Glutamyl-tRNA

5-Aminolevulinic acid

Protoporphyrinogen IX

Mg-Proto IXHeme

Mg-Proto IX ME

HEMA

GSA

PPO

FC

PORB

CHLD, CHLH, CHLI

Glutamate 1-semialdehyde

ALAD

Pchlide

Chlorophyll

Proto IX

ALA

Fig. S1. The tetrapyrrole pathway in plants showing intermediates and genes analyzed

in this study. Intermediates: Proto IX, protoporphyrin IX; Mg-Proto IX, Mg-protoporphyrin

IX; Mg-Proto IX ME, Mg-protoporphyrin IX methyl ester; Pchlide, protochlorophyllide.

Genes and enzymes that correspond to the gene names: HEMA, glutamyl-tRNA reductase;

GSA, glutamate 1-semialdehyde aminotransferase; ALAD, ALA dehydratase; PPO, protopor-

phyrinogen oxidase; FC, ferrochelatase; CHLD, D-subunit of Mg-chelatase; CHLH, H-sub-

unit of Mg-chelatase; CHLI, I-subunit of Mg-chelatase; PORB, protochlorophyllide oxidore-

ductase B.

Supplemental Data

0 10 20 30 40 50 60 70 80

up-regulateddown-regulated

Signal transduction mechanismsPosttranslational modification

Intracellular traffickingTranslation and ribosome

TranscriptionCarbohydrate metabolism

RNA processingLipid metabolism

Amino acid metabolismSecondary metabolismReplication and repair

CytoskeletonInorganic ion metabolism

Chromatin

Cell cycleCell wall

Nucleotide metabolismNuclear structure

Coenzyme metabolismExtracellular structure

Defence mechanisms

Energy production and conversion

Number of differentially expressed genes by ALA

Fig. S2. COG classes of the proteins encoded by the differentially expressed genes in

response to exogenous ALA. In each functional category, the genes are grouped

according to their regulation by ALA treatment (> 1.6-fold change). Red bars show ALA-

induced genes and cyan bars ALA-repressed genes.

Transcription factor

Translation initiation

Proteasome

Elongation factor

Ribosome

RNA polymerase

RibosomeRNA binding

Transcription factorCell cycle

rRNA processing

Signal transduction

Signal transduction

Fig. S3. Network of a significantly (P ≤ 0.05) altered genes in ALA-treated rice plants.

Genes in network are the corresponding Arabidopsis homologues of differentially expressed

rice genes in microarray analysis. Each node in this network represents differentially ex-

pressed genes. The red and blue nodes correspond to the up-regulated and down-regulated

states, respectively, according to the relative gene-expression levels between control and

ALA-treated rice plants. The lines between these nodes represent the protein-protein interac-

tions.

Rel

ativ

e m

RN

A le

vel

0

1

2

3

4

5

Cont-6ALA-6

Os07g

0545

400

Os10g

0396

300

Os01g

0888

700

Os11g

0700

500

Os03g

0803

900

Os01g

0607

800

Os01g

0179

600

Os07g

0656

800

 

Fig. S4. Validation of microarray results with the expression profiles of eight genes in

the ALA-treated rice plants as detected by qRT-PCR. The plants were subjected to the

same treatments as in Fig. 2. Treatment notations are the same as in Fig. 2. Total RNAs

were purified from plants and reverse transcribed. The resultant cDNAs were used as

templates for qRT-PCR using Actin as an internal control. The Cont-6 was used for

normalization, with the expression level of the sample set to 1. Error bars represent SE, and

representative data from three independent experiments are presented.