5/1833

1. 10kb 50kb inverted region 10kb (393) (84) (+365) 389 75 609 183 60 204 249 99 195 93 264 399 TC3F TC3R TB2R 0cM RE EP PP IF2 FIP OXI CAS ANK PUM MYB GAG PME REV PUM 26SP RHZF (77) LRRPK 575 69 621 183 60 204 249 99 195 93 267 607 P1301 10kb TB2R 1kb P1301 NifU PPR POL WD40 LRRPK DNABP 1kb a TM0105 TA1R TB7R TC3R TB2R TC3F 5/1833 3/1833 1/1833 0/1833 0/1833 0/1833 TB21R TA3F TB2R b G3 G5 G1 G4 G2 M1 M3 M4 M6 M2 M5 c d e B22Not c8 c16 P1301 TM0885 0/173 0/798 2/409 0/292 M7 M8 G7 G8 M9 f Supplementary Figure 1 2004-02-15137B/Kawasaki/Suppl.Fig.1ppt

2. Supplementary Figure 1 Positional cloning of CASTOR and POLLUX genes. a, Genetic map around the CASTOR locus. Positions of flanking markers are indicated together with the number of recombinant plants/total number of mapping individuals. b, Physical map of the CASTOR locus. Designations of large insert clones originating from parental ecotype B-129 are; G1, BAC156-1E; G2, BAC124-7B; G3, BAC324-1D; G4, BAC104-3F; G5, BAC33-3E; and from parental ecotype MG-20: M1, LjT17M09; M2, LjT62O06; M3, LjT02K14; M4, LjT45I15; M5, LjT20F11 and M6, LjT46G19. An inverted region of 145kb between the genomes of B-129 and MG-20 was detected, which is probably responsible for the observed suppression of recombination around the CASTOR locus (0 cM). BAC end markers within the inverted region are; open oval, TB21R; closed oval, TA3F; open rectangle, TB2R. The TB7R marker located in the north side delimits CASTOR to ~240kb. c, Features and candidate genes predicted within the target region. LRRPK, leucine-rich repeat receptor kinase; 26SP, 26S proteasome regulatory subunit 7; RE, retro-element; EP, Avr9/Cf-9 elicited protein; PUM, pumilio-family RNA-binding protein; RHZF, Ring H2 zinc finger protein; PME, pectin methyl esterase; IF2, initiation factor 2 subunit; OXI, oxido-reductase; PP, polyprotein; MYB, myb family protein; GAG, gag-pol polyprotein; FIP; VirF-interacting protein FIP1; ANK, ankyrin-like protein; REV; reverse transcriptase; unlabelled, hypothetical protein. d, Exon-intron structure of CASTOR. Exons are indicated by upper boxes and numbers indicate their length in nucleotides. Introns are represented by lower triangles. Several alternative-splice variants were identified by cDNA-sequencing; a 4-nt insertion at the 3’ end of the 1st exon, and a 9-nt insertion at the 5’ end of the 2nd exon. The resulting exon sizes are shown in brackets. Also, the 7th intron of 365 nucleotides (+365) was retained in 15 out of 17 cDNA clones, leading of a premature stop codon. e-f Genomic region around the POLLUX gene: e, Genetic and physical map of the POLLUX locus. Positions of the flanking markers and the number of recombinant plants in the mapping populations are indicated. Abbreviations G6, G7, M7, M8 and M9 refer to large insert clones BAC131-3b, BAC45-6C, LjT39N07, LjB22b22 and LjT45B09, respectively. Genes predicted on the cosegregating TAC clone LjT45B09; WD40, WD40 repeat protein; NifU, putatively involved in Fe-S cluster synthesis; PPR, pentatricopeptide repeat protein; DNABP, DNA binding protein; LRRPK, leucine-rich repeat receptor kinase; unlabelled, hypothetical protein. f, Exon-intron structure of POLLUX. One alternative splice site was detected at the beginning of the 10th exon resulted in a 16-bp deletion.