High-Quality Genome Sequence Resource of a Rice False Smut Fungus Ustilaginoidea virens Isolate, UV-FJ-1

Ustilaginoidea virens is the fungal pathogen causing rice false smut, resulting in not only yield lost but also grain pollution with toxic mycotoxins. Here we deployed PacBio Sequel II HIFI-read sequencing technology to generate a near-complete genome assembly for the U. virens isolate UV-FJ-1 (38.48 Mb), which was isolated from Fujian province, China. The genome assembly contains 116 contigs with N50 of 0.65 Mb and a maximum length of 2.10 Mb, and the genome completeness is ≥98% assessed by benchmarking universal single-copy orthologs (BUSCOs) and the mapping rate of Illumina short reads. Excluding 35.78% repeat sequences, we identified a total of 7,164 protein-coding genes, of which 5,818 were functionally annotated and 223 encode putative effector proteins. Moreover, 21 secondary metabolite biosynthesis gene clusters were found in UV-FJ-1 genome. Taken together, this high-quality genome assembly and gene annotation resource will provide a better insight for characterizing the biological and pathogenic mechanisms of U. virens.

Extraction of Areas of Rice False Smut Infection Using UAV Hyperspectral Data

Rice false smut (RFS), caused by Ustilaginoidea virens, is a significant grain disease in rice that can lead to reduced yield and quality. In order to obtain spatiotemporal change information, multitemporal hyperspectral UAV data were used in this study to determine the sensitive wavebands for RFS identification, 665–685 and 705–880 nm. Then, two methods were used for the extraction of rice false smut-infected areas, one based on spectral similarity analysis and one based on spectral and temporal characteristics. The final overall accuracy of the two methods was 74.23 and 85.19%, respectively, showing that the second method had better prediction accuracy. In addition, the classification results of the two methods show that the areas of rice false smut infection had an expanding trend over time, which is consistent with the natural development law of rice false smut, and also shows the scientific nature of the two methods.

Genetic diversity and population structure of the rice false smut pathogen Ustilaginoidea virens in Sichuan-Chongqing region

Rice false smut caused by Ustilaginoidea virens is currently one of the most devastating fungal diseases of rice panicles worldwide. In this study, two novel molecular markers derived from SNP-rich genomic DNA fragments and a previously reported molecular marker were used for analyzing the genetic diversity and population structure of 167 U. virens isolates collected from nine areas in Sichuan-Chongqing region, China. A total of 62 haplotypes were identified, and a few haplotypes with high frequency were found and distributed in two to three areas, suggesting gene flow among different geographical populations. All isolates were divided into six genetic groups. The groups Ⅰ and Ⅵ were the largest including 61 and 48 isolates, respectively. The pairwise FST values showed significant genetic differentiation among all compared geographical populations. AMOVA showed that intergroup genetic variation accounted for 40.17% of the total genetic variation, while 59.83% of genetic variation came from intragroup. The UPGMA dendrogram and population structure revealed that the genetic composition of isolates collected from ST (Santai), NC (Nanchong), YC (Yongchuan), and WS (Wansheng) dominated by the same genetic subgroup was different from those collected from other areas. In addition, genetic recombination was found in a few isolates. These findings will help to improve the strategies for rice false smut management and resistance breeding, such as evaluating breeding lines with different isolates or haplotypes at different elevations and landforms.

Transcription factor UvMsn2 is important for vegetative growth, conidiogenesis, stress response, mitochondrial morphology and pathogenicity in the rice false smut fungus Ustilaginoidea virens

Transcription factors (TFs) play critical roles in the control of development and pathogenicity of phytopathogens by directly or indirectly regulating the expression of downstream genes. Here, we identified and characterized a zinc finger TF UvMsn2 in Ustilaginoidea virens, a homolog of MoMsn2 from the rice blast fungus. Heterogenous complementation assay revealed that UvMsn2 fully restored the defects of the ∆Momsn2 mutant in vegetative growth, conidiation and pathogenicity. Deletion of UvMsn2 in U. virens led to a reduction of the pathogen in vegetative growth, aerial hyphae and conidiation. Additionally, the ∆Uvmsn2 mutant displayed defects in conidial morphology and germination, as well as mitochondrial morphology. Pathogenicity and toxicity assays revealed that the ∆Uvmsn2 mutant was non-pathogenic and less inhibitory to rice seed germination. The ∆Uvmsn2 mutant showed different sensitivity to various stresses. Further microscopic examination found that UvMsn2 was localized in both cytosol and nucleus, and translocated from cytosol to nucleus under the treatment of NaCl. Our results demonstrate that UvMsn2 is a critical TF that regulates the vegetative growth, conidiogenesis, stress response, mitochondrial morphology and virulence in the rice false smut fungus.

Quantitative Proteomics Analysis Reveals the Function of the Putative Ester Cyclase UvEC1 in the Pathogenicity of the Rice False Smut Fungus Ustilaginoidea virens

Rice false smut is a fungal disease distributed worldwide and caused by Ustilaginoidea virens. In this study, we identified a putative ester cyclase (named as UvEC1) as being significantly upregulated during U. virens infection. UvEC1 contained a SnoaL-like polyketide cyclase domain, but the functions of ketone cyclases such as SnoaL in plant fungal pathogens remain unclear. Deletion of UvEC1 caused defects in vegetative growth and conidiation. UvEC1 was also required for response to hyperosmotic and oxidative stresses and for maintenance of cell wall integrity. Importantly, ΔUvEC1 mutants exhibited reduced virulence. We performed a tandem mass tag (TMT)-based quantitative proteomic analysis to identify differentially accumulating proteins (DAPs) between the ΔUvEC1-1 mutant and the wild-type isolate HWD-2. Proteomics data revealed that UvEC1 has a variety of effects on metabolism, protein localization, catalytic activity, binding, toxin biosynthesis and the spliceosome. Taken together, our findings suggest that UvEC1 is critical for the development and virulence of U. virens.