1. Volume 7, Issue 2, Pages 356-368 (February 2014)
Suppression of the External Mitochondrial NADPH Dehydrogenase, NDB1, in Arabidopsis thaliana Affects Central Metabolism and Vegetative Growth 
Sabá V. Wallström, Igor Florez-Sarasa, Wagner L. Araújo, Mari Aidemark, María Fernández-Fernández, Alisdair R. Fernie, Miquel Ribas- Carbó, Allan G. Rasmusson 
Molecular Plant 
Volume 7, Issue 2, Pages (February 2014)
DOI: /mp/sst115
Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

2. Figure 1 Alignment of NDB cDNA Sequences from A. thaliana.
Sequences of NDB homologs are aligned to the NDB1 sequence used for the inverse repeat construct. Restriction sites added in the ends of the NDB1 construct are not shown. Black vertical bars denote bases that are identical to the NDB1 sequence. Unconserved DNA is denoted as horizontal lines. The alignment was made with ClustalW2 ( and shaded using MacBoxshade The following sequences, with accession numbers, were used in the alignment: NDB1, NM_ (99–202bp); NDB2, BT (18–130bp); NDB3, NM_ (18–130bp); NDB4, NM_ (80–200bp).
Molecular Plant 2014 7, DOI: ( /mp/sst115)
Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

3. Figure 2 NDB1 Transcript Levels in Different RNAi Lines.
RNA was extracted from leaves of 26-day-old soil-grown plants of the T3 generation (A) or rosettes from 35-day-old soil-grown plants of the T4 generation (B). Transcript levels were analyzed by real-time RT–PCR and normalized against those for the 28.5-kDa subunit of respiratory complex I. Means ± SD are shown for two or three independent RNA preparations. Significant differences to the WT are denoted with asterisks.
Molecular Plant 2014 7, DOI: ( /mp/sst115)
Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

4. Figure 3
External NAD(P)H Dehydrogenase Activities of Isolated Mitochondria.
Activity measurements were carried out on mitochondria from 10-day-old T4 seedlings, using decylubiquinone as electron acceptor, in the absence and presence of 1mM CaCl2. The figure shows the Ca2+-dependent and -independent components of the external NADPH (A) and NADH (B) oxidation. Means ± SE are shown for three independent experiments. For clarity, only the upward and downward error bar is shown for Ca2+-dependent and -independent activities, respectively.
Molecular Plant 2014 7, DOI: ( /mp/sst115)
Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

5. Figure 4 Rosette Diameters of Sterile-Grown Transgenic Seedlings.
Means ± SE for 32–55 10-day-old T4 seedlings from two or three separate plates are shown. Significant differences to the WT are denoted with asterisks.
Molecular Plant 2014 7, DOI: ( /mp/sst115)
Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

6. Figure 5
Evolution of 14CO2 from Seedlings of NDB1-Suppressing Lines in Light.
T4 seedlings were grown on glucose containing medium for 10 d. They were then incubated in 10mM MES–KOH solution, pH 6.5, containing 0.3mM glucose supplemented with 2.32 kBq ml–1 of [1-14C]-, [2-14C]-, [3,4-14C]-, or [6-14C]-glucose at an irradiance of 200 μmol m–2 s–1. The 14CO2 liberated was captured (at hourly intervals) in a KOH trap and the amount of radiolabel released was subsequently quantified by liquid scintillation counting. Values are presented as means ± SE from determinations on six individual liquid scintillation vials containing at least 10 seedlings per line.
Molecular Plant 2014 7, DOI: ( /mp/sst115)
Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

7. Figure 6 Dry Weight of NDB1-Suppressing Lines.
Plants of the T4 generation were grown in a 12-h light cycle. Rosettes were harvested 30 d after sowing, and the tissue was dried at 80°C for 5 d. Means ± SE for 10 individual samples, each containing two rosettes are shown. Significant differences to the WT are denoted with asterisks.
Molecular Plant 2014 7, DOI: ( /mp/sst115)
Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

8. Figure 7 Tissue NAD(P)(H) Levels of NDB1-Suppressing Lines.
Rosettes from 35-day-old T4 plants grown on soil without (A) or with a 2-h high-light treatment (B) were used. Means ± SE are shown for extractions on four to seven independent samples (each consisting of plant material from three different rosettes). Significant differences to the WT are denoted with asterisks. FW, fresh weight.
Molecular Plant 2014 7, DOI: ( /mp/sst115)
Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

9. Figure 8
Venn Diagrams Showing Overlaps between Transcript Changes in Different Genotypes.
(A) Transcript changes in the A. thaliana NDB1-suppressing lines (NDB1 RNAi) were compared to the 250 most affected genes in the A. thaliana abi4-102 mutant, as categorized in up- and down-regulated genes.
(B) A. thaliana NDB1-suppressing lines were compared to A. thaliana lines overexpressing HsfA2e (OsHsfA2e OE) and SMCP1 (OsSMCP1 OE) from rice, regarding up-regulated genes. Transcripts more than four-fold up-regulated in lines OsHsfA2eOE#02 and R07047 (OsSMCP1 overexpressor) were included (Yokotani et al., 2008, 2009). Transcripts changed in the NDB1-suppressing lines are as given in Supplemental Table 4. Chi-square tests revealed a significant overlap in each comparison, with p-values <10–60.
Molecular Plant 2014 7, DOI: ( /mp/sst115)
Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

10. Figure 9 Metabolic Profiling of NDB1-Suppressing Lines.
Rosettes from 35-day-old soil-grown T4 plants were used for the metabolite extractions. Data are shown for each metabolite that displayed a significant ≥10% difference between the WT values and the pool of values from both NDB1-supressing genotypes. The full data set is shown in Supplemental Table 5. Responsive metabolites are sorted as sugars and polyols (A), amino acids (B), and organic acids (C). Averages are shown for six or seven sets of three rosettes, with the level in WT set to unity and error bars denoting SE. Glc, glucose; Gal, galactose; Frc, fructose; Suc, sucrose.
Molecular Plant 2014 7, DOI: ( /mp/sst115)
Copyright © 2014 The Authors. All rights reserved. Terms and Conditions