TTTCCTGGCCATCAGATTTGT AAGGTAGCCAACCCAATGTCAOASTL3 TCTTCCCTAGTCACTCCATTTGC ATTCGAACTGTGGGTCCAGAAECS AACCCAACGGTTATCTCAGCAT TCCGATCTCCATCTGTGTCAAGGSHS GCTTCCCTTCCTTCGAATCTG CTGCCTTCTCGGTCTCCATTTCBL TTGGAAGCGTAAAGTCCCTCAT GGTATGCTCGCGTGTGACACGS GGGCCTAAAAAGTCATCCTGAA CACCACCAAAACCAGTCATCTGMeS1 GCAATCTTATGGTTCTCGTTGTGT GCTTGGGTCGACTCACATCAMeS2 GGGTCAACCCGGATTGTG CAGAGCCGGCTTGACTTCTCSAMS1 TTCGGGATACCTGCAGAGAGA TCAGCATCCAGGCCAACASAMS2 TGCCTCTGAGCCATGTCCTT TCTTGCGGACTTCGGTCAASAMDC AATTATACACGTGGGAGCTTCATCT GAAGCTACGATGCGGAAACGACO GAAGCTGTTGAGGGTGAAGTCA TCAGGGATTTCAGAGATGTTGGASAHH1 TCGACATGCTCGGGTTAGAGA GGTTTGTGGCTTGATCGTGATSAHH2 GAATTCAAAACGTCTGGGACAAT ACGATCTGGAACTCAGCATTATCADHAR1 GGGAAGCCGGCGAGTAA CGCCGACTTCGCCATTAATDHAR2 AGATCCCAATGATGGCTCAGA GATGCTCATCCAATGCTTTCAAGR1 GCCTGTTGCCTTGATGGAA TCGGCTGCTCACCAAAAACGR2 CCTTCGAAGCCCAAAAAGATT CCAGCAAACTCAACAGCGATATUBQ GGAAGCAGTTGGAAGATGGC GGCCAAAGTTCGTCCGTCCTC

Vector construction for protein expression

INV CCGCTCGAGAAAAGAATGCCATCCAAAAATTCAAAATCTC

TGCTCTAGAAGATAGGATCTAATGTCGGCGGAGTT

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631

Supplemental references Nakajima, M., Yamashita, T., Takahashi, M., Nakano, Y., Takeda, T. (2012). Identification, cloning, and characterization of β-glucosidase from Ustilago esculenta. Appl. Microbiol. Biotechnol. 93: 1989–1998.� Mangeon, A., Magioli, C., Menezes-Salgueiro, A.D., Cardeal, V., de Oliveira, C., Galvao, V.C., Margis, R., Engler, G., and Sachetto-Martins, G. (2009). AtGRP5, a vacuole-located glycine-rich protein involved in cell elongation. Planta 230: 253-265.

Supplemental Data. Takahashi et al. Plant Cell (2014) 10.1105/tpc.114.131631�