Map-Based Cloning of PDV1

2010/08/17 12:39:52

PDV1 and PDV2 Mediate Recruitment of the Dynamin-Related Protein ARC5 to the Plastid Division Site

The Plant Cell, 2006, 18: 2517-2530

Shin-ya Miyagishima, John E. Froehlich and Katherine W. Osteryoung

The pdv1-1 mutation was mapped with molecular markers based on a cleaved amplified polymorphic sequence (Konieczny and Ausubel, 1993) and simple sequence length polymorphisms (Bell and Ecker, 1994). We used some markers listed on The Arabidopsis Information Resource (TAIR; http://www.Arabidopsis.org); other markers were designed based on polymorphisms listed at TAIR (http://www.Arabidopsis.org/Cereon) in the Monsanto SNP and Ler Sequence Collection. The pdv1-1 homozygous mutant was crossed with Landsberg erecta wild-type plants to generate a mapping population. Analyses using 48 F2 progeny with the pdv1-1 phenotype showed that the mutation is located in a region of 0.7 Mb on chromosome 5 (between polymorphisms SGCSNP18686 and SGCSNP220). Using 500 F2 plants, we fine-mapped the pdv1-1 mutation to a region of 65 kb that includes 12 genes (between polymorphisms CER437355 and SGCSNP18689). Of these 12 genes, 6 were amplified from pdv1-1 and sequenced; a mutation was found in At5g53280. At5g53280 was then amplified from pdv1-2 plants, and a different mutation was found in the same gene.