title: Microsatellite isolation and marker development in carrot - genomic distribution, linkage mapping, genetic diversity analysis and marker transferability across Apiaceae.
abstract: BACKGROUND
  The Apiaceae family includes several vegetable and spice crop species among which carrot is the most economically important member, with ~21 million tons produced yearly worldwide. Despite its importance, molecular resources in this species are relatively underdeveloped. The availability of informative, polymorphic, and robust PCR-based markers, such as microsatellites (or SSRs), will facilitate genetics and breeding of carrot and other Apiaceae, including integration of linkage maps, tagging of phenotypic traits and assisting positional gene cloning. Thus, with the purpose of isolating carrot microsatellites, two different strategies were used; a hybridization-based library enrichment for SSRs, and bioinformatic mining of SSRs in BAC-end sequence and EST sequence databases. This work reports on the development of 300 carrot SSR markers and their characterization at various levels.
  
  
  RESULTS
  Evaluation of microsatellites isolated from both DNA sources in subsets of 7 carrot F2 mapping populations revealed that SSRs from the hybridization-based method were longer, had more repeat units and were more polymorphic than SSRs isolated by sequence search. Overall, 196 SSRs (65.1%) were polymorphic in at least one mapping population, and the percentage of polymophic SSRs across F2 populations ranged from 17.8 to 24.7. Polymorphic markers in one family were evaluated in the entire F2, allowing the genetic mapping of 55 SSRs (38 codominant) onto the carrot reference map. The SSR loci were distributed throughout all 9 carrot linkage groups (LGs), with 2 to 9 SSRs/LG. In addition, SSR evaluations in carrot-related taxa indicated that a significant fraction of the carrot SSRs transfer successfully across Apiaceae, with heterologous amplification success rate decreasing with the target-species evolutionary distance from carrot. SSR diversity evaluated in a collection of 65 D. carota accessions revealed a high level of polymorphism for these selected loci, with an average of 19 alleles/locus and 0.84 expected heterozygosity.
  
  
  CONCLUSIONS
  The addition of 55 SSRs to the carrot map, together with marker characterizations in six other mapping populations, will facilitate future comparative mapping studies and integration of carrot maps. The markers developed herein will be a valuable resource for assisting breeding, genetic, diversity, and genomic studies of carrot and other Apiaceae.
authors.affiliation: Dept. of Horticulture, University of Wisconsin-Madison, 1575 Linden Drive, Madison, WI 53706, USA.
authors.forename: Pablo F
authors.initials: PF
authors.lastname: Cavagnaro
authors.forename: Sang-Min
authors.initials: SM
authors.lastname: Chung
authors.forename: Sylvie
authors.initials: S
authors.lastname: Manin
authors.forename: Mehtap
authors.initials: M
authors.lastname: Yildiz
authors.forename: Aamir
authors.initials: A
authors.lastname: Ali
authors.forename: Maria S
authors.initials: MS
authors.lastname: Alessandro
authors.forename: Massimo
authors.initials: M
authors.lastname: Iorizzo
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Senalik
authors.forename: Philipp W
authors.initials: PW
authors.lastname: Simon
mesh_terms: D002874:Chromosome Mapping; D018552:Daucus carota / Q000235:genetics*; D020224:Expressed Sequence Tags / Q000378:metabolism; D005819:Genetic Markers / Q000235:genetics; D018745:Genome, Plant / Q000235:genetics; D023281:Genomics; D006824:Hybridization, Genetic; D018895:Microsatellite Repeats / Q000235:genetics*; D011110:Polymorphism, Genetic / Q000235:genetics*; D013045:Species Specificity
chemical_list: D005819:Genetic Markers