A SNP in Osmca1 Responding for a Plant Architecture Defect by Deactivationof Bioactive

Electronic supplementary material

A SNP in OsMCA1 responding for a plant architecture defect by deactivationof bioactive GA in rice

Plant molecular biology

Zhenwei Liu, Qin Cheng, Yunfang Sun, Huixia Dai, Gaoyuan Song, ZhibinGuo, 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:

Fig. S1The G to C mutation was confirmed by sequencing WT and pad.

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).

Fig. S3Schematicrepresentation of internode elongation patterns in the WT and pad.

Fig. S4 Expression profiles of the genes involve in GA metabolism in pad. QRT-PCR analysis of the genes involved in GAmetabolism 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.

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.

Fig. S6Regulation model of OsMCA1/PAD on plant architecture defect by deactivationof bioactive GA in rice.OsMCA1/PADupregulated 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

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 (pOsPMP619and pOsPMP617).