Obtaining DNA markers for diversity evaluation in Trifolium pratense cultivars
| Authors | |
|---|---|
| Year of publication | 2005 |
| Type | Article in Proceedings |
| Conference | Book of Abstracts, 6th International Symposium in the Series Recent Advances in Plant Biotechnology, 12.-16.9.2005 |
| MU Faculty or unit | |
| Citation | |
| Field | Plant cultivation |
| Keywords | DNA markers; microsatellites; Trifolium pratense |
| Description | Red clover (Trifolium pratense L.) is one of the main forage species of temperate regions. Cultivars of red clover are allogamous and heterogeneous which their genetic analysis makes difficult. The objective of this study was obtaining DNA markers (RAPD, SSR) for genetic diversity evaluation among red clover cultivars and verification of utility of SSR oligonucleotides from white clover (T. repens). RAPD (Random Amplified Polymorphic DNA) requires no knowledge of DNA sequences for primers designing. SSRs (Simple Sequence Repeats) are highly informative codominant molecular markers which are well known in white clover. Red clover cv. Kvarta and Start and white clover cv. Hájek and Jura were used for molecular analysis. Genomic DNA of red and white clover plants was extracted by bulking method from different plant numbers. CTAB-based method and GenEluteTMPlant Genomic DNA Kit (Sigma) were used for DNA isolation. RAPD analysis was performed with 20 primers and four PCR programmes from literature with modifications. No sequences for SSR markers are available and thus oligonucleotides designed for white clover in our work with red clover were used. Microsatellite loci were amplified using 20 primer pairs and we optimised individual PCR steps. The reliable DNA fragment amplification and visualisation required TaKaRa Taq polymerase (BioTech), electrophoresis in polyacrylamide gels and silver staining. In this investigation the availability of oligonucleotides for SSR markers of related species was evaluated in red clover. |
| Related projects: |