Studies were conducted in the presence of Verticillium wilt (Verticillium albo-atrum) to determine the effect of fall harvesting and grazing over time on plant stand and forage yield of alfalfa (Medicago sativa). Resistant and susceptible cultivars were tested on established and newly seeded fields. In the fall (experiments 1 and 2), cultivars were either: (i) cut (third time); (ii) grazed; (iii) cut and grazed; or (iv) left uncut and ungrazed. Although Verticillium was present, test sites for experiments 2 and 3 were sprayed with a spore suspension of V. albo-atrum immediately following the first cutting of each experiment to standardize disease pressure. In experiment 1, the moderately resistant cultivar Apollo II, harvested twice without a late third cutting or fall grazing, produced the highest forage yield the following year. Fall grazing reduced subsequent yields in both the 2- and 3-cut treatments. In experiment 2, a third cutting decreased plant density and forage yield in both resistant and susceptible cultivars, while grazing had no effect. Neither fall treatment affected incidence of Verticillium wilt. In experiment 3, application of the fungicide benomyl to plant stubble following each harvest decreased Verticillium wilt in Apollo but not in Arrow. Overall, with the resistant cultivar Arrow, harvesting twice annually and grazing after a killing frost in lieu of late fall cutting slowed disease development, prolonged stand life, and maximized forage yield and quality.
A rapid technique for identification and detection of Phoma sclerotioides, the causal agent of brown root rot of alfalfa, has been developed using polymerase chain reaction (PCR). Amplification products obtained from random amplified polymorphic DNA (RAPD) reactions were cloned and sequenced, and two extended primer sets were designed from the resulting data that were used to detect sequence-characterized DNA markers. A single 499-bp DNA amplification product was consistently obtained from primers PSB12499 that was specific for 19 isolates of P.sclerotioides but was not produced from Phoma medicaginis or Phoma betae, or from other soilborne pathogens including Aphanomyces euteiches, Rhizoctonia solani, Fusarium oxysporum, Pythium ultimum, or Phytophthora infestans. A 499-bp amplification product was also produced from root tissue known to be infected with the fungus as verified by microscopic examination. A similar PCR product was obtained from soil samples collected from fields with an established infection of P. sclerotioides on alfalfa. This PCR-based assay enables detection of P. sclerotioides from alfalfa root tissue and in soil samples in a single day, including extraction of DNA, compared with standard methods that require up to 100 days for identification using agar media.
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