Investigation of Using Hyperspectral Vegetation Indices to Assess Brassica Downy Mildew.

Downy mildew caused by Hyaloperonospora brassicae is a severe disease in Brassica oleracea that significantly reduces crop yield and marketability. This study aims to evaluate different vegetation indices to assess different downy mildew infection levels in the Brassica variety Mildis using hyperspectral data. Artificial inoculation using H. brassicae sporangia suspension was conducted to induce different levels of downy mildew disease. Spectral measurements, spanning 350 nm to 1050 nm, were conducted on the leaves using an environmentally controlled setup, and the reflectance data were acquired and processed. The Successive Projections Algorithm (SPA) and signal sensitivity calculation were used to extract the most informative wavelengths that could be used to develop downy mildew indices (DMI). A total of 37 existing vegetation indices and three proposed DMIs were evaluated to indicate downy mildew (DM) infection levels. The results showed that the classification using a support vector machine achieved accuracies of 71.3%, 80.7%, and 85.3% for distinguishing healthy leaves from DM1 (early infection), DM2 (progressed infection), and DM3 (severe infection) leaves using the proposed downy mildew index. The proposed new downy mildew index potentially enables the development of an automated DM monitoring system and resistance profiling in Brassica breeding lines.

A Rapid Glove-Based Inoculum Sampling Technique to Monitor Erysiphe necator in Commercial Vineyards.

Information on the presence and severity of grape powdery mildew (GPM), caused by Erysiphe necator, has long been used to guide management decisions. While recent advances in the available molecular diagnostic assays and particle samplers have made monitoring easier, there is still a need for more efficient field collection of E. necator. The use of vineyard worker gloves worn during canopy manipulation as a sampler (glove swab) of E. necator was compared with samples identified by visual assessment with subsequent molecular confirmation (leaf swabs) and airborne spore samples collected by rotating-arm impaction traps (impaction traps). Samples from United States commercial vineyards in Oregon, Washington, and California were analyzed using two TaqMan qPCR assays targeting the internal transcribed spacer regions or cytochrome b gene of E. necator. Based on qPCR assays, visual disease assessments misidentified GPM up to 59% of the time with a higher frequency of misidentification occurring earlier in the growing season. Comparison of the aggregated leaf swab results for a row (n = 915) to the row's corresponding glove swab had 60% agreement. The latent class analysis (LCA) indicated that glove swabs were more sensitive than leaf swabs in detecting E. necator presence. The impaction trap results had 77% agreement to glove swabs (n = 206) taken from the same blocks. The LCAs estimated that the glove swabs and impaction trap samplers varied each year in which was more sensitive for detection. This likely indicates that these methods have similar levels of uncertainty and provide equivalent information. Additionally, all samplers, once E. necator was detected, were similarly sensitive and specific for detection of the A-143 resistance allele. Together, these results suggest that glove swabs are an effective sampling method for monitoring the presence of E. necator and, subsequently, the G143A amino acid substitution associated with resistance to quinone outside inhibitor fungicides in vineyards. Glove swabs could reduce sampling costs due to the lack of need for specialized equipment and time required for swab collection and processing.

Environmental Effects on Taxonomic Turnover in Soil Fauna across Multiple Forest Ecosystems in East Asia.

The large-scale spatial variation in and causes of biotic turnover of soil fauna remain poorly understood. Analyses were conducted based on published data from 14 independent sampling sites across five forest ecosystems in East Asia. Jaccard and Sørensen's indices were used to measure turnover rates in soil fauna orders. A redundancy analysis was used to investigate multiple environmental controls of the composition of soil fauna communities. The results showed that both Jaccard's and Sørensen's index increased significantly with increasing latitude difference. The environment explained 54.1%, 50.6%, 57.3% and 50.9% of the total variance, and spatial factors explained 13.8%, 15.9%, 21.0% and 12.6% of the total variance in the orders' composition regarding overall, phytophagous, predatory and saprophagous fauna, respectively. In addition, climate factors in environmental processes were observed to have a stronger effect than soil factors on the orders' turnover rates. Our results support the hypothesis that the effect of environment factors on soil animal taxa turnover is more important than the effect of spatial factors. Climatic factors explained more variation in the turnover of phytophagic fauna, but soil and environment factors equally explained the variation in the turnover of predatory fauna. This study provides evidence to support both environmental filtering and dispersal limitation hypotheses at the regional and population scales.

Identification of Putative SDHI Target Site Mutations in the SDHB, SDHC, and SDHD Subunits of the Grape Powdery Mildew Pathogen Erysiphe necator.

Succinate dehydrogenase inhibitors (SDHIs) are fungicides used in control of numerous fungal plant pathogens, including Erysiphe necator, the causal agent of grapevine powdery mildew (GPM). Here, the sdhb, sdhc, and sdhd genes of E. necator were screened for mutations that may be associated with SDHI resistance. GPM samples were collected from 2017 to 2020 from the U.S. states of California, Oregon, Washington, and Michigan, and the Canadian province of British Columbia. Forty-five polymorphisms were identified in the three sdh genes, 17 of which caused missense mutations. Of these, the SDHC-p.I244V substitution was shown in this study to reduce sensitivity of E. necator to boscalid and fluopyram, whereas the SDHC-p.G25R substitution did not affect SDHI sensitivity. Of the other 15 missense mutations, the SDHC-p.H242R substitution was shown in previous studies to reduce sensitivity of E. necator toward boscalid, whereas the equivalents of the SDHB-p.H242L, SDHC-p.A83V, and SDHD-p.I71F substitutions were shown to reduce sensitivity to SDHIs in other fungi. Generally, only a single amino acid substitution was present in the SDHB, SDHC, or SDHD subunit of E. necator isolates, but missense mutations putatively associated with SDHI resistance were widely distributed in the sampled areas and increased in frequency over time. Finally, isolates that had decreased sensitivity to boscalid or fluopyram were identified but with no or only the SDHC-p.G25R amino acid substitution present in SDHB, SDHC, and SDHD subunits. This suggests that target site mutations probably are not the only mechanism conferring resistance to SDHIs in E. necator.

Spatiotemporal Distribution of Potato-Associated Alternaria Species in Wisconsin.

Potato early blight caused by Alternaria solani and potato brown spot caused by Alternaria alternata are two common potato foliar diseases. Fungicide applications in Wisconsin target early blight with the expectation of managing brown spot simultaneously. However, distributions of the two pathogens over time and space have not been previously reported in Wisconsin. In this study, six potato fields in Wisconsin were systematically sampled during 2014 to 2017 to investigate the incidences of both pathogens; incidences of the pathogens were compared by location, timing, and year. Incidence of A. alternata had been recorded starting in late June in every field every year, whereas incidence of A. solani started later in some of the fields. At the end of each growing season throughout the four years, the highest incidence (100% most of the years) of A. solani was observed in a field in Plover, located in a concentrated potato production area of central Wisconsin, and a field in Arena, located in an isolated potato production area in southern Wisconsin. The highest incidence (100% every year) of A. alternata was observed in a field in Madison, an isolated potato field that was managed organically. Such distribution patterns could be a result of the different host ranges of the pathogens and abundance of inoculum in different geographic locations. In 2016, disease severity was correlated with the incidence of both pathogens (P < 0.0001 to P = 0.0299), but in 2017, disease severity was correlated with the incidence of A. solani in most of the fields (P < 0.0001 to P = 0.0799). These findings suggest that both Alternaria spp. are important for considering disease management strategies in central and southern Wisconsin.

Distribution and Stability of Quinone Outside Inhibitor Fungicide Resistance in Populations of Potato Pathogenic Alternaria spp. in Wisconsin.

Quinone outside inhibitor (QoI) fungicides have been an important class in managing potato early blight caused by Alternaria solani and brown spot caused by A. alternata. Because of the single-site mode of action character of QoI fungicides, which are relied on for management of diseases in Wisconsin, and the abundant asexual conidia production of the Alternaria species, pathogen isolates with QoI resistance have been detected after just a few years of QoI fungicide usage in commercial production fields. Resistance to QoIs has been attributed to amino acid substitutions F129L and G143A in cytochrome b of A. solani and A. alternata, respectively, as a result of point mutations. The aim of this study was to assess Alternaria populations in Wisconsin for QoI resistance before and after fungicide applications in order to evaluate resistance stability. A TaqMan single nucleotide polymorphism genotyping assay was designed based on the sequences of the cytochrome b gene from Alternaria isolates collected in Wisconsin to profile QoI resistance in Alternaria populations as well as to explore factors that may influence frequency of QoI resistance in the pathogen populations. This assay successfully identified the mutations conferring QoI resistance in isolates collected from four locations each year from 2015 to 2017. During the course of this study, the frequency of A. solani isolates with the F129L mutation was consistently high and showed primarily the TTA mutation type. The frequency of A. alternata isolates with the G143A mutation started relatively low and increased at the end of the production season in each year (P = 0.0109, P = 0.2083, and P = 0.0159). A potato field managed without use of QoI fungicides showed a significantly lower (P < 0.05) frequency of A. alternata isolates carrying G143A than conventionally managed potato fields. The overall frequency of A. alternata isolates carrying G143A in the four locations was similar over the 3 years (P = 0.2971). The QoI resistance characteristics of the isolates were stable even when QoI selection pressure was removed for at least five subculture transfers, and the mutation types of codons 129 and 143 in the cytochrome b gene in A. solani and A. alternata, respectively, remained the same. This indicated that the application of QoIs in the field is not the sole factor responsible for the variation of the frequency of QoI resistance in the pathogen populations.

Aerial Concentrations of Pathogens Causing Early Blight and Brown Spot Within Susceptible Potato Fields.

Early blight caused by Alternaria solani and brown spot caused by A. alternata are two common foliar diseases of potato, with early blight typically predominating in incidence and severity. Renewed interest in these two diseases has arisen as a result of notable differences in fungicide resistance profiles of the pathogens and inconsistent outcomes of disease management tactics. The pathogens share similar disease cycles, but they differ in the shape and size of their conidia. A. solani has a host range that includes just the Solanaceae, whereas A. alternata has a broad host range spanning numerous plant families. Such differences may result in differences in dispersal of the pathogens and subsequently influence epidemiology and management outcomes. In the commercial potato fields investigated in this study, the aerial conidial concentrations of A. solani and A. alternata differed significantly, with those of A. alternata conidia being higher in number and more variable than those of A. solani. The aerial conidial concentration of A. solani almost always significantly decreased with height (0 to 3 m above the canopy), whereas such a decrease was only observed for 4 of 12 days for A. alternata. The atmospheric concentrations of A. alternata were higher than those of A. solani at both upwind and downwind edges of the field (P < 0.0001), indicating more potential for long-distance dispersal. A higher aerial concentration of conidia at the downwind than the upwind location was observed for A. solani (P < 0.05), whereas overall no such effect was observed for A. alternata. This indicated that the potato fields investigated were likely the source of A. solani conidia, but they may not be the sole source of A. alternata. Results are consistent with inoculum of A. solani coming primarily from within the potato crop, whereas that of A. alternata may be generated from diverse plant sources across the landscape.

Diversity and Virulence of Alternaria spp. Causing Potato Early Blight and Brown Spot in Wisconsin.

Early blight, caused by Alternaria solani, along with brown spot, caused by A. alternata, have the potential to reduce quality and yield in potato production globally. Prior to this study, the incidence, disease impact, and fungicide resistance attributes of A. alternata in Wisconsin were poorly understood. Potato pathogens were isolated from foliar lesions at three commercial locations in Wisconsin in 2012 and 2017 and were initially morphologically identified as A. solani (n = 33) and A. alternata (n = 40). Identifications were further corroborated with the phylogenetic analysis of the internal transcribed spacer (ITS), translation elongation factor 1 (TEF1), gapdh, Alt a 1, and OPA10-2. A multigene phylogeny of ITS, TEF1, gapdh, and Alt a 1 showed five genotypes of A. alternata and one single genotype of A. solani. We demonstrated that the A. alternata isolates were virulent on potato cultivars Russet Burbank (P < 0.013) and Atlantic (P < 0.0073), though they caused less disease than A. solani (P < 0.0001 and P < 0.0001, respectively). A. alternata caused little disease on the breeding line 24-24-12 (P = 0.9929), and A. solani caused fewer disease symptoms on 24-24-12 than on Russet Burbank (P < 0.0001) or Atlantic (P < 0.0001). Breeding line 24-24-12 may be a promising source of potential resistance for the two diseases. There was no significant difference in virulence of different A. alternata genotypes, and no significant difference in virulence or genotype clustering among isolates from the three locations. Isolates of A. alternata that induced chlorosis caused larger lesion areas than isolates that did not in Russet Burbank (P < 0.0001), Atlantic (P < 0.0001), and 24-24-12 (P = 0.0365). There was no significant difference in virulence between quinone outside inhibitor (QoI)-sensitive and QoI-resistant isolates of A. alternata. This study enhanced our understanding of potato early blight and brown spot in Wisconsin, and suggested that A. alternata in addition to A. solani should be carefully monitored and possibly uniquely managed in order to achieve overall disease control.