Barley Yellow Dwarf Research Articles

Non-Invasive Characterization of Single-, Double- and Triple-Viral Diseases of Wheat With a Hand-Held Raman Spectrometer.

Plant diseases can reduce crop yield by up to 100%. Therefore, timely and confirmatory diagnosis of plant diseases is strongly desired. Typical pathogen assaying methods include polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). These approaches are quite useful but are also time-consuming and destructive to the sample. Raman spectroscopy (RS) is a modern analytical technique that enables non-invasive plant disease detection. In this study, we report on Raman-based detection of wheat diseases caused by wheat streak mosaic virus (WSMV) and barley yellow dwarf virus (BYDV). Our results show that RS can be used to differentiate between healthy wheat and wheat infected by these two viruses. We also show that RS can be used to identify whether wheat is infected by these individual viruses or by a combination of WSMV and BYDV, as well as WSMV, BYDV, and Triticum mosaic virus (TriMV). We found that wheat spectra showed non-linear spectroscopic responses to coinfection by different viruses. These results suggest that RS can be used to probe pathogen-specific changes in plant metabolism. The portable nature of this approach opens the possibility of RS directly in the field for confirmatory diagnostics of viral diseases.

Host nutrition mediates interactions between plant viruses, altering transmission and predicted disease spread

Interactions among co-infecting pathogens are common across host taxa and can affect infectious disease dynamics. Host nutrition can mediate these among-pathogen interactions, altering the establishment and growth of pathogens within hosts. It is unclear, however, how nutrition-mediated among-pathogen interactions affect transmission and the spread of disease through populations. We manipulated the nitrogen (N) and phosphorus (P) supplies to oat plants in growth chambers and evaluated interactions between two aphid-vectored Barley and Cereal Yellow Dwarf Viruses: PAV and RPV. We quantified the effect of each virus on the other's establishment, within-plant density, and transmission. Co-inoculation significantly increased PAV density when N and P supplies were low and tended to increase RPV density when N supply was high. Co-infection increased PAV transmission when N and P supplies were low and tended to increase RPV transmission when N supply was high. Despite the parallels between the effects of among-pathogen interactions on density and transmission, changes in virus density only partially explained changes in transmission, suggesting that virus density-independent processes contribute to transmission. A mathematical model describing the spread of two viruses through a plant population, parameterized with empirically derived transmission values, demonstrated that nutrition-mediated among-pathogen interactions could affect disease spread. Interactions that altered transmission through virus density-independent processes determined overall disease dynamics. Our work suggests that host nutrition alters disease spread through among-pathogen interactions that modify transmission.

Rich dynamics of a simple delay host-pathogen model of cell-to-cell infection for plant virus

Viral dynamics within plant hosts can be important for understanding plant disease prevalence and impacts. However, few mathematical modeling efforts aim to characterize within-plant viral dynamics. In this paper, we derive a simple system of delay differential equations that describes the spread of infection throughout the plant by barley and cereal yellow dwarf viruses via the cell-to-cell mechanism. By incorporating ratio-dependent incidence function and logistic growth of the healthy cells, the model can capture a wide range of biologically relevant phenomena via the disease-free, endemic, mutual extinction steady states, and a stable periodic orbit. We show that when the basic reproduction number is less than \begin{document}$ 1 $\end{document} ( \begin{document}$ R_0 ), the disease-free steady state is asymptotically stable. When \begin{document}$ R_0>1 $\end{document} , the dynamics either converge to the endemic equilibrium or enter a periodic orbit. Using a ratio-dependent transformation, we show that if the infection rate is very high relative to the growth rate of healthy cells, then the system collapses to the mutual extinction steady state. Numerical and bifurcation simulations are provided to demonstrate our theoretical results. Finally, we carry out parameter estimation using experimental data to characterize the effects of varying nutrients on the dynamics of the system. Our parameter estimates suggest that varying the nutrient supply of nitrogen and phosphorous can alter the dynamics of the infection in plants, specifically reducing the rate of viral production and the rate of infection in certain cases.

Registration of oat cultivar ‘Natty’

AbstractThe development of oat (Avena sativa L.) cultivars with improved grain yield and excellent test weight is necessary to increase the profitability of oat production. ‘Natty’ (Reg. no. CV‐385, PI 676027) was developed by the South Dakota State University (SDSU) oat breeding program and was released in 2014. The pedigree of Natty is SD020701//SD030888/ND030349. Natty exhibited high grain yield potential and excellent test weight when evaluated in the South Dakota Crop Performance Testing Oat Variety Trials and in the Uniform Early Oat Performance Nursery. The grain of Natty is characterized by a large proportion of plump kernels and groats with low beta‐glucan and oil content. Natty is resistant to smut, moderately susceptible to susceptible to crown rust, and moderately tolerant to Barley yellow dwarf virus. Natty is a tall and early‐maturing cultivar suited for the production of oat grain for the feed market. Although its milling quality is good, the low beta‐glucan content of Natty is not desirable for the food market. Based on its height, Natty is also suited for straw production. Three of the oat cultivars released by SDSU were named after Lon Hall's grandchildren; Natty is one of them. Lon Hall was oat breeder at SDSU from 2001 to 2014.

Population growth of Rhopalosiphum padi under different thermal regimes: an agent‐based model approach

Rhopalosiphum padi (Linnaeus, 1758) is abundant and has a broad geographic distribution. It is one of the most important cereal pests. In Brazil, the economic losses associated with this aphid result mainly from the transmission of the barley yellow dwarf viruses. Decision‐making for the adoption of management measures must consider the initial population size, the potential for population increase, and the time when this population will reach levels at which the resulting damage is equal to the costs of control measures. Consequently, the establishment of management programmes and decision support systems should be based on models that estimate the potential population growth of this pest species. Temperature is one of the main factors that determine the growth rate of insect populations. Generally, controlled experiments are designed to examine the relationship of temperature at fixed intervals in relation to the development phases of insects. In nature, thermal regimes are not constant, and population growth is the result of a series of combined events. In this work, the effects of different thermal regimes on the population growth of R. padi were compared. An agent‐based model was used to estimate population growth, and the parameters defined in controlled regimes were compared with fluctuating temperatures under natural conditions. The temperature‐driven model presented here can serve as a tool to predict population growth and decision‐making for aphid management. The model structure and the proposed experimental design allow the addition of modules and layers of factors that can progressively affect the populations of aphids to gradually improve the model.

Simultaneous Increase in CO2 and Temperature Alters Wheat Growth and Aphid Performance Differently Depending on Virus Infection.

Climate change impacts crop production, pest and disease pressure, yield stability, and, therefore, food security. In order to understand how climate and atmospheric change factors affect trophic interactions in agriculture, we evaluated the combined effect of elevated carbon dioxide (CO2) and temperature on the interactions among wheat (Triticum aestivum L.), Barley yellow dwarf virus species PAV (BYDV-PAV) and its vector, the bird cherry-oat aphid (Rhopalosiphum padi L.). Plant traits and aphid biological parameters were examined under two climate and atmospheric scenarios, current (ambient CO2 and temperature = 400 ppm and 20 °C), and future predicted (elevated CO2 and temperature = 800 ppm and 22 °C), on non-infected and BYDV-PAV-infected plants. Our results show that combined elevated CO2 and temperature increased plant growth, biomass, and carbon to nitrogen (C:N) ratio, which in turn significantly decreased aphid fecundity and development time. However, virus infection reduced chlorophyll content, biomass, wheat growth and C:N ratio, significantly increased R. padi fecundity and development time. Regardless of virus infection, aphid growth rates remained unchanged under simulated future conditions. Therefore, as R. padi is currently a principal pest in temperate cereal crops worldwide, mainly due to its role as a plant virus vector, it will likely continue to have significant economic importance. Furthermore, an earlier and more distinct virus symptomatology was highlighted under the future predicted scenario, with consequences on virus transmission, disease epidemiology and, thus, wheat yield and quality. These research findings emphasize the complexity of plant–vector–virus interactions expected under future climate and their implications for plant disease and pest incidence in food crops.

Does the Infectious Status of Aphids Influence Their Preference Towards Healthy, Virus-Infected and Endophytically Colonized Plants?

Aphids (Hemiptera: Aphididae) cause significant damage and transmit viruses to various crop plants. We aimed to evaluate how the infectious status of aphids influences their interaction with potential hosts. Two aphid (Myzus persicae and Rhopalosiphum padi) and plant (Nicotiana tabacum and Triticum aestivum) species were used. The preferences of aphids towards healthy, virus-infected (Potato Leafroll Virus (PLRV) and Barley Yellow Dwarf virus (BYDV)), and endophytic entomopathogenic fungi (EEPF)-inoculated (Beauveria bassiana and Metarhizium acridum) plants were investigated in dual-choice tests. The headspace volatiles of the different plant modalities were also sampled and analyzed. Viruliferous and non-viruliferous aphids were more attracted to EEPF-inoculated plants compared to uninoculated plants. However, viruliferous aphids were more attracted to EEPF-inoculated plants compared to virus-infected plants, while non-viruliferous insects exhibited no preference. Fungal-inoculated plants released higher amounts of aldehydes (i.e., heptanal, octanal, nonanal and decanal) compared to other plants, which might explain why viruliferous and non-viruliferous aphids were more abundant in EEPF-inoculated plants. Our study provides an interesting research perspective on how EEPF are involved in behavior of virus vector, depending on the infectious status of the latter.

Cover Image, Volume 76, Issue 8

The cover image is based on the Review Identifying drivers of spatio‐temporal dynamics in barley yellow dwarf virus epidemiology as a critical factor in disease control by Renik Van den Eynde et al., https://doi.org/10.1002/ps.5851. image

Surveillance of Barley Yellow Dwarf Virus (BYDV) in All Barley Growing Areas of Ethiopia

Surveillance of Barley Yellow Dwarf Virus (BYDV) in All Barley Growing Areas of Ethiopia

Can a PCR assay of aphids caught in‐crop on yellow sticky traps inform field level barley yellow dwarf virus risk assessment?

AbstractInfection with barley yellow dwarf virus (BYDV), caused by strains of virus belonging to the family Luteovirideae including BYDV‐PAV, can result in significant yield losses in autumn sown cereals following transmission by aphid vectors such as Rhopalosiphum padi (Homoptera: Aphididae) and Sitobion avenae (Homoptera: Aphididae). Spatial and temporal variance in the infectivity of alate populations influences risk to crops from the disease, which is greatest on infection at early crop growth stages. A decision support system (DSS) to guide optimised integration of crop protection strategies through risk assessment would help avoid unnecessary application of synthetic insecticides. This study contributes to the development of a DSS by exploring the viability and relevance of a methodology to detect virus levels in individual aphids trapped in‐crop using yellow sticky traps. Using a reverse transcription polymerase chain reaction assay, the detectability of virus from a BYDV‐PAV‐positive control colony was found not to be reduced by the process of trapping, extraction and cold storage, but did drop significantly after between 3 and 7 days of exposure on trap. This method has potential to contribute to localised risk assessment and guide optimisation of crop protection strategies.

Aphid Feeding on Plant Lectins Falling Virus Transmission Rates: A Multicase Study.

Aphids are insect vectors that have piercing-sucking mouthparts supporting diversified patterns of virus-vector interactions. Aphids primarily retain circulative viruses in the midgut/hindgut, whereas noncirculative viruses tend to be retained in the stylet. Most viruses, and many proteins from animals, have carbohydrate or carbohydrate-binding sites. Lectins vary in their specificity, of which some are able to bind to viral glycoproteins. To assess the potential competition between lectins and viral particles in virus transmission by aphids, this study examined how feeding plant lectins to aphids affects the transmission efficiency of viruses. Sitobion avenae (F, 1794) (Homoptera: Aphididae) aphids fed with Pisum sativum lectin (PSL) transmitted Barley yellow dwarf virus with significantly lower efficiency (four-fold ratio). Pea enation mosaic virus was significantly reduced in Acyrthosiphon pisum Harris (Homoptera: Aphididae) aphids fed with the lectin Concanavalin A. In comparison, the transmission of Potato virus Y was significantly reduced when Myzus persicae Sultzer (Homoptera: Aphididae) aphids were fed with PSL. Thus, lectin could be used as a blocking agent of plant viruses, facilitating an alternative approach for crop protection.

Virome identification in wheat in the Czech Republic using small RNA deep sequencing

High-throughput deep-sequencing technology and bioinformatics analysis of the small RNA (sRNA) population isolated from plants allows universal virus detection and complete virome reconstruction for a given sample. In the present sRNA deep-sequencing analysis of virus-infected wheat samples in the Czech Republic, samples were firstly tested for barley yellow dwarf viruses (BYDVs), wheat streak mosaic virus (WSMV) and wheat dwarf virus (WDV) using ELISA, RT-PCR and PCR. Subsequent sRNA sequencing of these samples yielded more than ~60 million single-end 50-bp reads with high confidence for nine field samples of wheat. Overall, 16.5% of reads were virus-specific and 83.5% were mapped to the host. More 21-nt reads (~7.7E+06 reads) were found than 24-nt (~6.20E+06 reads) or 22-nt (~4.30E+06 reads) reads. De novo assembly of the high-quality contigs revealed the presence of three earlier reported viruses in the Czech Republic: BYDVs (31.48%), WSMV (24.23%) and WDV (26.66%). We also showed the presence of cereal yellow dwarf virus (14.33%; two species CYDV-RPS and CYDV-RPV (family Luteoviridae/Polerovirus) and wheat yellow dwarf virus (WYDV, 3.30%; Luteoviridae). Phylogenetic analysis showed CYDV and WYDV grouped separately from BYDVs. Furthermore, several recombination breakpoints were found among the groups of yellow dwarf viruses (BYDVs, CYDV, and WYDV). Using RNA deep sequencing, we confirmed the presence of the three known viruses (BYDVs, WSMV, and WDV) and the first record of two species of CYDV and WYDV in wheat in the Czech Republic.

Genetic diversity in exotic oat germplasm & resistance against barley yellow dwarf virus

Oat (Avena sativa L.) is an important fodder crop of Pakistan, though with low productivity. The present study was conducted to evaluate the performance and genetic diversity of exotic oat germplasm, with emphasis on cereal yellow dwarf virus resistance. A total of 16 exotic line (introduced from Aarhus University Denmark) and 1 local line (provided by The University of Agriculture Peshawar), were grown during the season 2017–18 in Completely Randomized Block Design with three replications across two locations of Khyber Pakhtunkhwa i.e., Peshawar and Kohat. Field testing enabled to collect the data on BYDV incidence, BYDV severity, aphid infestation, plant height, leaf area, panicle length, panicle weight, spikelets per panicle, 1000 grain weight (g), grain yield (g), biological yield (g) and harvest index (%). Prevalence of BYDV was variable across location and over time. Six weeks data showed high disease pressure at Peshawar (85%), with SA-O-01 genotype having AUDPC value of 95%. Almost all the varieties showed less tolerance towards the Aphids attack. Line SA-O-15 showed the maximum 1000 grain weight (42.6 g) at Kohat, while SA-O-4 showed the maximum 1000 grain weight (60.7 g) at Peshawar. Line SA-O-05 (3634 g per (0.9 m2) plot) gave the maximum biological yield at Kohat station, while Line SA-O-01 gave the maximum biological yield (2517 g) at Peshawar. Mean grain yield for Kohat was recorded 0.155 g per (0.9 m2) plot while for Peshawar it was 0.231 g per (0.9 m2) plot. At Kohat line SA-O-10 produced the maximum grain yield (0.229 g), while line SA-O-12 produced the maximum grain yield at Peshawar (0.288 g). Molecular genotyping with a set of 4 RAPD primers revealed substantial diversity among17 oat lines. A total of 23 loci were amplified showing a high level of variations and polymorphism among the proposed lines. The maximum number of loci was recorded for GLA-04 (8), while the minimum number of loci was recorded for GLD-18 (4). Among the tested RAPD primers the maximum gene diversity (0.529) was recorded for loci GLA-03B230, GLA-04B130, GLA-04B300, GLB-05B150 and GLA-18B100 while the minimum (0.118) genetic diversity was recorded for loci GLA-03B600, GLB-05B330 and GLA-18B500. A clear divergence was found between most of the exotic oat lines. The observed genetic diversity in exotic oat germplasm and its resistance towards Barley Yellow Dwarf virus could be useful for oat genetic improvement and broadening the genetic background of cultivated oat germplasm.

Occurrence and High-Throughput Sequencing of Viruses in Ohio Wheat.

Ohio is a leading producer of soft red winter wheat in the United States. Many viruses impact wheat production, but there is a lack of contemporary information on the distribution and potential impact of wheat viruses in Ohio. To address this knowledge gap, we created a comprehensive dataset of viruses identified by high-throughput sequencing (HTS) and their incidence in field sites sampled across the state. Samples were collected from 103 field sites in surveys conducted in 2012, 2016, and 2017 and subjected to RNA HTS, reverse transcription (RT) PCR, or enzyme-linked immunosorbent assay to assess virus sequence diversity, prevalence, and incidence within fields. Partial and complete virus sequences were assembled and detection validated by RT-PCR. Assembled sequences were compared with previously known virus sequences, and novel sequences were validated by Sanger sequencing. The viruses detected most often included barley yellow dwarf virus (BYDV), cereal yellow dwarf virus (CYDV), wheat streak mosaic virus (WSMV), and wheat spindle streak mosaic virus (WSSMV). These viruses were detected at 67, 69, 55, and 28% of the field sites sampled, with mean incidences of 18, 19, 20, and 49%, respectively, within fields where they were detected. Brome mosaic virus (BMV) and cocksfoot mottle virus (CfMV) were also viruses of potential importance detected in Ohio, found in 26 and 17% of the field sites sampled, respectively. Based on results from logistic regression analyses, the presence of BYDV, CYDV, WSMV, and WSSMV was associated with the presence of volunteer wheat, BYDV and CfMV with monocots as the previous crop, and BMV with the presence of nearby corn fields (P < 0.10). For six viruses, there was evidence of spatial clustering in at least one field site and the variance of mean incidence was higher at the county level than at the regional spatial level. This finding suggests that county- and site-specific factors influenced the incidence and spatial pattern of some viruses. The results of this study provide a snapshot of viruses present in Ohio wheat and insights into their biology, potential risks to wheat production, and possible management strategies.

Identifying drivers of spatio-temporal dynamics in barley yellow dwarf virus epidemiology as a critical factor in disease control.

Barley yellow dwarf virus (BYDV) is one of the most important viral diseases of small grains worldwide. An understanding of its epidemiology is crucial to control this disease in a sustainable way. The virus moves through the agricultural landscape via cereal aphids as vectors. Understanding movement of these aphids in space and time is of key importance and in doing so, the spatial and temporal variables that influence BYDV epidemiology can be identified. The presence of summer hosts, crop rotation, crop diversity, agricultural practices and climate variables are crucial. Through digitalization, spatial (e.g. land‐use) and temporal (e.g. weather) information is becoming more readily available. Including this information into a prediction model could improve decision support systems that will rationalize the decision‐making process towards a more integrated control of the disease. © 2020 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Barley yellow dwarf virus-PAV is the dominant species causing Barley yellow dwarf disease in South Dakota and Minnesota

Barley yellow dwarf (BYD), caused by ten different species of viruses from the Luteoviridae family, is an economically important viral disease of small grains in the US Northern Great Plains. Identification of virus species in the region associated with BYD is critical to deployment of specific effective resistance genes. However, a comprehensive survey of virus species using modern diagnostic tools has not been done before in the US Northern Great Plains. In this study, species composition of viruses associated with BYD in small grains of Minnesota and South Dakota were determined by sampling BYD infected tissue from a large geographical area over three years. Whole genomic RNA was extracted from leaf tissue and reverse transcription polymerase chain reaction (RT-PCR) was conducted to detect infection of BYD and classify into species using primers from a two-step multiplex PCR protocol. Out of 463 samples collected from Minnesota and South Dakota from 2013 to 2015, about 50% (n = 222) of the samples tested positive for BYD. The majority of virus infected samples (n = 146) were identified as the Barley yellow dwarf virus PAV. These results indicate that the RT-PCR protocol used in this study is a viable molecular diagnostic tool for detection of BYD causing viruses in field collected samples. Moreover, the results indicate that Barley yellow dwarf virus PAV is most the dominant species associated with BYD in the US Northern Great Plains. Due to the species-specific nature of resistance to BYD these results can also guide breeding strategies in small grains for BYD management.

Eukaryotic Initiation Factor 3 Interactions with 5′ and 3′ RNA structures of the Barley Yellow Dwarf Virus Facilitate a Non‐Canonical Translation Mechanism

Barley Yellow Dwarf Virus (BYDV) is a positive strand RNA plant virus that translates without using a 5′ 7‐methylguanosine cap or a 3′ poly‐adenosine tail that are required for the canonical translation mechanism used by most mRNA. BYDV’s non‐canonical translation relies on RNA structures in the 5′ and 3′ untranslated regions (UTRs) that recruit eukaryotic initiation factors (eIFs) and ribosomes. Using a combination of biophysical, biochemical and chemical probing techniques, we have measured direct binding of eukaryotic initiation factors (eIFs) to BYVD 5′ and 3′ UTRs and probed binding sites. Our data show that one of the initiation factors, eIF3, can bind tightly, specifically, and simultaneously to structures in both 5′ and 3′ UTRs. This suggests that eIF3’s mechanistic role includes stabilization of the long‐distance RNA‐RNA interaction and the transfer of 3′‐bound factors/complexes to the 5′ UTR. Further work with RNA chemical probing has identified regions of the BYDV UTRs that interact with eIF3. This data highlights commonalities between BYDV structures and structures found in other viruses (e.g. Hepatitis C virus) and cellular mRNAs (e.g. c_JUN mRNA).Support or Funding InformationNSF MCB‐1902054 to Dixie J. Goss, NSF IGERT‐0965983 to Hunter College

Incidence and Distribution of Insect-Transmitted Cereal Viruses in Wheat in China from 2007 to 2019.

Diseases caused by insect-transmitted viruses are the predominant constraint to wheat production worldwide. However, detailed knowledge of virus incidence and dynamics in China in recent years is very limited. Here, major wheat-growing regions of China were surveyed over 10 years for insect-transmitted viruses, and 2,143 samples were collected (in 2007 to 2015) and analyzed by molecular hybridization or multiplex reverse-transcription PCR for barley yellow dwarf viruses (BYDVs: BYDV-GAV, -GPV, and -PAV) and wheat dwarf virus (WDV). In a 4-year survey (2016 to 2019), the incidence of eight insect-transmitted viruses (BYDVs, WDV, wheat yellow striate virus [WYSV], barley yellow striate mosaic virus [BYSMV], northern cereal mosaic virus [NCMV], and rice black-streaked dwarf virus [RBSDV]) was investigated, and BYDVs and WDV were widely distributed across China. BYDV-GAV (29.0% of the tested sample) was the most abundant, followed by BYDV-PAV (23.2%) from 2007 to 2015. From 2016 to 2019, however, BYDV-PAV had become the predominant species (39.5% positive of 952 samples tested), while the incidence of BYDV-GAV (13.4%) had declined. During the entire survey, the incidence of BYDV-GPV was very low in some locations in northwestern and northern China, and all eight viruses caused only local epidemics, not large-scale outbreaks throughout China. Two new cereal-infecting rhabdoviruses, leafhopper-transmitted WYSV and planthopper-transmitted BYSMV, were also found in China in recent years.

Enhanced heat tolerance of viral-infected aphids leads to niche expansion and reduced interspecific competition

Vector-borne pathogens are known to alter the phenotypes of their primary hosts and vectors, with implications for disease transmission as well as ecology. Here we show that a plant virus, barley yellow dwarf virus, increases the surface temperature of infected host plants (by an average of 2 °C), while also significantly enhancing the thermal tolerance of its aphid vector Rhopalosiphum padi (by 8 °C). This enhanced thermal tolerance, which was associated with differential upregulation of three heat-shock protein genes, allowed aphids to occupy higher and warmer regions of infected host plants when displaced from cooler regions by competition with a larger aphid species, R. maidis. Infection thereby led to an expansion of the fundamental niche of the vector. These findings show that virus effects on the thermal biology of hosts and vectors can influence their interactions with one another and with other, non-vector organisms.

Maize lethal necrosis viruses and other maize viruses in Rwanda.

Maize lethal necrosis (MLN) is emergent in East Africa, first reported in 2011 in Kenya, and is devastating to maize production in the region. MLN is caused by coinfection of maize with the emergent maize chlorotic mottle virus (MCMV) and any of several maize‐infecting potyviruses endemic in East Africa and worldwide. Here, we examined the distribution of MCMV and sugarcane mosaic virus (SCMV), the major viruses contributing to MLN in Rwanda. These and other viruses in maize across Rwanda were further characterized by deep sequencing. When identified, MCMV had high titres and minimal sequence variability, whereas SCMV showed moderate titres and high sequence variability. Deep sequencing also identified maize streak virus and other maize‐associated viruses, including a previously described polerovirus, maize yellow mosaic virus, and barley yellow dwarf virus, diverse maize‐associated totiviruses, maize‐associated pteridovirus, Zea mays chrysovirus 1, and a maize‐associated betaflexivirus. Detection of each virus was confirmed in maize samples by reverse transcription polymerase chain reaction.