Potato early dying (PED), also known as Verticillium wilt, caused by Verticillium dahliae, is a seasonal yield-limiting disease of potato worldwide, and PED-resistant cultivars currently represent only a small percentage of potato production. In this study, we developed a real-time quantitative polymerase chain reaction (Q-PCR) approach to detect and quantify V. dahliae. The efficiency of the designed primer pair VertBt-F/VertBt-R, derived from the sequence of the beta-tubulin gene, was greater than 95% in monoplex Q-PCR and duplex (using Plexor technology) procedures with primers PotAct-F/PotAct-R, obtained from the sequence of the actin gene, designed for potato. As few as 148 fg of V. dahliae DNA were detected and quantified, which is equivalent to five nuclei. Q-PCR detected V. dahliae in naturally infected air-dried potato stems and fresh stems of inoculated plants. Spearman correlations indicated a high correlation (upward of 80%) between V. dahliae quantifications using Q-PCR and the currently used plating assays. Moreover, Q-PCR substantially reduced the variability compared with that observed in the plating assay, and allowed for the detection of V. dahliae in 10% of stem samples found to be pathogen free on the culture medium. The described Q-PCR approach should provide breeders with a more sensitive and less variable alternative to time-consuming plating assays to distinguish response of breeding lines to colonization by V. dahliae.

Creeping bentgrass (Agrostis stolonifera L.) is one of the most adapted bentgrass species for use on golf course fairways and putting greens because of its high tolerance to low mowing height. It is a highly outcrossing allotetraploid species (2n=4x=28, A(2) and A(3) subgenomes). The first linkage map in this species is reported herein, and it was constructed based on a population derived from a cross between two heterozygous clones using 169 RAPD, 180 AFLP, and 39 heterologous cereal and 36 homologous bentgrass cDNA RFLP markers. The linkage map consists of 424 mapped loci covering 1,110 cM in 14 linkage groups, of which seven pairs of homoeologous chromosomes were identified based on duplicated loci. The numbering of all seven linkage groups in the bentgrass map was assigned according to common markers mapped on syntenous chromosomes of ryegrass and wheat. The number of markers linked in coupling and repulsion phase was in a 1:1 ratio, indicating disomic inheritance. This supports a strict allotetraploid inheritance in creeping bentgrass, as suggested by previous work based on chromosomal pairing and isozymes. This linkage map will assist in the tagging and eventually in marker-assisted breeding of economically important quantitative traits like disease resistance to dollar spot (Sclerotinia homoeocarpa F.T. Bennett) and brown patch (Rhizoctonia solani Kuhn).

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Verticillium wilt (VW) of potato, caused primarily by the fungus Verticillium dahliae, results in yield loss and is therefore an important soil-borne disease. Resistance to VW exists in potato germplasm and is used by breeders during cultivar development. Breeders could make more rapid progress toward the development of VW resistant clones if they had an eVective early generation selection strategy. The purpose of this study was to determine whether selection for VW resistance could be carried out in the Wrst tuber generation on single hills. One hundred and Wfty-two clones from 19 families were planted as single hills on a V. dahliae-infested Weld. Each plant was scored for vine maturity, VW symptom expression, yield, stem colonization (colony forming units (cfu), in dried basal stem segments) and incidence (percent infected stems). In the second clonal generation, which consisted of replicated four-hill plots, stem colonization scores and incidence values were used to identify clones which were more resistant than a moderately resistant cultivar and others which were more susceptible than a susceptible cultivar. The eYciency and reliability of the single-hill selection strategy, based on symptoms and yield, was then determined by comparison to the four-hill results. We determined that the best single-hill selection strategy was negative selection (discard clones with the lowest performance) with low stringency, based on yield.