Supplemental Table1 Genotyping of Pikahei-1(t)-containing plant materials using co-segregating markers of rice blast R genes pi21, Pi34, and qBR12-1a

Marker / Chromosome # / Target gene / Kahei / R-8 / S-8 / Hitachi IL-1 / Koshihikari / Nipponbare
#43-2 / 4 / pi21 / A / B / B / B / B / B
ID01-37 / 4 / Pikakei-1(t) / A / A / B / A / B / B
RM21 / 11 / Pi34 / A / A / A / B / B / B
RM26871 / 11 / Pi34 / A / A / A / B / B / B
RM5196 / 12 / qBR12-1 / A / A / A / B / B / B
INDEL02-21 / 12 / qBR12-1 / A / A / A / B / B / B
INDEL01-41 / 12 / qBR12-1 / A / A / A / B / B / B
RM6973 / 12 / qBR12-1 / A / A / A / B / B / B

aA and B representsgenotypes containing and not-containing the target gene respectively. Primer sequences and genotyping methodsare as described by Fukuoka et al. (2012).

Supplemental Table 2Polymerase chain reaction primer sequences used for RGAamplification in the construction oftransgenicDNAs

Gene name / Primer sequences (5'→3')
RGA1 / TATAGGCGCGCCCGCTGCTTGGACTTGATGTA
GGCCTTAATTAATTCGTCTCGCGAATTAGTCC
RGA2 / TATAGGCGCGCCGAGCTCCAACCCTCCTCTCT
GGCCTTAATTAACTCGCAAGTTTTGCAACGTA
RGA3 / TATAGGCGCGCCTGGCGTATGCATTAGCACAT
GGCCTTAATTAATTGATGACAGTCCCGAAACA
RGA4 / TATAGGCGCGCCCGTTGCAGCAAGTGTGATCT GGCCTTAATTAATGAGTGCGACTAGCCAAATG
RGA5 / ACGCGTCGACATCCCGAATCCAAATCCTTC
ACGCGTCGACCCTATTCACCCGCACCTCTA
RGA6 / TATAGGCGCGCCGAGAAAAGAGGGGAGGGATG GGCCTTAATTAACCGTTTGACTCCGTTCAACT
RGA7 / TATAGGCGCGCCaagacgttcctgtgggtgtc
GGCCTTAATTAAccacacgtttggccttattt

Supplemental Table 3Primer sequences used for quantitative reverse transcriptase-polymerase chain reaction analysis

Gene name / Primer sequences (5'→3')
Rubq1 / GGAGCTGCTGCTGTTCTAGG
TTCAGACACCATCAAACCAGA
RGA1 / TCCCTATCACATCGAATGTTTG
TTCGTCTCGCGAATTAGTCC
RGA2 / ATGATCACAGCGAACATCCA
CTCGCAAGTTTTGCAACGTA
RGA3 / GCATTAAAAGCCTGCCAGAG
CCTGTAGCTTCTCCGTTGCT
RGA4 / ATCTGGACTGGAATGGCTTG
AAGTGTCGGAGGGAGTTGAG
GCTGTCCACTTCATGGGAAT
CAAGCCATTCCAGTCCAGAT
RGA5 / AGCAAGAGAGAGCGCTTCAG
TGAGGGAAAGAAGGCTGTGT
RGA6-1 / ACTCAACAAACGGCGAAAAT
CCGTTTGACTCCGTTCAACT
RGA6-2 / TGGAGCTTGATCCTCGAAAT
ATTTTCGCCGTTTGTTGAGT
RGA7 / GGTGGCTCGTTAAGCACACT
TGGTAAGTACCGCAGCTTGA
RGA1-NB / CGGTGGATTCAACAGTTAGTGA
CACATAAGGGGAAGCGATTG

SupplementalTable 4Annotation data of the putative open reading frames (ORFs) in the Pikahei-1(t) regiona

RGA no. / ORF no. / Protein coded by predicted genes
1 / unknown protein
2 / hypothetical protein
3 / putative kelch motif family protein
4 / putative Ser/Thr protein kinase
5 / hypothetical protein
6 / putative OTU domain-containing protein 6B
7 / putative protein kinase family protein
8 / putative F-box domain containing protein
9 / hypothetical protein
10 / putative ATBAG1 (ARABIDOPSIS THALIANA BCL-2-ASSOCIATED ATHANOGENE 1); protein binding
11 / putative MRP-like ABC transporter
12 / unknown protein
13 / unknown protein
14 / putative remorin family protein
15 / putative UDP-N-acetylenolpyruvoyl glucosamine reductase
16 / hypothetical protein
17 / putative SIT4 phosphatase-associated family protein
18 / putative nitrate-induced NOI protein
19 / putative tobacco nucleolin
1 / 20 / putative NBS-LRR disease resistance protein homologue
21 / putative RecName: Full = Adhesive plaque matrix protein; Alt Name: Full = Foot protein 1; Alt Name: Full = MCFP1; Flags: Precursor
2 / 22 / putative NB-ARC domain containing protein
23 / putative MuDR family transposase domain containing protein
3 / 24 / putative NBS-LRR disease resistance protein homologue
25 / putative epoxide hydrolase
26 / putative polyprotein
4 / 27 / putative XA1
5 / 28 / putative XA1
29 / putative bromodomain containing protein
30 / putative oligopeptide transport system permease protein OppB
31 / putative SWIM zinc finger family protein
32 / putative methyltransferase
33 / putative histidinol dehydrogenase
6 / 34 / putative XA1
35 / putative bromodomain containing protein
36 / unknown protein similar to OSJNBb0085C12.9
37 / putative CDR1 (CONSTITUTIVE DISEASE RESISTANCE 1); aspartic-type endopeptidase
7 / 38 / putative XA1
39 / putative bromodomain containing protein
40 / unknown protein similar to Os04g0623200
41 / hypothetical protein
42 / putative lysine-specific histone demethylase 1

a Gene prediction of the candidate genomic region was performed using RiceGAAS( by submitting all the sequences. Seven RGAs were indicated and numbered following the genomic sequence order.

Supplemental Table 5Blast fungus isolates used in this study

Isolate / Cultivar a / Ecosystem / Loacation / Year / J-race b / U-race c
Kyu9439013 / Natsuhikari / Irrigated lowland / Kouchi, Japan / 1994 / 047.0 / U13-i7-k100-z01-ta001
H10-168-1 / Hitachi IL1 / Irrigated lowland / Aichi, Japan / 2010 / NT / NT
Kyu89-246 / Nangokumochi / Irrigated lowland / Miyazaki, Japan / 1989 / 003.0 / U13-i0-k100-z00-ta001
H09-132-1 / NERICA1 / Upland / Namlonge, Uganda / 2009 / NT / U53-i7-k127-z00-ta031

aRice cultivar from which the blast strain was isolated;b Race numbers determined by virulent/avirulent reactions to 12 differentials; c Race numbers characterized by reactions to 26 Lijian-xintuan-heigu (LTH) monogenic lines for targeting 23 resistance genes.

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