Development of S-SAP markers based on an LTR-like sequence from Medicago sativa L

Molecular Genetics and Genomics : MGG
A PorcedduF Veronesi

Abstract

The Sequence-Specific Amplification Polymorphism (S-SAP) method, recently derived from the Amplified Fragment Length Polymorphism (AFLP) technique, produces amplified fragments containing a retrotransposon LTR sequence at one end and a host restriction site at the other. We report the application of this procedure to the LTR of the Tms1 element from Medicago sativa L. Genomic dot-blot analysis indicated that Tms1 LTRs represent about 0.056% of the M. sativa genome, corresponding to 16 x 10(3) copies per haploid genome. An average of 66 markers were amplified for each primer combination. Overall 49 polymorphic fragments were reliably scored and mapped in a F(1) population obtained by crossing diploid M. falcata with M. coerulea. The utility of the LTR S-SAP markers was higher than that of AFLP or SAMPL (Selective Amplification of Microsatellite Polymorphic Loci) markers. The efficiency index of the LTR S-SAP assay was 28.3, whereas the corresponding values for AFLP and SAMPL markers were 21.1 and 16.7, respectively. The marker index for S-SAP was 13.1, compared to 8.8 for AFLP and 9.5 for SAMPL. Application of the Tms1 LTR-based S-SAP to double-stranded cDNA resulted in a complex banding pattern, demonstrating the presence of Tm...Continue Reading

Citations

Feb 9, 2005·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Sheh May TamMarie-Angèle Grandbastien
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Jul 21, 2009·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Sheh May TamMarie-Angèle Grandbastien
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