Electronic supplementary material
A SNP in OsMCA1 responding for a plant architecture defect by deactivation of
bioactive GA in rice
Plant molecular biology
Zhenwei Liu, Qin Cheng, Yunfang Sun, Huixia Dai, Gaoyuan Song, Zhibin Guo,
Xuefeng Qu, Daiming Jiang, Chuan Liu, Wei Wang, Daichang Yang*
State Key Laboratory of Hybrid Rice and College of Life Sciences, Wuhan University,
430072, China
*Corresponding author: Daichang Yang
Address: State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan
University, Luojia Hill, Wuhan, Hubei Province 430072, China.
Tel: 86-27-6875-4680
Fax: 86-27-8757-0670
E-mail: dyang@whu.edu.cn
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Fig. S1 The G to C mutation was confirmed by sequencing WT and pad.
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Fig. S2 Domain prediction of the OsMCA1/PAD protein. Amino acids corresponding
to OsMCA1/ PAD. Different colors represent the domains and their locations. STYKc
domain (aa 6–70); coiled-coil domain (aa 189–217); PLAC8 domain (aa 298–396);
TM1 (aa 8–30); and TM2 (aa 339–360).
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Fig. S3 Schematic representation of internode elongation patterns in the WT and pad.
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Fig. S4 Expression profiles of the genes involve in GA metabolism in pad. QRT-PCR
analysis of the genes involved in GA metabolism in the uppermost (A) and second (B)
internodes of WT and pad. The means ± SD were calculated from three biological
replicates and values were determined by the Student’s t test.
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Fig. S5 Phylogenetic tree of the putative homologs of OsMCA1/PAD proteins.
Phylogenetic analysis of the putative OsMCA1/PAD homologs was performed using
the neighbor-joining (NJ) method of MEGA 5.2. The number of bootstrap replicates
was estimated at 1000, and gaps/missing sequences were deleted. The numbers in the
branches indicate bootstrap values (percent). The bar on the bottom indicates the
genetic distance based on 0.05 amino acid changes per residue.
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Fig. S6 Regulation model of OsMCA1/PAD on plant architecture defect by deactivation
of bioactive GA in rice. OsMCA1/PAD upregulated the expression of the genes for GA
deactivation, resulting in reduction of bioactive GA contents. The lower bioactive GA
contents lead to upregulate the expression of the genes for GA biosynthesis by feedback
regulation cycles
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Table S1. Primers used in this study
Gene
Locus
Primer name
Forward primer (5’→ 3’)
Reverse primer (5’→ 3’)
For plasmid construction
OsMCA1
Os03g0157300
Promoter-F/R
CCCAAGCTTAAGTGAGCCACACCTTAG
ACAGGAAAACCACCAAGAATCCAAGAC
For identification of transgenic lines
OsMCA1
Os03g0157300
618-1-F/R
GGGGAACGGAAATCTCAG
CGATGAAAATGATGGAGCAC
Sec18-F/Nos-R
AATCAAGAGCCCAGTAACCTATGCC
TTTATTGCCAAATGTTTGAACGATC
GUS-F/R
TCCGTCCTGTAGAAACCCC
ACGCTGACATCACCATTGG
For quantitative real-time RT-PCR
OsMCA1
Os03g0157300
OsMCA1RT-F/R
TTGATGTGTTGTTGCTGTGCG
ACTGAAATGAGGGTGGGCTAATC
OsCPS1
Os02g0278700
OsCPS1-F/R
TGTCAACAGGCACTGGACTG
AGGCGTAGTAGACGGAAAGC
OsKS1
Os04g0611800
OsKS1-F/R
GGGCGTCTCCTGAATGACA
CAGTGAGACACTGTTCAGCTTTCC
OsKO2
Os06g0570100
OsKO2-F/R
TGCTACCAGCGACTATTGTGATTT
GTGCAGAAGTACCCAACATGCTT
OsKAO
Os06g0110000
OsKAO-F/R
CTTCCTCCATCATTTTCTCC
AAGCAGTTGTCCACAGGC
OsGA20ox2
Os01g0883800
OsGA20ox2-F/R
CCAATTTTGGACCCTACCGC
GAGAGAAGCCCAACCCAACC
OsGA2ox1
Os05g0158600
OsGA2ox1-F/R
CGTCTTGTAGATGGTGGTGC
CCTGCCTGATGAGTTAGAAAAG
OsGA2ox3
Os01g0757200
OsGA2ox3-F/R
TGGTGGCCAACAGCCTAAAG
TGGTGCAATCCTCTGTGCTAAC
OsGA3ox2
Os01g0177400
OsGA3ox2-F/R
TCCTCCTTCTTCTCCAAGCTCAT
GAAACTCCTCCATCACGTCACA
OsActin
Os03g0718100
OsActin-F/R
GCCTTGGCAATCCACATC
AGCATGAAGATCAAGGTGGTC
Note: The primer pairs Sec18-F/Nos-R and GUS-F/R were designed according to the vector sequences (pOsPMP619 and pOsPMP617).
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