Supplemental data. Pommerrenig et al. (2011). Plant Cell /tpc
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1 Supplemental Figure 1. Prediction of phloem-specific MTK1 expression in Arabidopsis shoots and roots. The images and the corresponding numbers showing absolute (A) or relative expression levels (B) of MTK1 were calculated by the Translatome efp Browser ( for At1g49820.
2 Supplemental Figure 2. Predictions of the phloem-specific expression of MTN and ARD genes in Arabidopsis shoots. The images and the corresponding numbers showing the relative expression of (A) the MTN1 and MTN2 genes and (B) of the ARD1 to ARD4 genes in different tissues of Arabidopsis shoots were taken from the Translatome efp Browser ( and assembled. 2
3 Supplemental Figure 3. Predictions of the expression pattern of MTI1 and DEP1 in Arabidopsis shoots. The images and the corresponding numbers showing the relative expressions of MTI1 and DEP1 in different tissues of Arabidopsis shoots were taken from the Translatome efp Browser ( 3
4 Supplemental Figure 4. Polyamine biosynthesis and polyamine biosynthetic enzymes in Arabidopsis. Intermediates of polyamine biosynthesis, abbreviations of gene names, MIPSnumbers and images showing the relative expressions in different Arabidopsis leaf tissues (Translatome efp Browser; are presented. 4
5 Supplemental Figure 5. Predictions of the expression patterns of transaminases potentially involved in the formation of Met from KMTB in the Arabidopsis vasculature. The images and the corresponding numbers showing the relative expressions of AAT, ASP2, ASP3 and ASP5 in different tissues of Arabidopsis shoots were taken from the Translatome efp Browser ( 5
6 Metabolites Elemental composition Phase RT (min) Identified ion m/z Mass accuracy (mda) 1 MS/MS Fragments (m/z) Coelution Disaccharides C 12H 22O 11 unpolar 0.43 [M+HCO 2] , 89, 101, 113, 134, 179 Monosaccharides C 6H 12O 6 unpolar 0.43 [M+HCO 2] , 89, 101, 113, 119, 131, 143, 179 Arginine C 6H 14N 4O 2 polar 0.38 [M+H] , 116, 142, 175 Methionine C 5H 11NO 2S polar 0.52 [M+H] SAM C 15H 22N 6O 5S polar 0.46 [M+H] Collision energy Ratio of S32/S34 Isotopomer (%) MTA C 11H 15N 5O 3S polar 0.88 [M+H] (6.1) 2 1 Mass difference between calculated and experimental mass (as neutral mass) 2 Theoretical value for C 11H 15N 5O 3S calculated by the software Isotopident ( Supplemental Table 1. Data confirming identification of differentially accumulating metabolites. 6
7 AtARD1p-f AtARD1-r AtARD2p-f AtARD2-r AtARD3p-f AtARD3p-r AtARD4p-f AtARD4-r AtDEP1-1649f AtDEP1+3660r AtDEP1c+1f AtDEP1c+1524r AtMTI1-1925f AtMTI1+2078r AtMTI1c+1f AtMTI1c+1125r AtMTKp-f AtMTKp-r AtMTN1p-f AtMTN1p-r AtSAMDC2-1231f AtSAMDC2-1r PmACL5A-fwd PmACL5A-rev PmACL5B-fwd PmACL5B-rev PmACO1-fwd PmACO1-rev PmALDP1-fwd 5 -CAC CCA TTT TGC AGC CAA TCC AAT-3 (At ARD1 promoter-gene fragment) 5 -TTT AGC CGA GGC ATT GAT CAT GAA G-3 (At ARD1 promoter-gene fragment) 5 -CAC CGA GGA AAC TTG GCT TGC TTG-3 (ARD2 promoter-gene fragment) 5 -GGC TGA CGC GTC TAT GAC ACC A-3 (ARD2 promoter-gene fragment) 5 -CAC CAC AGG AAA CTT GGC CAA CAC-3 (At ARD3 promoter) 5 -GGT TAT TCT TCT TTT GTG TTC A-3 (At ARD3 promoter) 5 -CAC CGG CCG TTG AAA TCT TGG TTA-3 (At ARD4 promoter-gene fragment) 5 -ATG TGC TTT AAC GGT TTC TCC A-3 (At ARD4 promoter-gene fragment) 5 -CAC CAT CTG CGA CAC AAT CTG CTG-3 (At DEP1 promoter-gene fragment) 5 -CAC CAT CTG CGA CAC AAT CTG CTG-3 (At DEP1 promoter-gene fragment) 5 -TCT CTG CGG CCG CAA GCT TGT AAA AGA AAT GGC GGT GGC TGC AGC GG-3 (At DEP1) 5 -TCT CTG CGG CCG CCT AGA TTT GGG AGA ATG ATG TGA CAG TCT-3 (At DEP1) 5 -CAC CTG AAA GAG GCC TAG CGA AGA-3 (At MTI1 promoter-gene fragment) 5 -CCA AGA AGA TAA CAG GAA ACT CAT CTC GT-3 (At MTI1 promoter-gene fragment) 5 -TCT CTG AAT TCA AGC TTG TAA AAG AAA TGT CCG GCG AAG GAG ACA CGA C-3 (At MTI1) 5 -TCT CTG AAT TCT CAA CGA GAT GAG TTT CCT GTT ATC TTC TTG-3 (At MTI1) 5 -CAA ATC ATT TTT ATA CCT CGA TGC-3 (At MTK promoter) 5 -GGC TTT TGG TAC AAA TTT TCA GA-3 (At MTK promoter) 5 -CAC CAG CGA AAA CAC CTT TGA TGG-3 (At MTN1 promoter) 5 -AAC CTT ATC CCT CTC TTC CG-3 (At MTN1 promoter) 5 -CAC CCA CGG GAA GAT AAC AAG GTC A-3 (At SAMDC2 promoter) 5 -ATA ATC TCA TTC ACG CTT TTT C-3 (At SAMDC2 promoter) 5 -ATC TTG AAG CCC AAG CTG AA-3 (Pm ACL5A) 5 -CTT CGA TGC TGA GTG GTT GA-3 (Pm ACL5A) 5 -AAA TGG GAG TGG TCA TGG AG-3 (Pm ACL5B) 5 -TGG ATT TGG ATG ATT CAG CA-3 (Pm ACL5B) 5 -CAA AGG CCT TGA AGC TGT TC-3 (Pm ACO1) 5 -GGA GCT GAA GAC CAC TGA CC-3 (Pm ACO1) 5 -TCC AAG GGT TCA GAG T-3 (Pm ALDP1) 7
8 PmALDP1-rev 5 -TTC CTG CTG TTT TTT G-3 (Pm ALDP1) PmARD-fwd 5 -TGT GTT GCA GGG AGT GGT TA-3 (Pm ARD1) PmARD-rev 5 -GCA TCA ACA TCA CC-3 (Pm ARD1) PmCPA1-fwd 5 -TAG GGA TTT GCT GGG ATC AG-3 (Pm CPA1) PmCPA1-rev 5 -TTT TGC CAT GCT CAG TTT CA-3 (Pm CPA1) PmDEP1-fwd 5 -TCA CAG AAT CTC TTG CTG AAG CA-3 (Pm DEP1) PmDEP1-rev 5 -ATC AAA GCA TAT CCA AAA ACG ACC GCT-3 (Pm DEP1) PmDEP1/2c-f 5 -GCA GGG GCT GTT ATT CAT AGT CAT GG-3 (Pm DEP1/2) PmDEP1/2c-r 5 -TTA TGA TAC ATT CAG CCT G-3 (Pm DEP1/2) PmDEP1/2-fwd1 5 -CCW CTB TTT WTG AAG GCD TAT-3 (Pm DEP1/2 degenerate primer) PmDEP1/2-rev1 5 -TAA TGA TAA CAT TCA GCC TG-3 (Pm DEP1/2 degenerate primer) PmDEP2-fwd 5 -GTG GAG TCT TTA ACA GAA TCG-3 (Pm DEP2) PmDEP2-rev 5 -AGA TCC TAC CCA AAG GCA ACA-3 (Pm DEP2) PmMTI1c-f 5 -ACC CAC CGC AAC ACT GGC AGT-3 (Pm MTI1) PmMTI1c-r 5 - TCG AAG GCT GGG TTC CAC AC-3 (Pm MTI1) PmMTI1-fwd 5 -GCT GTG ATT GTT GGA GCT GA-3 (Pm MTI1) PmMTI1-fwd2 5 - ACM CAT TGT AAT ACT GGM AG-3 (Pm MTI1 degenerate primer) PmMTI1-rev 5 -GCT GGG TTC CAC ACA GAA AT-3 (Pm MTI1) PmMTI1-rev3 5 - TCG AAK GCK GGG TTC CAB AC-3 (Pm MTI1 degenerate primer) PmMTK-fwd 5 -CAA CAA CGC AGA ACT CCA GA-3 (Pm MTK) PmMTK-rev 5 -CCG GAA TTT GAT GCA AGA CT-3 (Pm MTK) PmSAMDC1-fwd 5 -TGT AGA GAT TGC CGC TTC CT-3 (Pm SAMDC1) PmSAMDC1-rev 5 -CTG CAT GTG CCA AGT CTG TT-3 (Pm SAMDC1) PmSAMDC2-fwd 5 -TCC TCA TCC TCG CTC TCA CT-3 (Pm SAMDC2) PmSAMDC2-rev 5 -TGG GTT ACC TGG TTG AGA GG-3 (Pm SAMDC2) PmSPMS1-fwd 5 -CAT CAG ATC CTG TGG GTC CT-3 (Pm SPMS1) PmSPMS1-rev 5- GAC CTT CCG TTG AGC ATA GC-3 (Pm SPMS1) PmTUA-5 5 -CAT CTG CCG TAA ATC TCT TGA TAT-3 (Pm TUA2) PmTUA-3 5 -AGC AAC AGA AAG CTG CTC ATG GTA-3 (Pm TUA2) TOPO-1 5 -TCG GAT CCA CTA GTA ACG-3 (TOPO-Zero blunt II vector) TOPO GTG TGA TGG ATA TCT GC-3 (TOPO-Zero blunt II vector) Supplemental Table 2. Oligonucleotide primers used in this study. Primer names and sequences are listed in alphabetical order; gene targets are given in brackets. 8
9 Supplemental Data Set 1. Alignment (ClustalW; PHYLIP format) of the sequences used to calculate the phylogenetic tree shown in Figure 5. Two-letter abbreviations of the organisms (e.g. Av = Anabaena variabilis) and partial accession or MIPS numbers (Arabidopsis) are given at the top of the figure (shown as separate txt file). Supplemental Data Set 2. Alignment (ClustalW; PHYLIP format) of the sequences used to calculate the phylogenetic tree shown in Figure 7B. Sequences of the the N-terminal part (residues 1 to 248) of the Arabidopsis DEP1 protein, of related plant proteins and of characterized methylthioribulose-1-phosphate dehydratases from non-plant organisms were aligned. Two-letter abbreviations of the organisms (e.g. Ur = Uncinocarpus resii) and accession or MIPS numbers (Arabidopsis) are given at the top of the figure (shown as separate txt file). Supplemental Data Set 3. Alignment (ClustalW; PHYLIP format) of the sequences used to calculate the phylogenetic tree shown in Figure 7C. Sequences the C-terminal part (residues 249 to 507) of the Arabidopsis DEP1 protein, of related plant proteins and of characterized enolase-phosphatases from non-plant organisms were aligned. Two-letter abbreviations of the organisms (e.g. Vv = Vitis vinifera) and accession or MIPS numbers (Arabidopsis) are given at the top of the figure (shown as separate txt file). 9
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