Figure 3

Chromosomal location of TaASY1 determined by Southern blot and PCR analysis. (A) Digested DNA from each of the 21 nullisomic-tetrasomic Chinese Spring wheat lines is shown, representing all 7 chromosome groups, which was hybridised with a full length TaASY1 clone labelled with α-32P dCTP. Absence of hybridisation signals in lanes 14–16, which contains DNA of lines null 5A-T5B, null 5B-T5D, null 5D-T5A respectively, illustrated that TaASY1 was located on chromosome group 5. (B) Amplification of the 446 bp fragment from nullisomic-tetrasomic lines with lanes 1 and 5 null 5A-T5D, lanes 2 and 6 null 5B-T5D, lanes 3 and 7 null 5D-T5A, and lanes 4 and 8 wild-type Chinese Spring. Lanes 1 to 4 contain products amplified using non-genome specific primers, and lanes 5 to 8 contain products amplified using 'A genome' specific primers, as illustrated by the lack of product in lane 5 (null 5A-T5D). (C) Amplification of the 446 bp fragment from DNA template of wild-type and various bread wheat deletion lines (lane 1 wild-type Chinese Spring, lane 2 null 5A-T5D, lane 3 mutant 5AL 4-1 (0.55 FL), lane 4 5AL 19-5 (0.35 FL) and lane 5 5AL 12-1 (0.32 FL)). The DNA molecular weight marker in (B) and (C) is a 100 bp ladder (Invitrogen, Japan).