RAPD Variation among Salt-selected and Nonselected Buffalograss [Buchloë dactyloides (Nutt.) Engelm.] Clones

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  • 1 Dept. of Horticulture, Colorado State Univ., Fort Collins, CO 80523

The use of random amplified polymorphic DNA (RAPD) markers has been shown to be a potentially useful technique for identifying buffalograss breeding lines. Analysis of RAPD markers has also revealed considerable variation within, as well as among, each of four natural buffalograss populations surveyed. Identification of genetic markers for quantitative traits, such as physiological components of tolerance to salt stress, can provide important information for plant improvement programs. The objectives of this study were to develop DNA fingerprints for buffalograss clones selected from an in vitro seedling screening program for survival at high NaCI (200–250 mM) levels, identify markers for future analysis, and assess the variability among the lines. DNA was extracted from leaves of 10 salt-selected and 15 non-selected buffalograss clones. Fifty-two 10-mer primers were screened for ability to produce bands with DNA from four clones as visualized on ethidium-stained agarose gels. Bands were most reproducible with a genomic template DNA concentration of 1 ngμl–1 reaction volume. Primers selected for ability to produce a moderate number of clear bands were used to produce RAPD profiles of the 25 clones. Abundant polymorphism to distinguish among clones was found. Four primers produced a total of 45 polymorphic markers. The primer 5′-CGGAGAGCCC-3′ produced 11 readily scored markers, allowing identification in 94.67% of pair-wise comparisons. As a group, RAPD profiles of salt-selected clones are more variable than non-selected clones from the same population; however, no unique pattern of markers generated by primers screened to date differentiates all salt-selected clones from the non-selected group.

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