Nitrogen management strategies on plant growth and severities of Sclerotinia stem rot of canola in eastern Canada

Effective nitrogen (N) management strategies are important for ensuring a balance between optimizing plant growth and minimizing disease damage. A field experiment was conducted for three years to (i) assess the effects of N fertilizer application on the growth and seed yield of canola, and severities of Sclerotinia stem rot (SSR), and (ii) determine a reasonable N-rate for optimizing plant growth and minimizing the loss from SSR in eastern Canada. The experiment was designed with factorial combinations of eight N treatments and two canola hybrids. All N-treatments reduced canola emergence with increasing preplant N application rates above 100 kg ha–1, but had a positive impact on plant height, fresh weight, dry weight and seed yield. The development of SSR showed differential responses to N application rates. Of all the treatments, the split application (50 kg N ha–1 at preplant plus 100 kg N ha–1 side-dressed at the 6-leaf stage) increased canola growth, and often produced the highest or similar seed yields to those of equivalent N rate applied as preplant. At the 150 kg ha–1 N rate, no severe development of SSR was observed in either preplant-only or split application. Overall, this study demonstrates that the split-N management strategy (50+100 kg ha–1) maintained a balance between enhancing plant growth and mitigating the negative impacts of SSR on canola.

Genetic Mapping and Genomic Prediction of Sclerotinia Stem Rot Resistance to Rapeseed/Canola (Brassica Napus L.) at Seedling Stage

The lack of complete host resistance and a complex resistance inheritance nature between rapeseed/canola and Sclerotinia sclerotiorum often limits the development of functional molecular markers that enable breeding for sclerotinia stem rot (SSR) resistance. However, genomics-assisted selection has the potential to accelerate the breeding for SSR resistance. Therefore, genome-wide association (GWA) mapping and genomic prediction (GP) was performed using a diverse panel of 337 rapeseed/canola genotypes. Three-weeks old seedlings were screened using the petiole inoculation technique (PIT). Days to wilt (DW) up to 2 weeks and lesion phenotypes (LP) at 3, 4, and 7 days post inoculation (dpi) were recorded. A strong correlation (r = -0.94) between DW and LP_4dpi implied that a single time point scoring at four days could be used as a proxy trait. GWA analyses using single-locus (SL) and multi-locus (ML) models identified a total of 35, and 219 significantly associated SNPs, respectively. Out of these, seventy-one SNPs were identified by a combination of the SL model and any of the ML models, at least two ML models, or two traits. These SNPs explained 1.4-13.3% of the phenotypic variance, and considered as significant, could be associated with SSR resistance. Eighty-one candidate genes with a function in disease resistance were associated with the significant SNPs. Six GP models resulted in moderate to high (0.45-0.68) predictive ability depending on SSR resistance traits. The resistant genotypes and significant SNPs will serve as valuable resources for future SSR resistance breeding. Our results also highlight the potential of genomic selection to improve rapeseed/canola breeding for SSR resistance.

Integration of Row Spacing, Seeding Rates, and Fungicide Application for Control of Sclerotinia Stem Rot in Glycine max

Soybean (Glycine max) farmers in the Upper Midwest region of the United States frequently experience severe yield losses due to Sclerotinia stem rot (SSR). Previous studies have revealed benefits of individual management practices on SSR. This study examined the integration of multiple control practices on the development of SSR, yield, and the economic implications of these practices. Combinations of row spacings, seeding rates, and fungicide applications were examined in multi-site field trials across the Upper Midwest from 2017-2019. These trials revealed that wide row spacing and low seeding rates individually reduced SSR levels but also reduced yields. Yields were similar across the three higher seeding rates examined. However, site-years where SSR developed showed the highest partial profits in the intermediate seeding rates. This indicates that partial profits in diseased fields were negatively impacted by high seeding rates, but this trend was not observed when SSR did not develop. Fungicides strongly reduced the development of SSR, while also increasing yields. However, there was a reduction in partial profits due to their use at a low soybean sale price, but at higher sale prices fungicide use was similar to not treating. Additionally, the production of new inoculum was predicted from disease incidence, serving as an indicator of increased risk for SSR development in future years. Overall, this study suggests the use of wide rows and low seeding rates could be useful in fields with a history of SSR, while reserving narrow rows and higher seeding rates for fields without a history of SSR.

Genetic and Proteomic Basis of Sclerotinia Stem Rot Resistance in Indian Mustard [Brassica juncea (L.) Czern & Coss.]

Sclerotinia stem rot is one of the utmost important disease of mustard, causing considerable losses in seed yield and oil quality. The study of the genetic and proteomic basis of resistance to this disease is imperative for its effective utilization in developing resistant cultivars. Therefore, the genetic pattern of Sclerotinia stem rot resistance in Indian mustard was studied using six generations (P1, P2, F1, F2, BC1P1, and BC1P2) developed from the crossing of one resistant (RH 1222-28) and two susceptible (EC 766300 and EC 766123) genotypes. Genetic analysis revealed that resistance was governed by duplicate epistasis. Comparative proteome analysis of resistant and susceptible genotypes indicated that peptidyl-prolyl cis-trans isomerase (A0A078IDN6 PPIase) showed high expression in resistant genotype at the early infection stage while its expression was delayed in susceptible genotypes. This study provides important insight to mustard breeders for designing effective breeding programs to develop resistant cultivars against this devastating disease.

Genome-wide association mapping and genomic prediction for adult stage sclerotinia stem rot resistance in Brassica napus (L) under field environments

Sclerotinia stem rot (SSR) is a fungal disease of rapeseed/canola that causes significant seed yield losses and reduces its oil content and quality. In the present study, the reaction of 187 diverse canola genotypes to SSR was characterized at full flowering stage using the agar plug to stem inoculation method in four environments. Genome-wide association study (GWAS) using three different algorithms identified 133 significant SNPs corresponding with 123 loci for disease traits like stem lesion length (LL), lesion width (LW), and plant mortality at 14 (PM_14D) and 21 (PM_21D) days. The explained phenotypic variation of these SNPs ranged from 3.6 to 12.1%. Nineteen significant SNPs were detected in two or more environments, disease traits with at least two GWAS algorithms. The strong correlations observed between LL and other three disease traits evaluated, suggest they could be used as proxies for SSR resistance phenotyping. Sixty-nine candidate genes associated with disease resistance mechanisms were identified. Genomic prediction (GP) analysis with all the four traits employing genome-wide markers resulted in 0.41–0.64 predictive ability depending on the model specifications. The highest predictive ability for PM_21D with three models was about 0.64. From our study, the identified resistant genotypes and stable significant SNP markers will serve as a valuable resource for future SSR resistance breeding. Our study also suggests that genomic selection holds promise for accelerating canola breeding progress by enabling breeders to select SSR resistance genotypes at the early stage by reducing the need to phenotype large numbers of genotypes.

Host-Induced Gene Silencing of a Multifunction Gene Sscnd1 Enhances Plant Resistance Against Sclerotinia sclerotiorum

Sclerotinia sclerotiorum is a devastating necrotrophic fungal pathogen and has a substantial economic impact on crop production worldwide. Magnaporthe appressoria-specific (MAS) proteins have been suggested to be involved in the appressorium formation in Magnaporthe oryzae. Sscnd1, an MAS homolog gene, is highly induced at the early infection stage of S. sclerotiorum. Knock-down the expression of Sscnd1 gene severely reduced the virulence of S. sclerotiorum on intact rapeseed leaves, and their virulence was partially restored on wounded leaves. The Sscnd1 gene-silenced strains exhibited a defect in compound appressorium formation and cell integrity. The instantaneous silencing of Sscnd1 by tobacco rattle virus (TRV)-mediated host-induced gene silencing (HIGS) resulted in a significant reduction in disease development in tobacco. Three transgenic HIGS Arabidopsis lines displayed high levels of resistance to S. sclerotiorum and decreased Sscnd1 expression. Production of specific Sscnd1 siRNA in transgenic HIGS Arabidopsis lines was confirmed by stem-loop qRT-PCR. This study revealed that the compound appressorium-related gene Sscnd1 is required for cell integrity and full virulence in S. sclerotiorum and that Sclerotinia stem rot can be controlled by expressing the silencing constructs of Sscnd1 in host plants.

Transcriptome Analysis Reveals the Complex Molecular Mechanisms of Brassica napus–Sclerotinia sclerotiorum Interactions

Sclerotinia stem rot caused by Sclerotinia sclerotiorum is a devastating disease for many important crops worldwide, including Brassica napus. Although numerous studies have been performed on the gene expression changes in B. napus and S. sclerotiorum, knowledge regarding the molecular mechanisms of B. napus–S. sclerotiorum interactions is limited. Here, we revealed the changes in the gene expression and related pathways in both B. napus and S. sclerotiorum during the sclerotinia stem rot (SSR) infection process using transcriptome analyses. In total, 1,986, 2,217, and 16,079 differentially expressed genes (DEGs) were identified in B. napus at 6, 24, and 48 h post-inoculation, respectively, whereas 1,511, 1,208, and 2,051 DEGs, respectively, were identified in S. sclerotiorum. The gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that most of the hormone-signaling pathways in B. napus were enriched, and thus, the hormone contents at four stages were measured. The DEGs and hormone contents revealed that salicylic acid was activated, while the jasmonic acid pathway was repressed at 24 h post-inoculation. Additionally, the expressional patterns of the cell wall-degrading enzyme-encoding genes in S. sclerotiorum and the hydrolytic enzymes in B. napus were consistent with the SSR infection process. The results contribute to a better understanding of the interactions between B. napus and S. sclerotiorum and the development of future preventive measures against SSR.

Silicon Alleviates the Disease Severity of Sclerotinia Stem Rot in Rapeseed

Sclerotinia stem rot, caused by Sclerotinia sclerotiorum, is a devastating disease in rapeseed. The objective of this study was to investigate the role and the mechanism of silicon (Si) in alleviating the disease severity of S. sclerotiorum in rapeseed. In the absorption assays, the rapeseed that absorbed 10 mM of K2SiO3 exhibited an 86% decrease in lesion size on infected leaves as compared with controls. In the spray assay, the lesion length on rapeseed stems was reduced by 30.5–32.9% with the use of 100 mM of a foliar Si fertilizer as compared with controls. In the pot assay, the lesion length on rapeseed stems was reduced by 34.9–38.3% when using the Si fertilizer as basal fertilizer. In the field assay, both the disease incidence and disease index of sclerotinia stem rot were significantly reduced with the usage of a solid Si fertilizer, Si foliar fertilizer, and the application of both, without negative affection on the main agronomic traits and seed quality of rapeseed. The transcriptome sequencing, quantitative reverse transcription PCR (qRT-PCR), and biochemical assays between K2SO4- and K2SiO3- treated rapeseed leaves revealed that Si promoted the biosynthesis of defense-related substances and enhanced the antioxidation and detoxification abilities of rapeseed after infection. Thus, this study concluded that Si can alleviate the disease severity of S. sclerotiorum in rapeseeds, partially due to the induced defense responses.

Baseline Sensitivity and Control Efficacy of Various Group of Fungicides against Sclerotinia sclerotiorum in Oilseed Rape Cultivation

Sclerotinia stem rot (SSR), caused by Sclerotinia sclerotiorum, is a devastating disease of oilseed rape that may cause significant yield losses if not controlled by cultural management strategies and fungicide applications. Studies were conducted to evaluate the efficacy of different group of fungicides as well as a biopesticide, including azoxystrobin, boscalid, fludioxonil, prothioconazole, tebuconazole, azoxystrobin/tebuconazole, boscalid/pyraclostrobin, prothioconazole/fluopyram and Bacillus amyloliquefaciens, on baseline sensitivity of S. sclerotiorum isolates under in-vitro conditions as well as control of SSR in the field. Artificial inoculation and mist irrigation prompt to reproducible SSR infection in oilseed rape cultivation. All compounds significantly reduced 36.7% to 86.9% SSR severity and increased 55.2% to 98.7% yield, 1.5% to 7.0% thousand grain-weight, 1.5% to 5.9% oil and 0.1% to 1.3% protein content. Fludioxonil, boscalid/pyraclostrobin and fluopyram/prothioconazole achieved strongest fungicidal activity against SSR. The biopesticide provided 36% of disease control. Under in vitro conditions, B. amyloliquefaciens not only strongly inhibited mycelial growth but also the formation of sclerotia in all concentrations. Boscalid and fludioxonil exhibited the highest level of fungicidal activity against S. sclerotiorum, with mean EC50 values of 1.23 and 1.60 μg a.s. mL−1. The highest variability of EC50 values between isolates was observed towards prothioconazole and azoxystrobin.