Phyllachora Maydis Research Articles

Foliar Inoculation of Phyllachora maydis into Corn Induces Infection and Local Spread in Field Environments.

Tar spot of corn (Zea mays L.) is a significant disease in the United States and Canada caused by Phyllachora maydis, an obligate biotroph fungus. However, field research critical for understanding and managing the disease has been hindered by a need for methods to inoculate corn with P. maydis in field environments. In this study, we developed and demonstrated the efficacy of a method to initiate tar spot in field settings using inoculations of corn leaves with P. maydis inoculum that had been stored at -20°C for 10 months. Stromata of P. maydis were observed 19 days after inoculations in two field experiments, and stromata resulting from secondary spread were initially observed 39 to 41 days after the initial inoculations. Tar spot was not present in the fields beyond the inoculated areas or localized spread area, signifying that the establishment of initial disease resulted solely from inoculations. This study enhances our understanding of inoculation and infection of corn with P. maydis and tar spot development in field environments. The results will aid new research into understanding the corn tar spot pathosystem and improving management strategies.

Genome-Informed Trophic Classification and Functional Characterization of Virulence Proteins from the Maize Tar Spot Pathogen Phyllachora maydis.

Phyllachora maydis is an ascomycete foliar fungal pathogen and the causal agent of tar spot in maize. Although P. maydis is considered an economically important foliar pathogen of maize, our general knowledge of the trophic lifestyle and functional role of effector proteins from this fungal pathogen remains limited. Here, we utilized a genome-informed approach to predict the trophic lifestyle of P. maydis and functionally characterized a subset of candidate effectors from this fungal pathogen. Leveraging the most recent P. maydis genome annotation and the CATAStrophy pipeline, we show that this fungal pathogen encodes a predicted carbohydrate-active enzymes (CAZymes) repertoire consistent with that of biotrophs. To investigate fungal pathogenicity, we selected 18 candidate effector proteins that were previously shown to be expressed during primary disease development. We assessed whether these putative effectors share predicted structural similarity with other characterized fungal effectors and determined whether any suppress plant immune responses. Using AlphaFold2 and Foldseek, we showed that one candidate effector, PM02_g1115, adopts a predicted protein structure similar to that of an effector from Verticillium dahlia. Furthermore, transient expression of candidate effector-fluorescent protein fusions in Nicotiana benthamiana revealed two putative effectors, PM02_g378 and PM02_g2610, accumulated predominantly in the cytosol, and three candidate effectors, PM02_g1115, PM02_g7882, and PM02_g8240, consistently attenuated chitin-mediated reactive oxygen species production. Collectively, the results presented herein provide insights into the predicted trophic lifestyle and putative functions of effectors from P. maydis and will likely stimulate continued research to elucidate the molecular mechanisms used by P. maydis to induce tar spot.

Uncovering the Infection Strategy of Phyllachora maydis During Maize Colonization: A Comprehensive Analysis.

Tar spot, a disease caused by the ascomycete fungal pathogen Phyllachora maydis, is considered one of the most significant yield-limiting diseases of maize (Zea mays) within the United States. P. maydis may also be found in association with other fungi, forming a disease complex that is thought to result in the characteristic fisheye lesions. Understanding how P. maydis colonizes maize leaf cells is essential for developing effective disease control strategies. Here, we used histological approaches to elucidate how P. maydis infects and multiplies within susceptible maize leaves. We collected tar spot-infected maize leaf samples from four different fields in northern Indiana at three different time points during the growing season. Samples were chemically fixed and paraffin-embedded for high-resolution light and scanning electron microscopy. We observed a consistent pattern of disease progression in independent leaf samples collected across different geographical regions. Each stroma contained a central pycnidium that produced asexual spores. Perithecia with sexual spores developed in the stomatal chambers adjacent to the pycnidium, and a cap of spores formed over the stroma. P. maydis reproductive structures formed around but not within the vasculature. We observed P. maydis associated with two additional fungi, one of which is likely a member of the Paraphaeosphaeria genus; the other is an unknown fungi. Our data provide fundamental insights into how this pathogen colonizes and spreads within maize leaves. This knowledge can inform new approaches to managing tar spot, which could help mitigate the significant economic losses caused by this disease.

Investigations into Economic Returns Resulting from Foliar Fungicides and Application Timing on Management of Tar Spot in Indiana Hybrid Corn.

Tar spot, caused by Phyllachora maydis, is the most significant yield-limiting disease of corn (Zea mays L.) in Indiana. Currently, fungicides are an effective management tool for this disease, and partial returns from their use under different disease severity conditions has not previously been studied. Between 2019 and 2021, two separate field experiments were conducted in each year in Indiana to assess the efficacy of nine foliar fungicide products and nine fungicide application timings based on corn growth stages on tar spot symptoms and stromata, canopy greenness, yield, and influence on partial returns. All fungicides evaluated significantly suppressed tar spot development in the canopy and increased canopy greenness over the nontreated control. Additionally, applications of mefentrifluconazole + pyraclostrobin, metconazole + pyraclostrobin, cyproconazole + picoxystrobin at tassel, and propiconazole + benzovindiflupyr + azoxystrobin between the tassel and dough growth stages were the most effective at significantly reducing disease severity, increasing canopy greenness, protecting yield, and offered the greatest partial return. Fungicide products varied in their ability to protect yield under low and high disease severity conditions relative to the nontreated control. Consistently, positive yield increases were observed when disease severity was high, which translated to greater profitability relative to low severity conditions. On average, the yield increases across foliar fungicide products and timed application treatments were 544.6 and 1,020.7 kg/ha greater, and partial returns using a grain value of $0.17/kg were $92.6/ha and $173.5/ha greater, respectively, when high severity conditions occurred. This research demonstrates that foliar fungicides and appropriately timed fungicide applications can profitably be used to manage tar spot in Indiana under high disease severity conditions.

First report of Phyllachora maydis causing tar spot on corn in Delaware.

In October 2023, lesions consistent with descriptions of tar spot (Phyllachora maydis) were observed on corn (Zea mays) in Kent and Sussex County, Delaware (DE). Black, raised stromata were observed on leaves of commercially grown corn hybrids. Plants were at physiological maturity and disease severity was low with symptoms present on 1 to 10% of plants. In collected tissue, individual leaf severities ranged from 1 to 3% of leaf area with lesions. Hyaline conidia measuring approximately 15.5 µm in length and 0.5 µm in width were observed microscopically (n=5). Stromata were excised and sterilized in a 0.825% sodium hypochlorite solution for 30 s, rinsed in sterile deionized water for 30 s, and dried on a sterile paper towel for 30 s. Tissues were ground in a 1.5 mL microcentrifuge tube with a sterile plastic pestle. DNA was extracted using a DNeasy Plant Mini Kit (Qiagen). DNA was amplified at the internal transcribed spacer (ITS) region with ITS4 and ITS5 primers using polymerase chain reaction (PCR). NCBI BLAST search results yielded 100% sequence homology and 100% query cover (350/515 bp) to P. maydis accession MG881848.1 (Moura et al. 2023). Koch's postulates could not be completed due to the obligate nature of P. maydis. Tarspot was initially discovered in the United States in 2016 in Indiana and Illinois (Ruhl et al. 2016).This is the first report of tar spot on corn in DE. Yield losses from P. maydis can range depending on time of infection, environmental factors, and hybrid susceptibility and have been recorded up to 100% (Rocco da Silva et al. 2021). Because the disease did not enter the area until the end of the season, no yield impact was observed for 2023. Monitoring for the progression of disease will be crucial for future seasons (Telenko et al. 2020). High humidity and moisture levels favor disease development. Approximately half of DE corn acreage is irrigated due to sandy soils, current irrigation timing strategies may need to be reevaluated. Fungicide efficacy trials for management of tar spot have been conducted in other regions, but continued research will be needed to assess management options and optimize application timing for farmers in DE and the Mid-Atlantic region.

Development of a qPCR Assay for Species-Specific Detection of the Tar Spot Pathogen Phyllachora maydis

Phyllachora maydis is a fungal plant pathogen that causes tar spot of corn ( Zea mays) in North and South America, causing devastating yield losses under favorable conditions. Although the causal agent is relatively easy to diagnose via macroscopic and microscopic observations, other diseases and conditions, such as insect frass, have been mistaken for tar spot of corn. Furthermore, conidia and ascospores in isolation can be difficult to visually distinguish from other fungi, and the development of signs and symptoms of the disease may not be observed until 12 to 20 days after infection. Therefore, we developed a TaqMan quantitative polymerase chain reaction (qPCR) assay for the detection and quantification of this pathogen to be used for diagnostics and airborne spore quantification. The assay was designed for the internal transcribed spacer region of P. maydis. The specificity of the assay was confirmed and tested against various nontarget Phyllachora species, corn pathogens, endophytes, and P. maydis samples from several states in the Midwest and from Mexico. The detection limit of this assay was determined to be 100 fg of genomic P. maydis DNA. To demonstrate the transferability of this technology, the assay was tested in different labs using various qPCR thermal cyclers. This assay can be used in downstream research involving latency period, disease prediction, and diagnostics. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Uncovering the environmental conditions required for Phyllachora maydis infection and tar spot development on corn in the United States for use as predictive models for future epidemics

Phyllachora maydis is a fungal pathogen causing tar spot of corn (Zea mays L.), a new and emerging, yield-limiting disease in the United States. Since being first reported in Illinois and Indiana in 2015, P. maydis can now be found across much of the corn growing regions of the United States. Knowledge of the epidemiology of P. maydis is limited but could be useful in developing tar spot prediction tools. The research presented here aims to elucidate the environmental conditions necessary for the development of tar spot in the field and the creation of predictive models to anticipate future tar spot epidemics. Extended periods (30-day windowpanes) of moderate mean ambient temperature (18–23 °C) were most significant for explaining the development of tar spot. Shorter periods (14- to 21-day windowpanes) of moisture (relative humidity, dew point, number of hours with predicted leaf wetness) were negatively correlated with tar spot development. These weather variables were used to develop multiple logistic regression models, an ensembled model, and two machine learning models for the prediction of tar spot development. This work has improved the understanding of P. maydis epidemiology and provided the foundation for the development of a predictive tool for anticipating future tar spot epidemics.

Is Microdochium maydis Associated with Necrotic Lesions in the Tar Spot Disease Complex? A Culture-Based Survey of Maize in Mexico and the Midwestern United States.

Tar spot, caused byPhyllachora maydis, is an emerging disease of corn in the United States. Stromata ofP. maydisare sometimes surrounded by necrotic lesions known as fisheyes and were previously reported to be caused by the fungus Microdochiummaydis. The association ofM. maydiswith fisheye lesions has not been well documented outside of initial descriptions from the early 1980s. The objective of this work was to assess and identify Microdochium-like fungi associated with necrotic lesions surrounding P. maydis stromata using a culture-based method. In 2018, corn leaf samples with fisheye lesions associated with tar spot stromata were collected from 31 production fields across Mexico, Illinois, and Wisconsin. Cultures of pure isolates collected from Mexico believed to be M. maydis were included in the study. A total of 101 Microdochium/Fusarium-like isolates were obtained from the necrotic lesions, and 91% were identified asFusariumspp., based on initial ITS sequence data.Multi-gene (ITS, TEF1-α, RPB1, and RPB2)phylogenies were constructed for a subset of 55 isolates;Microdochium,Cryptostroma, andFusariumreference sequences were obtained from GenBank. All the necrotic lesion isolates clustered withinFusariumlineages and were phylogenetically distinct from the Microdochiumclade. All Fusarium isolates from Mexico belonged to theF. incarnatum-equisetispecies complex, whereas >85% of the U.S. isolates grouped within theF. sambucinumspecies complex. Our study suggests that initial reports of M. maydis were misidentifications of resident Fusarium spp. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

A new and effective method to induce infection of Phyllachora maydis into corn for tar spot studies in controlled environments

BackgroundTar spot of corn is a significant and spreading disease in the continental U.S. and Canada caused by the obligate biotrophic fungus Phyllachora maydis. As of 2023, tar spot had been reported in 18 U.S. states and one Canadian Province. The symptoms of tar spot include chlorotic flecking followed by the formation of black stromata where conidia and ascospores are produced. Advancements in research and management for tar spot have been limited by a need for a reliable method to inoculate plants to enable the study of the disease. The goal of this study was to develop a reliable method to induce tar spot in controlled conditions.ResultsWe induced infection of corn by P. maydis in 100% of inoculated plants with a new inoculation method. This method includes the use of vacuum-collection tools to extract ascospores from field-infected corn leaves, application of spores to leaves, and induction of the disease in the dark at high humidity and moderate temperatures. Infection and disease development were consistently achieved in four independent experiments on different corn hybrids and under different environmental conditions in a greenhouse and growth chamber. Disease induction was impacted by the source and storage conditions of spores, as tar spot was not induced with ascospores from leaves stored dry at 25 ºC for 5 months but was induced using ascospores from infected leaves stored at -20 ºC for 5 months. The time from inoculation to stromata formation was 10 to 12 days and ascospores were present 19 days after inoculation throughout our experiments. In addition to providing techniques that enable in-vitro experimentation, our research also provides fundamental insights into the conditions that favor tar spot epidemics.ConclusionsWe developed a method to reliably inoculate corn with P. maydis. The method was validated by multiple independent experiments in which infection was induced in 100% of the plants, demonstrating its consistency in controlled conditions. This new method facilitates research on tar spot and provides opportunities to study the biology of P. maydis, the epidemiology of tar spot, and for identifying host resistance.

First report of tar spot on corn caused by Phyllachora maydis in Kentucky.

In September 2021, signs of black circular to oval shaped fungal structures (stromata) were observed on corn (Zea mays L.) leaves on a non-commercial inbred line in Todd County, Kentucky. Signs were only observed in a small pocket within the larger field, with disease levels ranging from 1- 5% incidence and 1-25% severity on individual leaves affected in the field. Corn leaves had senesced and only fungal structures were available to aid in diagnosis. Microscopic examination of stromata uncovered ascomata within the clypei/stromata. Further examination of ascomata revealed multiple asci containing eight hyaline, uniseriate, aseptate, oval to ovoid ascospores ranging in size from 8 to 12 µm x 5 to 7 µm. Observed signs were consistent with published reports of tar spot caused by Phyllachora maydis (Parbery 1967; Valle-Torres et al. 2020). For molecular confirmation of the causal agent, corn leaves were surface sterilized in diluted bleach (10%) for 30 seconds and stromata were excised from the leaves using a sterile scalpel. Five to seven stromata were placed into each microcentrifuge tube. Liquid nitrogen was added to the microcentrifuge tubes and the frozen stromata were ground using a sterilized pestle. The ground stromata tissue was used for DNA extraction using a Synergy 2.0 plant DNA extraction kit (OPS Diagnostics, Lebanon, NJ). A portion of the internal transcribed spacer (ITS) region was amplified by PCR utilizing ITS-4 and ITS-5 primers. Amplicons were subjected to Sanger sequencing to obtain a consensus sequence. Using the BLASTn algorithm the consensus sequence shared 100% similarity to three P. maydis Genbank accessions: MG881848.1, MG8814847.1, MG881846.1. A representative sequence was deposited in GenBank (accession no. OQ034699.1). Due to P. maydis being an obligate parasite, Koch's postulates were not attempted.

First Report of Tar Spot on Corn Caused by Phyllachora maydis in the Great Plains.

Tar spot caused by the fungus Phyllachora maydis Maubl. is a significant foliar disease of corn (Zea mays L.). Threatening corn production across the Americas, this disease can reduce the quality of silage and grain yield (Rocco da Silva et al. 2021; Valle-Torres et al. 2020). Lesions caused by P. maydis are usually black, glossy, and raised stromata on the leaf surface and occasionally on the husk. (Liu 1973; Rocco da Silva et al. 2021). Samples consistent with tar spot of corn were collected between September and October of 2022 from 6 fields in Kansas, 23 in Nebraska, and 6 in South Dakota. One sample was selected from each of the three states for further microscopic evaluation and molecular analysis. Signs of the fungus were visually and microscopically confirmed in eight Nebraska counties in October 2021; however, in the 2021, season tar spot sings were not found in Kansas and South Dakota. In the 2022 season disease severity varied by location; some fields in Kansas had <1% incidence, whereas in South Dakota disease incidence approached 1-2%, and in Nebraska between <1-5%. Stromata were present on both green and senescing tissues. Morphological characteristics of the pathogen were similar and consistent with the description of P. maydis (Parbery 1967) from all examined leaves across all locations. Asexual spores (conidia) were produced in pycnidial fruiting bodies ranging in size 1.29 to 2.82 x 8.84 to 16.95 µm (n = 40, average 1.98 × 13.30 μm). The pycnidial fruiting bodies were often found adjacent to perithecia within the stromata. For molecular confirmation, stromata were aseptically removed from leaves collected at each location and DNA extracted using a phenol chloroform method. The internal transcribed spacer (ITS) regions of the ribosomal RNA gene were sequenced using ITS1/ITS4 universal primers (Larena et al. 1999). Amplicons were Sanger sequenced (Genewiz, Inc., South Plainfield, NJ), and a consensus sequence for each sample was deposited in GenBank: Kansas (OQ200487), Nebraska (OQ200488), and South Dakota (OQ200489). Using the BLASTn, sequences from Kansas, Nebraska and South Dakota showed 100% homology with 100% query cover to other P. maydis GenBank accessions (MG881848.1; OL342916.1; OL342915.1). Koch's postulates were not performed given the obligate nature of the pathogen (Muller and Samuels 1984). This report documents the first confirmation of tar spot on corn in Kansas, Nebraska, and South Dakota (Great Plains).

Elucidating the obligate nature and biological capacity of an invasive fungal corn pathogen.

Tar spot is a devasting corn disease caused by the obligate fungal pathogen Phyllachora maydis. Since its initial identification in the United States in 2015, P. maydis has become an increasing threat to corn production. Despite this, P. maydis has remained largely understudied at the molecular level due to difficulties surrounding its obligate lifestyle. Here, we generated a significantly improved P. maydis nuclear and mitochondrial genome using a combination of long- and short-read technologies and also provide the first transcriptomic analysis of primary tar spot lesions. Our results show that P. maydis is deficient in inorganic nitrogen utilization, is likely heterothallic, and encodes for significantly more protein coding genes, including secreted enzymes and effectors, than previous determined. Furthermore, our expression analysis suggests that following primary tar spot lesion formation, P. maydis might reroute carbon flux away from DNA replication and cell division pathways and towards pathways previously implicated in having significant roles in pathogenicity, such as autophagy and secretion. Together, our results identified several highly expressed unique secreted factors that likely contribute to host recognition and subsequent infection, greatly increasing our knowledge of the biological capacity of P. maydis, which have much broader implications for mitigating tar spot of corn.

Potential Biocontrol Agents of Corn Tar Spot Disease Isolated from Overwintered Phyllachora maydis Stromata.

Tar spot disease in corn, caused by Phyllachora maydis, can reduce grain yield by limiting the total photosynthetic area in leaves. Stromata of P. maydis are long-term survival structures that can germinate and release spores in a gelatinous matrix in the spring, which are thought to serve as inoculum in newly planted fields. In this study, overwintered stromata in corn leaves were collected in Central Illinois, surface sterilized, and caged on water agar medium. Fungi and bacteria were collected from the surface of stromata that did not germinate and showed microbial growth. Twenty-two Alternaria isolates and three Cladosporium isolates were collected. Eighteen bacteria, most frequently Pseudomonas and Pantoea species, were also isolated. Spores of Alternaria, Cladosporium, and Gliocladium catenulatum (formulated as a commercial biofungicide) reduced the number of stromata that germinated compared to control untreated stromata. These data suggest that fungi collected from overwintered tar spot stromata can serve as biological control organisms against tar spot disease.

Assessment of symptom induction via artificial inoculation of the obligate biotrophic fungus Phyllachora maydis (Maubl.) on corn leaves

ObjectiveTar spot is a foliar disease of corn caused by Phyllachora maydis, which produces signs in the form of stromata that bear conidia and ascospores. Phyllachora maydis cannot be cultured in media; therefore, the inoculum source for studying tar spot comprises leaves with stromata collected from naturally infected plants. Currently, there is no effective protocol to induce infection under controlled conditions. In this study, an inoculation method was assessed under greenhouse and growth chamber conditions to test whether stromata of P. maydis could be induced on corn leaves.ResultsExperiments resulted in incubation periods ranging between 18 and 20 days and stromata development at the beginning of corn growth stage VT-R1 (silk). The induced stromata of P. maydis were confirmed by microscopy, PCR, or both. From thirteen experiments conducted, four (31%) resulted in the successful production of stromata. Statistical analyses indicate that if an experiment is conducted, there are equal chances of obtaining successful or unsuccessful infections. The information from this study will be valuable for developing more reliable P. maydis inoculation methods in the future.

The parental inbred lines of the Nested Association Mapping (NAM) population of corn show sources of resistance to tar spot in northern Indiana.

Tar spot is a major foliar disease of corn caused by the obligate fungal pathogen Phyllachora maydis, first identified in Indiana in 2015. Under conducive weather conditions, P. maydis causes significant yield losses in the U.S. and other countries constituting a major threat to corn production. Relatively little is known about resistance to tar spot other than a major quantitative gene that was identified in tropical maize lines. To test for additional sources of resistance against populations of P. maydis in North America, twenty-six parental inbred lines of the Nested Associated Mapping (NAM) population were evaluated for tar spot resistance in Indiana in replicated field trials under natural infection for 3 years. Tar spot disease severity was scored visually using a 0 to 100 % scale. Maximum disease severities (MDS) for tar spot scoring at reproductive growth stage ranged from 0 to 48.3 % with 0% being most resistant and 48.3% being most susceptible. Nine inbred lines were resistant to P. maydis with MDS ranging from 0 to 5.0%, six were moderately resistant (5.2 to 10.6% MDS), two were moderately susceptible (11.7 to 26.0% MDS) and the remaining eight inbred lines were rated as susceptible (30.0 to 48.3% MDS). There was some variability between years, due to higher disease pressure after 2019. Inbred B73, the common parent of the NAM populations, was rated as susceptible with MDS of 30.0%. The nine highly resistant lines provide a potential source of new genes for genetic analysis and mapping of tar spot resistance in corn.

Impact of Fungicides on Dual-Purpose and Brown Midrib Zea mays Hybrids Used for Silage in Wisconsin

Decisions about hybrid choice and fungicide application when growing corn ( Zea mays L.) for silage are important considerations to maximize sustainable production. The objective of this research was to evaluate how brown midrib hybrids compare with dual-purpose hybrids and how fungicides interact with these two classes of corn hybrids. To explore this, field research trials were conducted in Arlington, Wisconsin, in 2020 and 2021. In both years, the trials were arranged in a randomized complete block design and included one brown midrib hybrid, one dual-purpose hybrid, and seven fungicide application regimes. In-field disease ratings were made for tar spot (caused by Phyllachora maydis), ear rot, and stalk rot (caused by Gibberella zeae). Harvested silage was analyzed for yield, quality, and deoxynivalenol (DON) concentration. The selection of hybrid significantly ( P &lt; 0.01) influenced the silage quality parameters, such as starch and total tract neutral detergent fiber digestibility, a measure of fiber digested throughout the tract of a cow. Fungicide use influenced dry matter yield ( P &lt; 0.05), tar spot severity ( P &lt; 0.01), and DON concentration ( P = 0.05). Our findings suggest that using appropriate fungicides improves yield and reduces disease severity and DON contamination. Better quality silage could be obtained from brown midrib hybrids at the expense of yield. Therefore, Wisconsin farmers are encouraged to primarily utilize a dual-purpose class of corn hybrids to ensure optimal milk production and secondarily apply fungicides to reduce DON levels in corn chopped for silage.

Effects of Nitrogen Application Rate and Plant Density on Severity of Tar Spot of Corn

Tar spot of corn, caused by the fungus Phyllachora maydis, is an economically important foliar disease recently reported in the United States and Canada. Due to the recent introduction of Phyllachora maydis, the impacts of cultural management practices on disease development are still unknown. Separate field studies were conducted to determine the effects of nitrogen (N) application rate and plant density on disease development. Field trials were conducted across six site years in Michigan with two corn hybrids of differing disease susceptibility. The relative area under the disease progress curve was used to compare disease development between N application rates and plant densities. Nitrogen application rate had no significant effect on disease at any location. Plant density and disease had a significant ( P &lt; 0.05) inverse relationship at five of six site years, with an average 41% decrease in the relative area under the disease progress curve for every 1,000 plants per hectare increase. The economically optimal planting density ranged from 73 to 77 thousand plants per hectare for US$150 to 300 per metric ton corn prices, demonstrating that relatively low planting densities were more profitable despite greater disease. Therefore, other disease management practices including hybrid selection may be more effective at protecting yield than increasing plant density.

Effects of Disease Resistance and Fungicide Application on Management of Tar Spot on Corn in the North Central United States

Tar spot, caused by Phyllachora maydis, is the most significant disease of corn in the North Central United States. Elucidating an integrated management approach to protect grain yield is a high priority. The integration of tillage, hybrid, and fungicide application on tar spot severity, canopy greenness, and grain yield was assessed in Indiana in 2019, 2020, and 2021. A split-plot arrangement of two tillage treatments as main plots with factorial arrangement of subplots with three hybrids (tar spot susceptible and two partially resistant) and fungicide application (propiconazole + benzovindiflupyr + azoxystrobin) were evaluated. Furthermore, a multistate study comparing two hybrids (susceptible and partially resistant) and fungicide application (propiconazole + benzovindiflupyr + azoxystrobin) was conducted in Indiana, Michigan, and Wisconsin in 2020. This research demonstrates that partially resistant hybrids with a standardized area under the disease progress curve of 2.5 and 3.0 had significantly less tar spot than a susceptible hybrid with a standardized area under the disease progress curve of 10.1 and increased canopy greenness ratings of 48.1 and 51.5%, which were significantly higher than the susceptible hybrid at 13.0% at maturity. Tar spot severity was further reduced and canopy greenness increased with a fungicide application in the susceptible hybrid. These results suggest that partial resistance alone may be used to manage tar spot. Yields did not differ significantly across hybrids with or without a fungicide application. However, a general trend of preserved yield potential was observed in the treatments with fungicide, indicating that knowledge of genetics and yield potential will be necessary to achieve the greatest benefit from partially resistant hybrids and judicious fungicide applications.

Tar Spot of Corn: A Diagnostic and Methods Guide

Tar spot of corn is an emerging plant disease in the continental United States and Canada caused by the fungal pathogen Phyllachora maydis Maubl. Tar spot has been known to occur in Mexico, the Caribbean, and Central and South America since the early to mid-1900s. In 2015, it was reported for the first time in the continental United States. Since that time, tar spot has spread across corn-producing areas in the United States with epidemics as recent as 2021 resulting in significant yield losses. Although tar spot has been known to affect corn for over a century in the Americas, the biology of the pathogen, etiology, and epidemiology of the disease are not well understood. Additionally, symptoms and signs of tar spot resemble other foliar diseases and abiotic disorders of corn, which may lead to misdiagnosis. In this paper, we provide a brief description of current knowledge about tar spot of corn, including pathogen taxonomy, host range, symptoms and signs, specimen storage, pathogenicity testing, diagnostic protocols, and geographic distribution. This information will be useful to diagnosticians, researchers, and other professionals working with this disease.

Candidate Effector Proteins from the Maize Tar Spot Pathogen Phyllachora maydis Localize to Diverse Plant Cell Compartments.

Most fungal pathogens secrete effector proteins into host cells to modulate their immune responses, thereby promoting pathogenesis and fungal growth. One such fungal pathogen is the ascomycete Phyllachora maydis, which causes tar spot disease on leaves of maize (Zea mays). Sequencing of the P. maydis genome revealed 462 putatively secreted proteins, of which 40 contain expected effector-like sequence characteristics. However, the subcellular compartments targeted by P. maydis effector candidate (PmEC) proteins remain unknown, and it will be important to prioritize them for further functional characterization. To test the hypothesis that PmECs target diverse subcellular compartments, cellular locations of super yellow fluorescent protein-tagged PmEC proteins were identified using a Nicotiana benthamiana-based heterologous expression system. Immunoblot analyses showed that most of the PmEC-fluorescent protein fusions accumulated protein in N. benthamiana, indicating that the candidate effectors could be expressed in dicot leaf cells. Laser-scanning confocal microscopy of N. benthamiana epidermal cells revealed that most of the P. maydis putative effectors localized to the nucleus and cytosol. One candidate effector, PmEC01597, localized to multiple subcellular compartments including the nucleus, nucleolus, and plasma membrane, whereas an additional putative effector, PmEC03792, preferentially labelled both the nucleus and nucleolus. Intriguingly, one candidate effector, PmEC04573, consistently localized to the stroma of chloroplasts as well as stroma-containing tubules (stromules). Collectively, these data suggest that effector candidate proteins from P. maydis target diverse cellular organelles and could thus provide valuable insights into their putative functions, as well as host processes potentially manipulated by this fungal pathogen.