Isolation and characterization of microsatellite loci from the mulberry, Morus L.

Development of microsatellite markers for mulberry, Morus L. was performed according to the strong affinity between biotin and streptavidin. The method included ligation of 400-1000 bp enzyme-digested fragments and adaptor, affinity capture of microsatellite repeat using biotinylated oligoprobe attached to Dynabeads M-280 streptavidin beads. A (CA)15 enriched microsatellite library was constructed for mulberry. After sequence analysis on 96 randomly picked positive colonies, 24 (25%) of the colonies were found to contain microsatellite sequences, and 20 primer pairs were designed. For polymorphism assessment, 27 mulberry genotypes were checked and 10 of the primers tested were shown to be polymorphic. The novel polymorphic microsatellites produced an average of 4.9 alleles/locus, an average observed heterozygosity of 0.4296 and an average PIC (polymorphism information content) value of 0.4483. Cluster analysis using microsatellite markers was further carried out on the matrix of similarity coefficient with UPGMA method and the phenogram was constructed.