Supplement
Supplement 1.
Phylogenetic Relationship of SERK family in Arabidopsis and rice
Arabidopsis
thaliana:
AtSERK1
(At1g71830),
AtSERK2
(At1g34210),
AtSERK3/AtBAK1 (At4g33430), AtSERK4 (At2g13790), AtSERK5 (At2g13800),
At2g23950.
Oryza
(Os04g0457800),
sativa:
OsBAK1/OsSERK1
OsSERK3
(Os06g0225300),
(Os08g0174700),
OsSERK4
OsSERK2
(Os02g0283800),
Os03g0703200, Os08g0442700. Tree was generated by PHYML (ML) and MEGA
(NJ). Bootstrap values (ML/NJ) were shown.
Supplement 2
Alignment of amino acid sequence of SERK family in Arabidopsis and rice.
Proteins from the SERK family contain conserved construct, including a signal
peptide, leucine zipper, five leucine-rich repeats (LRR1~LRR5), a Ser-Pro-Pro (SPP)
motif, a hydrophobic transmembrane domain and a kinase domain. Amino acid
residues conserved among all SERKs listed are shaded. The conserved activation loop
located in the kinase domain is boxed. A Thr residue in the activation loop is essential
for BAK1 function and is marked by ★. Percentages indicate amino acid similarity
to AtBAK1 and OsSERK1. WG represents whole gene; KD represents kinase domain.
Supplement 3.
The expression of OsBRI1 and OsBZR1 in the OsBAK1 transgenic rice detected by
real-time PCR.
WT, Wild-type Zhonghua 10; d61-1, the weak rice BR-insensitive mutant; OE
1
(OsBAK1-OE), transgenic plants overexpressing OsBAK1; AS (OsBAK1-AS),
transgenic plants suppressing OsBAK1 by antisense RNA; ICDΔ (OsBAK1-ICDΔ),
transgenic plants overexpressing the truncated intracellular domain of OsBAK1; ECD
(OsBAK1-ECD), transgenic plants overexpressing the extracellular domain of
OsBAK1.
2
Supplement 1
3
Supplement 2
4
Supplement 3
5
Supplement 4
gene
Use
OsBAK1
OsBAK1-ICDΔ
Gene isolation
Rice transformation
Arabidopsis
transformation
Gene isolation
Arabidopsis
transformation
Gene isolation
Arabidopsis
transformation
Gene isolation
Arabidopsis
transformation
Rice transformation
OsBAK1-AS
Rice transformation
OsBAK1-ECD
Rice transformation
BIFC
GFP fusion
OsSERK2
OsSERK3
OsSERK4
OsBAK1
Forward primer (restriction enzyme site
underlined)
5'TCCCCCGGGAGGGTGGTGCTGATTTGG
TGT3' (SmaI)
Reverse primer (restriction enzyme site
underlined)
5'GGGGTACCGTTCCTTGGGCTCCTGCTG
TT 3' (KpnI)
5'GCTCTAGATTGGGTCCACTTGGCAGCG
GTGAG3' (XbaI)
5'GGGGTACCAGGGCGAGGAAGCCGTCG
AGCAAA3' (KpnI)
5'GGGGTACCCCCCACTCTGTTTACTCCC3
' (KpnI)
5'CGAGCTCGGAATGGCAAGTGTTCTGTC
T3' (SacI)
5'GGGGTACCTTCTTCTCCTCCTCCTCTT 3' 5'CGAGCTCGCAGAGTCATGGCTCTTTAG
(KpnI)
T3' (SacI)
5'GAAGATCTTCAGGGTGGTGCTGATTTG 5'GGGGTACCCCTCCTTGGGCTCCTGCTG
GTGT 3' (BglII) used to amplified, BamHI TT 3' (KpnI)
inside this gene used to ligase vector
The same primers to that of OsBAK1-ICDΔ, KpnI (in primer) and SacI (inside this gene) was used
to ligase the vector in antisense orientation
5'CGCGGATCCATGGCGGCGCATCGGTGG 5'CGGGGTACCATACCATGCAAAACCAAT
GCGGTG3' (BamHI)
AGCAGG 3' (KpnI)
5'CGGGGTACCAGAGGGTGGTGCTGATTT 5'CCGCTCGAGCCTCGGCCCTGATAGCTC
GGTG 3' (KpnI)
AAC 3' ( XhoI)
6
OsBRI1
OsBAK1
OsBAK1-ICD
OsBAK1-ICDΔ
AtPP2A
AtCPD
OsSERK2
OsSERK3
OsSERK4
OsBAK1-A
OsBAK1-B
OsBRI1
OsBZR1
OsD2
OsACTIN
BIFC
5'GCTCTAGAATGGTCGTGAGGCAGTGAG
C 3'' (XbaI)
BIFC
5'GGACTAGTAATGCGTGATGTGTGAGAT
C 3' (SpeI)
BIFC
5'TGCTCTAGAATGTTCAGCAATAAAAAC
ATTCT3' (XbaI)
BIFC
5'TGCTCTAGAATGTTCAGCAATAAAAAC
ATTCT3' (XbaI) used to amplified, BamHI
inside this gene used to ligase vector
Real-time PCR
5’GCTGTTGAGGGTTGTGCTG 3'
Real-time PCR
5'TTACCGCAAAGCCATCCAA 3'
Real-time PCR
5’CGGAGGGTGATGCCCTAT3'
Real-time PCR
5’AAATCTTGAAGTGCTGGACTTG3'
Real-time PCR
5’TGTCCTGTCCTTGGCTGGT3'
Real-time PCR
5'GAGTTGATCTTGGGAATGCTGC 3'
Real-time PCR specific 5' TCTTTGATGTGCCTGCTGA 3'
for
endogenous
OsBAK1
level
in
OsBAK1-ECD
Real-time PCR
5' AGCCTCAACTACATCAATGGGTC 3'
Real-time PCR
5' ACCTACCGCAAGGGATGT 3'
Real-time PCR
5' TCGGTGGTGTTCGAAATCCT 3'
Real-time PCR
5'CGTATGAGCAAGGAGATCAC3'
7
5'CGGACTAGTATCCTTCTCCTCCTTGGCT
T 3' (SpeI)
5'CGGGGTACCCCTCGGCCCTGATAGCTC
AA 3 (KpnI)
5'CGGGGTACCCCTCGGCCCTGATAGCTC
AA 3' (KpnI)
5'CGGGGTACCCCTCGGCCCTGATAGCTC
AA 3' (KpnI)
5’GGCTGGCAGTAACGATTGTG 3'
5’TCATCACCACCACCGTCAAC 3'
5’CACGCTGTTGTCAGGGTTAC3'
5’GGTTATTGGAGAAACTGATTGG3'
5’GGCCAAGAGAAGCTGGTATTTC3'
5'CACTAGGTATCGTTCCGCTTATGTT3'
5' TTTATTGCTGAAGGTATCTGTTG 3'
5' GGTATCTCGCCCTCCAGCT 3'
5' CTCAGCAGCTGCGTTGAC 3'
5' ATGAGGCCGACAATAAATTCCT 3'
5'CACATCTGTTGGAAGGTGCT3'