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Supplemental Figure 1. Half-tunicate phenotype of single copy Tu1-md and
Tu1-rec heterozygous mutants.
(A) A plant heterozygous for Tu1-md having Tu1-A allele showed half-tunicate
phenotype in mature ear.
(B) Similar ear phenotype was detected from a plant heterozygous for Tu1-rec
that has also only one copy of Tu1-A allele.
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
1
Supplemental Figure 2. A 30 kb tandem duplication at Tu1.
(A) Schematic representation of duplicated Tu1-A and Tu1-B loci spanning
GRMZM2G006297. Black arrows indicate the position of primers used for (B).
(B) PCR showed that 3’ end of Tu1-B is about 5 kb apart from 5’ upstream of
GRMZM2G006297. Sequencing and assembly suggests the presence of gypsy-
like and copia-like retrotransposons and non-autonomous dSpm-like and hAT-
type DNA transposons in this 5kb region. The gel image was inverted for better
contrast and 1 Kb Plus DNA ladder (Invitrogen) was used to determine the size.
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
2
Supplemental Figure 3. Upregulation of Zmm19 expression in Tu1 reproductive
tissues.
(A) and (B) Zmm19 expression in immature tassels (1-2 cm long) (A) and
immature ears (0.5-0.9 cm long) (B) was determined by qRT-PCR. Quantitative
RT-PCR of three biological replicates for each tissue. Results are plotted as the
ratio to the wild-type level of Zmm19 and are represented as mean +/- SEM.
Zmm19 is normalized to Ubiquitin levels. Tu1/+ indicates heterozygous Tu1
plants and Tu1/Tu1 homozygous Tu1.
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
3
Supplemental Figure 4. Tu1-A:YFP and Tu1-B:RFP transgenic tassels
phenocopy Tu1 in a dosage-dependent manner.
(A) T2 transgenic tassels harboring one Tu1 transgene appear to have relatively
thick rachises and lateral branches with elongated glumes. The main rachis of
the double transgenic plant (Tu1-A:YFP/+; Tu1-B:RFP/+) was even thicker than
that of the single transgenic plants. The order of each tassel in (A) corresponds
to that in (B) to (F).
(B) to (F) A close-up view of (A), showing the additive effect of the glume
phenotype.
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
4
Supplemental Figure 5. TU1-A:YFP is expressed in vegetative and reproductive
tissues.
(A) to (C) YFP expression was clearly detected in nuclei from not only vegetative
tissue, i.e., the husk (A), but also reproductive tissues, i.e., trichomes (B), and
the floral meristem (FM, [C]) in 1-cm-long immature ears. Scale bar, 10 µm (A),
20 µm (B), and 50 µm (C).
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
5
Supplemental Figure 6. TU1-A:YFP and TU1-B:RFP proteins are co-localized in
nuclei.
(A) and (B) Transgenic lines harboring both Tu1-A:YFP and Tu1-B:RFP
displayed co-localization of TU1-A:YFP and TU1-B:RFP in the nuclei of glume
(A) and trichome (B) epidermis. Merged images show that TU1-A is co-
expressed with TU1-B in the same nuclei. Scale bar, 10 µm (A) and (B).
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
6
Supplemental Figure 7. Gene expression data of GRMZM2G370777 and
GRMZM2G006297.
(A) and (B) The maize electronic Fluorescent Pictograph Browser (eFP browser;
bar.utoronto.ca/maizeefp) displays the expression level of target genes,
GRMZM2G370777 (A) and GRMZM2G006297 (B), during maize development.
The expression data are based on a report by Sekhon et al. (2011).
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
7
Supplemental Table 1. Primers used in fine-mapping and qRT-PCR.
Forward Primer Reverse Primer
Fine Mapping
umc1847
GRMZM2G386088
GRMZM2G006297
IDP7898
umc1194
IDP8018
GRMZM2G370777
GRMZM2G081318
IDP4760
GCCCAAGGTAGATTTTTACTCTCCA
ATATTTAAGGGCACCGTGGAGAAT
GCTGCTACAGTTCGCGTTC
ATGATCTCCGTGGTCTTTGG
ACCACCAGACATGGGAAACTTCT
GAGACCTTTGACGCACTTCC
AGATGCATTGATCGGTGATG
GTGAACCATGGCCGAGAAAC
GCATACCCAGATTTGTGTCC
AAGTCGTAGAGCTCGTCGTGATG
CCAGACTCCCCATTGTAAAACTTG
CGTGCTGCTTGTCAAGTTGT
TGCAAAGCACCTATACACGC
AAGGCGGACACTACTCTACCCTCT
ACATGAGAAGGCGATTCTGG
ACATATCCCCCACGTTGTGT
CAGCGCACAGTGATCTAGGA
TTGCGATCACTCAGAAGACG
qRT-PCR
Zmm19
Ubiquitin
GCTCATCGTCTTCTCCTCCA
TAAGCTGCCGATGTGCCTGCGTCG
TTGCTTCCACAAGTTGCTCA
TGAAAGACAGAACATAATGAGCACAG
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
8
Supplemental Table 2. Primers used in transgene construction.
The att sites required for Gateway cloning are underlined.
Tu1AB
_attB1
Tu1A_attB
5(R)
Tu1A_attB
5
Tu1AB
_attB4
Tu1AB
_attB3
Tu1AB
_attB2
Tu1B_attB
5(R)
Tu1B_attB
5
YFP
_attB4(R
)
YFP
_attB3(R
)
RFP
_attB4(R
)
RFP
_attB3(R
)
Prim
er
Nam
e G
GG
GA
CA
AG
TTTGTA
CA
AA
AA
AG
CA
GG
CTTA
TCG
CC
AA
GA
CTTTG
TGA
AG
A
GG
GG
AC
AA
CTTTTG
TATA
CA
AA
GTTG
TGA
TGG
AG
AC
AC
CG
ATG
AC
TTTC
GG
GG
AC
AA
CTTTG
TATA
CA
AA
AG
TTGC
AA
CTTG
TTCG
GA
CA
CTTG
TCA
CA
GG
GG
AC
AA
CTTTG
TATA
GA
AA
AG
TTGG
GTG
CTTC
CA
TGC
AA
CA
CA
AG
GC
GG
GG
AC
AA
CTTTG
TATA
ATA
AA
GTTG
CA
TGA
AA
AA
AC
AA
GA
TGA
TCTG
TGTTG
GG
GG
AC
CA
CTTTG
TAC
AA
GA
AA
GC
TGG
GTA
TCTA
AG
GA
TCC
AA
AC
GA
TGG
GG
GG
AC
AA
CTTTTG
TATA
CA
AA
GTTG
TGTA
GG
TCG
TTTTCA
TGG
GTTTC
GG
GG
AC
AA
CTTTG
TATA
CA
AA
AG
TTGC
AA
CA
AC
GTG
GG
GG
ATA
TGTTG
GG
GG
AC
AA
CTTTTC
TATA
CA
AA
GTTG
CA
TAA
GG
CC
GG
CC
TGG
AG
GTG
GA
GG
TG
GA
GC
T
GG
GG
AC
AA
CTTTA
TTATA
CA
AA
GTTG
TGG
CC
CC
AG
CG
GC
CG
CA
GC
AG
CA
CC
AG
CA
GG
ATC
GG
GG
AC
AA
CTTTTC
TATA
CA
AA
GTTG
CA
GG
CC
GG
CC
TGG
AG
GTG
GA
GG
TGG
AG
CTG
CC
TCC
TCC
GA
GG
AC
GTC
ATC
GG
GG
AC
AA
CTTTA
TTATA
CA
AA
GTTG
TGG
CC
CC
AG
CG
GC
CG
CA
GC
AG
CA
CC
AG
CA
GG
ATC
GG
CG
CC
GG
TGG
AG
TGG
CG
GC
C
Prim
er Sequence
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
9
Supplemental Table 3. Primers used in allele-specific and long-range PCRs.
Primer Sequence Figure
Allele-specific
Zmm19 5’ F14
Tu1 5’ F10
Tu1 int2 R6
Tu1 3’ F35
Tu1 3’ F39
2G006297 R11
TTGCCCAGGACTGACTGATA
GATGCATGAAGAAGCCATGA
ACATATCCCCCACGTTGTGT
TCTGTGTTGATCTGGGTGGA
CGTCGAAGCTATCGATTATGAAG
CACTCTGATACCCAGCAATCG
2C
2D
2C, 2D, 2E
S2B
S2B
S2B
Long-range
2G006297 F44
Tu1 Ex6 F34
2G006297 R21
GCTTCCATGGTTGACCTTCT
GGAGAACAGGCAACTGAGGA
AAGCAGTCGATTCCTCCTCA
2E
2F, S2B
2F
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
10
Supplemental Reference Sekhon, R.S., Lin, H., Childs, K.L., Hansey, C.N., Buell, C.R., de Leon, N., and
Kaeppler, S.M. (2011). Genome-wide atlas of transcription during maize
development. Plant J. 66, 553-563
!
Supplemental Data. Han et al. (2012). Plant Cell 10.1105/tpc.112.100537
11