Estimating the Climate Niche of Sclerotinia sclerotiorum Using Maximum Entropy Modeling.

Sclerotinia sclerotiorum, a fungal pathogen, causes world-wide crop losses and additional disease management strategies are needed. Modeling the climate niche of this fungus may offer a tool for the selection of biological control organisms and cultural methods of control. Maxent, a modeling technique, was used to characterize the climate niche for the fungus. The technique requires disease occurrence data, bioclimatic data layers, and geospatial analysis. A cross-correlation was performed with ArcGIS 10.8.1, to reduce nineteen bioclimatic variables (WorldClim) to nine variables. The model results were evaluated by AUC (area under the curve). A final model was created with the random seed procedure of Maxent and gave an average AUC of 0.935 with an AUC difference of -0.008. The most critical variables included annual precipitation (importance: 14.1%) with a range of 450 mm to 2500 mm and the mean temperature of the coldest quarter (importance: 55.6%) with a range of -16 °C to 24 °C, which contributed the most to the final model. A habitat suitability map was generated in ArcGIS 10.8.1 from the final Maxent model. The final model was validated by comparing results with another occurrence dataset. A Z-Score statistical test confirmed no significant differences between the two datasets for all suitability areas.

Host selectivity and genetic variation of Discula umbrinella isolates from two oak species: analyses of intergenic spacer region sequences of ribosomal DNA.

Discula umbrinella, a fungal endophyte of oak species, colonizes and reproduces on leaves of Quercus alba and Q. rubra in forest ecosystems. Twenty-nine isolates collected from leaves of both oak species (16 from Q. alba and 13 from Q. rubra) were assayed for oak species preference and genetic variation based on primer-specific polymerase chain reactions for the intergenic spacer region (IGS) of ribosomal DNA. DNA sequencing of the polymerase chain reaction products revealed a 10-bp insertion (237-247 bp) at the 3' end of the IGS region present in nine isolates and absent in 20 of the isolates. Phylogenetic analysis of the IGS region using the neighbor-joining method identified IGS groups (groups I-V) based on single nucleotide sequence differences. Host selectivity and geographic origin of isolates were correlated in some instances with the IGS groups. Isolates within each IGS group were further analyzed for nucleotide polymorphisms to confirm genotype identity and genotype diversity. Ten different genotypes (Va-Vj) were identified among the isolates analyzed. Genotype diversity was greatest in IGS groups I, IV, and V. Seventy percent of the genotypes (Vc, Vd, Ve, Vf, Vg,Vi, and Vj) contained isolates with single tree species preferences.

A protocol for direct sequencing of multiple gene specific PCR products from Discula umbrinella, a fungal endophyte, utilizing bufferless precast electrophoresis.

A protocol for direct sequencing of polymerase chain reaction (PCR) products from mycelia of Discula umbrinella, a fungal endophyte, using bufferless electrophoresis is described. This improved method allows researchers to conduct high-capacity screening of multiple gene regions for fungal endophytes applicable to microbial ecology and population genetic studies.