Simple sequence repeats (SSRs or microsatellites) are frequently used as a robust, rapid, and relatively inexpensive means of genotyping. Recent reports in cultivated strawberry (Fragaria 9 ananassa) have identified a widely applicable set of SSRs that permits discrimination of closely-related genotypes. In the present study this same set of SSRs is analyzed in cultivars commonly grown in the state of Florida, as well as advanced breeding selections from the University of Florida program. Nine primer pairs have been used to produce discrete SSR patterns for all lines tested, including diagnostic sets that distinguish between closely-related cultivars and/or breeding selections. A comparison of markers to known pedigrees is presented, along with analysis of relative relatedness between lines studied. We also detail important technical considerations for transferability and limitations of these technologies between laboratories. The resulting genotype data for cultivars commonly used in Florida are accessible as supplemental data in graphic format and allow comparison with other cultivars. Users can thus select the most diagnostic SSRs to test the lines in question. These resources provide new tools for breeders and nurseries to authenticate genotypes, follow inheritance patterns and enforce patent protection. As important, this report underscores the strengths and weaknesses of applying the original methodologies to a different plant population in independent laboratories.

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Graft-indexing of an advanced selection from the University of Florida strawberry breeding program produced virus-like symptoms on Fragaria vesca. However; RT-PCR testing of the material did not detect the presence of any of 16 strawberry virus species or members of virus groups for which strawberries are routinely indexed. Large scale sequencing of the material revealed the presence of an isolate of Strawberry necrotic shock virus. The nucleotide sequence of this isolate from Florida shows a significant number of base changes in the annealing sites of the primers compared to the primers currently in use for the detection of SNSV thereby explaining the most probable reason for the inability to detect the virus in the original screening. RT-PCR and Taqman(®) qPCR assays were developed based on conserved virus sequences identified in this isolate from Florida and other sequences for SNSV currently present in GenBank. The two assays were applied successfully on multiple samples collected from several areas across the United States as well as isolates from around the world. Comparison between the RT-PCR and the qPCR assays revealed that the qPCR assay is at least 100 times more sensitive than conventional PCR.