Supporting Information Tables S1–S3 and Figs S1–S3
Table S1 Primers used in this study.
Name
Sequence
Application
qPCR GCHS1_F
5’-AATGAGAAAGAAGTCGTCGGAGAAC
Quantitative RT-PCR
qPCR GCHS1_R
5’-CGATTGACAGAGGATGTATGATGACT
Quantitative RT-PCR
qPCR G2PS1_F
5’-TGACGAGGTGAGGAAGAGATCTATG
Quantitative RT-PCR
qPCR G2PS1R
5’-ATTGGCAACCGCAGCAGTAA
Quantitative RT-PCR
qPCR GCHS3 _F
5’-GGTCTCAAAATCATATTTAAACGTGC
Quantitative RT-PCR
qPCR GCHS3_R
5’-CCATTGATATGTTTTCAAAGAAAACC
Quantitative RT-PCR
qPCR GCHS4_F
5’-GCTACCGATGGGCTCAAGA
Quantitative RT-PCR
qPCR GCHS4_R
5’-AACACACGAAGCCCAAACCA
Quantitative RT-PCR
qPCR GMYB10_F
5’-ACTAACAATATTTCCGCGCCC
Quantitative RT-PCR
qPCR GMYB10_R
5’-CGTCTGCTGGAGTAAATGACCAC
Quantitative RT-PCR
qPCR GGAPDH_F
5’-CCAGGAACCCAGAGGAGATACC
Quantitative RT-PCR
qPCR GGAPDH_R
5’-GGAGCGGATATGATGACCTTCTTG
Quantitative RT-PCR
In situ GCHS1_F
5’-AGATAACAATGGCGTCCTCCGT
In situ probe
In situ GCHS1_R
5’-TTTCATGGCGGCTTCCTTG
In situ probe
In situ GCHS4_F
5’-TTTCGAAAAGCGCAACGAG
In situ probe
In situ GCHS4_R
5’-TAGCGGCTTCTTTACCGAGCT
In situ probe
Att-GCHS1 L_F
5’-AAAAAGCAGGCTCGGAATTGCGTCTATCAAGCGGAT
VIGS vector
Att-GCHS1 L_R
5’-AGAAAGCTGGGTCGTTGGTACATCATGAAGCGTTTG
VIGS vector
Att-GCHS4 L_F
5’-AAAAAGCAGGCTCGCACAGCAGTTACTTTTCGTGGG
VIGS vector
Att-GCHS4 L_R
5’-AGAAAGCTGGGTCTGGCACGTAGTTTATCTGGCTC
VIGS vector
Att-GCHS1 S_F
5’-AAAAAGCAGGCTAGAAAGAAGTCGTCGGAGAACG
VIGS vector
Att-GCHS1 S_R
5’-AGAAAGCTGGGTTTAACAAACGTACATTCATTCCAACA
VIGS vector
Att-GCHS4 S_F
5’-AAAAAGCAGGCTGAGCCAGATAAACTACGTGCCA
VIGS vector
Att-GCHS4 S_R
5’-AGAAAGCTGGGTAGTTCGCGTACACCCAAAGAA
VIGS vector
attB1
5’-GGGGACAAGTTTGTACAAAAAAGCAGGCT
VIGS vector
attB1
5’-GGGGACCACTTTGTACAAGAAAGCTGGGT
VIGS vector
CHS GCHS1_F
5’-CGTGGTTCCCATGGCATGGCGTCCTCCGTTGACATG
Expression vector
CHS GCHS1_R
5’-TCGAATTCGGATCCTTAGACGGCAACCGTCACGG
Expression vector
CHS GCHS3_F
5’-CGTGGTTCCCATGGCATGGCCACCTCTCCGGCAGT
Expression vector
CHS GCHS3_R
5’-TCGAATTCGGATCCTCAATTTTGGGTGGCAACTGAAATAGTAGC
Expression vector
CHS GCHS4_F
5’-CGTGGTTCCCATGGCATGGTTAATATTGAGGAGTTTCG
Expression vector
CHS GCHS4_R
5’-TCGAATTCGGATCCTTAAATAGGCACGCTGTGAA
Expression vector
Table S2 Open reading frames (ORFs), the number of amino acids, molecular weights
and isoelectric points (pIs) of the deduced polypetides for the CHS-like genes in
Gerbera hybrida
Enzyme
GCHS1
GCHS3
GCHS4
G2PS1
ORF (bp)
1197
1212
1170
1209
Amino acids
398
403
389
402
Molecular weight (kDa)
43.5
44.1
42.9
43.7
pI
6.8
6.6
6.2
6.8
References
Helariutta et al. (1995b)
Helariutta et al. (1995b)
This work
Helariutta et al. (1995b)
Table S3 Identities between CHS-like proteins based on amino acid sequences (%)
GCHS1
GCHS3
GCHS4
Arabidopsis CHS
M. sativa CHS2
GCHS3
GCHS4
88
-
82
80
-
Arabidopsis
CHS
84
82
83
-
M. sativa CHS2
G2PS
82
79
82
82
-
73
73
69
68
69
Fig. S1 Cyanidin biosynthesis in leaf petioles and inflorescence scapes is GMYB10
dependent. In cv. Terraregina (left panel), leaf petioles and inflorescences scapes are
naturally red, and contain primarily cyanidin derived pigments. In the GMYB10 antisense
transgenic lines of cv. Terraregina, leaf petioles and inflorescence scapes are nonpigmented (right panel, one out of three lines where both GMYB10 transcript levels and
vegetative anthocyanin pigmentats were strongly reduced).
Fig. S2 Comparison of nucleotide sequence of the inserts in vectors TRV:GCHS1 and
TRV:GCHS4 to the corresponding sequence in GCHS4 and GCHS1, respectively.
Fig. S3 Close view of petals in cv. President after silencing of GCHS1 or GCHS4 by VIGS.
By the silencing of GCHS1, inflorescences of cv. President developed white petals (top
panel), whereas, by the silencing of GCHS4, petals became only slightly paler (middle
panel) compared with those infected with empty TRV control (bottom panel, from the same
shot as the middle panel).