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Suplemental Data
Figure S1. Alignment of deduced amino acid sequence of CaGA2ox1 and other
GA2ox homologs from the pepper EST database. Dark shading indicates amino
acid residues that are conserved in CaGA2ox1. The asterisks indicate amino acid
residues presumed to bind Fe at the active site. The closed triangles show the
putative 2-oxoglutarate binding site.
CaActin
Cadhn
CaGA2ox1
100μM ABA
H2O
0 1 3 6 12 24 48 0 1 3 6 12 24 48H2O
Figure S2. Expression level of the CaGA2ox1 gene in pepper plants inoculated with a non-host
bacterial pathogen, Xag8ra, was detected by RT-PCR The CaGA2ox1 transcripts in pepper plants
treated with ABA and wound. The expression of Cadhn in ABA-treated leaves were monitored
as positive controls.
0 1.5 6 12 24 0 1.5 6 12 24MgCl2 Xag8ra
CaActin
CaPR1
CaGA2ox1
0 0.5 1.5 3 6 9 12 18 24 72
Wound
CaActin
CaGA2ox1
pET
pET:CaG
A2ox1
pGEX
75503725
kDa M
75503725
kDa
CaGA2ox1
SoGA2ox
GST
U I U I U I U I
α-His ab
pGEX:S
oGA2o
x
Figure S3. Expression of recombinant CaGA2ox1 protein in BL21 pLysS.
The recombinant fusion protein CaGA2ox1 was detected by western blot
analysis with an anti-His antibody. U: uninduced, I: IPTG-induced.
0
0.5
1
1.5
2
2.5
3
0
0.01
0.02
0.03
0.04
0
0.01
0.02
0
0.05
0.1
0.15
0.2
0.25
CaPR1 CaACCox
Rel
ativ
e m
RN
AR
elat
ive
mR
NA
0
0.02
0.04
0.06
0.08
0.1
0.12
6h 12h 6h 12h
CaGA2ox1 CaKS25000C15 cacn18210
MgCl2 Xag8ra
Figure S4. Expression patterns of GA-responsive genes and several defense-related genes in
pepper plants using Q-RT-PCR analysis. Total RNA was extracted from leaf tissues of pepper
plants at 6 h and 12 h after MgCl2 or Xag8ra inoculation, respectively.
0
0.5
1
1.5
CaPR4
6h 12h
Primers Sequences
CaGA2ox1 F GTCTAATATGGTAGTGGCAACTC
CaGA2ox1 R GGTAGCTAAGACTGCTAAGTACTGC
cacn18210 F AAGAGCAACCATGGTGATCTTGTC
cacn18210 R AATTCGGAGTTTGTCGATACGAGG
CaKS25002C15 F ACTACCCACCATATTGTAGTGATG
CaKS25002C15 R CCCTATGCTTTACACTCTTAAACC
CaPR1 F ACTTGCAATTATGATCCACCCaPR1 R ACTCCAGTTACTGCACCATTCadhn F AGTGATCATTCTTTGCTTTATTCTTACCadhn R AACATTCATTCCCATGCTATCCaActin F TTGGACTCTGGTGATGGTGTG
CaActin R AACATGGAGCCACCACTG
Table S1. Primers used in a RT-PCR analysis.
Primers Sequences
CaGA2ox1 F CGTAAGTGCAGTTAGGAAGCTGGCCaGA2ox1 R GCTCAGATGATGGGTCCCAATGATCaPR1 F CAAGCCCAAAATTCTCCCCAAGACaPR1 R TTGCATCCTGCAGTCTCCAATTCCaGA3ox F TAAATTCTTACCCGGCTTGTCCGCaGA3ox R TTAACAACTAACGCGCCACGAAGCaGA20ox F GCGATACATTTATGGCGCTTTCACaGA20ox R CATGTGAGGTCGGGATACATTCGCaDhn F GACAAGTTGCCAGGTGGAGGACCaDhn R CCTTCTTCTCCTTCCCCTCCGCacn18210 F CCGCTGACCCTTTCGGCTATCacn18210 R ATTTACCGCAGCCCGGATGTCaKS25002C15 F CAGGTGATGACAAATGGGAGGTTTCaKS25002C15 R CAAGCTGTCTTGATCCCCTTTCACaACCox F GCCCCAAACCCGATTTGATTCaACCox R TGGAGTGGCGCATAGGAGGA
Table S2. Primers Used for Q-RT-PCR.
Gene Gene Bank Accession # (or EST #) Reference
CaGA2ox1 DQ465393 in this paper
cacn18210 http://genepool.kribb.re.kr/new/
CaKS25002C15 http://genepool.kribb.re.kr/new/CaGA3ox (CaKS13043D06) http://genepool.kribb.re.kr/new/
CaGA20ox (cacn5286) http://genepool.kribb.re.kr/new/
CaPR1 AF053343 Lee et al., 2004
CaActin GQ339766 Lee et al., 2004
CaDhn AY225438 Chung et al., 2003
Table S3. Pepper genes in this paper
REFERENCE
Chung E, Kim SY, Yi SY, Choi D. 2003. Capsicum annuum dehydrin, an osmotic-stress
gene in hot pepper plants. Mol Cells 15: 327-332.
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