Sclerotinia sclerotiorum, the causal agent of white stem rot, is responsible for significant losses in crop yields around the globe. While our understanding of S. sclerotiorum infection is becoming clearer, genetic control of the pathogen has been elusive and effective control of pathogen colonization using traditional broad-spectrum agro-chemical protocols are less effective than desired. In the current study, we developed species-specific RNA interference-based control treatments capable of reducing fungal infection. Development of a target identification pipeline using global RNA sequencing data for selection and application of double stranded RNA (dsRNA) molecules identified single gene targets of the fungus. Using this approach, we demonstrate the utility of this technology through foliar applications of dsRNAs to the leaf surface that significantly decreased fungal infection and S. sclerotiorum disease symptoms. Select target gene homologs were also tested in the closely related species, Botrytis cinerea, reducing lesion size and providing compelling evidence of the adaptability and flexibility of this technology in protecting plants against devastating fungal pathogens.

Pasmo is one of the most widespread diseases threatening flax production. To identify genetic regions associated with pasmo resistance (PR), a genome-wide association study was performed on 370 accessions from the flax core collection. Evaluation of pasmo severity was performed in the field from 2012 to 2016 in Morden, MB, Canada. Genotyping-by-sequencing has identified 258,873 single nucleotide polymorphisms (SNPs) distributed on all 15 flax chromosomes. Marker-trait associations were identified using ten different statistical models. A total of 692 unique quantitative trait nucleotides (QTNs) associated with 500 putative quantitative trait loci (QTL) were detected from six phenotypic PR datasets (five individual years and average across years). Different QTNs were identified with various statistical models and from individual PR datasets, indicative of the complementation between analytical methods and/or genotype × environment interactions of the QTL effects. The single-locus models tended to identify large-effect QTNs while the multi-loci models were able to detect QTNs with smaller effects. Among the putative QTL, 67 had large effects (3–23%), were stable across all datasets and explained 32–64% of the total variation for PR in the various datasets. Forty-five of these QTL spanned 85 resistance gene analogs including a large toll interleukin receptor, nucleotide-binding site, leucine-rich repeat (TNL) type gene cluster on chromosome 8. The number of QTL with positive-effect or favorite alleles (NPQTL) in accessions was significantly correlated with PR (R2 = 0.55), suggesting that these QTL effects are mainly additive. NPQTL was also significantly associated with morphotype (R2 = 0.52) and major QTL with positive effect alleles were present in the fiber type accessions. The 67 large effect QTL are suited for marker-assisted selection and the 500 QTL for effective genomic prediction in PR molecular breeding.

Although many Bacillus species are known to be good antibiotic producers capable of acting as biocontrol agents, the underlying antimicrobial mechanisms are often poorly understood. In this study, 21 Bacillus strains, demonstrating over 50% mycelial inhibition against Sclerotinia sclerotiorum as well as significant control in plant assays, were examined for the presence of antibiotic biosynthetic genes. Primers specific for bacillomycin D, iturin A, surfactin, mycosubtilin, fengycin, and zwittermicin A were used to amplify biosynthetic genes from these bacteria using PCR. The majority of strains harbored surfactin (21/21) and iturin A (20/21) biosynthetic genes. Three strains (Bacillus subtilis 3057, Bacillus amyloliquefaciens BS6, and Bacillus mycoides 4079) were positive for bacillomycin D, whereas 4 strains (B. subtilis H-08-02, B. subtilis 3057, B. amyloliquefaciens BS6, and B. mycoides 4079) showed the presence of the fengycin biosynthetic gene. The zwittermicin A gene was detected in B. mycoides S, Bacillus thuringiensis BS8, and B. amyloliquefaciens BS6. Sequence analysis of purified PCR products revealed homology with corresponding genes from other Bacillus sp. in the GenBank database. Production of particular antibiotics in strains BS6, H-08-02, 3057, and 4079 was confirmed through matrix-assisted laser desorption ionization - time of flight - mass spectroscopy (MALDI-TOF-MS). This study revealed the equivalent capability of different Bacillus strains from various microhabitats to produce the above-mentioned antibiotics and highlights the possibility of using some strains as potential biocontrol agents under different microhabitats distant from their original habitat. Furthermore, it will enable researchers to develop rational strategies for the application of the antagonists and their metabolites within an agroecosystem. To the best of our knowledge, this is the first report of a B. mycoides strain that carries biosynthetic genes and produces fengycin and surfactin.

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