First report of Triticum mosaic virus infecting oat crop in the United States.

Triticum mosaic virus (TriMV, genus Poacevirus, family Potyviridae) was first reported in 2006 (Seifers et al. 2008) to infect wheat, and since then, it has been established as a constraint for US wheat production (Byamukama et al. 2013). In the field, TriMV often exists as a coinfection with wheat streak mosaic virus (WSMV), and these two viruses interact synergistically to produce severe symptoms and greater yield loss (Byamukama et al. 2012; Tatineni et al. 2022). Both TriMV and WSMV are transmitted by wheat curl mites (Aceria tosichella Keifer) (McMechan et al. 2014). Wheat is the primary host reported for TriMV in the field, but Seifers et al. (2010) established oat, rye, barley, and several other cereals and grasses as hosts under controlled conditions. However, there are no documented cases of TriMV infecting oats in the field. Between 10-25 June, 2023, a total of 273 field oat plants showing foliar yellowing, yellow flecking, and streaking symptoms were collected from four different fields in Nebraska (Big Springs: 41.1029° N, 102.1451° W; Mead: 41.2292° N, 96.4938° W; Odell: 40.0459° N, 96.7984° W; Stumf: 40.5048° N, 101.4223° W). Total RNA was extracted using the MagMax Plant RNA Isolation kit (Thermo Fisher Scientific) and the KingFisher Flex Magnetic Particle Processor (Thermo Fisher Scientific) (Mondal et al. 2023). Sample RNA was assayed with a single-step multiplex reverse transcription polymerase chain reaction (RT-PCR) to determine presence of WSMV and TriMV. Out of 273 symptomatic oat plants, 254 (93.04%) tested positive for at least one virus. Out of total positive samples, 238 were positive for WSMV (93.70 %), 12 plants tested positive for both TriMV and WSMV (4.70%), and 4 plants were infected with TriMV alone (1.60%). As a secondary confirmation, amplified fragments from the TriMV single infection were gel purified using a gel extraction kit (QIAquick) and sequenced (Eurofins Genomics). The nucleotide sequences were analysed using the BlastN program, compiled, and edited in the BioEdit software (Hall 1999). Sequences were deposited in the NCBI GenBank database (accession number PP475806). Nucleotide BLAST searches of the target coat protein (CP) gene showed > 98% identity to the corresponding sequences in TriMV accession MK318274. For further validation, virus inoculum was prepared by grinding field-collected plant material from plants with only TriMV present in 20 mM sodium phosphate buffer, pH 7.0, and then mechanically inoculating two-week-old oats (cv. Shaw n=8) and wheat (cv. Sattler, n=8) plants. Three weeks post-inoculation, all the eight wheat plants exhibited mild yellowing and streaking symptoms, while oat plants did not show obvious foliar symptoms. All wheat and oat plants were further tested positive with DAC-ELISA (antibodies produced against TriMV CP at the USDA-ARS facility in Lincoln, NE) and with RT-PCR. The specific attribution of these symptoms to TriMV in oats is not possible as none produced prominent symptoms. Asymptomatic oat infection from symptomatic field-collected oat samples could be due to oat cultivar differences. Although the prevalence of TriMV in wheat has been established across the Great Plains of the United States, to our knowledge, this is the first report of TriMV infection in US oat fields. Our finding warrant further investigation into the incidence and impact of the virus in oat crop and its potential for serving as a asymptomatic virus reservoir.

Higher-order species interactions cause time-dependent niche and fitness differences: Experimental evidence in plant-feeding arthropods.

Species interact in different ways, including competition, facilitation and predation. These interactions can be non-linear or higher order and may depend on time or species densities. Although these higher-order interactions are virtually ubiquitous, they remain poorly understood, as they are challenging both theoretically and empirically. We propose to adapt niche and fitness differences from modern coexistence theory and apply them to species interactions over time. As such, they may not merely inform about coexistence, but provide a deeper understanding of how species interactions change. Here, we investigated how the exploitation of a biotic resource (plant) by phytophagous arthropods affects their interactions. We performed monoculture and competition experiments to fit a generalized additive mixed model to the empirical data, which allowed us to calculate niche and fitness differences. We found that species switch between different types of interactions over time, including intra- and interspecific facilitation, and strong and weak competition.

Molecular characterization of a divergent genetic variant of wheat Eqlid mosaic virus from a Texas wheat field.

Investigations conducted during the spring 2020 season to diagnose the associated viral agent of a severe mosaic disease of wheat in a Texas Panhandle field revealed the presence of wheat Eqlid mosaic virus (WEqMV; genus Tritimovirus, family Potyviridae) in the analyzed samples. The complete genome sequences of two WEqMV isolates were determined, and each was found to be 9,634 nucleotides (nt) in length (excluding the polyA tail) and to contain 5' and 3' untranslated regions of 135 nt and 169 nt, respectively, based on rapid amplification of cDNA ends (RACE) assays. Both sequences contained an open reading frame (ORF) of 9,330 nt encoding a polyprotein of 3,109 amino acids (aa). The ORF sequences of the two isolates were 100% identical to each other, but only 74.7% identical to that of the exemplar WEqMV-Iran isolate, with 85.7% aa sequence identity in the encoded polyprotein. The Texas WEqMV isolates also diverged significantly from WEqMV-Iran in the individual proteins at the nt and aa levels. This is the first report of WEqMV in the United States and the first report of this virus outside of Iran, indicating an expansion of its geographical range.

Effective specialist or jack of all trades? Experimental evolution of a crop pest in fluctuating and stable environments.

Understanding pest evolution in agricultural systems is crucial for developing effective and innovative pest control strategies. Types of cultivation, such as crop monocultures versus polycultures or crop rotation, may act as a selective pressure on pests' capability to exploit the host's resources. In this study, we examined the herbivorous mite Aceria tosichella (commonly known as wheat curl mite), a widespread wheat pest, to understand how fluctuating versus stable environments influence its niche breadth and ability to utilize different host plant species. We subjected a wheat-bred mite population to replicated experimental evolution in a single-host environment (either wheat or barley), or in an alternation between these two plant species every three mite generations. Next, we tested the fitness of these evolving populations on wheat, barley, and on two other plant species not encountered during experimental evolution, namely rye and smooth brome. Our results revealed that the niche breadth of A. tosichella evolved in response to the level of environmental variability. The fluctuating environment expanded the niche breadth by increasing the mite's ability to utilize different plant species, including novel ones. Such an environment may thus promote flexible host-use generalist phenotypes. However, the niche expansion resulted in some costs expressed as reduced performances on both wheat and barley as compared to specialists. Stable host environments led to specialized phenotypes. The population that evolved in a constant environment consisting of barley increased its fitness on barley without the cost of utilizing wheat. However, the population evolving on wheat did not significantly increase its fitness on wheat, but decreased its performance on barley. Altogether, our results indicated that, depending on the degree of environmental heterogeneity, agricultural systems create different conditions that influence pests' niche breadth evolution, which may in turn affect the ability of pests to persist in such systems.

Occurrence of Wheat Curl Mite and Mite-Vectored Viruses of Wheat in Colorado and Insights into the Wheat Virome.

The wheat curl mite (WCM) is a vector of three important wheat viruses in the U.S. Great Plains: wheat streak mosaic virus (WSMV), triticum mosaic virus (TriMV), and High Plains wheat mosaic virus (HPWMoV). This study was conducted to determine the current profile of WCM and WCM-transmitted viruses of wheat and their occurrence in Colorado, including novel wheat viruses via virome analysis. There was a high rate of virus incidence in symptomatic wheat samples collected in 2019 (95%) and 2020 (77%). Single infection of WSMV was most common in both years, followed by coinfection with WSMV + TriMV and WSMV + HPWMoV. Both type 1 and type 2 mite genotypes were found in Colorado. There was high genetic diversity of WSMV and HPWMoV isolates, whereas TriMV isolates showed minimal sequence variation. Analysis of WSMV isolates revealed novel virus variants, including one isolate from a variety trial, where severe disease symptoms were observed on wheat varieties carrying Wsm2, a known virus resistance locus. Virome analysis identified two to four sequence variants of all eight RNA segments of HPWMoV, which suggests co-occurrence of multiple genotypes within host populations and presence of a variant of HPWMoV. A possible novel virus in the family Tombusviridae and several mycoviruses were identified. Overall, the data presented here highlight the need to define the effect of novel WCM-transmitted virus variants on disease severity and the role of novel viruses.

Hitchhiking or hang gliding? Dispersal strategies of two cereal-feeding eriophyoid mite species.

Dispersal shapes the dynamics of populations, their genetic structure and species distribution; therefore, knowledge of an organisms' dispersal abilities is crucial, especially in economically important and invasive species. In this study, we investigated dispersal strategies of two phytophagous eriophyoid mite species: Aceria tosichella (wheat curl mite, WCM) and Abacarus hystrix (cereal rust mite, CRM). Both species are obligatory plant parasites that infest cereals and are of economic significance. We investigated their dispersal success using different dispersal agents: wind and vectors. We hypothesised that in both mite species the main mode of dispersal is moving via wind, whereas phoretic dispersal is rather accidental, as the majority of eriophyoid mite species do not possess clear morphological or behavioural adaptations for phoresy. Results confirmed our predictions that both species dispersed mainly with wind currents. Additionally, WCM was found to have a higher dispersal success than CRM. Thus, this study contributes to our understanding of the high invasive potential of WCM.

High Plains wheat mosaic virus: An enigmatic disease of wheat and corn causing the High Plains disease.

BRIEF HISTORY: In 1993, severe mosaic and necrosis symptoms were observed on corn (maize) and wheat from several Great Plains states of the USA. Based on the geographical location of infections, the disease was named High Plains disease and the causal agent was tentatively named High Plains virus. Subsequently, researchers renamed this virus as maize red stripe virus and wheat mosaic virus to represent the host and symptom phenotype of the virus. After sequencing the genome of the pathogen, the causal agent of High Plains disease was officially named as High Plains wheat mosaic virus. Hence, High Plains virus, maize red stripe virus, wheat mosaic virus, and High Plains wheat mosaic virus (HPWMoV) are synonyms for the causal agent of High Plains disease. TAXONOMY: High Plains wheat mosaic virus is one of the 21 definitive species in the genus Emaravirus in the family Fimoviridae. VIRION: The genomic RNAs are encapsidated in thread-like nucleocapsids in double-membrane 80-200 nm spherical or ovoid virions. GENOME CHARACTERIZATION: The HPWMoV genome consists of eight single-stranded negative-sense RNA segments encoding a single open reading frame (ORF) in each genomic RNA segment. RNA 1 is 6,981-nucleotide (nt) long, coding for a 2,272 amino acid protein of RNA-dependent RNA polymerase. RNA 2 is 2,211-nt long and codes for a 667 amino acid glycoprotein precursor. RNA 3 has two variants of 1,439- and 1,441-nt length that code for 286 and 289 amino acid nucleocapsid proteins, respectively. RNA 4 is 1,682-nt long, coding for a 364 amino acid protein. RNA 5 and RNA 6 are 1,715- and 1,752-nt long, respectively, and code for 478 and 492 amino acid proteins, respectively. RNA 7 and RNA 8 are 1,434- and 1,339-nt long, code for 305 and 176 amino acid proteins, respectively. BIOLOGICAL PROPERTIES: HPWMoV can infect wheat, corn (maize), barley, rye brome, oat, rye, green foxtail, yellow foxtail, and foxtail barley. HPWMoV is transmitted by the wheat curl mite and through corn seed. DISEASE MANAGEMENT: Genetic resistance against HPWMoV in wheat is not available, but most commercial corn hybrids are resistant while sweet corn varieties remain susceptible. Even though corn hybrids are resistant to virus, it still serves as a green bridge host that enables mites to carry the virus from corn to new crop wheat in the autumn. The main management strategy for High Plains disease in wheat relies on the management of green bridge hosts. Cultural practices such as avoiding early planting can be used to avoid mite buildup and virus infections.

Temperature-dependent development and survival of an invasive genotype of wheat curl mite, Aceria tosichella.

Quantifying basic biological data, such as the effects of variable temperatures on development and survival, is crucial to predicting and monitoring population growth rates of pest species, many of which are highly invasive. One of the most globally important pests of cereals is the eriophyoid wheat curl mite (WCM), Aceria tosichella, which is the primary vector of several plant viruses. The aim of this study was to evaluate temperature-dependent development and survival of WCM at a wide range of constant temperatures in the laboratory (17-33 °C). The development time of each stage depended significantly on temperature and it was negatively correlated with temperature increase. At high temperatures (27-33 °C), individuals had shorter developmental times, with the shortest (6 days) at 33 °C, whereas at the lowest tested temperatures (17-19 °C), developmental time was almost 3× longer. Moreover, temperature had a clear effect on survival: the higher the temperature, the lower the survival rate. These data provide information promoting more efficient and effective manipulation of WCM laboratory colonies, and further our understanding of the ramifications of temperature change on WCM physiology and implications for the growth and spread of this globally invasive pest.

Reactive oxygen species metabolism and photosynthetic performance in leaves of Hordeum vulgare plants co-infested with Heterodera filipjevi and Aceria tosichella.

KEY MESSAGE: Defence responses of cyst nematode and/or wheat curl mite infested barley engage the altered reactive oxygen species production, antioxidant machinery, carbon dioxide assimilation and photosynthesis efficiency. The primary aim of this study was to determine how barley responds to two pests infesting separately or at once; thus barley was inoculated with Heterodera filipjevi (Madzhidov) Stelter (cereal cyst nematode; CCN) and Aceria tosichella Keifer (wheat curl mite; WCM). To verify hypothesis about the involvement of redox metabolism and photosynthesis in barley defence responses, biochemical, photosynthesis efficiency and chlorophyll a fluorescence measurements as well as transmission electron microscopy were implemented. Inoculation with WCM (apart from or with CCN) brought about a significant suppression in the efficiency of electron transport outside photosystem II reaction centres. This limitation was an effect of diminished pool of rapidly reducing plastoquinone and decreased total electron carriers. Infestation with WCM (apart from or with CCN) also significantly restricted the electron transport on the photosystem I acceptor side, therefore produced reactive oxygen species oxidized lipids in cells of WCM and double infested plants and proteins in cells of WCM-infested plants. The level of hydrogen peroxide was significantly decreased in double infested plants because of glutathione-ascorbate cycle involvement. The inhibition of nitrosoglutathione reductase promoted the accumulation of S-nitrosoglutathione increasing antioxidant capacity in cells of double infested plants. Moreover, enhanced arginase activity in WCM-infested plants could stimulate synthesis of polyamines participating in plant antioxidant response. Infestation with WCM (apart from or with CCN) significantly reduced the efficiency of carbon dioxide assimilation by barley leaves, whereas infection only with CCN expanded photosynthesis efficiency. These were accompanied with the ultrastructural changes in chloroplasts during CCN and WCM infestation.

A comprehensive and cost-effective approach for investigating passive dispersal in minute invertebrates with case studies of phytophagous eriophyid mites.

Dispersal is a fundamental biological process that operates at different temporal and spatial scales with consequences for individual fitness, population dynamics, population genetics, and species distributions. Studying this process is particularly challenging when the focus is on microscopic organisms that disperse passively, whilst controlling neither the transience nor the settlement phase of their movement. In this work we propose a comprehensive approach for studying passive dispersal of microscopic invertebrates and demonstrate it using wind and phoretic vectors. The protocol includes the construction of versatile, modifiable dispersal tunnels as well as a theoretical framework quantifying the movement of species via wind or vectors, and a hierarchical Bayesian approach appropriate to the structure of the dispersal data. The tunnels were used to investigate the three stages of dispersal (viz., departure, transience, and settlement) of two species of minute, phytophagous eriophyid mites Aceria tosichella and Abacarus hystrix. The proposed devices are inexpensive and easy to construct from readily sourced materials. Possible modifications enable studies of a wide range of mite species and facilitate manipulation of dispersal factors, thus opening a new important area of ecological study for many heretofore understudied species.

Wheat streak mosaic virus: a century old virus with rising importance worldwide.

Wheat streak mosaic virus (WSMV) causes wheat streak mosaic, a disease of cereals and grasses that threatens wheat production worldwide. It is a monopartite, positive-sense, single-stranded RNA virus and the type member of the genus Tritimovirus in the family Potyviridae. The only known vector is the wheat curl mite (WCM, Aceria tosichella), recently identified as a species complex of biotypes differing in virus transmission. Low rates of seed transmission have been reported. Infected plants are stunted and have a yellow mosaic of parallel discontinuous streaks on the leaves. In the autumn, WCMs move from WSMV-infected volunteer wheat and other grass hosts to newly emerged wheat and transmit the virus which survives the winter within the plant, and the mites survive as eggs, larvae, nymphs or adults in the crown and leaf sheaths. In the spring/summer, the mites move from the maturing wheat crop to volunteer wheat and other grass hosts and transmit WSMV, and onto newly emerged wheat in the fall to which they transmit the virus, completing the disease cycle. WSMV detection is by enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR) or quantitative RT-PCR (RT-qPCR). Three types of WSMV are recognized: A (Mexico), B (Europe, Russia, Asia) and D (USA, Argentina, Brazil, Australia, Turkey, Canada). Resistance genes Wsm1, Wsm2 and Wsm3 have been identified. The most effective, Wsm2, has been introduced into several wheat cultivars. Mitigation of losses caused by WSMV will require enhanced knowledge of the biology of WCM biotypes and WSMV, new or improved virus detection techniques, the development of resistance through traditional and molecular breeding, and the adaptation of cultural management tactics to account for climate change.

Population Dynamics of the Wheat Curl Mite (Acari: Eriophyidae) During the Heading Stages of Winter Wheat.

The wheat curl mite (Aceria tosichella Keifer) is the only known vector of three viruses in wheat-Wheat streak mosaic virus, Wheat mosaic virus, and Triticum mosaic virus. The economic impact of this disease complex is linked to the presence of suitable hosts prior to winter wheat maturing in early summer and the movement of wheat curl mite from wheat to oversummering hosts prior to wheat harvest. Previous research has documented the prevalence and density of mite populations on maturing wheat heads; however, these studies were limited to a few late stages of wheat. A study was conducted to evaluate mite population densities across all stages of head development to determine when wheat curl mites are most abundant and the relative increase in abundance over time. In addition, a study was conducted to evaluate the impact of rainfall on mite populations during wheat heading. A final study was conducted to determine the potential for direct infestation of seedlings germinating from wheat curl mite-infested wheat heads. Results showed a rapid buildup in mite populations from low densities in early heading and peaking at the hard dough stage, with nearly all wheat heads having some mite presence. In addition, high mite populations resulted in direct infestation of germinated seedlings from the early through hard dough stages. Rainfall applications had no observable impact on mite population densities in wheat heads. These results demonstrate the increased potential for mites to infest hosts prior to winter wheat maturing and illustrate the increased risk for these hosts to serve as oversummering hosts.

Planting Date and Variety Selection for Management of Viruses Transmitted by the Wheat Curl Mite (Acari: Eriophyidae).

Wheat is an important food grain worldwide, and it is the primary dryland crop in the western Great Plains. A complex of three viruses (Wheat streak mosaic, Wheat mosaic, and Triticum mosaic viruses) is a common cause of loss in winter wheat production in the Great Plains. All these viruses are transmitted by the wheat curl mite (Aceria tosichella Keifer). Once these viruses are established, there are no curative actions; therefore, prevention is the key to successful management. A study was designed to evaluate preventative management tactics (planting date, resistant varieties) for reducing the impact from this virus complex. The main plot treatments were three planting dates, and split-plot treatments were three wheat varieties. Varieties were planted at three different times during the fall to simulate early, recommended, and late planting dates. The varieties evaluated in this study were Mace (virus resistant), Millennium (mild tolerance), and Tomahawk (susceptible). Measurements of virus symptomology and yield were used to determine virus impact. Results consistently showed that the resistant Mace yielded more than Millennium or Tomahawk under virus pressure. In some years, delayed planting improved the yields for all varieties, regardless of their background; however, under the most severe virus pressure the combination of both management strategies was not sufficient to provide practical control of this complex. These results illustrate the importance of using a combination of management tactics for this complex, but also reinforce the importance for producers to use additional management strategies (e.g., control preharvest volunteer wheat) to manage this complex.