Fukushima et al., Supplemental Figure S1 A Col - 0 +MV0.01 M - - PDF document

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Fukushima et al., Supplemental Figure S1 A Col - 0 +MV0.01 M - - PDF document

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pad2-1 pad2-1 Fukushima et al., Supplemental Figure S1 A Col - 0 +MV0.01 M +MV0.05 M +MV0.1 M MS B +MV0.01 M MS pad2-1 Col - 0 +MV0.05 M +MV0.03 M Fukushima et al., Supplemental Figure S2 1) Control (no-stress) Col-0

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SLIDE 1

Fukushima et al., Supplemental Figure S1

pad2-1 pad2-1

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SLIDE 2

A B pad2-1 Col-0

MS +MV0.01 µM +MV0.05 µM +MV0.1 µM

Col-0

MS +MV0.01 µM +MV0.03 µM +MV0.05 µM Fukushima et al., Supplemental Figure S2

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SLIDE 3

Col-0 pad2-1 cad2-1

1) Control (no-stress) 3) P-lim 2) MS with MV

Fukushima et al., Supplemental Figure S3

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SLIDE 4

WT 7-5 13-6

mg FW per plant nmol/g FW nmol/g FW

50 100 150 200 2 4 6 8 10 12 14 0.0 10.0 20.0 30.0 40.0 50.0 60.0 70.0 50 100 150 200 250 300 WT 7-5 13-6

No-stress P-lim MV

WT 7-5 13-6 WT 7-5 13-6

* * a a a a * * ** * a b b b b b b b * b b b

A B C D E

Fukushima et al., Supplemental Figure S4

No-stress MV P-lim Shoot biomass GSH content GSSG content GSH:GSSG ratio

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SLIDE 5
  • 60
  • 40
  • 20

20 40 60

  • 100
  • 80
  • 60
  • 40
  • 20

20 40 60 80 100 t[3] t[2]

Control WT Control cad2-1 Control pad2-1 MV WT MV cad2-1 MV pad2-1 P-lim WT P-lim cad2-1 P-lim pad2-1

  • 60
  • 40
  • 20

20 40 60

  • 100
  • 50

50 100 t[3] t[1]

PC1 (17%) PC3 (6%) PC2 (10%) PC3 (6%)

A B

Fukushima et al., Supplemental Figure S5

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SLIDE 6

Increased metabolite set Decreased metabolite set Increased metabolite set cad2 vs WT

(FDR < 0.05, |log2fold-change| >= 1)

Decreased metabolite set pad2 vs WT

(FDR < 0.05, |log2fold-change| >= 1)

A B

Fukushima et al., Supplemental Figure S6

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SLIDE 7

No-stress P-lim MV pad2-1/WT cad2-1/WT

PG 34:3 Glucose G6P F6P F6PP 3PGA Leucine PEP Valine Alanine Threonine Pyruvate Isoleucine Cou-agmatine Sinapate HomoSerine Oxaloacetate Lysine β−Alanine Malate cis-Aconitate Aspartate Dihydrouracil Fumarate Isocitrate Asparagine Succinate 2-Oxoglutarate Arginine Succinyl-CoA Ornithine Glutamate Glutamine Putrescine Proline 4-Amino- butyrate Spermidine Urea

  • 2
  • 1 1 2

Quer F5 Quer F6 Kaem F1 Kaem F2 Kaem F3 Kaem F4 Antho A5 Antho A8 Antho A10 Antho A11

Log2fold-change

4-MSOB Galacturonate Raffinose Garactinol Sucrose Mannose Ascorbate Threonate Xylose Dehydroascorbate 5-MSOP Arabinose Fructose 6-MSOH Glycerol-3P Maltose 7-MSOH Trehalose Glycerol Inositol 8-MSOO GSH Glycine Serine Glycerate Inositol-1P Tryptophan Shikimate Tyrosine α-Tocopherol 4-MTB Cysteine Phenylalanine 5-MTP 6-MTH Methionine Caffeate 7-MTH 8-MTO β-Sitosterol Octadecatrienoate Campesterol 3-MTP 3-MSOP 1H-I3M 4H-I3M Gluconate TAG 52:5 TAG 54:7 TAG 54:8 TAG 54:9 DGDG 34:2 DGDG 36:3 PG 34:2 PE 36:2 PI 34:2 PI 34:3 PC 34:6 PC 36:1 PC 36:2

Dolichol undecaprenol Dolichol dehydro- decaprenol

Acetyl-CoA Citrate

MV/no-stress P-lim/no-stress

A B

PG 34:3 Glucose G6P F6P F6PP 3PGA Leucine PEP Valine Alanine Threonine Pyruvate Isoleucine Cou-agmatine Sinapate HomoSerine Oxaloacetate Lysine β−Alanine Malate cis-Aconitate Aspartate Dihydrouracil Fumarate Isocitrate Asparagine Succinate 2-Oxoglutarate Arginine Succinyl-CoA Ornithine Glutamate Glutamine Putrescine Proline 4-Amino- butyrate Spermidine Urea

  • 2
  • 1 1 2

Quer F5 Quer F6 Kaem F1 Kaem F2 Kaem F3 Kaem F4 Antho A5 Antho A8 Antho A10 Antho A11

WT cad2-1 pad2-1 Log2fold-change

4-MSOB Galacturonate Raffinose Garactinol Sucrose Mannose Ascorbate Threonate Xylose Dehydroascorbate 5-MSOP Arabinose Fructose 6-MSOH Glycerol-3P Maltose 7-MSOH Trehalose Glycerol Inositol 8-MSOO GSH Glycine Serine Glycerate Inositol-1P Tryptophan Shikimate Tyrosine α-Tocopherol 4-MTB Cysteine Phenylalanine 5-MTP 6-MTH Methionine Caffeate 7-MTH 8-MTO β-Sitosterol Octadecatrienoate Campesterol 3-MTP 3-MSOP 1H-I3M 4H-I3M Gluconate TAG 52:5 TAG 54:7 TAG 54:8 TAG 54:9 DGDG 34:2 DGDG 36:3 PG 34:2 PE 36:2 PI 34:2 PI 34:3 PC 34:6 PC 36:1 PC 36:2

Dolichol undecaprenol Dolichol dehydro- decaprenol

Acetyl-CoA Citrate * * * * * * * * * * * * *

Treatment comparison Genotype comparison

Fukushima et al., Supplemental Figure S7

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SLIDE 8
  • A. Genotype comparison under MV
  • B. Genotype comparison under P-lim

pad2-specific term under MV pad2-specific term under control Link in pad2 vs WT under control Link in pad2 vs WT under MV pad2-specific term under P-lim pad2-specific term under control Link in pad2 vs WT under control Link in pad2 vs WT under P-lim

Iron ion homeostatis Metal ion ROS Organism biotic Iron ion homeostasis Response metal ion inorganic Hormone stimulus response salicylic ROS Response salt stress

(FDR < 1E-04, Enrichment Map)

Fukushima et al., Supplemental Figure S8

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SLIDE 9

pad2 MV vs. Control A P-lim vs. Control WT MV P-lim

FDR = 8.0E-7

Control B

FDR = 1.5E-8 FDR = 1.3E-3 FDR = 1.0E-2 FDR = 2.6E-3 FDR = 1.5E-2 FDR = 7.8E-2 FDR = 8.2E-2 FDR = 3.0E-2

Hormone metabolism

Fukushima et al., Supplemental Figure S9

Treatment comparison Genotype comparison (pad2 vs. WT)

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SLIDE 10

pad2 MV vs. Control A P-lim vs. Control WT MV P-lim Genotype comparison (pad2 vs. WT) Control B pad2 WT Treatment comparison

Glucosinolate metabolism (1.7E-10) (3.6E-12) (3.2E-3) (1.9E-5) (1.2E-3) (5.7E-3) (1.1E-2) (3.0E-3) (1.5E-6) Isoprenoids (5.0E-3) (5.0E-3)

Secondary metabolism

Fukushima et al., Supplemental Figure S10

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SLIDE 11

pad2 MV vs. Control A P-lim vs. Control WT MV P-lim Genotype comparison (pad2 vs. WT) Control B Treatment comparison

LRR (7.6E-2) HRGP (9.1E-2) CS (3.1E-2) Pectin*esterases (2.2E-4) Modification (8.1E-4) CS (3.1E-2) Precursor synth. (2.2E-3) 10.6.2 (2.6E-2) AGPs (3.7E-2)

Cell wall metabolism

Fukushima et al., Supplemental Figure S11

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SLIDE 12

A Treatment comparison

Down-regulated genes under MV Down-regulated genes under P-lim

Included: WRKY40 WRKY70 MYB77 MYBL2 RD29A SAUR78 Included: WRKY33 UGT74E2 GSTU9 GSTU24 UGT73C1 UGT73C6 CYP81D8 HSFA2 HSP70 HSP101 HSP23.6-MITO HSP17.6 Hsp90-1 Included: ERF-1 ERF6 ERF104 ERF105 bZIP1 BZIP9 LBD37 LBD39 Included: ERF6 ERF104 PYE WRKY70 BHLH039 BHLH101 BHLH136 Included: GSTU9 UGT74E2 CYP81D8 HSFA2 DJC77 HSP70 HSP101 HSP23.6-MITO HSP17.4 HSP17.6II HSP17.6 Hsp90-1 Included: ERF105 DJC23 MYBL2 ATCTH

Down-regulation in pad2-1

Included: FAMT Included: GSTF3 CSLA10 CNGC11 PYD4 ARK3 CRK5 RLK1 Included: JAZ7 TAT Included: HMT3

B Genotype comparison

Fukushima et al., Supplemental Figure S12

(log2fold-change <= 1 and FDR < 0.05, LIMMA)

under no-stress MV-treatment P-lim pad2-1 WT pad2-1 WT