Supporting Information legends
Figure S1. Fatty acid composition of monogalactosyldiacylglycerol (MGDG),
digalactosyldiacylglycerol (DGDG), sulfoquinovosyldiacylglycerol (SQDG), phosphatidylglycerol (PG),
and phosphatidylcholine (PC) in mgd1-2 grown under Pi-sufficient (+Pi) or Pi-deficient (–Pi) conditions.
Values are the means ± SE from six independent experiments for +Pi plants and three experiments for
–Pi plants.
Figure S2. Cellular images of leaf tissues from wild type and mgd1-2 grown under Pi-deficient
conditions. Plastids in mesophyll cells were observed with a fluorescence microscope (BZ-8000;
Keyence, Osaka, Japan). Merged images show the overlay of chlorophyll fluorescence and optical image.
Red fluorescence from chlorophylls was granulated in the mgd1-2 plastids. Bars = 20 m.
Figure S3. Multiple electron micrographs of plastids from mgd1-2 leaves grown under Pi-sufficient (a)
and -deficient (b) conditions. Bars = 1.0 m.
Figure S4. Envelope invagination observed in plastids from mgd1-2 leaves grown under Pi-sufficient
(a,b) and –deficient (c-f) conditions. Subset images correspond to labeled areas of the original picture.
Yellow arrowheads indicate the sites of envelope invagination. Scale bars = 1.0 m.
Figure S5. Diameter histogram of plastid nucleoids in protoplasts from wild type, mgd1-2, and chlh
leaves grown under Pi-sufficient (+Pi) and -deficient (–Pi) conditions.
Figure S6. Characterization of the hema1 mutant. (a) Schematic diagram showing the T-DNA insertion
site in hema1 (SALK_053036). Exons and untranslated regions are represented by filled and open boxes,
respectively. Lines between the boxes correspond to introns. The T-DNA insertion site is represented by
the inverted triangle. Location and direction of primers used for PCR analyses are indicated by arrows (F,
R, and LB). ATG, translation initiation codon; Stop, translation termination codon. (b) PCR analysis of
genomic DNA from three independent seedlings of the homozygous hemea1 line and wild type. PCR
amplification products between the F and the R primer and the LB and the R primer are shown in the
upper and the lower panel, respectively. DNA from the heterozygous hema1 seedling (Hetero) was used
as a positive control for both reactions. (c) RT-PCR analysis of the HEMA1 gene in the homozygous
hema1 and wild-type seedlings (upper panel). The F and R primer set was used for amplification of the
HEMA1 gene. HEMA2 transcripts were analyzed as a control (lower panel). Three independent
replicates are shown in both lines. PCR products from wild-type genomes (Genome), that are larger in
molecular size than those from cDNAs due to the existence of introns, are displayed as a control for
genomic amplification (leftmost lane). (d) Chlorophyll contents in hema1 and wild type seedlings grown
for 7 days on MS media containing 1% (w/v) sucrose. Values are the means ± SD from three
independent experiments. (e) Phenotypes of the hema1 mutant and wild type grown for 21 days on MS
media containing 1% (w/v) sucrose.
Table S1. Primers used for qRT-PCR analysis
Appendix S1. Supplemental analysis for fatty acid composition of membrane lipids in mgd1-2 under
Pi-controlled conditions