The necrotrophic fungus Rhizoctonia cerealis is the causal agent of devastating diseases of cereal crops including wheat (Triticum aestivum). We present a high-quality genome assembly of R. cerealis Rc207, a virulent strain causing wheat sharp eyespot. The assembly (56.36 Mb) is composed of 17.87% repeat sequences and 14, 433 predicted protein-encoding genes. The Rc207 genome encodes a large and diverse set of genes involved in pathogenicity, especially rich in those encoding secreted proteins, carbohydrate-active enzymes (CAZymes), peptidases, nucleases, cytochrome P450, and secondary metabolism-associated enzymes. Most secretory protein-encoding genes, including CAZymes, peroxygenases, dehydrogenases, and cytochrome P450, were up-regulated during fungal infection of wheat. We identified 831 candidate secretory effectors and validated the functions of 10 up-regulated candidate effector proteins. Of them, nine were confirmed as necrotrophic pathogen's effectors promoting fungal infection. Abundant potential mobile or plastic genomic regions rich in repeat sequences suggest their roles in fungal adaption and virulence-associated genomic evolution. This study provides valuable resources for further comparative and functional genomics on important fungal pathogens, and provides essential tools for development of effective disease control strategies.

Wheat (Triticum aestivum) is necessary for global food security. The necrotrophic fungus Rhizoctonia cerealis is the causal agent of sharp eyespot, a devastating disease of wheat. Although the Elongator complex, composed of six subunits, has been implicated in growth, development, and innate immunity in Arabidopsis, little is known about its functions in wheat or the involvement of Elongator subunit 4 in histone acetylation. In this study, we identified the Elongator subunit 4-encoding gene TaELP4 in wheat resistance response to R. cerealis, and verified that TaELP4 increased histone acetylation in regions of defense-associated genes and regulated immune response to R. cerealis. TaELP4 was more highly expressed in resistant than in susceptible wheat cultivars and was induced in resistant wheat after infection by R. cerealis. Silencing of TaELP4 in wheat not only impaired resistance to R. cerealis, but also repressed both histone acetylation levels and the expression of a subset of defense-associated genes, including TaAGC1, TaCPK7-D, TaPAL5, Defensin, and Chitinase2. Ectopic expression of TaELP4 in Arabidopsis increased histone acetylation levels in coding and promoter regions of defense genes and increased their transcription, leading to increased resistance to infection by the pathogen Botrytis cinerea. These results suggest that TaELP4 positively regulates innate immune responses of wheat and Arabidopsis to R. cerealis and B. cinerea by increasing histone acetylation levels of defense-associated genes and increasing their transcription. This study has shed light on the involvement of TaELP4 in histone acetylation and resistance response against R. cerealis. TaELP4 may potentially be used to improve wheat resistance against sharp eyespot.

Sharp eyespot, mainly caused by the soil-borne fungus Rhizoctonia cerealis, affects wheat (Triticum aestivum L.) production worldwide. In this study, we isolated TaCML36 gene encoding a wheat calmodulin-like protein, and studied its defense role in protection against R. cerealis. Transcription of TaCML36 was significantly elevated by both R. cerealis infection and exogenous ethylene treatment. Transcription was higher in resistant wheat lines than in susceptible ones. There were copies of TaCML36 on chromosomes 5A, 5B, and 5D. The TaCML36 protein is composed of 183 amino acids and contains two calcium-binding EF-hand domains. Subcellular localization assays in wheat indicated that TaCML36 localizes in both the cytoplasm and nucleus. Virus-induced gene silencing and disease assessment indicated that compared to the controls, TaCML36-silenced wheat plants displayed significantly reduced resistance to R. cerealis and had greater fungal biomass, suggesting that knockdown of TaCML36 impaired host resistance. Knockdown of TaCML36 also significantly repressed expression of pathogenesis-related genes such as Chitinase 1, PDF35, and PR17C, the ethylene response factor-encoding gene TaPIE1, and ethylene biosynthesis gene ACO2. Collectively, our results suggest that TaCML36 positively participates in the innate immune response to R. cerealis infection by modulating expression of defense-associated genes possibly in the ethylene signaling pathway.

Barley yellow dwarf virus (BYDV) can infect wheat (Triticum aestivum L.), leading to yield loss. Among four BYDV strains (GAV, GPV, PAV, and RMV) identified in China, BYDV-GAV is the prevailing isolate. YW642, a wheat–Thinopyrum intermedium translocation line, is resistant to BYDV isolates at both seedling and adult stages. Zhong 8601 is the wheat recurrent parent of YW642 and is susceptible to BYDV. In this study, we investigated the adult-stage resistance mechanism of YW642, measured BYDV titer and hydrogen peroxide (H₂O₂) in adult-stage leaves of YW642 and Zhong 8601 inoculated with BYDV-GAV, and identified transcriptional differences between YW642 and Zhong 8601 using microarray-based comparative transcriptomics. Enzyme-linked immunosorbent assay and H₂O₂ assay showed that both BYDV titer and H₂O₂ content were markedly lower in YW642 than in Zhong 8601 at 21, 28, 35, and 40days post-inoculation (dpi). The transcriptomic comparison revealed that many types of genes were significantly up-regulated at 35dpi in adult-stage leaves of YW642 compared to Zhong 8601. The important up-regulated genes associated with the adult-stage resistance encoded 15 resistance-like proteins, pathogenesis-related proteins (such as defensin and lipid transfer proteins), protein kinase homologs, transcription factors, reactive oxygen species scavenging-related proteins, and jasmonic acid and gibberellic acid biosynthesis enzymes. These results suggest that precise expression regulation of these proteins plays a crucial role in adult-stage resistance of YW642 against BYDV infection.

Wall-associated kinases (WAKs) play an important role in plant defense and development. Considerable progress has been made in understanding WAK genes in Arabidopsis thaliana. However, much less is known about these genes in common wheat. Here, we isolated a novel wheat WAK gene TaWAK5 from sharp eyespot disease-resistant wheat line CI12633, based on a differentially-expressed sequence identified by microarray analysis. The transcript abundance of TaWAK5 was rapidly increased following inoculation with the pathogen Rhizoctonia cerealis. TaWAK5 in resistant wheat lines was induced to higher levels than in susceptible lines at 7 days post inoculation with R. cerealis. The expression of TaWAK5 was also induced by treatments with exogenous salicylic acid, abscisic acid, and methyl jasmonate. The deduced TaWAK5 protein contained a signal peptide, two epidermal growth factor (EGF)-like repeats, a transmembrane domain, and a serine/threonine protein kinase catalytic domain. Subcellular localization analyses in onion epidermal cells indicated that the TaWAK5 protein was localized to the plasma membrane. Virus-induced gene silencing of TaWAK5 in CI12633 plants showed that the silencing of TaWAK5 did not obviously impair wheat resistance to R. cerealis, suggesting that TaWAK5 may be not the major gene in wheat defense response to R. cerealis, or that it is functionally redundant with other genes. This study paves the way for further research into WAK functions in wheat stress physiology.