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.