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Gauthier Crouch posted an update 1 year, 7 months ago
Blackleg of oilseed rape is a damaging invasive disease caused by the species complex Leptosphaeria maculans (Lm)/L. biglobosa (Lb), which are composed of at least two and seven phylogenetic subclades, respectively. Generally, Lm is more virulent than Lb, however, under certain conditions, Lb can cause a significant yield loss in oilseed rape. Lb ‘brassicae’ (Lbb) has been found to be the causal agent for blackleg of oilseed rape in China, whereas Lm and Lb ‘canadensis’ (Lbc) were frequently detected in imported seeds of oilseed rape, posing a risk of spread into China. In order to monitor the blackleg-pathogen populations, a diagnostic tool based on loop-mediated isothermal amplification (LAMP) was developed using a 615-bp-long DNA sequence from Lbb that was derived from a randomly amplified polymorphic DNA assay. The LAMP was optimized for temperature and time, and tested for specificity and sensitivity using the DNA extracted from Lbb, Lbc, Lm, and 10 other fungi. The results showed that the optimal temperature and time were 65°C and 40 min, respectively. The LAMP primer set was specific to Lbb and highly sensitive as it detected the Lbb DNA as low as 132 fg per reaction. The LAMP assay was validated using the DNA extracted from mycelia and conidia of a well-characterized Lbb isolate, and its utility was evaluated using the DNA extracted from leaves, stems, pods and seeds of oilseed rape. The LAMP assay developed herein will help for monitoring populations of the blackleg pathogens in China and developing strategies for management of the blackleg disease.Corynespora cassiicola, which causes Corynespora leaf spot, results in considerable yield loss of cucumber grown in greenhouses. Frequent reports of reduced efficacy and control failure of fungicides warrant new, efficient alternative chemistries. In the present study, the sensitivity of C. cassiicola to benzovindiflupyr was evaluated using a collection of 81 isolates collected from Shandong, China. The mean EC50 values for mycelial growth, spore germination and germ tube elongation of C. cassiicola were 0.69 ± 0.44, 0.12 ± 0.063 and 0.13 ± 0.076 µg mL-1, respectively. Benzovindiflupyr treatment led to a reduced respiration rate and ATP production of C. cassiicola and decreased spore pathogenicity by 21.9% on average. Additionally, detached cucumber leaves sprayed with fungicides before or after inoculation were used to assess the efficacy of benzovindiflupyr against C. cassiicola. Benzovindiflupyr (150 µg mL-1) exhibited preventive and curative efficacies of 86.9% and 77.1%, respectively. Benzovindiflupyr at 150 g a.i. ha-1 provided over 70% efficacy in field trials performed in 2018 and 2019, which was significantly higher than that of the reference fungicides fluopyram and fluxapyroxad at the same dose. Furthermore, the yield of commercial cucumber increased as disease incidence decreased. Our findings pave the way for the introduction of benzovindiflupyr in the integrated management of Corynespora leaf spot.Fusarium root rot, caused by Fusarium solani and F. tricinctum, is a major soybean disease in the North Central United States. This study investigated the effects of the macroconidia density and the additive effects of soybean cyst nematode (SCN), Heterodera glycines, on the severity of Fusarium root rot. To determine the effect of spore density on severity, experiments were conducted in La Prairie silt loam soil in a greenhouse using conidial suspensions ranging from 101 to 106 macroconidia/ml soil. Root discoloration and lesion lengths on taproots increased as spore numbers increased, with significant effects of spore densities starting at 104 and 105 macroconidia/ml soil for F. solani and F. tricinctum, respectively. A non-linear sigmoid model was fitted to root discoloration against density, while a linear regression model was fitted to root lesion length against density. The interaction between the nematode at different egg densities with the two Fusarium species at 105 macroconidia/ml soil was investigated. In the greenhouse, root discoloration and lesion length were significantly greater in plants inoculated with Fusarium spp. and H. glycines at 10 eggs/ml soil or greater, compared to Fusarium spp. Phospho(enol)pyruvic acid monopotassium alone. In field trials, co-infestation of soil with the two Fusarium spp. and H. glycines significantly increased root rot severity at an egg density of 16.7 eggs/ml soil. The results indicated that the presence of SCN can increase severity of root rot caused by F. solani and F. tricinctum and egg density in the soil is an important factor in the interaction.Phytophthora ramorum (Werres, De Cock & Man in’t Veld) was recovered from symptomatic foliage of periwinkle at a botanical garden in WA in March 2015. Symptoms were tan colored lesions with a dark brown margin visible on both surfaces of the leaf and were found on wounds or around leaf margins. Periwinkle is native to Europe and is commonly used for ground cover in ornamental landscapes. It is known to be invasive in US forests near the urban/wildland interface. Potential spread of P. ramorum into WA forests is of regulatory concern, as well as long distance spread to other states via nursery stock (7 CFR §301.92-2). Phytophthora ramorum was isolated from symptomatic foliage by excising leaf pieces 4-6 mm in diameter and surface-sterilizing in 0.6% sodium hypochlorite followed by two rinses in sterile water. Leaf pieces were plated on PARP medium (Ferguson and Jeffers 1999) and after 2-3 days at 20°C, slow-growing dense colonies with coralloid hyphae were isolated onto V8 agar. Colony morphology and chlamydosof P. ramorum confirmed by observation of colony morphology and chlamydospore production. The presence of P. ramorum was also confirmed with DNA extraction from symptomatic foliage from plants from each of the two trials followed by PCR and sequencing of the COX1 gene (EU124920, 100%) (Figure S2). None of the water-inoculated controls were positive for P. ramorum. Low isolation success could be attributed to reduced pathogenicity due to being a non-wildtype isolate. Acknowledgements This work was supported by the USDA National Institute of Food and Agriculture, McIntire-Stennis project 1019284 and USDA APHIS Cooperative Agreement AP17PPQS&T00C070.

