Supplemental Data. Candat et al. Plant Cell (2014) /tpc Cytosol. Nucleus. Mitochondria. Plastid. Peroxisome. Endomembrane system
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Transcription
1 Cytosol Nucleus PSORT MultiLoc YLoc SubLoc BaCelLo WoLF PSORT PSORT MultiLoc YLoc SubLoc BaCelLo WoLF PSORT Mitochondria Plastid Peroxisome Endomembrane system PSORT MultiLoc YLoc SubLoc TargetP Predotar PProwler MITOPRED BaCelLo WoLF PSORT PSORT MultiLoc YLoc TargetP Predotar PProwler BaCelLo WoLF PSORT PSORT MultiLoc YLoc PProwler BaCelLo WoLF PSORT PSORT MultiLoc YLoc SubLoc TargetP Predotar PProwler BaCelLo WoLF PSORT Probability in % < Supplemental Figure 1. Heatmap of subcellular localization predictions of the 51 Arabidopsis LEA proteins. The heatmap illustrates probability over 50 % of LEA protein targeting to various cellular compartments given by the different software indicated on the right. LEA proteins are numbered from 1 to 51, and the color coding represents the targeting probability below 50 % (yellow) or between 50 to 100% (red). The two qualitative predictors BaCelLo and WoLF PSORT were included with black square indicating a positive prediction.
2 LEA-GFP NLS-RFP Merge GFP-LEA Supplemental Figure 2. Transient expression of cytosolic LEA protein in Arabidopsis protoplasts. Representative examples of LEA-GFP and GFP-LEA fusion proteins localized either in the cytosol only or both in the cytosol and the nucleus were co-expressed with a RFP nuclear marker in Arabidopsis mesophyll protoplasts. Numbers refer to the corresponding LEA proteins. Green, GFP; Red, RFP; Blue, chlorophyll. Bar = 10 µm.
3 LEA-GFP NLS-RFP Merge GFP-LEA Supplemental Figure 2. (continued).
4 LEA-GFP NLS-RFP Merge GFP-LEA Supplemental Figure 2. (continued).
5 LEA-GFP NLS-RFP Merge GFP-LEA Supplemental Figure 2. (continued).
6 LEA-GFP NLS-RFP Merge GFP-LEA Supplemental Figure 2. (continued).
7 LEA-GFP NLS-RFP Merge GFP-LEA 51 Supplemental Figure 2. (continued).
8 Supplemental Figure 3. Diffusion of cytosolic LEA-GFP proteins in the nucleus. The graph shows the distribution of LEA protein fusions between cytosolic only (C) or cytosolic-nuclear (C+N) compartments as a function of their molecular mass. Each box encloses 50% of the data with the median value of the variable displayed as a line. The mean is indicated as a dashed line. The top and bottom of the box mark the limits of ± 25% of the variable population. The lines extending from the top and bottom of each box mark the minimum and maximum values within the data set that fall within an acceptable range.
9 GFP Supplemental Figure 4. Western blot analysis of LEA protein expressed in protoplasts. Total proteins extracted from protoplasts expressing LEA-GFP fusions were separated by SDS-PAGE and transferred on a nylon membrane. LEA-GFP were immuno-detected with a specific anti-gfp antibody. Each LEA protein is indicated by its number. The molecular mass in the scale is in kda. Lanes between black bars (2, 4, 5, 21, 26, 30, 36, 38, 41, 47 and 49) were obtained from a second western blot analysis and were merged in the figure for more clarity. Arrows indicate the bands of interest.
10 Supplemental Figure 5. Cytosolic localization of GFP-LEA fusions for proteins localized in mitochondria or plastid when expressed as LEA-GFP fusions (see Figures 3 and 4). Representative images of GFP-LEA fusion proteins expressed in wild type Arabidopsis mesophyll protoplasts. Numbers refer to different LEA proteins for which LEA-GFP fusions were localized in plastid or mitochondria. Green, GFP; Blue, chlorophyll. Bar = 10 µm.
11 LEA-GFP mt-rfp Merge Supplemental Figure 6. Leaf cells from Arabidopsis transgenic lines expressing LEA proteins targeted to mitochondria or dual targeted to mitochondria and plastid. Representative images of LEA-GFP fusion proteins expressed in transgenic lines expressing a RFP mitochondrial marker. In the merged images fro LEA42 and 48, chlorophyll autofluorescence is also shown (in blue). Numbers refer to different LEA proteins. Green, GFP; Red, RFP; Blue, chlorophyll. Bar = 10 µm.
12 LEA-RFP Golgi-GFP Merge 13 Supplemental Figure 7. LEA13-RFP does not co-localize with Golgi in Arabidopsis protoplasts. LEA13-RFP fusion protein was expressed in Arabidopsis mesophyll protoplasts carrying a Golgi marker (GFP). Bar = 10 µm.
13 LEA-RFP ER-YFP Merge LEA-RFP Perox-YFP Merge 9 Supplemental Figure 8. Arabidopsis leaf cells from transgenic lines expressing LEA proteins targeted to the secretory pathway or the pexophagosome. Representative images of LEA-RFP fusion proteins expressed in transgenic lines expressing a YFP ER marker (ER-YFP) or a YFP peroxisome marker (Perox-YFP). Numbers refer to different LEA proteins. Green, YFP; Red, RFP. Bar = 10 µm.
14 Supplemental Figure 9. Transient expression in Arabidopsis protoplasts of GFP-LEA fusions for proteins which are targeted to the secretory pathway as LEA-GFP fusions. Representative images of GFP-LEA fusion proteins expressed in wild type Arabidopsis mesophyll protoplasts. Numbers refer to different LEA proteins for which LEA-GFP fusions were targeted to the secretory pathway. Green, GFP; Blue, chlorophyll. Bar = 10 µm.
15 LEA-RFP Golgi-GFP Merge 21 Supplemental Figure 10. Vacuolar and Golgi localization of LEA 21 in Arabidopsis protoplasts. LEA21-RFP fusion protein was expressed in Arabidopsis mesophyll protoplasts carrying a Golgi marker (GFP). Bar = 10 µm.
16 LEA21-GFP LEA22-GFP Supplemental Figure 11. Localization of LEA 21-GFP and LEA22-GFP in Arabidopsis protoplasts. GFP fusion protein was expressed in wild type Arabidopsis mesophyll protoplasts. In the case of LEA22-GFP, the fluorescence signal was very weak. Bar = 10 µm.
17 Supplemental Figure 12. LEA11 and LEA12 sequence alignment. Sequences were aligned using the MultAlin software ( using default parameters. The targeting peptide cleavage site of LEA11 is indicated with a black arrow.
18 ER (6) PProwler False positive 0 6 MultiLoc 0 5 Yloc 0 5 TargetP 0 5 Predotar 0 5 PSORT 1 3 SubLoc 2 2 Mitochondria (5) PProwler False positive 2 4 Yloc 2 4 TargetP 3 PSORT 1 2 MITOPRED 1 2 Predotar 1 1 MultiLoc 0 SubLoc 6 Plastid (6) PProwler False positive 0 5 TargetP 2 5 Predotar 0 3 Yloc 1 3 PSORT 2 2 MultiLoc 2 Correct prediction Correct prediction Correct prediction Cytosol (36) False positive Correct prediction Yloc 1 PSORT 2 14 SubLoc 3 7 MultiLoc Supplemental Figure 13. Estimation of prediction software efficiency. The bar graph shows the number of proteins with correct or false predictions by the different software. For each compartment, the total number of experimentally identified proteins is indicated between brackets. Correct predictions were achieved when the experimental subcellular localization matched with the prediction. False positives correspond to erroneous subcellular compartment attribution with a strong prediction.
19 LEA-RFP ER-GFP Merge 7 29 Supplemental Figure 14. Cytosolic localization of LEA7 and LEA29 in Arabidopsis protoplasts. LEA-RFP fusion proteins were expressed in Arabidopsis mesophyll protoplasts carrying a ER marker (GFP). Bar = 10 µm.
20 LEA# Loc ID Cell culture Flowers Leaves Pollen Roots Seedling shoots Seeds Siliques 1 C At1g C At1g C At1g C At1g C At1g C At1g C At1g C At1g Pxp At1g C At1g P At2g P At2g ER At2g C At2g C At2g C At2g C At2g C At2g C At2g C At2g V At2g V At2g P At2g P At2g C At2g C At2g C At2g C At3g C At3g ER At3g C At3g C At3g C At3g C At3g C At3g C At3g M At3g M At4g C At4g C At4g M At4g MP At4g ER At4g C At4g C At4g C At5g C At5g MP At5g C At5g C At5g C At5g Supplemental Figure 15. Proteomic quantification of LEA proteins in different organs of Arabidopsis. Proteomic data were obtained from the pep2pro database ( and the subcellular localizations were added from our experimental observations: C, cytosol ; P, plastid ; M, mitochondria ; MP, mitochondria and plastid ; ER, Endoplasmic Reticulum ; V, vacuole ; Pxp, pexophagosome. Values are expressed as Log2 of spectral counts and normalized (single hits were previously removed). Results were color coded as a function of values from yellow to red with Excel.
21 a b Supplemental Figure 16. LEA2 and LEA38 sequence alignment. Sequences were aligned using the MultAlin software ( using default parameters. The targeting peptide cleavage sites of LEA38 shown as black arrows were predicted using the MitoProt (a) and TargetP (b) software.
22 PsLEAM ( ) Mitochondria LEA36 ( ) Cytosol LEA9 ( ) Pexophagosome LEA11( ) Plastid LEA30 ( ) ER LEA42 (93-128) Mitochondria-Plastid Supplemental Figure 17. Modeling of class A α-helix motif of LEA proteins. Helical projections of α-helices were obtained using the HeliQuest webserver ( ). Each wheel was obtained with a 36 amino acids window. Residues are color coded with blue for positively (K, R) and red (D, E) for positively charged residues. Non polar residues are shown in yellow or grey, and others in purple, light blue or light pink colors. The arrow shows the hydrophobic moment.
23 LEA23 (61-96) Plastid LEA23 ( ) Plastid LEA24 (61-96) Plastid LEA24 (87-122) Plastid Supplemental Figure 18. Modeling of class A-like α-helix motif of LEA23 and LEA24. Helical projections of α-helices were obtained using the HeliQuest webserver ( ). Each wheel was obtained with a 36 amino acids window. Residues are color coded with blue for positively (K, R) and red (D, E) for positively charged residues. Non polar residues are shown in yellow or grey, and others in purple, light blue or light pink colors. The arrow shows the hydrophobic moment.
24 Supplemental Table 1. Theoretical and apparent molecular masses of LEA and fusion proteins. LEA protein Molecular weight (kda) Molecular weight with GFP (kda) Molecular weight with GFP deduced from Western blot (kda)
25 Supplemental Table 2. Quantitative data about transiently transformed protoplasts. Exp, independent transformation; FP, fluorescent protoplast number protoplasts Exp 1 Exp 2 Exp 3 Exp 4 Exp 5 LEA number of Protopl. Protopl. Protopl. Protopl. Protopl. Total independant FP FP FP FP FP observed observed observed observed observed FP transformation
26 Supplemental Table 3. Arabidopsis LEA templates used for PCR amplification. LEA# Accession number (TAIR) Template Origin 1 At1g01470 cdna Arabidopsis seeds MPIMP 2 At1g02820 pentr/sd/d-topo (+CDS) MPIMP 3 At1g03120 cdna Arabidopsis seeds MPIMP 4 At1g20440 pentr/sd/d-topo (+CDS) MPIMP 5 At1g20450 puni51 (U24382) ABRC 6 At1g32560 pentr/sd/d-topo (+CDS) MPIMP 7 At1g52690 pentr/sd/d-topo (+CDS) MPIMP 8 At1g54410 pentr/sd/d-topo (+CDS) MPIMP 9 At1g72100 pentr/sd/d-topo (+CDS) MPIMP 10 At1g76180 pentr/sd/d-topo (+CDS) MPIMP 11 At2g03740 pentr/sd/d-topo (+CDS) MPIMP 12 At2g03850 pentr/sd/d-topo (+CDS) MPIMP 13 At2g18340 cdna Arabidopsis seeds MPIMP 14 At2g21490 pentr/sd/d-topo (+CDS) MPIMP 15 At2g23110 pentr/sd/d-topo (+CDS) MPIMP 16 At2g23120 pentr/sd/d-topo (+CDS) MPIMP 17 At2g33690 puni51 (U63047) ABRC 18 At2g35300 cdna Arabidopsis seeds MPIMP 19 At2g36640 cdna Arabidopsis seeds MPIMP 20 At2g40170 pentr/sd/d-topo (+CDS) MPIMP 21 At2g41260 puc57 (synthetic gene) GenScript 22 At2g41280 pentr/sd/d-topo (+CDS) MPIMP 23 At2g42530 pentr/sd/d-topo (+CDS) MPIMP 24 At2g42540 pentr/sd/d-topo (+CDS) MPIMP 25 At2g42560 pentr/sd/d-topo (+CDS) MPIMP 26 At2g44060 puni51 (U83572) ABRC 27 At2g46140 pentr/sd/d-topo (+CDS) MPIMP 28 At3g02480 pentr/sd/d-topo (+CDS) MPIMP 29 At3g15670 pentr/sd/d-topo (+CDS) MPIMP 30 At3g17520 pentr/sd/d-topo (+CDS) MPIMP 31 At3g22490 pentr/sd/d-topo (+CDS) MPIMP 32 At3g22500 pentr/sd/d-topo (+CDS) MPIMP 33 At3g50970 pentr/sd/d-topo (+CDS) MPIMP 34 At3g50980 cdna Arabidopsis seeds MPIMP 35 At3g51810 pentr/sd/d-topo (+CDS) MPIMP 36 At3g53040 pentr/sd/d-topo (+CDS) MPIMP 37 At3g53770 pentr221 (pentr221-at3g53770) ABRC 38 At4g02380 pentr/sd/d-topo (+CDS) MPIMP 39 At4g13230 cdna Arabidopsis bud MPIMP 40 At4g13560 pentr/sd/d-topo (+CDS) MPIMP 41 At4g15910 puni51 (U23419) ABRC 42 At4g21020 puni51 (U20045) ABRC 43 At4g36600 puni51 (S63786) ABRC 44 At4g38410 pentr/sd/d-topo (+CDS) MPIMP 45 At4g39130 cdna Arabidopsis seeds MPIMP 46 At5g06760 pentr/sd/d-topo (+CDS) MPIMP 47 At5g27980 puc57 (synthetic gene) GenScript 48 At5g44310 puni51 (U66244) ABRC 49 At5g53260 puc57 (synthetic gene) GenScript 50 At5g53270 cdna Arabidopsis seeds MPIMP 51 At5g66400 pentr/sd/d-topo (+CDS) MPIMP
27 Supplemental Table 4. Primers used for the construction of LEA gene fusions and markers. Sequence Name Forward primer Length Name Reverse primer Length LEA1 LEA2 LEA3 LEA4 LEA5 LEA6 LEA7 LEA8 LEA9 LEA10 LEA11 LEA12 LEA13 LEA14 LEA15 LEA16 LEA17 LEA18 LEA19 LEA20 LEA21 LEA22 LEA23 LEA24 LEA25 LEA26 LEA27 LEA28 LEA29 LEA30 LEA31 LEA32 LEA33 LEA34 LEA35 LEA36 FL1 CACCATGGCGAGCTTGCTAGAT 22 RL1 GAAGAAATCTTTAAAAGTAGG 21 FnoL1 CACCGCGAGCTTGCTAGATAAA 22 RsL1 TCAGAAGAAATCTTTAAAAGT 21 FL2 CACCATGGCTCGTTCTCTCGCT 22 RL2 TTGCTTGTTGTTCAAGAG 18 FnoL2 CACCGCTCGTTCTCTCGCTAAC 22 RsL2 TTATTGCTTGTTGTTCAA 18 FL3 CACCATGGCACAGCATCAGCATTC 24 RL3 GAGTTGTTGATTGAGCCT 18 FnoL3 CACCGCACAGCATCAGCATTCTCC 24 RsL3 CTAGAGTTGTTGATTGAGC 19 FL4 CACCATGGCTGAGGAGTACAAG 22 RL4 ATCATCAGACTCTTTTTCTTTC 22 FnoL4 CACCGCTGAGGAGTACAAGAAC 22 RsL4 TTAATCATCAGACTCTTTTTCT 22 FL5 CACCATGGCAGAAGAGTACAAGAAC 25 RL5 ATCAGACACTTTTTCTTTCTTC 22 FnoL5 CACCGCAGAAGAGTACAAGAACACC 25 RsL5 TTAATCAGACACTTTTTCTTTC 22 FL6 CACCATGCAATCGGCGAAACAG 22 RL6 GTAGTGATGATGATTATGATGTCC 24 FnoL6 CACCCAATCGGCGAAACAGAAG 22 RsL6 TTAGTAGTGATGATGATTATGATG 24 FL7 CACCATGGCGTCTCATCAAGAA 22 RL7 CTTCCTCTGTGTCTCACG 18 FnoL7 CACCGCGTCTCATCAAGAACAG 22 RsL7 TCACTTCCTCTGTGTCTC 18 FL8 CACCATGGCAGGACTCATCAAC 22 RL8 ATCGCTGTCGCTGTCACT 18 FnoL8 CACCGCAGGACTCATCAACAAG 22 RsL8 TTAATCGCTGTCGCTGTC 18 FL9 CACCATGACGAATCTTTTGGCC 22 RL9 ACAAGCGGCGCCGAGTCTCTG 21 FnoL9 CACCACGAATCTTTTGGCCTTG 22 RsL9 TCAACAAGCGGCGCCGAGTCT 21 FL10 CACCATGGCTGAGGAAATCAAG 22 RL10 TTCTTTATCTTTCTTCTCCTCC 22 FnoL10 CACCGCTGAGGAAATCAAGAAT 22 RsL10 TTATTCTTTATCTTTCTTCTCC 22 FL11 CACCATGTCGATCTCCGGAGCT 22 RL11 TGCATCAGTTTTTGGAGGC 19 FnoL11 CACCTCGATCTCCGGAGCTGTG 22 RsL11 CTATGCATCAGTTTTTGGA 19 FL12 CACCATGGCGATGTCGATCTCC 22 RL12 AGTTTTTGGAGGCATTATAGC 21 FnoL12 CACCGCGATGTCGATCTCCGGA 22 RsL12 TTAAGTTTTTGGAGGCATTAT 21 FL13 CACCATGATGGAGAGAAGAAGAAC 24 RL13 GAGCTCAGCATAACGGTC 18 FnoL13 CACCATGGAGAGAAGAAGAACGG 23 RsL13 TTAGAGCTCAGCATAACG 18 FL14 CACCATGGCGGATTTGAGGGAC 22 RL14 TGGGTGGTTGTGGTTATG 18 FnoL14 CACCGCGGATTTGAGGGACGAA 22 RsL14 TCATGGGTGGTTGTGGTT 18 FL15 CACCATGGAGGATCAGAAAAAGCC 24 RL15 CGGAACGCCCTGACGGTT 18 FnoL15 CACCGAGGATCAGAAAAAGCCACC 24 RsL15 TCACGGAACGCCCTGACG 18 FL16 CACCATGGAGGCCGGGAAAACA 22 RL16 CGGAGCTTTCGCATCGGT 18 FnoL16 CACCGAGGCCGGGAAAACACCA 22 RsL16 TCACGGAGCTTTCGCATC 18 FL17 CACCATGTCGAAGAGTGAAGAGAA 24 RL17 CTTCTTAGCTTTTTGGTTAGCG 22 FnoL17 CACCTCGAAGAGTGAAGAGAAACA 24 RsL17 TCACTTCTTAGCTTTTTGGTTA 22 FL18 CACCATGCAGTCGGCGAAGGAA 22 RL18 GATCTGTCCCGGCGGGTA 18 FnoL18 CACCCAGTCGGCGAAGGAAAAG 22 RsL18 TTAGATCTGTCCCGGCGG 18 FL19 CACCATGGCGTCAGACAAACAAAA 24 RL19 CAGCTTTCCCTTATCTTTCC 20 FnoL19 CACCGCGTCAGACAAACAAAAGGC 24 RsL19 TCACAGCTTTCCCTTATCTT 20 FL20 CACCATGGCGTCTCAACAAGAG 22 RL20 GGTCTTGGTCCTGAATTTGG 20 FnoL20 CACCGCGTCTCAACAAGAGAAG 22 RsL20 TTAGGTCTTGGTCCTGAATT 20 FL21 CACCATGGGAAACCTCAAGTCTC 23 RL21 TGGCTTAGCTTGTTGGGC 18 FnoL21 CACCGGAAACCTCAAGTCTCTCG 23 RsL21 TCATGGCTTAGCTTGTTGG 19 FL22 CACCATGGGAAACCTCATGTCT 22 RL22 TGGCTTAGCTTCTTCCTT 18 FnoL22 CACCGGAAACCTCATGTCTCTC 22 RsL22 TCATGGCTTAGCTTCTTCC 19 FL23 CACCATGGCGATGTCTTTATCAGG 24 RL23 GGACTTTGTGGCATTCTTAG 20 FnoL23 CACCGCGATGTCTTTATCAGGAGC 24 RsL23 TCAGGACTTTGTGGCATTCT 20 FL24 CACCATGGCGATGTCTTTCTCAGG 24 RL24 CTTTGTGGCATCCTTAGC 18 FnoL24 CACCGCGATGTCTTTCTCAGGAGC 24 RsL24 CTACTTTGTGGCATCCTT 18 FL25 CACCATGGCGTCAGAGCAAGCA 22 RL25 ACGTTGTCCATGTTCCCG 18 FnoL25 CACCGCGTCAGAGCAAGCAAGG 22 RsL25 TCAACGTTGTCCATGTTC 18 FL26 CACCATGTCGACATCTGAGGATAA 24 RL26 TTCCTCATCGTCGTCATC 18 FnoL26 CACCTCGACATCTGAGGATAAACC 24 RsL26 TTATTCCTCATCGTCGTC 18 FL27 CACCATGGCATCAGCGGATGAA 22 RL27 AAAGAAGTCGCGAAGGGA 18 FnoL27 CACCGCATCAGCGGATGAAAAG 22 RsL27 TTAAAAGAAGTCGCGAAG 18 FL28 CACCATGGACAACAAGCAAAACGC 24 RL28 GTGGCTTTTGTTCATGCC 18 FnoL28 CACCGACAACAAGCAAAACGCGAG 24 RsL28 TTAGTGGCTTTTGTTCAT 18 FL29 CACCATGGCATCCAACCAACAG 22 RL29 CTTCCTCTGATAAGTCTGATGA 22 FnoL29 CACCGCATCCAACCAACAGAGC 22 RsL29 TCACTTCCTCTGATAAGTCTGA 22 FL30 CACCATGGGGTTAGAGAGGAAA 22 RL30 GAGCTCAGCATCATCGTC 18 FnoL30 CACCGGGTTAGAGAGGAAAGTG 22 RsL30 TCAGAGCTCAGCATCATC 18 FL31 CACCATGAGTCAAGAAGAACAACC 24 RL31 TATATCAGCTCTCTCGTTAAGC 22 FnoL31 CACCAGTCAAGAAGAACAACCAAA 24 RsL31 TCATATATCAGCTCTCTCGTTA 22 FL32 CACCATGAGCCAAGAGCAACCA 22 RL32 TATATCAACTCTCTCGTTAAGC 22 FnoL32 CACCAGCCAAGAGCAACCAAGG 22 RsL32 TCATATATCAACTCTCTCGTTA 22 FL33 CACCATGAATTCTCACCAGAATCA 24 RL33 GTGATGACCACCGGGAAG 18 FnoL33 CACCAATTCTCACCAGAATCAAAC 24 RsL33 CTAGTGATGACCACCGGG 18 FL34 CACCATGGAGTCTTACCAAAACC 23 RL34 ATGATGACCACCCGGAAG 18 FnoL34 CACCGAGTCTTACCAAAACCAG 22 RsL34 CTAATGATGACCACCCGG 18 FL35 CACCATGGCGTCAAAGCAACTG 22 RL35 CTTGTTGGTGAACTTTGACTC 21 FnoL35 CACCGCGTCAAAGCAACTGAGC 22 RsL35 TCACTTGTTGGTGAACTTTGA 21 FL36 CACCATGGCATCAGGACAACGG 22 RL36 CAGCTTTTTCTCTCCCAC 18 FnoL36 CACCGCATCAGGACAACGGGAG 22 RsL36 CTACAGCTTTTTCTCTCC 18
28 LEA37 FL37 CACCATGTCTCAATCACTTTTCAA 24 RL37 GCTCACTACGTACGTCTTTTG 21 FnoL37 CACCTCTCAATCACTTTTCAATCT 24 RsL37 TTAGCTCACTACGTACGTCTT 21 LEA38 FL38 CACCATGGCTCGTTCTATCTCTAA 24 RL38 CTGCTTGTTGTTCAAGAGAGC 21 FnoL38 CACCGCTCGTTCTATCTCTAACGT 24 RsL38 TCACTGCTTGTTGTTCAAGAG 21 LEA39 FL39 CACCATGACAAGCTTCGCGGTC 22 RL39 TTTGAGGTTCTTGGTGTTC 19 FnoL39 CACCACAAGCTTCGCGGTCGTTG 23 RsL39 TTATTTGAGGTTCTTGGTG 19 LEA40 FL40 CACCATGTCGCAACAACAATTC 22 RL40 TTTCTTCTCGTTCATGCC 18 FnoL40 CACCTCGCAACAACAATTCAAC 22 RsL40 TCATTTCTTCTCGTTCAT 18 LEA41 FL41 CACCATGGCCGCTCGTTCACTC 22 RL41 GAAAGACTTTGCTTTGTTTTTC 22 FnoL41 CACCGCCGCTCGTTCACTCTCC 22 RsL41 TCAGAAAGACTTTGCTTTGTTT 22 LEA42 FL42 CACCATGGCGGCCATGCAACTAAC 24 RL42 GTTAAATGTTATGAAATCGTCC 22 FnoL42 CACCGCGGCCATGCAACTAACAAG 24 RsL42 TCAGTTAAATGTTATGAAATCGTC 24 LEA43 FL43 CACCATGATGCTTACGACGGTG 22 RL43 AAGCTCAGCGCTACGGTC 18 FnoL43 CACCATGCTTACGACGGTGGTG 22 RsL43 CTAAAGCTCAGCGCTACG 18 LEA44 FL44 CACCATGGCGGATCATCCTCGT 22 RL44 GGTTTCCTTCTTCTTCTCATC 21 FnoL44 CACCGCGGATCATCCTCGTTCT 22 RsL44 CTAGGTTTCCTTCTTCTTCTC 21 LEA45 FL45 CACCATGGCGGATCTGAAAGAC 22 RL45 AAGATCATTATGGTGGCC 18 FnoL45 CACCGCGGATCTGAAAGACGAA 22 RsL45 TCAAAGATCATTATGGTG 18 LEA46 FL46 CACCATGCAGTCGATGAAAGAA 22 RL46 TCCAGTATATCCCCCGCC 18 FnoL46 CACCCAGTCGATGAAAGAAACA 22 RsL46 TTATCCAGTATATCCCCC 18 LEA47 FL47 CACCATGAGCGAAGAACAGCTG 22 RL47 TTTGGACTGATTGATCCG 18 FnoL47 CACCAGCGAAGAACAGCTGCAG 22 RsL47 TCATTTGGACTGATTGAT 18 LEA48 FL48 CACCATGGCGGCTATGCAGTTAACG 25 RL48 GAACCTCTTGAAATCATCATCC 22 FnoL48 CACCGCGGCTATGCAGTTAACGAG 24 RsL48 TTAGAACCTCTTGAAATCATCATC 24 LEA49 FL49 CACCATGGGTTCATCAAAAGATAG 24 RL49 AAGAGACGATGGATCGTG 18 FnoL49 CACCGGTTCATCAAAAGATAGTGC 24 RsL49 TCAAAGAGACGATGGATCG 19 LEA50 FL50 CACCATGATGTTCGGGTTCGGC 22 RL50 AAGAGATACATTGCATGG 18 FnoL50 CACCATGTTCGGGTTCGGCCTT 22 RsL50 TCAAAGAGATACATTGCA 18 LEA51 FL51 CACCATGGCGTCTTACCAGAAC 22 RL51 ACGGCCACCACCGGGAAG 18 FnoL51 CACCGCGTCTTACCAGAACCGT 22 RsL51 TTAACGGCCACCACCGGG 18 U16034 (At3g10920) MSD1-F CACCATGGCGATTCGTTGTGTAGC 24 MSD1-R GTTGTTTTCCTTCTCATAAACCTC 24 U12392 (At4g05180) PsbQ-F CACCATGGCTCAAGCAGTGACTTCG 25 PsbQ-R ACCGAGCTTGGCAAGAACATTGTTC 25 U13389 (At5g38430) RBCS1B-F CACCATGGCTTCCTCTATGCTCT 23 RBCS1B-R AGCATCAGTGAAGCTTGG 18 p2fgw7 FpGX GCGAAACCCTATAAGAACC 19 RpGX ACCACTACCAGCAGAACA 18 p2gwf7 FpXG GTGGTGCAGATGAACTTCAG 20 RpXG GCACAATCCCACTATCCTTC 20 RevEGFP TTACTTGTACAGCTCGTCCAT 21 p2gwr7 RFP-F CACCATGGAGGGCTCCGTGAACGG 24 RevRFP TTAGGCGCCGGTGGAGTGGC 20 NLS-F CACCATGCCACCAAAAAAGAAAAGAAAGGTT 31 NLS-R AACCTTTCTTTTCTTTTTTGGTGGCAT 27 PDHA1 (NM_000284) PDHE1-F ATGAGGAAGATGCTCGCC 18 PDHE1t-R CCTGGTGAGCACTGTTGTGA 20
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