Sclerotinia sclerotiorum is a fungal plant pathogen and the causal agent of lettuce drop, an economically important disease of California lettuce. The structure of the S. sclerotiorum mating type locus MAT has previously been reported and consists of two idiomorphs that are fused end-to-end as in other homothallics. We investigated the diversity of S. sclerotiorum MAT using a total of 283 isolates from multiple hosts and locations, and identified a novel MAT allele that differed by a 3.6-kb inversion and was designated Inv+, as opposed to the previously known S. sclerotiorum MAT that lacked the inversion and was Inv-. The inversion affected three of the four MAT genes: MAT1-2-1 and MAT1-2-4 were inverted and MAT1-1-1 was truncated at the 3’-end. Expression of MAT genes differed between Inv+ and Inv- isolates. In Inv+ isolates, only one of the three MAT1-2-1 transcript variants of Inv- isolates was detected, and the alpha1 domain of Inv+ MAT1-1-1 transcripts was truncated. Both Inv- and Inv+ isolates were self-fertile, and the inversion segregated in a 1∶1 ratio regardless of whether the parent was Inv- or Inv+. This suggested the involvement of a highly regulated process in maintaining equal proportions of Inv- and Inv+, likely associated with the sexual state. The MAT inversion region, defined as the 3.6-kb MAT inversion in Inv+ isolates and the homologous region of Inv- isolates, was flanked by a 250-bp inverted repeat on either side. The 250-bp inverted repeat was a partial MAT1-1-1 that through mediation of loop formation and crossing over, may be involved in the inversion process. Inv+ isolates were widespread, and in California and Nebraska constituted half of the isolates examined. We speculate that a similar inversion region may be involved in mating type switching in the filamentous ascomycetes Chromocrea spinulosa, Sclerotinia trifoliorum and in certain Ceratocystis species.

A recent study on fungal mating type genes revealed two MAT alleles within homothallic Sclerotinia sclerotiorum differentiated by an inversion, Inv− (inversion negative) and Inv+ (inversion positive). An analysis of mating type in closely related S. minor was conducted to shed light on the evolution of this MAT inversion. Inv− and Inv+ MAT alleles were identified in S. minor and were characterized. Both MAT alleles in S. minor were flanked by APN2 and SLA2, and consisted of two idiomorphs fused as in other homothallic ascomycetes. However, in the Inv+ MAT, the 3·6 kb MAT region was inverted relative to the Inv− MAT. Except for the inversion, both Inv− and Inv+ MAT in S. minor were equal in size and identical in nucleotide sequence. The MAT inversion in Inv+ S. minor was at exactly the same place as in Inv+ S. sclerotiorum and affected three of four MAT genes: MAT1‐1‐1 was truncated and MAT1‐2‐4 and MAT1‐2‐1 were inverted. Unlike S. sclerotiorum, expression of MAT genes did not differ between Inv− and Inv+ S. minor. The 250 bp inverted repeat motif that flanked the inverted MAT region in S. sclerotiorum and believed responsible for the MAT inversion was also found in S. minor, but was 256 bp. Depending on the MAT genes, 93–96% nucleotide identity was observed between Sclerotinia species. Both Inv+ and Inv− MAT S. minor and S. sclerotiorum isolates were commonly found in lettuce fields of Arizona along with MAT heterokaryons.

Field experiments were conducted over 2 years in Yuma County, AZ, and Imperial County, CA, to determine the efficacy of several biocontrol agents for the management of lettuce drop caused by Sclerotinia spp. Commercial formulations of Trichoderma harzianum (Plantshield, Supersivit), Gliocladium virens (Soilgard), Coniothyrium minitans (Contans), and Bacillus subtilis (Companion) were evaluated and compared with the chemical fungicide iprodione (Rovral) against Sclerotinia sclerotiorum and S. minor. A single application of biocontrol products or of Rovral did not reduce lettuce drop caused by either Sclerotinia species. However, two applications of Contans, one at planting and one at post-thinning, significantly reduced the incidence of lettuce drop caused by S. sclerotiorum and increased yield but had no effect on S. minor at both locations in both years. Two applications of other biocontrol products did not significantly reduce disease incidence despite medium to high recovery following application. In contrast, Contans was only sporadically recovered following application. In vitro fungicide sensitivity evaluation revealed that both Trichoderma and Gliocladium species were tolerant to iprodione, dicloran (Botran), and vinclozolin (Ronilan) up to 1,000 ppm a.i., whereas both Sclerotinia spp. and C. minitans were sensitive to all three fungicides above 1 ppm. In summary, Contans was the most effective treatment for the control of lettuce drop caused by S. sclerotiorum, but no treatment was effective against S. minor in the desert lettuce production systems.

The Fusarium solani species complex (FSSC) includes important root pathogens of soybean in the United States, but the evolutionary lineages associated with soybean root rot are unknown. A multilocus phylogeny based on 93 isolates from soybean and pea roots from North Dakota and Minnesota revealed that root rot was associated with three known phylogenetic species, FSSC 3 + 4 (=Fusarium falciforme) (3 % of isolates), FSSC 5 (60 %), FSSC 11 (34 %), and one unknown species, FSSC X (2 %). Of these species FSSC 5 and FSSC 3 + 4 are clinically important while FSSC 11 is a plant pathogen. Isolates from FSSC 11 were pathogenic on soybean, dry bean, pea and lentil, and did not grow at 37 °C. However, isolates from FSSC 5 were weakly to non-pathogenic, but grew at 37 °C. Isolates from both FSSC 5 and FSSC 11 were highly resistant to fludioxonil in vitro. This is the first study revealing the pathogenic robustness of FSSC 11 in causing root rot among Fabaceae crops and also the association of clinically important members of the FSSC with roots of a widely grown field crop in the United States.

Lettuce drop caused by Sclerotinia spp. is an economically important disease of lettuce (Lactuca sativa L.), and cultivars with resistance to mycelial infection by Sclerotinia sclerotiorum (Lib.) de Bary and S. minor Jagger as well as to S. sclerotiorum ascospores are needed. Assessing resistance in field experiments can be complicated by fast bolting or small stature lettuce lines that may escape rather than resist the pathogens. Therefore, methods to select resistant lines from morphologically variable populations are needed. We used S. sclerotiorum and S. minor-infested field experiments, regression analysis, field experiments with artificially high plant densities, and S. sclerotiorum ascospore inoculations to identify lettuce lines with resistance to both pathogens. Three replicated experiments in S. sclerotiorum-infested fields were conducted in Yuma, AZ, and three replicated experiments in a S. minor-infested field were conducted in Salinas, CA, using diverse populations of iceberg, romaine, leaf, butterhead, Latin, oilseed lettuce, and wild relatives of lettuces; and genetic variation for the incidence of lettuce drop from mycelial infections was identified. In two S. minor field experiments, a quadratic regression model was developed that related rapid bolting with reduced lettuce drop. Regression residuals were calculated, and eight cultivars or PIs had negative residuals in two independent field experiments, indicating higher resistance than predicted by their rate of bolting. Eruption, a small-statured Latin cultivar, had significantly lower disease levels than susceptible cultivars in experiments with high plant densities, indicating that its small size did not facilitate disease escape. Ascospore inoculations confirmed resistance in ‘Eruption’ and L. virosa SAL012, whereas the oilseed lettuce PI 251246 may have partial resistance to infection. These lines will likely be useful for development of Sclerotinia spp.-resistant lettuce cultivars.

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Field experiments were conducted over 2 years in Yuma, AZ, and Holtville, CA, to establish the relationship between soil sclerotium density of Sclerotinia sclerotiorum and the incidence of lettuce drop on different lettuce (Lactuca sativa) types under different irrigation systems, and to determine the efficacy of the biocontrol agent Coniothyrium minitans (Contans) against S. sclerotiorum on crisphead lettuce at varied sclerotium densities under different irrigation systems. There was no significant interaction of irrigation (overhead sprinkler versus furrow) with either sclerotium density or with biocontrol treatment. Lettuce drop incidence was lowest in romaine lettuce compared with crisphead or leaf lettuce at all soil sclerotium densities. There was a significant positive correlation between the sclerotial density and the percent disease incidence. Disease incidence in plots infested with 2 sclerotia/m2 of bed was not significantly higher than in control plots regardless of lettuce type. However, plots infested with 40 or 100 sclerotia/m2 of bed revealed a significantly higher disease incidence over the control in all lettuce types. A single application of Contans at planting significantly reduced the incidence of lettuce drop in all lettuce types even under high disease pressure. There were no significant differences between recommended (2.2 kg/ha) and high (4.4 kg/ha) application rates of Contans or between one or two applications of the product.