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Figure S1
mrn1
FKF1
PHYA
TOC1
PHYB
EMF1
CRY1
ZTL
CRY2
GI
CO
FLK
LD
SOC1
FPA
FT
FLC
elF4A
GAI
ACTIN7
RGA
Photoreceptors genes
WT
Autonomous genes
mrn1
GA genes
Photoperiod genes
Clock genes
WT
Figure S1. Expression of genes related to flowering time in wild-type and mrn1
leaves. Total RNA was isolated form 3-week-old wild-type and mrn1 leaves and
converted into first-strand cDNAs that were used as templates to PCR amplify
transcripts of FKF1, TOC1, EMF1, ZTL, GI, CO, SOC1, FT, PHYA, PHYB,
CRY1, CRY2, FLK, LD, FPA, FLC, GAI, and RGA with primers listed in Lim et
al (2004). The elf4A (At3g13920) and ACTIN7 (At5g09810) genes were used
as a control.
Lim, M.H., Kim, J., Kim, Y.S., Chung, K.S., Seo, Y.H., Lee, I., Kim, J., Hong, C.B.,
Kim, H.J., Park, C.M. (2004) A new Arabidopsis gene, FLK, encodes an RNA binding
protein with K homology motifs and regulates flowering time via FLOWERING LOCUS
C. Plant Cell , 16, 731–740.
Figure S2
(a)
At5g42600
(MRN1)
At5g42590
At5g42610
At5g42620
1kb
4729
1
T-DNA
LB
RB
Salk_152492
WT
(b)
mrn1
MRN1
ACTIN2
Figure S2. Mapping of T-DNA insertion sites and expression in MRN1 gene. (a)
Structure of MRN1 (At5g42600) and T-DNA insertion site. Boxes and lines
indicate exon and intron of the MRN1 gene, respectively. Arrows indicate the
sites for specific primers, which were used in RT-PCR analysis. (b) RT-PCR
analysis of MRN1 gene in wild-type and mrn1 roots. Total RNA was isolated
form wild-type and mrn1 roots and 100 ng of total RNA was used in RT-PCR
analysis. The ACTIN2 (At3g18780) gene was used as a control.
Figure S3
WT
mrn1
KRP1
KRP2
KRP3
KRP4
KRP5
CycB1;2
CycD3;1
ACTIN7
Figure S3. Expression of cell-cycle related genes in Arabidopsis wild-type and
mrn1 leaves. Total RNA was isolated from 3-week-old wild-type and mrn1
leaves and were converted into first-strand cDNAs that were used as templates
to PCR amplify transcripts of KRP1, KRP2, KRP3, KRP4, KRP5, KRP6, KRP7,
CycB1;2, and CycD3 genes with specific primers listed in Table S2 online. The
Actin7 (At5g09810) gene was used as a control.
Figure S4
WT
(a)
FK
SMT2
mrn1
WT
(b)
mrn1
FK
SMT2
DWF7
DWF7
DWF5
DWF5
DWF1
DWF1
DWF6
DWF6
DWF4
DWF4
DWF3
DWF3
BR6ox1
BR6ox1
CYP710A1
CYP710A1
CYP710A2
CYP710A2
ACTIN7
ACTIN7
Figure S4. Expression of BR and sterol metabolic genes in wild-type and mrn1
roots (a) and leaves (b). Total RNA was isolated from 2~3 weeks old roots and
leaves in Arabidopsis wild-type and mrn1 plants and converted into first-strand
cDNAs that were used as templates to PCR amplify transcripts of BR and sterol
metabolic genes (FK, SMT2, DWF7, DWF5, DWF1, DWF6, DWF4, DWF3,
BR6ox1, CYP710A1, CYP710A2) with specific primers listed in Table S3. The
ACTIN7 (At5g09810) gene was used as a control.
Figure S5
HMG-CoA
Acetyl-CoA
squalene
MVA
HMGR
SQE
O
2,3-Oxidosqualene
MRN1
CAS1
LUP1
AS1
LAS1
H
H
(Marneral)
H
H
H
HO
HO
H
H
H
H
Lanosterol
Cycloartenol
HO
Marnerol
H
HO
H
OH
H
Lupeol
H
OH
H
+
H
HO
H
H
HO
H
-Sitosterol
HO
H
H
O
H
O
Brassinolide
Primary metabolism
H
HO
H
-Amyrin
Secondary
metabolism
Figure S5. The simplified biosynthetic pathways of sterol, steroid hormone, and
nonsteroidal triterpenoids in plants. Abbreviations: HMG-CoA, 3-hydroxy-3methylglutaryl-CoA; HMGR, 3-hydroxy-3-methylglutaryl-CoA reductase; LUP1,
lupeol synthase; MRN1, marneral synthase; MVA, mevalonate; SQE, squalene
epoxidase; AS1, -amyrin synthase.