Four populations of Sclerotinia sclerotiorum in North America were inferred previously, based on analyses of both rapidly evolving markers (DNA fingerprint and mycelial compatiblity), and multilocus DNA sequence spanning the range between fast and slow evolution. Each population was defined as an interbreeding unit of conspecific individuals sharing a common recent ancestor and arising in a unique evolutionary event. The present study applies this standard to extend characterization of S. sclerotiorum populations to the Western United States. Isolates of S. sclerotiorum (N = 294) were determined to represent three genetically differentiated populations: California (CA, lettuce), Washington (WA, pea/lentil), and Ontario (ON, lettuce). CA was the most diverse population yet sampled in North America. Clonality was detected in ON and WA. No DNA fingerprints were common among the populations. The index of association (I(A)), based on fingerprint, was closer to zero (0) for CA than it was for the other populations. High diversity and lack of association of markers in California are consistent either with genetic exchange and recombination, or with large population size and high standing genetic variation. Intra- and interlocus conflict among three DNA sequence loci was consistent with recombination. The coalescent IGS genealogy confirmed subdivision and showed CA to be older than WA or ON. The Nearest Neighbor statistic on combined data confirmed subdivision among all present and previously defined populations. All isolates had both MAT1-1 and MAT1-2, consistent with uniform homothallism.

Experiments were conducted in field plots to evaluate the effects of broccoli residue on population dynamics of Verticillium dahliae in soil and on Verticillium wilt development on cauliflower under furrow and subsurface-drip irrigation and three irrigation regimes in 1994 and 1995. Treatments were a factorial combination of three main plots (broccoli crop grown, harvested, and residue incorporated in V.dahliae-infested plots; no broccoli crop or residue in infested plots; and fumigated control plots), two subplots (furrow and subsurface-drip irrigation), and three sub-subplots (deficit, moderate, and excessive irrigation regimes) arranged in a split-split-plot design with three replications. Soil samples collected at various times were assayed for V. dahliae propagules using the modified Anderson sampler technique. Incidence and severity of Verticillium wilt on cauliflower were assessed at 7- to 10-day intervals beginning a month after cauliflower transplanting and continuing until harvest. Number of propagules in all broccoli plots declined significantly (P < 0.05) after residue incorporation and continued to decline throughout the cauliflower season. The overall reduction in the number of propagules after two broccoli crops was approximately 94%, in contrast to the fivefold increase in the number of propagules in infested main plots without broccoli after two cauliflower crops. Disease incidence and severity were both reduced approximately 50% (P < 0.05) in broccoli treatments compared with no broccoli treatments. Differences between furrow and subsurface-drip irrigation were not significant, but incidence and severity were significantly (P < 0.05) lower in the deficit irrigation regime compared with the other two regimes. Abundance of microsclerotia of V. dahliae on cauliflower roots about 8 weeks after cauliflower harvest was significantly (P < 0.05) lower in treatments with broccoli compared with treatments without broccoli. Rotating broccoli with cauliflower and incorporating broccoli residues into the soils is a novel means of managing Verticillium wilt on cauliflower and perhaps on other susceptible crops. This practice would be successful regardless of the irrigation methods or regimes followed on the susceptible crops.

Polygalacturonic acid (PGA) is an important constituent of Sorensen's NP-10 medium (NP-10) for estimating the population density of Verticillium dahliae in soil. Different types of PGA are available, but not all of them favor the growth of V. dahliae. Unavailability of PGA sodium salt from orange (P-1879) has created an unprecedented problem for the quantification of microsclerotia (MS) of V. dahliae in soil. The PGA from orange (P-3889) that is now available does not support the growth of V. dahliae. Therefore, experiments were conducted to optimize the use of NP-10 prepared with P-3889 and various concentrations of NaOH. NP-10 with P-3889 amended with eight concentrations of NaOH were compared with NP-10 prepared from PGA sodium salt from orange (P-1879, now discontinued) and citrus (P-3850) along with cellophane and Napectate media for recovery of MS from soil and growth of V. dahliae on the media. Seven soils were assayed for MS, and eight isolates of V. dahliae were evaluated for growth and production of MS. Concentrations of NaOH >0.035N and <0.02N in NP-10 with P-3889 reduced mycelial growth, microsclerotial production, and recovery of MS from soils. Similarly, NP-10 with P-3850 alone, cellophane, and Na-pectate media had significantly reduced growth on media and recovery of V. dahliae from soils. The NP-10 with P-3889 and 0.025N NaOH consistently yielded numbers of V. dahliae MS from soil samples and supported the growth and production of MS similar to the NP-10 with P-1879. The medium developed in this study can serve as a direct replacement for the original NP-10 that was developed nearly three decades ago, an important component of which is no longer available.

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