title: Microsatellite cross-species amplification and utility in southern African elasmobranchs: A valuable resource for fisheries management and conservation.
abstract: BACKGROUND
  Similarly to the rest of the world, southern Africa's diverse chondrichthyan fauna is currently experiencing high fishing pressures from direct and non-direct fisheries to satisfy market demands for shark products such as fins and meat. In this study, the development of microsatellite markers through cross-species amplification of primer sets previously developed for closely related species is reported as an alternative approach to de novo marker development. This included the design of four microsatellite multiplex assays and their cross-species utility in genetic diversity analysis of southern African elasmobranchs. As this study forms part of a larger project on the development of genetic resources for commercially important and endemic southern African species, Mustelus mustelus was used as a candidate species for testing these multiplex assays in down-stream applications.
  
  
  RESULTS
  Thirty five microsatellite primer sets previously developed for five elasmobranch species were selected from literature for testing cross-species amplification in 16 elasmobranch species occurring in southern Africa. Cross-species amplification success rates ranged from 28.6%-71.4%. From the successfully amplified microsatellites, 22 loci were selected and evaluated for levels of polymorphism, and four multiplex assays comprising of the 22 microsatellites were successfully constructed, optimised and characterised in a panel of 87 Mustelus mustelus individuals. A total of 125 alleles were observed across all loci, with the number of alleles ranging from 3-12 alleles. Cross-species amplification of the four optimised multiplex assays was further tested on 11 commercially important and endemic southern African elasmobranch species. Percentage of polymorphism ranged from 31.8%-95.5% in these species with polymorphic information content decreasing exponentially with evolutionary distance from the source species.
  
  
  CONCLUSIONS
  Cross-species amplification of the 35 microsatellites proved to be a time- and cost-effective approach to marker development in elasmobranchs and enabled the construction of four novel multiplex assays for characterising genetic diversity in a number of southern African elasmobranch species. This study successfully demonstrated the usefulness of these markers in down-stream applications such as genetic diversity assessment and species identification which could potentially aid in a more integrative, multidisciplinary approach to management and conservation of commercially important cosmopolitan and endemic elasmobranch species occurring in southern Africa.
authors.forename: Simo N
authors.initials: SN
authors.lastname: Maduna
authors.forename: Charné
authors.initials: C
authors.lastname: Rossouw
authors.forename: Rouvay
authors.initials: R
authors.lastname: Roodt-Wilding
authors.affiliation: Department of Genetics, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa. aeb@sun.ac.za.
authors.forename: Aletta E
authors.initials: AE
authors.lastname: Bester-van der Merwe
mesh_terms: D000465:Algorithms; D000483:Alleles; D000818:Animals; D003247:Conservation of Natural Resources / Q000379:methods; D018940:Elasmobranchii / Q000145:classification / Q000235:genetics*; D005398:Fisheries / Q000379:methods; D014644:Genetic Variation; D005838:Genotype; D018895:Microsatellite Repeats / Q000235:genetics*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D013019:South Africa; D013045:Species Specificity