(A) Tu1-md Tu1-A allele showed half-tunicate · Plant Cell 10.1105/tpc.112.100537 1. Supplemental...

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.


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.


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.


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).


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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).


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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).


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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


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


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


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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

!


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(A) Tu1-md Tu1-A allele showed half-tunicate · Plant Cell 10.1105/tpc.112.100537 1. Supplemental...

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