Genetic analysis of the anther-derived progeny and isolated pollen grains of a culture-responsive Solanum clone

An understanding of the detailed genetic basis of agronomically important traits requires an integration of plant physiology, molecular biology and genetics. The present study integrates the use of anther culture, molecular markers analysis and genetic linkage assessment to demonstrate the value of this approach for potato. To develop a population for subsequent genetic linkage analysis, 23 diploid (2n = 2x = 24) potato clones were screened for their responses to four different anther culture media. A higher level of exogenous cytokinin (6-benzyladenine) to that of auxin (3-indoleacetic acid) proved to be favourable for the induction of calli/embryos. Anther-derived material (roots and plantlets) originating from clones 9507-04 and 6028-02 revealed differences in growth vigor. Flow cytometric estimation of ploidy in the initial regenerated roots and plantlets revealed that clone 9507-04 produced 44% haploid roots and 77% haploid plantlets. Seventy-three percent of anther-derived plantlets from clone 6028-02 were haploids.;Putatively haploid anther-derived roots and plantlets from Solanum clone 9507-04 were used for the random amplified polymorphic DNA (RAPD) analysis. Of 56 RAPD markers assayed, 44.6% did not follow the expected 1:1 Mendelian segregation ratio ( c2≥ 4.26, P < 0.05) in the anther-derived progeny. The results may indicate a greater anther culture related genetic selection pressure during plantlet regeneration when compared to that in root regeneration.;Pollen grains from the clone 9507-04 were also used for the RAPD analysis via primer UBC291 and UBC504. RAPD products were discernible from single pollen gains without requiring any DNA extraction procedures. The three UBC291-amplified RAPD markers of the parent segregated in the selected sample of 60 pollen grains. Potential applications of the linkage map and the RAPD analysis of single pollen grains are described. (Abstract shortened by UMI.).