SUPPLEMENTARY INFORMATION - Nature Research...T1W2 3 4567 091 0 20 40 60 80 100 TE-siR815b-oe8 T 1 plants TE-siR815b-oe12 T 1 plants 8 11 12 a b TE-siR815 0 10 20 50 Relative expression

SUPPLEMENTARY INFORMATIONDOI: 10.1038/NPLANTS.2016.16

NATURE PLANTS | www.nature.com/natureplants 1

Supplementary Figure 1. The osa-miR815 small RNA family in the microRNA database miRBase (http://www.mirbase.org). a, The predicted loci for generating osa-miR815 in rice genome. Arrow indicates a protein-encoding gene and transcription orientation. b, The predicted secondary RNA structures of the osa-miR815 loci.

25Mb

0Mb

10Mb

15Mb

20Mb

5Mb

Chr7

LOC_Os07g10110osa-miR815c

Chr5

LOC_Os05g51800osa-miR815a

osa-miR815b/TE-siR815bWRKY45-1

a

osa-miR815a

osa-miR815c

b

osa-miR815b/TE-siR815b

Transposon-derived small RNA is responsible for modified function of WRKY45 locus

Transposon-derived small RNA is responsible formodified function of WRKY45 locus

http://dx.doi.org/10.1038/nplants.2016.16

2 NATURE PLANTS | www.nature.com/natureplants

SUPPLEMENTARY INFORMATION DOI: 10.1038/NPLANTS.2016.16


osa-miR815a

osa-miR815c

Supplementary Figure 2. Comparison of rice osa-miR815 precursor sequences of osa-miR815a, osa-miR815b/TE-siR815b, and osa-miR815c. The osa-miR815 is underlined based on miRBase(http://www.mirbase.org). a, The predicted fold-back structures of rice precursors osa-miR815a, osa-miR815b/TE-siR815b, and osa-miR815c. b, Comparison of the DNA sequences of osa-miR815 precursors with WANDERER_OS -type transposon sequence from RepeatMasker.

a

b




WRKY45-1ATG TGA

3’5’

5.52

WANDERER_OSosa-miR815b precursorUTRexon

intron

Small RNAs in the callus of rice variety Nipponbare

5.154.785.15

3.683.6816.5

4.416.997.72

27.66.99

0.0380.038

0.038

0.0380.038

0.0380.038

0.038

0.038 Small RNAs in the leaves of rice variety Mudanjiang 8 at four-leaf stage (before bacterial inoculation)

2425

142619276

2829

303132

6

1

23

45

78

9

0.0985

0.09880.098 9

Small RNAs in the leaves of rice variety Mudanjiang 8 at four-leaf stage (1 day after inoculation of Xoo strain PXO61)

0.033 100.033 11

0.033 120.033 13

0.033140.033 15

0.065 160.033 17

0.06518

0.033190.0336

0.033 200.033 21

0.03322

0.03323

Supplementary Figure 3. Small RNAs identified in osa-miR815b/TE-siR815b locus by deep sequencing. Orange, red, blue, light blue, purple, grey, and green arrows indicate 25-, 24-, 23-, 22-, 21-, 20-, and 19-nt small RNA, respectively. Left-pointing arrows indicate small RNA or PCR amplification region in which sequences are identical to the bottom sequence of the fold-back precursor. Some (1, 2, 10, 11, and 12) right-pointing arrows indicate small RNA in which sequences are identical to the top sequence of the fold-back precursor, and the other right-pointing arrows indicate small RNA or PCR amplification region in which sequences are complementary to the bottom sequence of the fold-back precursor. The figures on the arrow tails are reads per million (RMP) sequences in the high-throughput sequencing data. The figures on arrow heads indicate different small RNA or PCR amplification region with different sequences. The data of Nipponbare callus were collected from miRBase (http://www.mirbase.org) and only small RNA with RPM larger than 3.5 are presented.

osa-miR815 in miRbase33

Predicted quantitative PCR amplification region33

34





Supplementary Figure 4. Accumulation of osa-miR815/TE-siR815 was increased in Dongjin but not in Minghui 63 after Xoo infection. Rice variety Dongjin carries osa-miR815/TE-siR815 locus and variety Minghui 63 does not carry osa-miR815/TE-siR815 locus. Data represent mean (3 replicates) standard deviation.

0

2

4

6

8

10

ck 12 24 48

WRKY45-1

Time after infection (hour)

0.0

0.5

1.0

1.5

2.0

ck 12 24 48

WRKY45-2R

elat

ive

expr

essio

n le

vel

0

2

4

6osa-miR815/TE-siR815

0.0

0.5

1.0

1.5

Dongjin Minghui 63

Time after infection (hour)

osa-miR815/TE-siR815




24nt21nt

TE-siR815(probe 1)

callus panicle leafshoot

tRNA5S rRNA

TE-siR815(probe 3)

24nt21nt

Supplementary Figure 5. Expression of TE-siR815 in different tissues of rice variety Minghui 63 analyzed by PAGE-northern. Tissues other than callus were collected at the booting stage. Minghui 63 only carries TE-siR815a and TE-siR815c loci. Probe information is presented in Fig. 1a.




Supplementary Figure 6. Expression of WRKY45-1 and WRKY45-2 in transgenic plants. a, The WRKY45-1-oe and WRKY45-2-oe family plants showed segregation of overexpressing WRKY45-1 and WRKY45-2, respectively. Expression was examined by Northern blot analysis. Leaf samples were collected at the booting stage. WT, wild-type Mudanjiang 8; N, negative transgenic plants segregated from the families; hybridization probe W45-CDS1, a 632-bp cDNA fragment flanking the first intron of WRKY45-1obtained by PstI digestion of WRKY45-1 cDNA clone; hybridization probe W45-CDS2, a 437-bp cDNA fragment harboring partial coding sequence and partial 3’-untranslated region obtained by PstI digestion of WRKY45-1 cDNA clone; hybridization probe W45-intron, a 79-bp fragment amplified from the first intron of WRKY45-1; rRNA, ribosomal RNA. b, Control analysis of hybridization probe W45-intron by Southern blot method. WRKY45-1, WRKY45-2, and WRKY45-1.2, PCR fragments used for generating WRKY45-1-oe, WRKY45-2-oe, and WRKY45-1.2-oe plants, respectively.

WT

1 2 N

WRKY45-2-oe4 T2 family

WRKY45-1-oe10 T2

family

4 5 N 2 7 N1 2 N 1 N

WRKY45-1-oe11 T2

family

WRKY45-2-oe6 T2 family

WRKY45-1.2-oe4 T2

family

WRKY45-1/WRKY45-2(Hybridization probe W45-CDS2)

6000500040003000

2000

1000

500

rRNA

Hyb

ridiz

atio

n pr

obe W

45-in

tron

Marker/Base

2000

1000

500

34724254

2690

1882

WRK

Y45

-1

WRK

Y45

-2

WRK

Y45

-1.2

Hybridization probe W45-intron

(Southern blot)

Marker/Base pair

6000500040003000

a

b

Hyb

ridiz

atio

n pr

obe W

45-in

tron

2000

1000

500

6000500040003000

1 N

WRKY45-1.2-oe13 T2

family

WRKY45-1/WRKY45-2(Hybridization probe W45-CDS1)

2000

1000

500




Supplementary Figure 7. Enhanced resistance of WRKY45-1.2-oe plants was associated with significantly increased expression of WRKY45-1 and ST1, but not TE-siR815. Rice plants were inoculated with Xoo strain PXO61 at the booting stage. Data represent mean (3–5 replicates for disease area and 3 replicates for gene expression) standard deviation. A significant difference was detected between wild-type (WT) plants and WRKY45-1.2-oe plants at **P < 0.01 or *P < 0.05.

02468

WRKY45-1

0

20

40

60ST1

00.5

11.5

2TE-siR815

02468

Rel

ativ

e ex

pres

sion

leve

lWRKY45-1

05

10152025

ST1

00.5

11.5

2 TE-siR815

WT 5 6 9 10 14 15 WT 1 2 3 9 11 125

WRKY45-1.2-oe4 T1 family (D220UM4)

OsWRKY45-1.2-oe13T1 family (D220UM13)

Dise

ase

area

(%)

0

20

40

60

80

0

20

40

60

80PXO61 infection PXO61 infection

** ** **** ** ** **

****

** ** ** **

**

**

**

****

*

*

* * * *

****

** **

**

**

**

****

** **

**

**




Supplementary Figure 8. Activation of TE-siR815b did not change rice response to Xoo. Plants were inoculated with Xoo at the booting stage. Data represent mean (3–5 replicates for disease area and 3 replicates for expression) standard deviation. a, Overexpressing TE-siR815b did not influence rice response to Xoo strain PXO61. WT, wild-type Mudanjiang 8. b, Overexpressing TE-siR815b did not influence rice plant response to Xoo strain PXO347. WT, wild-type Zhonghua 11.

0

20

40

60

80

WT 1 2 3 4 5 7 8 9 10

Dise

ase

area

(%)

WT 1 2 3 4 5 6 7 9 10

0

20

40

60

80

100

TE-siR815b-oe8 T1 plants TE-siR815b-oe12 T1 plants

8 11 12

a

b

TE-siR815

0

10

20

50

60

Rel

ativ

e ex

pres

sion

leve

l

PXO61 infection PXO61 infection

TE-siR815

0

4

8

12

16

TE-siR815

0

10

20

30

40

Rel

ativ

e ex

pres

sion

leve

ls

WT 2 3 6 7 8 11 12 2 3

TE-siR815b-oe6 (ZH11) T1 plants

4 6 7 8 9 10

TE-siR815b-oe8 (ZH11) T1 plants

***

**

**

** **

**

** ** **

****

** **

** **

*

* *

***

*

***

**

* *

*

*

**

**

****

**** **

0

20

40

60

80

100

Dise

ase

area

(%) PXO347 infection

TE-siR815b-oe

GUS




Supplementary Figure 9. Overexpression of WRKY45-1 and WRKY45-2 influenced rice response to Xoo. Plants were inoculated with Xoo strain PXO347 at the booting stage. Bar represents mean (3–5 replicates) standard deviation. A significant difference was detected between wild-type (WT) Zhonghua 11 (ZH11) and transgenic plants at **P < 0.01 or *P < 0.05. WT carries endogenous WRKY45-1. Positive transgenic plants were determined by PCR amplification of WRKY45-1-oe or WRKY45-2-oe construct using two pairs of primers, TEV/W45-R6 and W45-F6/W45-R6 (Supplementary Table 1). a, Increased susceptibility to Xoo was associated with the existence of WRKY45-1-oe construct in WRKY45-1-oe (ZH11) T1 families. b, Enhanced resistance to Xoo was associated with the existence of WRKY45-2-oe construct in WRKY45-2-oe (ZH11) T1 families.

0

20

40

60

80

100

Lesio

n ar

ea (%

)

WRKY45-1-oe7 (ZH11) T1 family

WT

1 2 4 5 6 7 8 9 11 12 13 14 15

0

20

40

60

80

100

Lesio

n ar

ea (%

)

WRKY45-2-oe4 (ZH11) T 1 family

WT

1 2 3 4 5 6 7 8 9 10 12 13 15 16 17 18 19 20

0

20

40

60

80

100

Lesio

n ar

ea (%

)


WT

1 2 3 4 5 6 7 8 10 12 13 14

b

**

**

****

**


WT

1 2 3 4 5 7 8 9 11 13 14 15

0

20

40

60

80

100

Lesio

n ar

ea (%

)

a** ** ** **

**

*

WRKY45-1_oe

WRKY45-1_oe

* * * **** *

** ** ****

****** ****

**

*

***

****

**

*

** ****

**** **

**

WRKY45-2_oe

WRKY45-2_oe




Supplementary Figure 10. Overexpressing TE-siR815b blocked resistance to Xoo conferred by WRKY45-1.2 and WRKY45-2. Plants were inoculated with Xoo PXO61 or PXO347 at the booting stage. Data represent mean (3–5 replicates for disease area and 3 replicates for expression) standard deviation. A significant difference was detected between wild-type (WT) and other plants at **P < 0.01 or *P < 0.05. a, The F2 plants (1, 2, 3, 6, 8, and 9) overexpressing both WRKY45-1.2and TE-siR815b had increased susceptibility to Xoo compared with WRKY45-1.2-oe plants. WT, Mudanjiang 8. b, The F2 plants (1, 3, 4, 7, 9 and 11) overexpressing both WRKY45-2 and TE-siR815b had increased susceptibility to Xoo compared to WRKY45-2-oe plants. WT, Zhonghua 11 (ZH11).

1 2 3 4 5 6 7 8 9 10

WRKY45-1.2-oe4/TE-siR815b-oe8-1 F2 plants

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20

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100

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(%)

TE-siR815b-oe8 (ZH11)/WRKY45-2-oe12 (ZH11)-2 F2 plants

Dise

ase

area

(%)

TE-siR815b-oe

WRKY45-1.2-oe

PXO61 infection

PXO347 infection

a

b

WT

WRK

Y45

-1.2

-oe4

TE-s

iR81

5b-o

e8

*

**

**

**

****

*

TE-siR815b-oe

WRKY45-2-oe

WT

TE-s

iR81

5b-o

e8 (Z

H11

)

WRK

Y45

-2-o

e12

(ZH

11)

** ** **




WRKY45-1-oeWRKY45-2-oe

1 7 8 9

WRKY45-1-oe7 (ZH11)/WRKY45-2-oe12 (ZH11)

F1 plants

0

20

40

60

80

Supplementary Figure 11. The positive F1 plants (1 and 9) overexpressing both WRKY45-1 and WRKY45-2 were susceptible to Xoo as WRKY45-1-oe plants. WT, wild-type Zhonghua 11.

Dise

ase

area

(%)

WT

WRK

Y45

-1-o

e7 (Z

H11

)

WRK

Y45

-2-o

e12

(ZH

11)

****

0

6

12

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24ST1

****

**

WRKY45

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** ** ** **R

elat

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expr

essio

n le

vel

0

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18

24TE-siR815

** ******

PXO347 infection




Supplementary Figure 12. Comparison of the DNA sequences of TE-siR815 precursor TE-siR815b with the predicted WANDERER_OS-type transposon in the second intron of ST1. Blue color indicates ST1 region showing changed DNA methylation. MT01-ME2F and MT01-ME1R are primers (Supplementary Table 1) used in DNA methylation analysis of ST1.

ST1 intron

ST1 intron

ST1 intron

ST1 intron

ST1 intron

ST1 intron

TE-siR815b

ST1 intron

TE-siR815b

ST1 intron

ST1 intron

MT01-ME2F

MT01-ME1R




a CG

020406080

100

Met

hyla

ted

(%)

WT WT1 10 11 4 6WRKY45-1-oe WRKY45-2-oe

CHG

0

10

20

30

WRKY45-2-oe

WT

WT1

10

11

4

WRKY45-1-oe

6

Supplementary Figure 13. DNA methylation in the intron region of ST1. WT, wild-type Mudanjiang 8; WT1, negative plant from WRKY45-1-oe family. a, WRKY45-1-oe and WRKY45-2-oe plants showed similar levels of CG and CHG contexts of DNA methylation levels. b, The CHH context of DNA methylation level was differently regulated in WRKY45-1-oe and WRKY45-2-oe plants. The circle and solid circle represent non-methylated and methylated cytosine, CG (red), CHG (blue), and CHH (green), respectively.

b




Supplementary Figure 14. Homozygous OsDRM2-knockout (/) mutant showed markedly reduced CHH, CHG, and CG methylation in the intron of ST1. The circle and solid circle represent non-methylated and methylated cytosine, CG (red), CHG (blue), and CHH (green), respectively.

+/+

+/+

Wild type

+/

/

OsDRM2mutant




0

20

40

60

80

100D

iseas

e ar

ea (%

)

WT 1 2 3 4 5 6 7 9 10 11 12 13 14 15 16 17 18 19 20

ST1-RNAi plants (T0 generation)

PXO61 infectiona

0

0.3

0.6

0.9

1.2ST1

Rel

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pres

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leve

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WT 1 3 9 10 18

b

Supplementary Figure 15. Suppressing ST1 did not change rice response to Xoo.Data represent mean (3–5 replicates for disease area and 3 replicates for expression) standard deviation. a, ST1-RNAi plants showed a similar level of susceptibility to Xoo compared to wild-type (WT) Mudanjiang 8. Plants were inoculated with Xoo strain PXO61 at the booting stage. ST1-RNAi, ST1-RNAi construct. b, ST1 expression was partially suppressed in ST1-RNAi plants.

ST1-RNAi plants (T0 generation)

ST1-RNAi




b

Supplementary Figure 16. WRKY45-1 and WRKY45-2 interacted with the promoter region of ST1. a, The promoter region of ST1 harbors cis-elements of W and W-like boxes for WRKY transcription factor binding. P3F and P4R are PCR primers used for amplification of promoter fragment for yeast one-hybrid assay. b,Both WRKY45-1 and WRKY45-2 bound to the promoter region of ST1 in yeast one-hybrid assays. The pHIS2:p53 and pGADT7-Rec2:53 are positive controls.

pHIS2:pST1

pGADT7-Rec2:OsWRKY45-1

pGADT7-Rec2:OsWRKY45-2

pGADT7-Rec2

pHIS2:pST1

pHIS2:pST1

pHIS2:p53

pGADT7-Rec2:53

SD –His/–Leu/–Trp

30 mM 60 mM

3-AT concentration

pHIS2:pST1

pGADT7-Rec2:53

SD –His/–Leu

ST1

+1 transcriptiona -2050 PST1

W-Box W-like Box

-472

P3F P4R




Supplementary Figure 17. Model of how the WRKY45 alleles functions differently in rice–Xoo interaction. Solid line indicates direct action and dotted line indicates indirect or potential action.

ST1

WRKY45-1

Resistance to Xoo

WRKY45-2TE-siR815

Rice

ST1




Supplementary Table 1. PCR primers used for plasmid construction and gene and small RNA expression analysis Gene name (GenBank accession number)

Primer name Forward primer (5’-3’) Reverse primer (5’-3’) Product size (bp)

Use

WRKY45 (CX109127)

W45-RF/W45-RR

TTCCTTGTTGATGTGTCGTCTCA

CCCCCAGCTCATAATCAAGAAC

101 Quantitative RT-PCR for WRKY45-1 or WRKY45-2

W45-F6/W45-R6

ATCACAAAGCATAGCATCATCT

CTCAGCACCTCCTCCTGGTCGG

645 (WRKY45-2,

WRKY45-1.2); 1147

(WRKY45-1)

Amplifying WRKY45-1, WRKY45-2 or WRKY45-1.2 for plasmid construction and detecting positive transgenic plants

W45-F4/R4 ATGGTACCaGCCTACGCATCATCTCCTTC

CAGGATCCbTTATGGCACAACATTTAGCA

1762 Amplifying WRKY45-1.2 for plasmid construction

W45-R9 ACAGATGATGCTATGCTTTGTGAT

Amplifying WRKY45-1.2 for plasmid construction

W45-F10 TGTGTGTACGTATTGCAGGACG

Amplifying WRKY45-1.2 or for plasmid construction

W45-R10 TCCCGGACGATTGCTGC

Amplifying WRKY45-1.2 or TE-siR815b for detecting positive transgenic plants

pU1301 TEV CGAATCTCAAGCAATCAAGCAT

Detecting positive transgenic plants

ST1 (KR063250)

150-R1F/150-R1R

GAGAATGGTAGCCGTGAAGCA

GGTTTCGATGTTGAACTGCAGAT

101 Quantitative RT-PCR for ST1

MT01-ME1F/MT01-ME2R

ATAGTATTTGATATGAGTATTTAAGGTTTT

CCAACAACATTTARTRCAAATACTTTCC

377 Analysis for DNA methylation in the second intron of ST1

MT01-ME2F/MT01-ME1R

GTTATGTGGAGATAGGAGAGATTGAGT

AATTTATTAATTTTAACAAATATCATACAC


MT01-6F/6R ACTAGTcGGTACCaAGGGACATCAAAACCAGCAA

GAGCTCdGGATCCbTCGCATTTGCTTCCACTTCA

479 Amplifying ST1 fragment for RNAi plasmid construction

MT01-P3F/P4R

CGTGAATTCAGCACGTAGAGCCCACAAAGA

ACGGAGCTCGTCGTCGACACAGCACTGCA

393 Amplifying ST1 promoter for RNAi plasmid construction

Actin (X15865)

Actin-F/R TGTATGCCAGTGGTCGTACCA

CCAGCAAGGTCGAGACGAA

121 Quantitative RT-PCR

TE-siR815

815-RT GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCCCAAT

Reverse transcription

815-RF/miR-RR

GATCCAAGGGGATTGAGGAG

CAGTGCAGGGTCCGAGGT

62 Quantitative RT-PCR for TE-siR815

M815F/R CGAGGTACCCGGGAAGAGGAAAGCC

CGAGGATCCCGTAGCATACCCCAAACAACT

842 TE-siR815b overexpressing plasmid construction

U6 snRNA (AC137507.

U6-F/R TACAGATAAGATTAGCATGGCCCC

GGACCATTTCTCGATTTGTACGTG

60 Quantitative RT-PCR for U6 snRNA




Supplementary Table 1. PCR primers used for plasmid construction and gene and small RNA expression analysis Gene name (GenBank accession number)

Primer name Forward primer (5’-3’) Reverse primer (5’-3’) Product size (bp)

Use

WRKY45 (CX109127)

W45-RF/W45-RR

TTCCTTGTTGATGTGTCGTCTCA

CCCCCAGCTCATAATCAAGAAC

101 Quantitative RT-PCR for WRKY45-1 or WRKY45-2

W45-F6/W45-R6

ATCACAAAGCATAGCATCATCT

CTCAGCACCTCCTCCTGGTCGG

645 (WRKY45-2,

WRKY45-1.2); 1147

(WRKY45-1)

Amplifying WRKY45-1, WRKY45-2 or WRKY45-1.2 for plasmid construction and detecting positive transgenic plants

W45-F4/R4 ATGGTACCaGCCTACGCATCATCTCCTTC

CAGGATCCbTTATGGCACAACATTTAGCA

1762 Amplifying WRKY45-1.2 for plasmid construction

W45-R9 ACAGATGATGCTATGCTTTGTGAT

Amplifying WRKY45-1.2 for plasmid construction

W45-F10 TGTGTGTACGTATTGCAGGACG

Amplifying WRKY45-1.2 or for plasmid construction

W45-R10 TCCCGGACGATTGCTGC

Amplifying WRKY45-1.2 or TE-siR815b for detecting positive transgenic plants

pU1301 TEV CGAATCTCAAGCAATCAAGCAT

Detecting positive transgenic plants

ST1 (KR063250)

150-R1F/150-R1R

GAGAATGGTAGCCGTGAAGCA

GGTTTCGATGTTGAACTGCAGAT

101 Quantitative RT-PCR for ST1

MT01-ME1F/MT01-ME2R

ATAGTATTTGATATGAGTATTTAAGGTTTT

CCAACAACATTTARTRCAAATACTTTCC


MT01-ME2F/MT01-ME1R

GTTATGTGGAGATAGGAGAGATTGAGT

AATTTATTAATTTTAACAAATATCATACAC


MT01-6F/6R ACTAGTcGGTACCaAGGGACATCAAAACCAGCAA

GAGCTCdGGATCCbTCGCATTTGCTTCCACTTCA

479 Amplifying ST1 fragment for RNAi plasmid construction

MT01-P3F/P4R

CGTGAATTCAGCACGTAGAGCCCACAAAGA

ACGGAGCTCGTCGTCGACACAGCACTGCA

393 Amplifying ST1 promoter for RNAi plasmid construction

Actin (X15865)

Actin-F/R TGTATGCCAGTGGTCGTACCA

CCAGCAAGGTCGAGACGAA

121 Quantitative RT-PCR

TE-siR815

815-RT GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCCCAAT

Reverse transcription

815-RF/miR-RR

GATCCAAGGGGATTGAGGAG

CAGTGCAGGGTCCGAGGT

62 Quantitative RT-PCR for TE-siR815

M815F/R CGAGGTACCCGGGAAGAGGAAAGCC

CGAGGATCCCGTAGCATACCCCAAACAACT

842 TE-siR815b overexpressing plasmid construction

U6 snRNA (AC137507.

U6-F/R TACAGATAAGATTAGCATGGCCCC

GGACCATTTCTCGATTTGTACGTG

60 Quantitative RT-PCR for U6 snRNA




3) aThe underlined nucleotides are the digestion site of KpnI. bThe underlined nucleotides are the digestion site of BamHI. cThe underlined nucleotides are the digestion site of SpeI. dThe underlined nucleotides are the digestion site of SacI.




3) aThe underlined nucleotides are the digestion site of KpnI. bThe underlined nucleotides are the digestion site of BamHI. cThe underlined nucleotides are the digestion site of SpeI. dThe underlined nucleotides are the digestion site of SacI.

Supplementary Table 2. Probes used for detection of small RNAs

Small RNA name Probe name Sequence (5’-3’) TE-siR815 probe 1 CCCAATCTCCTCAATCCCCTT

probe 2 AAGGGGATTGAGGAGATTGGG probe 3 CTAATGGCTCACCTCGTTTTACGTATCTTCCCAATCTCCTC probe 4 GAGGAGATTGGGAAGATACGTAAAACGAGGTGAGCCATTAG

miR156 miR156 GTGCTCACTCTCTTCTGTCA


Documents

SUPPLEMENTARY INFORMATION - Nature Research...T1W2 3 4567 091 0 20 40 60 80 100 TE-siR815b-oe8 T 1 plants TE-siR815b-oe12 T 1 plants 8 11 12 a b TE-siR815 0 10 20 50 Relative expression