Figure S1 FKF1 WT TOC1 EMF1 ZTL GI CO SOC1 FT elF4A PHYA PHYB CRY1 CRY2 RGA FLK LD FPA FLC GAI ACTIN7 mrn1 WT mrn1 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 Arabidops is gene, FLK, encodes an RNA binding protein with K homology mot ifs and regulates flowering time via FLOWERING LOCUS C. Plant Cell , 16, 731–740. Clock genes Photoperiod genes Photoreceptors genes Autonomous genes GA genes
Transcript
Figure S1
FKF1
WT
TOC1
EMF1
ZTL
GI
CO
SOC1
FT
elF4A
PHYA
PHYB
CRY1
CRY2
RGA
FLK
LD
FPA
FLC
GAI
ACTIN7
mrn1 WT mrn1
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.
Clo
ck
gen
esP
ho
top
erio
d
gen
es
Ph
oto
rece
pto
rs
gen
esA
uto
no
mo
us
g
enes
GA
gen
es
At5g42590 At5g42610 At5g42620At5g42600
(MRN1)
1kb
(a)
RB LBT-DNA
1 4729
(b)
Salk_152492
Figure S2
MRN1
ACTIN2
mrn1WT
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.
KRP1
KRP2
KRP3
KRP4
KRP5
CycB1;2
CycD3;1
ACTIN7
mrn1WT
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 S3
Figure S4
FK
SMT2
DWF7
DWF5
CYP710A1
DWF1
DWF6
DWF4
DWF3
BR6ox1
CYP710A2
ACTIN7
mrn1WT
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.
( b )
FK
SMT2
DWF7
DWF5
CYP710A1
DWF1
DWF6
DWF4
DWF3
BR6ox1
CYP710A2
ACTIN7
mrn1WT( a )
-Sitosterol
2,3-Oxidosqualene
Cycloartenol
Brassinolide
Marnerol
MRN1
Lanosterol
LAS1CAS1 LUP1AS1
Lupeol
+
-Amyrin
Acetyl-CoA HMG-CoA MVA squalene
SQE
HMGR
Primary metabolism Secondary metabolism
O
HO
H
H H
H
H
HO HOH
H
H
H
H
HHO
HO
HO
OH
OH
HH
H
H
O
O
H
H
H
H
HO
HOH
H
H
(Marneral)
Figure S5. The simplified biosynthetic pathways of sterol, steroid hormone, and nonsteroidal triterpenoids in plants. Abbreviations: HMG-CoA, 3-hydroxy-3-methylglutaryl-CoA; HMGR, 3-hydroxy-3-methylglutaryl-CoA reductase; LUP1, lupeol synthase; MRN1, marneral synthase; MVA, mevalonate; SQE, squalene epoxidase; AS1, -amyrin synthase.
