14 dap 25 dap - the plant cell · 2012. 7. 3. · morohashi et al. (2012). plant cell...

Morohashi et al. (2012). Plant Cell 10.1105/tpc.111.098004

Supplemental Figure 1. P1-rr and P1-ww pericarps at different developmental stages. P1-rr and P1-ww pericarps were collected at 14 and 25 days after pollination (DAP), showing the increasing level of pigmentation as P1-rr pericarps maturation proceeds, compared to lack of pigmentation observed in P1-ww pericarps.

P1-ww

P1-rr

14 DAP 25 DAP 5mm

GRMZM2G068917 (CCR)

Relative expression to actin DAP

14 25 P1-ww

P1-rr 14 25

* GRMZM2G068917 (CCR)

P1-ww P1-rr

Relative expression to actin 0

* ** GRMZM2G167336 (F2H1) 14

25 P1-ww

P1-rr 14 25

** GRMZM2G167336 (F2H1)

P1-ww P1-rr

0

*

GRMZM2G334660 (PAL) 14 25 P1-ww

P1-rr 14 25

GRMZM2G334660 (PAL)

P1-ww P1-rr

0

GRMZM2G441347 (PAL)

14 25 P1-ww

P1-rr 14 25

GRMZM2G441347 (PAL)

P1-ww P1-rr

0

** GRMZM2G170692 (PAL)

14 25 P1-ww

P1-rr 14 25 GRMZM2G170692 (PAL)

P1-ww P1-rr **

0

** GRMZM2G025832 (Pr1) 14

25 P1-ww

P1-rr 14 25

GRMZM2G025832 (Pr1)

P1-ww P1-rr

0

**

GRMZM2G162755 (3-GT) 14 25 P1-ww

P1-rr 14 25

* GRMZM2G162755 (3-GT)

P1-ww P1-rr

0

** ** GRMZM2G083130 (3-GT) 14

25 P1-ww

P1-rr 14 25

* GRMZM2G083130 (3-GT)

P1-ww P1-rr

0

GRMZM2G063550 (3-GT) 14 25 P1-ww

P1-rr 14 25 ** ** GRMZM2G063550 (3-GT)

P1-ww P1-rr

0

**

Pericarps Silks

P-value14= 0.013 P-value25= 0.178

P-value= 0.319

P-value14= 0.085 P-value25= 0.298

P-value= 0.107

P-value14< 0.01 P-value25= 0.108

P-value < 0.01


** GRMZM2G180283 (3-GT) 14

25 P1-ww

P1-rr 14 25

Relative expression to actin

** GRMZM2G180283 (3-GT)

P1-ww P1-rr

0

***

14 25 P1-ww

P1-rr 14 25

GRMZM2G162783 (3-GT)

GRMZM2G162783 (3-GT)

P1-ww P1-rr

0

**

** *

GRMZM2G151227 (WHP1)

0

14 25 P1-ww

P1-rr 14 25 ** GRMZM2G151227 (WHP1)

P1-ww P1-rr

0

**

GRMZM2G140527 (LAC10) 14 25 P1-ww

P1-rr 14 25

GRMZM2G140527 (LAC10)

P1-ww P1-rr

0

**

**

GRMZM2G079554 (MATE) 14 25 P1-ww

P1-rr 14 25 GRMZM2G079554 (MATE)

P1-ww P1-rr

0

** **

GRMZM2G031311 (RHM) 14 25 P1-ww

P1-rr 14 25

GRMZM2G162739 (NAC) *

14 25 P1-ww

P1-rr 14 25

0

GRMZM2G314769 (JAC1)

** **

0

14 25 P1-ww

P1-rr 14 25

* **

GRMZM2G177928 (STRS) 14 25 P1-ww

P1-rr 14 25

GRMZM2G162739 (NAC)

P1-ww P1-rr

0

* GRMZM2G314769 (JAC1)

P1-ww P1-rr

0

GRMZM2G031311 (RHM)

P1-ww P1-rr

0

* GRMZM2G177928 (STRS)

P1-ww

P1-rr

0

Relative expression to actin DAP

Pericarps Sillks

Supplemental Figure 2. RT-qPCR validation of genes identified as up-regulated in P1-rr by RNA-Seq. Genes (18) that showed up-regulation in P1-rr (pericarps and silks) RNA-Seq experiments were chosen for validation. Left, relative expression of genes to actin in P1-rr and P1-ww pericarps at 14 and 25 DAP. Right, RT-qPCR expression of the same genes relative to actin in P1-rr and P1-ww silks. The results obtained by RT-qPCR correlate to the expression values obtained by RNA-Seq in 16/18 for pericarps and 12/18 for silks. P-values for the difference in gene expression for biological triplicates were included for some of the genes described in more detail in the text. Single or double asterisks indicate significant (P < 0.05) or very significant (P < 0.01) differences between P1-rr and P1-ww.


Root Shoot Leaf P1-rr

Silks

P1-ww

14 DAP 25 DAP

P1-rr P1-ww P1-rr P1-ww

Pericarps

A

0 0.2 0.4 0.8 0.6

PCC Value

Supplemental Figure 3. Global gene expression analysis from RNA-Seq experiments conducted from root, shoot, leaf, silk and pericarp tissues. A. Box plot of gene expression distribution from RNA-Seq conducted from root, shoot, leaf, silk and pericarp tissues. The x-axis corresponds to the tissue and y-axis corresponds to the log10 transformed FPKM values. The width of the boxes is proportional to the square-roots of the number of gene expressed per sample. Expression of P1 (blue “!” signs), A1 (orange “+” signs) and two housekeeping genes, Ubiquitin (red “-” signs) and Actin1 (green “*” signs), are shown. B. Pearson correlation coefficient-based heat-map matrix of all the RNA-Seq expression analyses. The results of the hierarchical clustering are illustrated as a dendrogram, according to expression pattern similarities.

log 1

0(FP

KM

)

B

P1-w

w 2

5 D

AP!

P1-rr

25

DAP

!

P1-w

w 1

4 D

AP!

P1-rr

14

DAP

!

P1-w

w!

P1-rr!

Leaf!

Shoo

t!

Roo

t!

Silks

Pericarps

P1-ww 25 DAP

P1-rr 25 DAP

P1-ww 14 DAP

P1-rr 14 DAP

P1-ww

P1-rr

Leaf

Shoot

Root

Silks Pericarps


P1-rr/P1-ww > 1

Pericarps 25 DAP

Pericarps 14 DAP

54 0

Silks

4

1 2

43

68

P1-rr/P1-ww < 1

55

0

Pericarps 25 DAP

Pericarps 14 DAP

Silks

5 72

1 4

52

Pericarps 25 DAP

Pericarps 14 DAP

109 0

Silks

20

7

120

8

115

P1-rr/P1-ww P-value < 0.02

A

B C

D

Supplemental Figure 4. Control of the expression of transcription factor genes by P1. A. Venn diagram representation showing comparisons of differentially expressed transcription factors between P1-rr and P1-ww pericarps (14 and 25 DAP) and silks. B-C. Venn diagram representations showing comparisons of higher (B) or lower (C) levels of expression of expression of transcription factors between P1-rr and P1-ww pericarps (14 and 25 DAP) and silks. D. Frequency histogram of the observed range of expression of transcription factors in 14 DAP pericarps at 14 DAP, selecting the expression value observed between P1-rr and P1-ww. The x-axis corresponds to the log10 transformed FPKM. The P1 FPKM corresponds to 7.91 (log10 = 0.899).

Freq

uenc

y

Log10(FPKM)


K1 K2 K3 K4

K5 K6 K7 K8

Silk

s P

1-rr

/P1-

ww

Per

icar

ps 1

4 D

AP

P1-

rr/P

1-w

w

Per

icar

ps 2

5 D

AP

P1-

rr/P

1-w

w

Silk

s P

1-rr

/P1-

ww

Per

icar

ps 1

4 D

AP

P1-

rr/P

1-w

w

Per

icar

ps 2

5 D

AP

P1-

rr/P

1-w

w

Silk

s P

1-rr

/P1-

ww

Per

icar

ps 1

4D A

P P

1-rr

/P1-

ww

Per

icar

ps 2

5D A

P P

1-rr

/P1-

ww

Silk

s P

1-rr

/P1-

ww

Per

icar

ps 1

4 D

AP

P1-

rr/P

1-w

w

Per

icar

ps 2

5 D

AP

P1-

rr/P

1-w

w

log 2

(P1-

rr/P

1-w

w)

log 2

(P1-

rr/P

1-w

w)

Supplemental Figure 5. Box plots corresponding to expression distribution within the K1 - K8 clusters. Box plots depict the activation and repression patterns corresponding to clusters K1 to K8 for silks, pericarps 14 DAP and pericarps 25 DAP. Relative expression for each cluster was obtained by calculating the log2 of the ratio between P1-rr and P1-ww FPKM values.


A

B

C

Luc -1249 +10 Luc

-1500 +1

Luc -438 +1

pC2:Luc

pWHP1:Luc

pA1:Luc

pA1*:Luc

Roo

t

Sho

ot

Leaf

P1-ww

Silk

s

Per

icar

ps

14 D

AP

Per

icar

ps

25 D

AP

P1-rr

Silk

s

Per

icar

ps

14 D

AP

Per

icar

ps

25 D

AP

D

CCAACC GGTTGG -1062 -1057 -1090 -1020 -95 -89

+ - + -

-171 -71

WHP1

Supplemental Figure 6. Control of flavonoid biosynthetic genes by P1. A. Heat map showing the expression patterns of the chalcone synthase genes C2 and WHP1, the chalcone isomerase gene CHI1 and other putative CHI genes, and the dihydroflavonol reductase genes A1 and A1* across various tissues and genotypes. B. Diagrams of the constructs used for transient expression experiments in maize protoplasts for C2 (pC2:Luc), WHP1 (pWHP:Luc), A1 (pA1:Luc) and A1* (pA1*:Luc). C. Transient expression assays using maize protoplasts transformed with pA1:Luc, pA1*:Luc, pWHP:Luc and pC2:Luc. For each experiment, protoplasts were also transformed with p35S:P1 or p35S:C1 + p35S::R constructs in addition to p35S:Renilla. Each treatment was done in triplicate, and the data were normalized for Renilla activity as described (Grotewold et al., 2000). The fold activation was calculated as the ratio between each particular treatment and the treatment with the promoter constructs without activator. The average values are shown and the error bars indicate the standard deviation of the samples. D. Electrophorectic mobility shift assays (EMSA) showing binding of the MYB domain of P1 (+) expressed and affinity-purified from bacteria as a poly-histidine fusion (NHis6-P1MYB, (Xie et al., 2010)) to the indicated binding sites on the WHP1 5’ regulatory region. Lanes indicated by “-“ correspond to DNA-binding reactions performed in the absence of NHis6-P1MYB.

GRMZM2G422750 GRMZM2G151227 GRMZM2G155329 GRMZM2G175076 GRMZM2G119186 GRMZM2G026910 GRMZM2G013726

C2 WHP1 CHI1 Putative CHI-like gene Putative CHI-like gene A1 A1*

-423 +1 Luc

!" #!" $!!" $#!" %!!" %#!" &!!" &#!"

'()*$++,-."

'/%++,-."

'0$1++,-."

'0$++,-."

Fold Activation

*$"

/$23"

0 300 FPKM


pA1:Luc

pA1*:Luc

pC2:Luc

pWHP1:Luc

A

B m/z


D

C

Supplemental Figure 7. LC-MS analysis of Zm F2H1 activity products and phylogenetic analyses of amino acid sequences of cytochrome P450 enzymes. A. Fragmentation of peaks 1, 2 and 3 produced by Zm F2H1. For comparison, the fragmentation patterns of standards are shown on the middle top. Naringenin, apigenin and 2-hydroxynaringenin produced molecular ions of m/z+ 273, 271 and 289, respectively. B. Proposed fragmentation sites on the structure of 2-hydroxynaringenin. C. Diagram of the molecular structure of Zm F2H1, CYP93G1 (Oryza sativa), CYP93G3 and CYP93G4 (Sorghum bicolor) genes. Grey boxes indicate coding regions and black lines represent non-coding 5’ and 3’ untranslated regions (UTRs). Triangles represent the position of introns. D. Phylogenetic tree of deduced amino acid sequences of selected CYP93 family proteins. The tree was constructed using MEGA 4.0 Software with the Neighbor-Joining method based on ClustalW multiple alignments. The numbers indicate bootstrap values (10,000 replicates). Bar = 0.1 amino acid substitutions per site. The sequences used are shown in Supplemental method.


ATG TGA CYP93G5 (Zm F2H)

CYP93G3

CYP93G1

CYP93G4 0.4 kb

ATG TGA

ATG TGA

ATG TGA

P1

Histone H3

37 kDa

15 kDa

P1-rr

14 DAP 25 DAP

P1-ww

14 DAP 25 DAP

Supplemental Figure 8. P1 protein expression is highest in P1-rr 14 DAP pericarps. Immunoblot performed on P1-rr and P1-ww nuclear protein extracts from 14 and 25 DAP pericarps using the P1 polyclonal antibodies (!P1344) that recognize the non-conserved C-terminal region of P1 {Falcone Ferreyra, 2010 #5564}. The image shows higher levels of P1 protein accumulation at 14 DAP. Histone H3 antibodies were used as a loading control.


P1-ww P1-rr

Input !P1 Input !P1 Input !P1 Input !P1 14 25 14 25

P1-rr P1-ww

A C A C

Chr 2: 94,364,710 – 94,364,910

Chr 10: 270,299,65-27,030,165

Chr 1: 146,119,994-146,120,194

Chr 1: 137,410,708-137,410,908

Chr 1: 71,793,632-71,793,832

Chr 6: 39,234,585-39,234,785

Chr 4: 82,121,654-82,121,943

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

Chr 9: 84,480,315-84,480,515

Chr 7: 87,955,374-87,955,777

Chr 4: 198,110,639-19,8111,042

Chr 4: 96,716,525-96,716,725

Chr 5: 90,862,987-90,863,187

Chr 4: 100,034,141-100,034,341

Chr 1: 285,671,225-285,671,425

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

MACS_peak_5326

MACS_peak_3102

MACS_peak_1502

MACS_peak_1390

MACS_peak_698

MACS_peak_14124

MACS_peak_9774

MACS_peak_14201

MACS_peak_11756

MACS_peak_7980

MACS_peak_7157

MACS_peak_8948

MACS_peak_7196

MACS_peak_1995

A


P1-ww P1-rr

Input !P1 Input !P1 Input !P1 Input !P1 14 25 14 25

Chr 1: 215,569,996-215570,196

Chr 1: 119,327,747-119,328,373

Chr 1: 58,068,957-58,069,157

Chr 9: 135,600,120-135,600,320

Chr 7: 97,066,017-97,066,279

Chr 4: 109,863,422-109,863,966

Chr 2: 89,914,635-89,915,179

Chr 1: 299,214,928-299,215,138

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

P1-rr P1-ww

A C A C

MACS_peak_1533

MACS_peak_834

MACS_peak_370

MACS_peak_14517

MACS_peak_11806

MACS_peak_7320

MACS_peak_3780

MACS_peak_2075


A (continued)

Supplemental Figure 9. Validation of P1 ChIP-Seq results by ChIP-qPCR. Genome viewer representation of random peaks (MACS_peak_xxxx) chosen for validation are represented in a genome viewer showing their coordinate locations in the genome, and the position of the peaks as obtained from the ChIP-Seq A and C experiments, comparing P1-rr and P1-ww samples. A. Peaks were validated by ChIP-qPCR and images of the agarose gels compare input and !P1 for P1-rr and P1-ww at 14 and 25 DAP. B. Peaks were validated by ChIP-qPCR, and the fold enrichment was calculated using the Copia and Actin genomic regions as controls. Error bar indicates standard error.

P1-rr

P1-ww

A C A C

P1-rr

P1-ww

A C A C

P1-rr

P1-ww

A C A C

P1-rr

P1-ww

A C A C

P1-rr

P1-ww

A C A C

P1-rr

P1-ww

A C A C

P1-rr

P1-ww

A C A C

P1-rr

P1-ww

A C A C

MACS_13868

MACS_3153

MACS_1674

MACS_13868

MACS_17371

MACS_1148

MACS_16278

Actin

Copia P1-ww 14 DAP P1-rr 14 DAP

Fold enrichment to P1-ww

n=3, *; P < 0.05

*

*

*

*

B


Supplemental Figure 10. Motif enrichment in genome-wide location analysis peaks in P1-controlled genes. A. Top motif identified as enriched by MEME in peaks from genes up- (UP) or down- (DOWN) regulated by P1. B. Frequency distribution of sequences fitting the P1-binding consensus CC(T/A)ACC in experimental peaks from genes up- (UP) or down- (DOWN) regulated by P1. The frequency bars show the number of peaks fitting the consensus in 1,000 permutations of randomly chosen sequences; number and length corresponding to those in each of the experimental peak sets (227 for UP and 471 for DOWN). The P-value reflects the statistical likelihood that the observed number is different than the expected number.

A

P = 1.68x10-3

Number of P1 binding consensus motif

Observed number=75

Freq

uenc

y

P=7.36x10-5

Number of P1 binding consensus motif

Observed number=155

Freq

uenc

y

UP DOWN

B

UP

E-value = 3.0 x 10-41

DOWN

E-value = 8.0 x 10-41


MGF, Maize Genome FGS 5b60 RP, RNA-Seq Pericarps RS, RNA-Seq Silks MMF, Maize Genome FGS 5b60 Microarray MP, Microarray Pericarps MS, Microarray Silks

7909

2314

3901

17322

3741

4886

MGF

RP MMF

MP

7545

3287

3484

17950

2504

4886

MGF

RS MMF

MS

A. Pericarps

B. Silks

Supplementary Figure 11. Venn-diagrams summarizing the overlap between the genes expressed in pericarp and silks as detected with RNA-Seq and microarrays. Overlap between RNA-Seq and microarray (Sekhon et al., 2011) in silks and pericarps tissues. A. In pericarps, 2,314 genes detected by microarrays were not present in the RNA-Seq experiments, and 7,642 genes were detected by RNA-Seq, but not by microarrays. B. In silks, 3,287 genes detected in microarray experiments failed to be detected by RNA-Seq, and 5,988 genes were detected only by RNA-Seq.


Supplemental Figure 12. Phylogenetic analyses of UDP-sugar:glycosyltransferases from higher plants. The tree was constructed using MEGA 4.0 Software with the Neighbor-Joining method based on ClustalW multiple alignments (Tamura et al. 2007). The numbers indicate bootstrap values (10000 replicates). Bar = 0.1 amino acid substitutions per site. The following sequences, in addition to Zm UGTs, were analyzed: UGT78D2 (Arabidopsis thaliana, NP_197207), UGT78D3 (Arabidopsis thaliana, NP_197205), UGT78D1 (Arabidopsis thaliana, NP_197205), Vv GT1 (Vitis vinifera, AAB81683), UGT78A2 (Ariata cordata, AB103471), Ph F3GlcT (Petunia hybrida, AAD55985), Vv GT5 (Vitis vinifera, BAI22846), Vv GT6 (V. vinifera, BAI22847), UGT78G1 (Medicago truncatula, A6XNC6) Zm 3GlcT (Zea mays, X13501), Th 5GT (Torenia hybrida, AB076698), Pf 5GlcT (Perilla frutescens, BAA36421), UGT75C1 (Arabidopsis thaliana, Q0WW21), Ph A5GT (Petunia x hybrida BAA89009.1), Vh 5GlcT (Verbena x hybrida, BAA36423), UGT72B1 (OsCGT (Oryza sativa, ABC94602.1) UGT72B1 (A. thaliana, Q9M156), UGT706D1 (O. sativa, BAB68093), UGT707A3 (Oryza sativa, BAC83989) UGT71G1 (Medicago truncatula, AAW56092), UGT71F1 (Beta vulgaris, AY526081), Fa GT6 (Fragaria x ananassa, DQ289587), UGT89C1 (A. thaliana, Q9LNE6), UGT73J1 (Allium cepa, AY62063), Ac UGT73G1 (A. cepa, AY62062), UGT73C6 (A. thaliana, AEC09298), UGT73C8 (Medicago truncatula, DQ875459), Sb UF7GT, (Scutellaria baicalensis, AB031274),Nt 7GlcT (Nicotiana tabacum, AF346431), Fa GT7 (Fragaria x ananassa DQ289588), Db 7GlcT (Dorotheanthus bellidiformis, Y18871), UGT73A4 (Beta vulgaris, AY526080), Ph 1-6RhaT (P. hybrida, CAA50376), UGT79G16 (Ipomoea purpura, AB192315), Cm 1-2RhaT (Citrus maxima, AY048882), UGT94B1 (Bellis perennis, AB190262), Cr UGT3 (Catharanthus roseus, AB443870), UGT94D1 (Sesamum indicum, BAF99027). Different glycosyltransferase groups are clustered in circles based on the glycosylation position in the acceptors.


Fold enrichment to P1-ww

F2H1 (GRMZM2G167336)

A1 (GRMZM2G026930)

P1-ww 14 DAP P1-rr 14 DAP

n=6, **; P < 0.01, *; P < 0.05

Copia

Actin

**

**

* C3H

(GRMZM2G138074)

Supplemental Figure 13. P1 binds in vivo to F2H1 and CYP98A. ChIP-qPCR was performed as described in Supplemental Figure 9, with the A1 gene used as a positive control. Error bar indicates standard error(n=6).


Morohashi et al. (2012). P

lant Cell 10.1105/tpc.111.098004

Supplem

ental Data Table 1 S

umm

ary of RN

A-S

eq

Total #readsA

ligned to genesA

ligned to intergenic regions

Alignm

ent %N

onalignedC

ounting N

onaligned

(for aligned reads) S

equence R

edundancy

Redundancy

%

Pericarp

P1-rr at 14 D

AP

4,266,6413,345,592

360,18086.9

560,869757,020

3,317,36477.8

P1-rr at 25 D

AP

5,201,2864,198,256

517,42290.7

485,608765,901

4,198,05880.7

P1-w

w at 14 D

AP

5,455,2754,443,582

542,91591.4

468,778768,466

4,224,72177.4

P1-w

w at 25 D

AP

4,310,1423,413,903

410,50488.7

485,735703,424

3,390,71978.7

Silks

P1-rr

7,864,6395,847,662

478,84580.4

1,538,1321,706,601

3,589,90045.6

P1-w

w6,865,542

4,830,732417,880

76.41,616,930

1,775,4643,793,081

55.2


Supplemental Data Table 2 Summary of ChIP-Seq

Replicate P1-ChIP Total reads Unique-match Peak (P<10-4) A P1-ww 14 DAP 851,391 33,121

P1-rr 14 DAP 1,953,507 316,134C P1-ww 14 DAP 4,287,415 1,286,130

P1-rr 14 DAP 3,006,796 797,594Total P1-ww 14 DAP 5,138,806 1,319,251

P1-rr 14 DAP 4,960,303 1,113,728All 10,099,109 2,432,979

35,143

20,424

14,719


Supplemental Data Table 3 Summary of validated peaks

Replicate Peaks in either A or C Peaks in both A and CA 86% (6/7) C 80% (8/10) 100% (5/5)

Supplemental Data Table 4 Primers used in this paperName Sequence (5' to 3')STRS1-F CAGGTGCTTGACATCTTGGASTRS1-R AGCTTCCCGTCTTTCTCCTCJAC1-F CCTGCTCTAAAACCGACGACJAC1-R GCCTTCCCATTCTGTTGATGWHP-F GACCCGACCTACGAATAATGWHP-R AACTTGAGCTGGTGCACTGNAC-F CAAGAAGAACGAGTGGGAGAAGNAC-R GTCTGAGTGGGAGTGCGAGAGMATE-F CTCTGCCTCGAGACCTGGTAMATE-R GACAGAGAGTGAGGCCAAGGLAC10-F GGTAGCCGTTTTCATCGTAGAGLAC10-R AGGCCTCCTTTGTTAGATCCACPR1-F GGTAGGAGGAGCAGCTAGATTGPR1-R ACTGGTGGCGAGGGTGTAGTF2H-F CGGTCCATCCAAATTCAGF2H-R ACCAACATCGAACGGGTARHM-F ACTTTACTTTTGGGCTGTCGRHM-R TCGACCTCCTCTGCTGTTCCCR-F CTCCTGCTCCTGTCCTACCAGCCR-R CCTCCTCTCACCTTGTTCAGPAL1-F GTCTCGACTCTCCACACCACPAL1-R GGAGAGAACCAGCAGCAGTGPAL2-F GCCTCAGTGCCTCACCTAAGPAL2-R GGGCGAGGCGGTTATATAGGPAL3-F GGCTGCCATCCTATCCTATCCPAL3-R CCACCACTCACCTTGCTACAGUGT1-F GAGGAGCAGATTCGGTGAGCUGT1-R CGACTGACGACAGTGTCTGGUGT2-F CTCGTGAGGGAGATGATGACCUGT2-R CTGACTTGAGCCAGCCTCTTCUGT3-F ACTGGGCCTAGGCTAGACTGCUGT3-R GACCACCACAGTGGGGTATGUGT4-F AGTACGGCCATCCTCTGGTCUGT4-R CACCTTGACTCCGACCTTCCUGT5-F TGATTTCTGCGAGCCTGTUGT5-R AAGAAACGAGTGCGTGGARHM1-F GCACCAACCCTCTCTCTGAARHM1-R ATAAATGCAAGCGGTGATGGWHP1-F AAACCCACGTACTGGCAAACWHP1-R TGGCCATTATCCTGAACACAWHP2-F GCATCAGTGAGGTGCACAATWHP2-R GACATCAATTGACCCGCAAGP1_peak_7157-B1 AGTTGGTGGACGACCTGAAGP1_peak_8948-A1 GCTGATGATATCGGCGTTTTP1_peak_8948-B1 CCGACATCACCATCAGTCACP1_peak_7196-A1 GTTTCGGCTTTTGACTGCTCP1_peak_7196-B1 CCGGCCTAGGTACATTGAAAP1_peak_1995-A1 TGGTGCAGTCTCTCCAAGTGP1_peak_1995-B1 CACCCTTTTTCTTGCGATGTP1_peak_1533-A1 AACCCCATCTATGCAAGCACP1_peak_1533-B1 TCATCCATTCTTACTCGCCCP1_peak_834-A1 TCCTGTTTCTTGGAGGATGGP1_peak_834-B1 ATTGCCTTCGATTCATCCAGP1_peak_370-A1 TAAGGTGGCGAAAATTGAGGP1_peak_370-B1 ATTGAGCAATAAAATGGCCGP1_peak_14517-A1 ATAGCGGGATAGCTGCAGAAP1_peak_14517-B1 GAGTCCTACCTCACCACCCAP1_peak_11806-A1 AGGCAAATACAACCACCACCP1_peak_11806-B1 GACGACAGTGGTTCTAGGGCP1_peak_7320-A1 CTGCAAAAGTGGCAGCAATAP1_peak_7320-B1 GAGATCTTCAAGGGCCTTCCP1_peak_3780-A1 AAGTTGTCCATGGCTTCACCP1_peak_3780-B1 ACGTGACGGGTAAAATGCTCP1_peak_2075-A1 CAACCCTTCCGTGATTGAGTP1_peak_2075-B1 AAGACGTGCTACGACGAGGTqrt MACS_13868-A1 GGCGAACGGGCCAAAAACGGqrt MACS_13868-B1 CTTCCACGTCTCACGCGGGGqrt MACS_3153-A1 TGCAGCCGAATGACCACATGTCGqrt MACS_3153-B1 AGGTGCAGTTGAAGTGGCTTTCTTqrt MACS_1674-A1 TGATCAATTGCCACACAACCCCCA

Primers for probe construction

Primers for qRT-PCR

Primers for ChIP-PCR

Primers for ChIP-qPCR


qrt MACS_1674-B1 TCCGTAGGTGTTAGCCTTCGGTqrt MACS_13868-A1 GGCCGTGGCTGAACCGTTGTqrt MACS_13868-B1 GCCGTTCGGCCTTGCGTTTGqrt MACS_17371-A1 GAAGCCCTCTGTGAGCAGACqrt MACS_17371-B1 CACAACCACCCCACTGTGACqrt MACS_1148-A1 GTCCTGCAGCAACACCTGTCqrt MACS_1148-B1 GCTCCAGGGACTAGCGGTATqrt MACS_16278-A1 GGGTCTACGAGCAACAGGAGqrt MACS_16278-B1 GGCCAGCTCTCTGCAGGTTAqrt A1peak-A1 ACCCAAACCAAGCAAACAAGqrt A1peak-B1 ATCCGATCAAAGTCCCCAATqChIP-ZmCopia-F CGATGTGAAGACAGCATTCCTqChIP-ZmCopia-R CTCAAGTGACATCCCATGTGTqChIP-ZmAct1-5UTR-F TTTAAGGCTGCTGTACTGCTGTAGAqChIP-ZmAct1-5UTR-R CACTTTCTGCTCATGGTTTAAGGMIC_F2H_ChIP_L-5 CTCACAACGAGTAGGCATCAMIC_F2H_ChIP_R-5 TCAAGGTGACCAAAGTACAGG qrt CYP98A_MACS8458-A5 GTACAAGGGTGCTGAGTGCCqrt CYP98A_MACS8458-B5 ATCCGCCACAACATTTAAGGSLX-R_P1 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTSLX-R_P2 GATCGGAAGAGCTCGTATGCCGTCTTCTGCTTGSLX-11_P1 AATGATACGGCGACCACCGACACTCTTTCCCTACACGACGCTCTTCCGATCTAATCGTSLX-11_P2 CGATTAGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTCGGTGGTCGCCGTATCATTSLX-12_P1 AATGATACGGCGACCACCGACACTCTTTCCCTACACGACGCTCTTCCGATCTAACACTSLX-12_P2 GTGTTAGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTCGGTGGTCGCCGTATCATTSLX-13_P1 AATGATACGGCGACCACCGACACTCTTTCCCTACACGACGCTCTTCCGATCTAACGTTSLX-13_P2 ACGTTAGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTCGGTGGTCGCCGTATCATTSLX-14_P1 AATGATACGGCGACCACCGACACTCTTTCCCTACACGACGCTCTTCCGATCTAACTGTSLX-14_P2 CAGTTAGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTCGGTGGTCGCCGTATCATT

Primers for ChIP-qPCR

Primers for ChIP-Seq library

14 dap 25 dap - the plant cell · 2012. 7. 3. · morohashi et al. (2012). plant cell...

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