Applying early divergent characters in higher rank taxonomy of Melampsorineae (Basidiomycota, Pucciniales).

Rust fungi in the order Pucciniales represent one of the largest groups of phytopathogens, which occur on mosses, ferns to advanced monocots and dicots. Seven suborders and 18 families have been reported so far, however recent phylogenetic studies have revealed para- or polyphyly of several morphologically defined suborders and families, particularly in Melampsorineae. In this study, a comprehensive phylogenetic framework was constructed based on a molecular phylogeny inferred from rDNA sequences of 160 species belonging to 16 genera in Melampsorineae (i.e. Chrysomyxa, Cerospora, Coleopuccinia, Coleosporium, Cronartium, Hylospora, Melampsora, Melampsorella, Melampsoridium, Milesina, Naohidemyces, Pucciniastrum, Quasipucciniastrum, Rossmanomyces, Thekopsora, Uredinopsis). Our phylogenetic inference indicated that 13 genera are monophyletic with strong supports, while Pucciniastrum is apparently polyphyletic. A new genus, Nothopucciniastrum was therefore established and segregated from Pucciniastrum, with ten new combinations proposed. At the family level, this study further demonstrates the importance of applying morphologies of spore-producing structures (basidia, spermogonia, aecia, uredinia and telia) in higher rank taxonomy, while those traditionally applied spore morphologies (basidiospores, spermatia, aeciospores, urediniospores and teliospores) represent later diverged characters that are more suitable for the taxonomy at generic and species levels. Three new families, Hyalopsoraceae, Nothopucciniastraceae and Thekopsoraceae were proposed based on phylogenetic and morphological distinctions, towards a further revision of Pucciniales in line with the phylogenetic relationships.

Temporal, compositional, and functional differences in the microbiome of Bangkok subway air environment.

With the growing numbers of the urban population, an increasing number of commuters have relied on subway systems for rapid transportation in daily life. Analyzing the temporal distribution of air microbiomes in subway environments is crucial for the assessment and monitoring of air quality in the subway system, especially with regard to public health. This study employed culture-independent metabarcode sequencing to analyze bacterial diversity and variations in bacterial compositions associated with bioaerosols collected from a subway station in Bangkok over a four-month period. The bacteria obtained were found to consist primarily of Proteobacteria, Firmicutes, and Actinobacteria, with variations at the family, genus, and species levels among samples obtained in different months. The vast majority of these bacteria are most likely derived from outside environments and human body sources. Many of the bacteria found in Bangkok subway station were also identified as "core microorganisms" of subway environments around the world, as suggested by the MetaSUB Consortium. The diversity of bacterial communities was shown to be influenced by several air quality variables, especially ambient temperature and the quantity of particulate matters, which showed positive correlations with several bacterial species such as Acinetobacter lwoffii, Staphylococcus spp., and Moraxella osloensis. In addition, metabolic profiles inferred from metabarcode-derived bacterial diversity showed significant variations across different sampling times and sites and can be used as a starting point to further explore the functional roles of specific groups of bacteria in the subway environment. This study thus introduced the information required for surveillance of microbiological impacts and their contributions to the well-being of subway commuters in Bangkok.

Haplotype-phased and chromosome-level genome assembly of Puccinia polysora, a giga-scale fungal pathogen causing southern corn rust.

Rust fungi are characterized by large genomes with high repeat content and have two haploid nuclei in most life stages, which makes achieving high-quality genome assemblies challenging. Here, we described a pipeline using HiFi reads and Hi-C data to assemble a gigabase-sized fungal pathogen, Puccinia polysora f.sp. zeae, to haplotype-phased and chromosome-scale. The final assembled genome is 1.71 Gbp, with ~850 Mbp and 18 chromosomes in each haplotype, being currently one of the two giga-scale fungi assembled to chromosome level. Transcript-based annotation identified 47,512 genes for the dikaryotic genome with a similar number for each haplotype. A high level of interhaplotype variation was found with 10% haplotype-specific BUSCO genes, 5.8 SNPs/kbp, and structural variation accounting for 3% of the genome size. The P. polysora genome displayed over 85% repeat contents, with genome-size expansion and copy number increasing of species-specific orthogroups. Interestingly, these features did not affect overall synteny with other Puccinia species having smaller genomes. Fine-time-point transcriptomics revealed seven clusters of coexpressed secreted proteins that are conserved between two haplotypes. The fact that candidate effectors interspersed with all genes indicated the absence of a "two-speed genome" evolution in P. polysora. Genome resequencing of 79 additional isolates revealed a clonal population structure of P. polysora in China with low geographic differentiation. Nevertheless, a minor population differentiated from the major population by having mutations on secreted proteins including AvrRppC, indicating the ongoing virulence to evade recognition by RppC, a major resistance gene in Chinese corn cultivars. The high-quality assembly provides valuable genomic resources for future studies on disease management and the evolution of P. polysora.

Cross-kingdom synthetic microbiota supports tomato suppression of Fusarium wilt disease.

The role of rhizosphere microbiota in the resistance of tomato plant against soil-borne Fusarium wilt disease (FWD) remains unclear. Here, we showed that the FWD incidence was significantly negatively correlated with the diversity of both rhizosphere bacterial and fungal communities. Using the microbiological culturomic approach, we selected 205 unique strains to construct different synthetic communities (SynComs), which were inoculated into germ-free tomato seedlings, and their roles in suppressing FWD were monitored using omics approach. Cross-kingdom (fungi and bacteria) SynComs were most effective in suppressing FWD than those of Fungal or Bacterial SynComs alone. This effect was underpinned by a combination of molecular mechanisms related to plant immunity and microbial interactions contributed by the bacterial and fungal communities. This study provides new insight into the dynamics of microbiota in pathogen suppression and host immunity interactions. Also, the formulation and manipulation of SynComs for functional complementation constitute a beneficial strategy in controlling soil-borne disease.

Phylogenetic Relationships, Speciation, and Origin of Armillaria in the Northern Hemisphere: A Lesson Based on rRNA and Elongation Factor 1-Alpha.

Armillaria species have a global distribution and play various roles in the natural ecosystems, e.g., pathogens, decomposers, and mycorrhizal associates. However, their taxonomic boundaries, speciation processes, and origin are poorly understood. Here, we used a phylogenetic approach with 358 samplings from Europe, East Asia, and North America to delimit the species boundaries and to discern the evolutionary forces underpinning divergence and evolution. Three species delimitation methods indicated multiple unrecognized phylogenetic species, and biological species recognition did not reflect the natural evolutionary relationships within Armillaria; for instance, biological species of A. mellea and D. tabescens are divergent and cryptic species/lineages exist associated with their geographic distributions in Europe, North America, and East Asia. While the species-rich and divergent Gallica superclade might represent three phylogenetic species (PS I, PS II, and A. nabsnona) that undergo speciation. The PS II contained four lineages with cryptic diversity associated with the geographic distribution. The genus Armillaria likely originated from East Asia around 21.8 Mya in early Miocene when Boreotropical flora (56-33.9 Mya) and the Bering land bridge might have facilitated transcontinental dispersal of Armillaria species. The Gallica superclade arose at 9.1 Mya and the concurrent vicariance events of Bering Strait opening and the uplift of the northern Tibetan plateau might be important factors in driving the lineage divergence.

Population Genetic Structures of Puccinia striiformis f. sp. tritici in the Gansu-Ningxia Region and Hubei Province, China.

Wheat stripe rust, caused by the fungal pathogen Puccinia striiformis f. sp. tritici (Pst), is a destructive wheat disease in China. The Gansu-Ningxia region (GN) is a key area for pathogen over-summering in China, and northwestern Hubei (HB) is an important region for pathogen over-wintering, serving as a source of inoculum in spring epidemic regions. The spatiotemporal population genetic structure of Pst in HB and the pathogen population exchanges between GN and HB are important for estimating the risk of interregional epidemics. Here, 567 isolates from GN and HB were sampled from fall 2016 to spring 2018 and were genotyped using simple sequence repeat markers. The genotypic and genetic diversity of Pst subpopulations in HB varied among seasons and locations. Greater genetic diversification levels were found in the spring compared with fall populations using principal coordinate analysis and Bayesian assignments. In total, there were 17 common genotypes among the 208 determined, as shown by a small overlap of genotypes in the principal coordinate analysis and dissimilar Bayesian assignments in both regions, which revealed the limited genotype exchange between the populations of GN and HB.

Leptosphaerulina species isolated from golf turfgrass in China, with description of L. macrospora, sp. nov.

Leptosphaerulina leaf blight occurs on most turfgrasses. Hitherto, Leptosphaerulina species associated with this disease include L. americana, L. argentinensis, L. australis, and L. trifolii. However, following Koch's postulates, L. australis was confirmed as saprobes but not pathogens, and the other three species have not been tested. The pathogenicity of Leptosphaerulina spp. is still questionable. In this study, we isolated 19 Leptosphaerulina strains from diseased golf turfgrasses in China, and they were identified as L. gaeumannii, L. saccharicola, and a new species, L. macrospora, through multilocus (ITS, 28S, rpb2, and tub2) phylogenetic analyses and morphological observations. Pathogenicity test revealed that the three Leptosphaerulina species identified in this study cannot infect live/healthy turfgrass tissues of Poa pratensis and Agrostis stolonifera and only produced pseudothecia on the dead leaves of stressed seedlings. Considering the results of pathogenicity tests in this and previous studies, we speculate that most Leptosphaerulina species isolated from diseased turfgrass are not pathogens but saprobes. Applying proper management practices to prevent severe turfgrass stress is a key measure to reduce or eliminate the effects of Leptosphaerulina on golf turfgrass.

Genetic Structure and Asymmetric Migration of Wheat Stripe Rust Pathogen in Western Epidemic Areas of China.

Puccinia striiformis f. sp. tritici causes severe global epidemics of wheat stripe rust primarily by airborne urediniospores. Understanding long-distance migration patterns of P. striiformis f. sp. tritici is critical for disease forecasting and management. Although the western epidemic areas in China have been considered as the source of inoculum spread eastward across the country, migration pathways among different populations within the western epidemic areas are poorly understood. In this study, we investigated the population genetics of 200 P. striiformis f. sp. tritici isolates from western epidemic areas using amplified fragment length polymorphism and simple sequence repeat markers. A coalescent approach was also used to calculate the migration rates among populations. Data analyses with both marker systems indicated high genetic diversity in each regional population. The Mantel test revealed significant positive correlation between genetic and geographic distances. Both discriminant analysis of principal components and STRUCTURE analysis supported moderate population structure shaped by seasonality and geography. The calculated migration rates indicated the presence of asymmetric migration between major populations in western epidemic areas, with greater migration rates from high elevation, oversummering areas to low elevation, overwintering areas. Sichuan Basin, one of the low elevation, overwintering areas, sampled in both fall and spring, was inferred as a recipient in fall but a donor in spring. Migration among P. striiformis f. sp. tritici populations may be partly attributable to terrace farming and prevailing wind direction in different seasons. Our study provides a better understanding of fine-scale population structure and the interregional migration pattern of P. striiformis f. sp. tritici in northwestern China and will inform future rust management.

Myrothecium-like new species from turfgrasses and associated rhizosphere.

Myrothecium sensu lato includes a group of fungal saprophytes and weak pathogens with a worldwide distribution. Myrothecium s.l. includes 18 genera, such as Myrothecium, Septomyrothecium, Myxospora, all currently included in the family Stachybotryaceae. In this study, we identified 84 myrothecium-like strains isolated from turfgrasses and their rhizosphere. Five new species, i.e., Alfariapoae, Alf.humicola, Dimorphisetaacuta, D.obtusa, and Paramyrotheciumsinense, are described based on their morphological and phylogenetic distinctions. Phylogenies were inferred based on the analyses of sequences from four DNA loci (ITS, cmdA, rpb2 and tub2). The generic concept of Dimorphiseta is broadened to include a third type of seta, i.e. thin-walled, straight with obtuse apices.

A new leaf blight disease of turfgrasses caused by Microdochium poae, sp. nov.

A novel species of Microdochium was identified as the causal agent of a leaf blight of Poa pratensis (Kentucky blue grass) and Agrostis stolonifera (Creeping bentgrasses), two cold-season turfgrasses widely grown on golf courses in northern China. This disease first appears as small, water-soaked, and scattered leaf spots. Under conditions of high temperatures and successive days of rain, the infected leaves rapidly lose their integrity and large diseased patches appear. Fungal strains were isolated from blighted leaf spots. A phylogenetic analysis based on the nuc rDNA internal transcribed spacer regions and 5.8S rRNA gene (ITS1-5.8S-ITS2 = ITS) and parts of the ß-tubulin (TUB2) and RNA polymerase II second largest subunit (RPB2) genes strongly supported that these isolates are a distinct evolutionary lineage in Microdochium (Microdochiaceae, Xylariales) that represents a new taxonomic species, herein named as M. poae. Microscopic characters confirmed that these strains were morphologically distinct from known Microdochium species. The pathogenicity of M. poae was confirmed by inoculating spore suspension on both grasses and reisolation of the pathogen from symptomatic tissues. The optimal growth temperature suggests that the occurrence of the new leaf blight disease caused by M. poae was significantly different from the microdochium patch disease caused by M. nivale.

Population Genetic Structure and the Migration of Puccinia striiformis f. sp. tritici Between the Gansu and Sichuan Basin Populations of China.

Puccinia striiformis f. sp. tritici is the causal pathogen of interregional epidemics of wheat stripe rust in China via long-distance migration. Gansu Province serves as putative inoculum center providing oversummering inoculum, while Sichuan Basin area serves as a region providing huge amounts of overwintering inoculum. Thus, the relationship between these two regions in population exchange and migration become important in prediction of interregional epidemics. In this study, we compared the population genetic structure and race composition between Gansu and Sichuan Basin populations to infer their migration relationships. A total of 526 isolates, spanning 3 years, were genotyped using eight pairs of amplified fragment length polymorphism markers, and a subset of 98 isolates were inoculated onto 19 Chinese differentials to perform the race analysis. Twenty-three common races and 26 shared genotypes supplied molecular evidence for migration between Gansu and Sichuan Basin populations. Bayesian assignment and principal component analysis revealed that the genetic group assignment of the Sichuan Basin populations (10SB and 11SB) changed in the spring to align with the fall Gansu populations in the prior seasons (09GS and 10GS), which indicated an asymmetric migration from Gansu Province to the Sichuan Basin area. The linkage disequilibrium and the parsimony tree length permutation test revealed a strong annual recombination signal in the Gansu populations and an inconsistent signal in the Sichuan Basin populations.

Population Subdivision of Fusarium graminearum from Barley and Wheat in the Upper Midwestern United States at the Turn of the Century.

Fusarium graminearum, the causal agent of Fusarium head blight (FHB) in wheat and barley, is one of the most economically destructive pathogens of these grains worldwide. Recent population genetic studies of the pathogen obtained from wheat in North America supported population subdivision in part correlated with the spectrum of trichothecene mycotoxins (chemotype) produced by individuals within each population. In contrast, a recent study of F. graminearum obtained from diseased barley in the upper Midwestern United States concluded that only a single population was present, consisting of individuals with various chemotypes. To test whether strains derived from different hosts potentially have different population dynamics, we obtained the barley strains used in the previous study and compared them with wheat strains isolated at a similar time and geographic origin. A total of 247 F. graminearum isolates from barley were assigned firmly into two clusters using a Bayesian clustering method. Subdivision within the barley population corresponded to the previously described NA1 (correlated with the 15ADON chemotype) and NA2 (correlated with the 3ADON chemotype) populations from wheat. However, in both sampling periods the barley population exhibited a higher level of genetic differentiation between NA1 and NA2 populations, fewer admixed individuals and evidence of unidirectional gene introgression (15ADON strains with NA2 genetic backgrounds). These results suggest less recombination between NA1 and NA2 populations on barley compared with wheat. The frequency of 3ADON chemotype strains in the most recently surveyed barley population suggests a latitudinal cline from the northern (49%), central (40%) to the southern (29%) sampling area. The potential to produce a novel trichothecene, 3α-acetoxy,7α,15-dihydroxy-12,13-epoxytrichothe-9-ene (NX-2), was not detected in the barley population but occurred at a low rate (2.4%) in the wheat population.

Population Genetic Structure of Puccinia striiformis in Northwestern China.

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most important fungal diseases of wheat in China. Xinjiang, Qinghai, and Gansu Provinces, located in the northwest of China, are the key regions for interregional epidemics of wheat stripe rust due to their geographic as well as crop-planting features, in relation to pathogen's life cycle, reproduction, and population genetics. To study the population genetic structures of the pathogen in these areas, 217 isolates of P. striiformis f. sp. tritici were collected from different geographic locations at various elevations in the three provinces. The amplified fragment length polymorphism (AFLP) genotypes and virulence phenotypes were analyzed for Xinjiang, Qinghai, and Gansu populations. Frequent genotypic exchanges between Xinjiang and Qinghai and between Qinghai and Gansu populations were detected, demonstrating that the populations of Xinjiang and Gansu may not be completely isolated. Genotypic diversity in Gansu Province was much higher than that in Xinjiang or Qinghai Province. The Xinjiang population was genetically closer to the Qinghai population than to the Gansu population. The race dynamic patterns in Qinghai were consistent with those in Gansu but the similar component pattern of the race dynamics in Xinjiang appeared late, following those in Gansu and other interior epidemic regions of China. No significant correlation between the AFLP genetic distance and the virulence distance in the Xinjiang, Qinghai, and Gansu populations was determined. A possible recombination signature of the pathogen population was detected in Gansu population and some subpopulations in Qinghai but not in Xinjiang population.

Population Genetic Structures of Puccinia striiformis in Ningxia and Gansu Provinces of China.

Wheat stripe rust, caused by the fungal pathogen Puccinia striiformis f. sp. tritici, is one of the most destructive plant diseases in China. Gansu and Ningxia Provinces are considered to be the key areas for over-summering of this pathogen in China. In this study, 283 P. striiformis f. sp. tritici isolates were collected in these two provinces. Amplified fragment length polymorphism (AFLP) was used to analyze the population genetics and to infer the chance of population exchanges between different geographic locations and seasons. The genotypic diversity of the Gansu population (0.514) was slightly higher than that of the Ningxia population (0.489). The occurrence of frequent population exchanges between these two regions was observed, showing that 40 AFLP genotypes were shared by the populations of the two provinces. Gene flow between these two regions in autumn and spring subpopulations was also detected. The genotype distribution in three populations of Ningxia from opposite sides of the Liupan Mountains revealed possible significant effects of the mountains on limiting gene flow and population exchange. Phylogenetic analysis confirmed the possibility of recombination in some of the studied subpopulations in both provinces.