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.
Several paper mills in Wisconsin have programs for spreading paper mill residuals (PMR) on land. A growing number of vegetable farmers recognize the agronomic benefits of PMR applications, but there have been no investigations on the use of PMR for control of vegetable crop diseases. Our objective was to determine the effect of PMR amendments on soilborne and foliar diseases of cucumber and snap bean grown on a sandy soil. Raw PMR, PMR composted without bulking agent (PMRC), or PMR composted with bark (PMRBC) were applied annually in a 3-year rotation of potato, snap bean, and pickling cucumber. Several naturally occurring diseases were evaluated in the field, along with in situ field bioassays. All amendments suppressed cucumber damping-off and Pythium blight and foliar brown spot of snap bean. Both composts reduced the incidence of angular leaf spot in cucumber. In a separate field experiment planted with snap bean for two consecutive years, all amendments reduced common root rot severity in the second year. In a greenhouse experiment, the high rate of PMRBC suppressed anthracnose of snap bean. These results suggest that the application of raw and composted PMR to sandy soils has the potential to control several soilborne and foliar diseases.
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