SUPPLEMENTAL DATA
The supplemental data contains three supplemental figures (3 black-and-white figures) and five supplemental tables.
Identification of a canonical SCFSLF complex involved in S-RNase-based self-incompatibility of Pyrus (Rosaceae)
Chi Xu1,2,†, Maofu Li1,5,†, Junkai Wu3,Han Guo1,2, Qun Li1, Yu’e Zhang1, Jijie Chai4, Tianzhong Li3,* and Yongbiao Xue1,*
1State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology,Chinese Academy of Sciences and National Center for Plant Gene Research, Beijing 100101, China
2 University of Chinese Academy of Sciences, Beijing 100190, China
3Laboratoryof Fruit Tree Cell and Molecular Breeding, China Agricultural University, Beijing 100193, China
4School of Life Sciences, Tsinghua University, Beijing100084, China
5Current address: State Key Laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
†These authors contributed equally to this work.
* Corresponding authors: Yongbiao Xue and Tianzhong Li
Address: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences and National Center for Plant Gene Research, Beijing 100101, China
Tel.: 86-10-62552880
Fax: 86-10-62537814
E-mail:
Address: Laboratoryof Fruit Tree Cell and Molecular Breeding, China Agricultural University, Beijing100193, China
E-mail:
Supplemental Figure Legends:
Figure S1. Linkage analysis of PbSLFs and PbS-RNases.
(a) Genotype identificationof an F1 population of 38 progeny from a cross "YaLi" (S21S34) × "JinFeng" (S19S34) was performed. PbS21-RNase, PbS34-RNase and PbS19-RNase were detected by genomic DNA PCR using gene-specific primers. The parents "YaLi" (S21S34)and "JinFeng" (S19S34) were used as controls.
(b) Linkage analysis of PbSLF6-S21 and PbS21-RNase. CAPS procedure was conducted using the F1 population from the "YaLi" (S21S34) × "JinFeng" (S19S34) cross.
(c) Linkage analysis of PbSLF3-S34 and PbS34-RNase. dCAPS procedure was conducted using the F1 population from the "YaLi" (S21S34) × "JinFeng" (S19S34) cross.
Figure S2. A neighbor-joining phylogenetic tree of PbSLF proteins and other SLF/SFBB proteins.The deduced amino acid sequences are from Pyrus bretschneideri (6 PbSLF proteins), Pyrus pyrifolia (49 SFBB proteins), Malus domestica (2 SFBB proteins), Antirrhinum hispanicum (2 SLF proteins), Petunia hybrida (PhSLF-S1) and Petunia inflata (PiSLF1).The neighbor-joining tree is generated with 1000 bootstrap replicates, and bootstrap values of 50% and above are given at branch nodes. AhSLF-S2, AhSLF-S5, PhSLF-S1 and PiSLF1 are used as the outgroup. SLF1-8 represent eight types of the SLF proteins classified byKakui et al. (2011).
Figure S3. GST pull-down assays of GST with His6-SUMO and His6-SUMO-SLFs. Purified GST was used as bait against purified His6-SUMO and His6-SUMO-SLFs. Bound proteins were examined with anti-His antibody.
Supplemental Figure S1
Supplemental Figure S2
Supplemental Figure S3
Supplemental Tables
Supplemental Table 1. Amino acid percent identity of SSK proteins among pear, apple, cherry, strawberry, snapdragon and Petunia (%).
PbSSK1 / PbSSK2 / PavSSK1 / AhSSK1 / PhSSK1 / FvSSK1-like1 / MdSSK1-like1 / MdSSK1-like2 / MdSSK1-like3PbSSK1 / 69.1 / 75.1 / 39.0 / 30.0 / 66.9 / 95.3 / 66.0 / 40.0
PbSSK2 / 70.1 / 34.8 / 29.4 / 65.7 / 66.3 / 95.5 / 53.8
PavSSK1 / 32.3 / 31.6 / 70.9 / 74.6 / 68.9 / 41.4
AhSSK1 / 47.6 / 39.0 / 38.4 / 34.1 / 23.4
PhSSK1 / 34.3 / 30.0 / 28.3 / 24.1
FvSSK1-like1 / 68.0 / 66.3 / 35.9
MdSSK1-like1 / 65.8 / 40.0
MdSSK1-like2 / 50.3
MdSSK1-like3
Supplemental Table 2. Primers used in this paper.
Gene name / Purpose / Sequencea(from 5’ to 3’)PbS21-RNase / Genotype identification / F:ATATTGCAGGACAAGGAATCG
R: ATATGGTGATCCGGGTAGAAAG
RT-PCR / F:ATGGGGATTACGAGGATGATATAC
R:ATACTGGATACTGATGTTGGGGCAC
PbS34-RNase / Genotype identification
&RT-PCR / F: ATGGGGATGACGGGGATGAT
R: ATACTGAATACTATTGTTTGGGCAA
PbS19-RNase / Genotype identification / F: GACCCAAAATATTGCAAGGCG
R: TGGTTCTGTATTGGGGAAGACG
PbSLF3-S34 / dCAPS & RT-PCR / F: TTATCTTTCCATTTATAGTGACTAG
R:ACTATGATAAAATGTTCG
PbSLF6-S21 / CAPS & RT-PCR / F: TGTTCTTTTATGTGTATATACATG
R: CACTTTCAAATAGAATAACATTAC
PbSSK1 / RT-PCR / F: ATGTCGACTGAGAAAGAGGAT
R:TCAGTCTTCATCGACTCCT
PbSSK2 / RT-PCR / F: CTGAGGAAAATCAGCACC
R:TCAGTCTTCATCAACCCCC
PbCUL1 / RT-PCR / F: ATGGCGATCCACGAGCGGAAGACCATTG
R:TCATGCCAAGTACCTAAACAAATTGGG
Actin / RT-PCR / F: GGTGTCATGGTTGGTATGGGTC
R: TTCCGCAACCGCTTGAATAGA
a,F: forward primer; R: reverse primer.
Supplemental Table 3. Segregation analyses of an F1 population of 38 progeny from a cross "YaLi" (S21S34) × "JinFeng" (S19S34)
Cross / Number of progenya / Progeny / Expectedsegregation ratiod / Observed
segregation ratio / χ2 test eχ20.01=6.635
χ20.05=3.841
YaLi×JinFeng / 38 / NS19S21b:NS19S34c / 1:1 / 18:20 / 0.026, P>0.05
aThe number of F1population used for segregation analysis.
bThe number of the plants of S19S21 genotype.
cThe number of the plants of S19S34 genotype.
dThe segregation ratio expected if the segregation is random.
eThe chi-squared goodness-of-fit test of the segregation ration observed; if the χ2sampleχ20.01, the value of P<0.01, which means there was a significant difference between the segregation ratio observed and that expected. Otherwise, if the χ2sampleχ20.05, the value of P>0.05, which means the segregation ratio observed is fit to the segregation ratio expected.
Supplemental Table 4. CAPS/dCAPS makers
Gene / Method / Restriction enzyme / Restriction profile (fragment sizes in bp)PbSLF3-S34 / dCAPS / SpeI / 21-331 (352)
PbSLF6-S21 / CAPS / KpnI / 1152-241 (1393)
Supplemental Table 5.Accession numbers and locus names of genes used in thisstudy
Gene / Accession number/Locus name / Gene / Accession number/Locus nameOSK1 / Os11g26910 / SeSSK1-like1 / Solyc11g008400.1.1
OSK2 / Os10g30200 / SeSSK1-like2 / Solyc11g008230.1.1
OSK3 / Os02g13180 / SeSSK1-like3 / Solyc11g008240.1.1
OSK4 / Os09g10200 / SeSSK1-like4 / Solyc11g008420.1.1
OSK5 / Os09g10260 / SeSKP1-like1 / Solyc03g112710.1.1
OSK6 / Os07g05180 / SeSKP1-like2 / Solyc06g009870.1.1
OSK7 / Os09g10300 / SeSKP1-like3 / Solyc06g009880.1.1
OSK9 / Os07g05150 / SeSKP1-like4 / Solyc11g042930.1.1
OSK10 / Os06g02360 / SeSKP1-like5 / Solyc01g111650.2.1
OSK11 / Os06g02350 / SeSKP1-like6 / Solyc01g111640.2.1
OSK12 / Os09g10270 / SeSKP1-like7 / Solyc10g055610.1.1
OSK13 / Os09g10230 / SeSKP1-like8 / Solyc10g055580.1.1
OSK15 / Os08g28820 / StSSK1-like1 / PGSC0003DMP400013146
OSK16 / Os07g05160 / StSSK1-like2 / PGSC0003DMP400063332
OSK17 / Os07g43180 / StSSK1-like3 / PGSC0003DMP400067628
OSK18 / Os07g43200 / StSSK1-like4 / PGSC0003DMP400013182
OSK19 / Os07g43260 / StSKP1-like1 / PGSC0003DMP400065013
OSK20 / Os09g36830 / StSKP1-like2 / PGSC0003DMP400062284
OSK21 / Os07g22680 / StSKP1-like3 / PGSC0003DMP400060495
OSK22 / Os07g43250 / StSKP1-like4 / PGSC0003DMP400026071
OSK23 / Os07g43270 / StSKP1-like5 / PGSC0003DMP400065248
OSK25 / Os08g28800 / StSKP1-like6 / PGSC0003DMP400068914
OSK26 / Os07g43220 / StSKP1-lik7 / PGSC0003DMP400059895
OSK27 / Os07g43230 / StSKP1-like8 / PGSC0003DMP400065094
OSK28 / Os07g43240 / StSKP1-like9 / PGSC0003DMP400059369
OSK29 / Os08g28780 / StSKP1-like10 / PGSC0003DMP400067913
OSK30 / Os09g10020 / StSKP1-like11 / PGSC0003DMP400061158
MdSSK1-like1 / MDP0000259001 / StSKP1-like12 / PGSC0003DMP400064207
MdSSK1-like2 / MDP0000497762 / StSKP1-like13 / PGSC0003DMP400062388
MdSSK1-like3 / MDC005459.192 / StSKP1-likw14 / PGSC0003DMP400057904
MdSKP1 / ACB87913 / StSKP1-like15 / PGSC0003DMP400063974
MdSKP1-like2 / MDP0000306021 / StSKP1-like16 / PGSC0003DMP400016936
MdSKP1-like3 / MDP0000505840 / StSKP1-like17 / PGSC0003DMP400046438
MdSKP1-like4 / MDP0000273940 / StSKP1-like18 / PGSC0003DMP400010969
MdSKP1-like5 / MDP0000656961 / StSKP1-like19 / PGSC0003DMP400010971
MdSKP1-like6 / MDP0000140395 / StSKP1-like20 / PGSC0003DMP400046434
MdSKP1-like7 / MDP0000278773 / StSKP1-like21 / PGSC0003DMP400046439
MdSKP1-like8 / MDP0000177274 / PbS21-RNase / DQ494532
MdSKP1-like9 / MDP0000304806 / PbS19-RNase / EF643638
MdSKP1-like10 / MDP0000643505 / PbS34-RNase / DQ414813
FvSSK1-like1 / DV438525 / Actin / AF386514