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Abstract Fusarium head blight (FHB) is one of the most destructive wheat diseases worldwide. To understand the impact of human migration and changes in agricultural practices on crop pathogens, here population genomic analysis with 245 representative strains from a collection of 4,427 field isolates of Fusarium asiaticum, the causal agent of FHB in Southern China is conducted. Three populations with distinct evolution trajectories are identifies over the last 10,000 years that can be correlated with historically documented changes in agricultural practices due to human migration caused by the Southern Expeditions during the Jin Dynasty. The gradual decrease of 3ADON‐producing isolates from north to south along with the population structure and spore dispersal patterns shows the long‐distance (>250 km) dispersal of F. asiaticum. These insights into population dynamics and evolutionary history of FHB pathogens are corroborated by a genome‐wide analysis with strains originating from Japan, South America, and the USA, confirming the adaptation of FHB pathogens to cropping systems and human migration.

Abstract Background Advanced cancer patients with good Eastern Cooperative Oncology Group (ECOG) performance status (score 0–1) are underrepresented in current qualitative reports compared with their dying counterparts. Aim To explore the experiences and care needs of advanced cancer patients with good ECOG. Design A qualitative phenomenological approach using semi-structured interview was employed. Data was analyzed using the Colaizzi’s method. Setting/Participants Purposive sample of terminal solid cancer patients on palliative care aged 18–70 years with a 0–1 ECOG score were recruited from a tertiary general hospital. Results Sixteen participants were interviewed. Seven themes were generated from the transcripts, including experiencing no or mild symptoms; independence in self-care, decision-making, and financial capacity; prioritization of cancer growth suppression over symptom management; financial concerns; hope for prognosis and life; reluctance to discuss death and after-death arrangements; and use of complementary and alternative medicine (CAM) and religious coping. Conclusions Advanced cancer patients with good ECOG have distinct experiences and care needs from their dying counterparts. They tend to experience no or mild symptoms, demonstrate a strong sense of independence, and prioritize cancer suppression over symptom management. Financial concerns were common and impact their care-related decision-making. Though being hopeful for their prognosis and life, many are reluctant to discuss death and after-death arrangements. Many Chinese patients use herbal medicine as a CAM modality but need improved awareness of and accessibility to treatment options. Healthcare professionals and policy-makers should recognize their unique experiences and needs when tailoring care strategies and policies.

Common bunt caused by Tilletia laevis Kühn is one of the most serious fungal diseases of wheat. The root–microbial associations play key roles in protecting plants against biotic and abiotic factors. Managing these associations offers a platform for improving the sustainability and efficiency of agriculture production. Here, by using high throughput sequencing, we aimed to identify the bacterial and fungal associations in wheat, alfalfa, and oat crops cultivated in different years in the Gansu province of China. Soil samples (0–6 cm below the surface) from infected wheat by T. laevis had significantly more bacterial and fungal richness than control samples as per the Chao1 analysis. We found some dominant fungi and bacterial phyla in infected wheat by T. laevis, such as Proteobacteria, Acidobacteria, Actinobacteria, Chloroflexi, Ascomycota, Basidiomycota, and Mortierello mycota. We also analyzed the chemical and enzymatic properties of soil samples after T. laevis inoculation. The total nitrogen, total kalium (TK), ammonium nitrogen, available kalium, organic carbon, invertase, phosphatase, and catalase were more in T. laevis-infected samples as compared to the control samples, while pH, total phosphorus, nitrate nitrogen, available phosphorus, and urease were more in control samples compared to T. laevis-infected samples. The results of this study will contribute to the control of wheat common bunt by candidate antagonistic microorganisms and adverse properties of soil.

Tilletia controversa J. G. Kühn is the causal agent of wheat dwarf bunt (DB), a destructive disease causing tremendous economic losses. Small cysteine-rich secreted proteins (SCPs) of plant fungi are crucial in modulating host immunity and promoting infection. Little is known about the virulence effectors of T. controversa. Here, we characterized TcSCP_9014, a novel effector of SCPs, in T. controversa which suppressed programmed cell death triggered by BAX without relying on its signal peptide (SP). The SP in the N-terminus of TcSCP_9014 was functional in the secretory process. Live-cell imaging in the epidermal cells of Nicothiana benthamiana suggested that TcSCP_9014 localized to the plasma membrane, cytoplasm, and nucleus. Furthermore, yeast cDNA library screening was performed to obtain the interacting proteins in wheat. Yeast two-hybrid and BiFC assays were applied to validate the interaction of TcSCP_9014 with TaMTAN and TaGAPDH. Our work revealed that the novel effector TcSCP_9014 is vital in modulating plant immunity, which opens up new avenues for plant-pathogen interactions in the T. controversa infection process.

Abstract Wheat leaf rust is caused by Puccinia triticina (Pt), leading to serious wheat yield loss in the world. To study the population structure and reveal the transmission routes of Pt in eastern China, leaf samples were collected from the main wheat-producing areas from April to June 2020. Total of 372 Pt strains were amplified by 13 SSR makers and a high level of genetic diversity was revealed with 289 multi-locus genotypes (MLG) identified. STRUCTURE analysis suggests that all Pt strains were assigned to 3 clusters, and 11 populations were further defined by considering geographic locations. All 55 pairwise populations had number of migration (Nm) values > 1, indicating moderate genetic differentiation and frequent exchanges among populations. The genetic structure was significant different among populations in the northern and southern regions bounded by the Qinling Mountains-Huaihe River line. Pt strains in the southern regions, such as Jiangsu, Anhui and Zhejiang provinces, had higher level of genetic diversity and genetic variation, and Jiangsu might play an important role in the epidemic and population structure of Pt. Both genetic communication and horizontal wind field analyses showed that Pt had higher level of gene flow from the southern to northern regions than that of the reverse direction. The demonstrated genetic structure and dispersal route of Chinese eastern Pt populations would provide valuable information for epidemiological studies and disease control.

Wheat (Triticum aestivum L.) diseases are major factors responsible for substantial yield losses worldwide, which affect global food security. For a long time, plant breeders have been struggling to improve wheat resistance against major diseases by selection and conventional breeding techniques. Therefore, this review was conducted to shed light on various gaps in the available literature and to reveal the most promising criteria for disease resistance in wheat. However, novel techniques for molecular breeding in the past few decades have been very fruitful for developing broad-spectrum disease resistance and other important traits in wheat. Many types of molecular markers such as SCAR, RAPD, SSR, SSLP, RFLP, SNP, and DArT, etc., have been reported for resistance against wheat pathogens. This article summarizes various insightful molecular markers involved in wheat improvement for resistance to major diseases through diverse breeding programs. Moreover, this review highlights the applications of marker assisted selection (MAS), quantitative trait loci (QTL), genome wide association studies (GWAS) and the CRISPR/Cas-9 system for developing disease resistance against most important wheat diseases. We also reviewed all reported mapped QTLs for bunts, rusts, smuts, and nematode diseases of wheat. Furthermore, we have also proposed how the CRISPR/Cas-9 system and GWAS can assist breeders in the future for the genetic improvement of wheat. If these molecular approaches are used successfully in the future, they can be a significant step toward expanding food production in wheat crops.

ABSTRACT Wheat dwarf bunt leads to the replacement of seeds with fungal galls containing millions of teliospores of the pathogen Tilletia controversa Kühn. As one of the most devastating internationally quarantined wheat diseases, wheat dwarf bunt spreads to cause distant outbreaks by seeds containing teliospores. In this study, based on a combination of amplicon sequencing and isolation approaches, we analyzed the seed microbiome signatures of endophytes between resistant and susceptible cultivars after infection with T. controversa. Among 310 bacterial species obtained only by amplicon sequencing and 51 species obtained only by isolation, we found 14 overlapping species by both methods; we detected 128 fungal species only by amplicon sequencing, 56 only by isolation, and 5 species by both methods. The results indicated that resistant uninfected cultivars hosted endophytic communities that were much more stable and beneficial to plant health than those in susceptible infected cultivars. The susceptible group showed higher diversity than the resistant group, the infected group showed more diversity than the uninfected group, and the microbial communities in seeds were related to infection or resistance to the pathogen. Some antagonistic microbes significantly suppressed the germination rate of the pathogen’s teliospores, providing clues for future studies aimed at developing strategies against wheat dwarf bunt. Collectively, this research advances the understanding of the microbial assembly of wheat seeds upon exposure to fungal pathogen (T. controversa) infection. IMPORTANCE This is the first study on the microbiome signature of endophytes in wheat seed response to wheat dwarf bunt caused by Tilletia controversa Kühn. Some antagonistic microbes suppressed the germination of teliospores of the pathogen significantly, which will provide clues for future studies against wheat dwarf bunt. Collectively, this research first advances the understanding of the microbial assembly of wheat seed upon exposure to the fungal pathogen (T. controversa) infection.

ABSTRACT The folding and dynamics of three-dimensional (3D) genome organization are fundamental for eukaryotes executing genome functions but have been largely unexplored in nonmodel fungi. Using high-throughput sequencing coupled with chromosome conformation capture (Hi-C) data, we generated two chromosome-level assemblies for Puccinia striiformis f. sp. tritici, a fungus causing stripe rust disease on wheat, for studying 3D genome architectures of plant pathogenic fungi. The chromatin organization of the fungus followed a combination of the fractal globule model and the equilibrium globule model. Surprisingly, chromosome compartmentalization was not detected. Dynamics of 3D genome organization during two developmental stages of P. striiformis f. sp. tritici indicated that regulation of gene activities might be independent of the changes of genome organization. In addition, chromatin conformation conservation was found to be independent of genome sequence synteny conservation among different fungi. These results highlighted the distinct folding principles of fungal 3D genomes. Our findings should be an important step toward a holistic understanding of the principles and functions of genome architecture across different eukaryotic kingdoms. IMPORTANCE Previously, our understanding of 3D genome architecture has mainly come from model mammals, insects, and plants. However, the organization and regulatory functions of 3D genomes in fungi are largely unknown. In this study, we comprehensively investigated P. striiformis f. sp. tritici, a plant fungal pathogen, and revealed distinct features of the 3D genome, comparing it with the universal folding feature of 3D genomes in higher eukaryotic organisms. We further suggested that there might be distinct regulatory mechanisms of gene expression that are independent of chromatin organization changes during the developmental stages of this rust fungus. Moreover, we showed that the evolutionary pattern of 3D genomes in this fungus is also different from the cases in mammalian genomes. In addition, the genome assembly pipeline and the generated two chromosome-level genomes will be valuable resources. These results highlighted the unexplored distinct features of 3D genome organization in fungi. Therefore, our study provided complementary knowledge to holistically understand the organization and functions of 3D genomes across different eukaryotes.

Tilletia laevis Kühn (syn. Tilletia foetida (Wallr.) Liro.) causes wheat common bunt, which is one of the most devastating plant diseases in the world. Common bunt can result in a reduction of 80% or even a total loss of wheat production. In this study, the characteristics of T. laevis infection in compatible wheat plants were defined based on the combination of scanning electron microscopy, transmission electron microscopy and laser scanning confocal microscopy. We found T. laevis could lead to the abnormal growth of wheat tissues and cells, such as leakage of chloroplasts, deformities, disordered arrangements of mesophyll cells and also thickening of the cell wall of mesophyll cells in leaf tissue. What’s more, T. laevis teliospores were found in the roots, stems, flag leaves, and glumes of infected wheat plants instead of just in the ovaries, as previously reported. The abnormal characteristics caused by T. laevis may be used for early detection of this pathogen instead of molecular markers in addition to providing theoretical insights into T. laevis and wheat interactions for breeding of common bunt resistance.

Abstract Dwarf bunt caused by the pathogen Tilletia controversa Kühn is one of the most serious quarantine diseases of winter wheat. Metabolomics studies provide detailed information about the biochemical changes at the cell and tissue levels of plants. In the present study, a liquid chromatography/mass spectrometry (LC/MS) metabolomics approach was used to investigate the changes in the grain metabolomics of infected and noninfected with T. controversa samples. PCA suggested that T. controversa-infected and noninfected samples were separated during the interaction. LC/MS analysis showed that 62 different metabolites were recorded in the grains, among which a total of 34 metabolites were upregulated and 28 metabolites were downregulated. Prostaglandins (PGs) and 9-hydroxyoctadecadienoic acids (9-HODEs) are fungal toxin-related substances, and their expression significantly increased in T. controversa-infected grains. Additionally, the concentrations of cucurbic acid and octadecatrienoic acid changed significantly after pathogen infection, which play a large role in plant defense. The eight different metabolic pathways activated during T. controversa and wheat plant interactions included phenylalanine metabolism, isoquinoline alkaloid biosynthesis, starch and sucrose metabolism, tyrosine metabolism, sphingolipid metabolism, arginine and proline metabolism, alanine, aspartate, and glutamate metabolism, and tryptophan metabolism. In conclusion, we found differences in the metabolic profiles of wheat grains after T. controversa infection. To our knowledge, this is the first study to evaluate the metabolites in wheat grains after T. controversa infection.

Rhizosphere microbes play a vital role in plant health and defense against soil-borne diseases. Previous studies showed that maize-soybean relay strip intercropping altered the diversity and composition of pathogenic Fusarium species and biocontrol fungal communities in the soybean rhizosphere, and significantly suppressed soybean root rot. However, whether the rhizosphere bacterial community participates in the regulation of this intercropping on soybean root rot is not clear. In this study, the rhizosphere soil of soybean healthy plants was collected in the continuous cropping of maize-soybean relay strip intercropping and soybean monoculture in the fields, and the integrated methods of microbial profiling, dual culture assays in vitro, and pot experiments were employed to systematically investigate the diversity, composition, and function of rhizosphere bacteria related to soybean root rot in two cropping patterns. We found that intercropping reshaped the rhizosphere bacterial community and increased microbial community diversity, and meanwhile, it also recruited much richer and more diverse species of Pseudomonas sp., Bacillus sp., Streptomyces sp., and Microbacterium sp. in soybean rhizosphere when compared with monoculture. From the intercropping, nine species of rhizosphere bacteria displayed good antagonism against the pathogen Fusarium oxysporum B3S1 of soybean root rot, and among them, IRHB3 (Pseudomonas chlororaphis), IRHB6 (Streptomyces), and IRHB9 (Bacillus) were the dominant bacteria and extraordinarily rich. In contrast, MRHB108 (Streptomyces virginiae) and MRHB205 (Bacillus subtilis) were the only antagonistic bacteria from monoculture, which were relatively poor in abundance. Interestingly, introducing IRHB3 into the cultured substrates not only significantly promoted the growth and development of soybean roots but also improved the survival rate of seedlings that suffered from F. oxysporum infection. Thus, this study proves that maize-soybean relay strip intercropping could help the host resist soil-borne Fusarium root rot by reshaping the rhizosphere bacterial community and driving more beneficial microorganisms to accumulate in the soybean rhizosphere.

Seed-borne pathogens cause diverse diseases at the growth, pre- and post-harvest stage of soybean resulting in a large reduction in yield and quality. The physiological and metabolic aspects of seeds are closely related to their defense against pathogens. Recently, Fusarium fujikuroi has been identified as the dominant seed-borne fungi of soybean seed decay, but little information on the responses of soybean seeds induced by F. fujikuroi is available. In this study, a time-course symptom development of seed decay was observed after F. fujikuroi inoculation through spore suspension soaking. The germination rate and the contents of soluble sugar and soluble protein were significantly altered over time. Both chitinase and β-1,3-glucanase as important fungal cell wall–degrading enzymes of soybean seeds were also rapidly and transiently activated upon the early infection of F. fujikuroi. Metabolic profile analysis showed that the metabolites in glycine, serine, and threonine metabolism and tryptophan metabolism were clearly induced by F. fujikuroi, but different metabolites were mostly enriched in isoflavone biosynthesis, flavone biosynthesis, and galactose pathways. Interestingly, glycitein and glycitin were dramatically upregulated while daidzein, genistein, genistin, and daidzin were largely downregulated. These results indicate a combination of physiological responses, cell wall–related defense, and the complicated metabolites of soybean seeds contributes to soybean seed resistance against F. fujikuroi, which are useful for soybean resistance breeding.

Abstract Dwarf bunt of wheat, which is caused by Tilletia controversa J.G. Kühn, is a soil-borne disease which may lead up to an 80% loss of yield together with degradation of the quality of the wheat flour by production of a fishy smell. In this study, high-throughput sequencing technology was employed to characterize the microbial composition of wheat tissues (roots, spikes, first stem under the ear, and stem base) and rhizosphere soil of wheat varieties that are resistant and susceptible to T. controversa. We observed that the soil fungal community abundance and diversity were higher in resistant varieties than in susceptible varieties in both inoculated and uninoculated wheat, and the abundances of Sordariomycetes and Mortierellomycetes increased in the resistant varieties infected with T. controversa, while the abundances of Dothideomycetes and Bacteroidia increased in the susceptible varieties. Regarding the bacteria present in wheat tissues, the abundances of Chloroflexi, Bacteroidetes, Gemmatimonadetes, Verrucomicrobia and Acidobacteria in the ear and the first stem under the ear were higher than those in other tissues. Our results indicated that the abundances of Sordariomycetes, Mortierellomycetes, Leotiomycetes, Chryseobacterium and Massilia were higher in T. controversa-infected resistant varieties than in their controls, that Dothideomycetes, Bacteroidia, Nocardioides and Pseudomonas showed higher abundances in T. controversa-infected susceptible varieties, and that Curtobacterium, Exiguobacterium, Planococcus, and Pantoea may have higher abundances in both T. controversa-infected susceptible and resistant varieties than in their own controls.

Abstract Background Dwarf bunt, which is caused by Tilletia controversa Kühn, is a soilborne and seedborne disease that occurs worldwide and can lead to 70% or even total losses of wheat crops. However, very little information is available about the histological changes that occur in dwarf bunt-resistant and dwarf bunt-susceptible wheat plants at the tillering stage (Z21). In this study, we used scanning electron microscopy and transmission electron microscopy to characterize the histological changes at this stage in resistant and susceptible wheat cultivars infected by T. controversa. Results Using scanning electron microscopy, the root, stem, and leaf structures of resistant and susceptible cultivars were examined after T. controversa infection. The root epidermal and vascular bundles were more severely damaged in the susceptible T. controversa-infected plants than in the resistant plants. The stem cell and longitudinal sections were much more extensively affected in susceptible plants than in resistant plants after pathogen infection. However, slightly deformed mesophyll cells were observed in the leaves of susceptible plants. With transmission electron microscopy, we found that the cortical bundle cells and the cell contents and nuclei in the roots were more severely affected in the susceptible plants than in the resistant plants; in the stems and leaves, the nuclei, chloroplasts, and mesophyll cells changed significantly in the susceptible plants after fungal infection. Moreover, we found that infected susceptible and resistant plants were affected much more severely at the tillering stage (Z21) than at the seedling growth stage (Z13). Conclusion Histological changes in the wheat roots, stems and leaves were much more severe in T. controversa-infected susceptible plants than in infected resistant plants at the tillering stage (Z21).

Abstract Wheat dwarf bunt is caused by Tilletia controversa Kühn, which is one of the most destructive diseases of wheat worldwide. To explore the interaction of T. controversa and wheat, we analysed the transcriptome profile of spikes of the susceptible wheat cultivar Dongxuan 3, which was subjected to a T. controversa infection and a mock infection. The results obtained from a differential expression analysis of T. controversa-infected plants compared with mock-infected ones showed that 10,867 out of 21,354 genes were upregulated, while 10,487 genes were downregulated, and these genes were enriched in 205 different pathways. Our findings demonstrated that the genes associated with defence against diseases, such as PR-related genes, WRKY transcription factors and mitogen-activated protein kinase genes, were more highly expressed in response to T. controversa infection. Additionally, a number of genes related to physiological attributes were expressed during infection. Three pathways were differentiated based on the characteristics of gene ontology classification. KEGG enrichment analysis showed that twenty genes were expressed differentially during the infection of wheat with T. controversa. Notable changes were observed in the transcriptomes of wheat plants after infection. The results of this study may help to elucidate the mechanism governing the interactions between this pathogen and wheat plants and may facilitate the development of new methods to increase the resistance level of wheat against T. controversa, including the overexpression of defence-related genes.

Exploiting wheat cultivars with stable resistance to Fusarium Head blight (FHB) and toxin accumulation is a cost-effective and environmentally friendly strategy to reduce the risk of yield losses and contamination with mycotoxins. To facilitate the deployment of stable cultivar resistance, we evaluated FHB resistance and resistance to mycotoxin accumulation in 410 wheat lines bred by local breeders from four major wheat growing regions in China after natural infection at three distinct locations (Hefei, Yangzhou and Nanping). Significant differences in disease index were observed among the three locations. The disease indexes (DI’s) in Nanping were the highest, followed by Yangzhou and Hefei. The distribution of DI’s in Yangzhou showed the best discrimination of FHB resistance in cultivars. Growing region and cultivar had significant effect on DI and mycotoxins. Among the climate factors, relative humidity and rainfall were the key factors resulting in the severe disease. Even though most cultivars were still susceptible to FHB under the strongly conducive conditions applied, the ratio of resistant lines increased in the Upper region of the Yangtze River (UYR) and the Middle and Lower Region of the Yangtze River (MLYR) between 2015 and 2019. Deoxynivalenol (DON) was the dominant mycotoxin found in Hefei and Yangzhou, while NIV was predominant in Nanping. Disease indexes were significantly correlated with DON content in wheat grain.

Abstract Common bunt of wheat caused by Tilletia laevis and/or T. caries (syn. T. tritici), is a major disease in wheat-growing regions worldwide that could lead to 80% or even total loss of production. Even though T. laevis can be distinguished from T. caries on the bases of morphology of teliospores using microscopy technique. However, molecular methods could serve as an additional method to quantify the pathogen. To develop a rapid diagnostic and quantify method, we employed the ISSR molecular marker for T. laevis in this study. The primer ISSR857 generated a polymorphic pattern displaying a 1385 bp T. laevis-specific DNA fragment. A pair of specific primers (L57F/L57R) was designed to amplify a sequence-characterized amplified region (SCAR) (763 bp) for the PCR detection assay. The primers amplified the DNA fragment in the tested isolates of T. laevis but failed in the related species, including T. caries. The detection limit of the primer set (L57F/L57R) was 5 ng/µl of DNA extracted from T. laevis teliospores. A SYBR Green I real-time PCR method for detecting T. laevis with a 100 fg/µl detection limit and droplet digital PCR with a high sensitivity (30 fg/µl detection limit) were developed; this technique showed the most sensitive detection compared to the SCAR marker and SYBR Green I real-time PCR. Additionally, this is the first study related the detection of T. laevis with the droplet digital PCR method.

Tilletia laevis Kühn [synonym T. foetida (Wallr.) Liro] can lead to a wheat common bunt, which is one of the most serious diseases affecting kernels, a serious reduction in grain yield, and losses can reach up to 80% in favorable environments. To understand how wheat tassels respond to T. laevis, based on an RNA-Seq technology, we analyzed a host transcript accumulation on healthy wheat tassels and on tassels infected by the pathogen. Our results showed that 7,767 out of 15,658 genes were upregulated and 7,891 out of 15,658 genes were downregulated in wheat tassels. Subsequent gene ontology (GO) showed that differentially expressed genes (DEGs) are predominantly involved in biological processes, cellular components, and molecular functions. Additionally, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that 20 pathways were expressed significantly during the infection of wheat with T. laevis, while biosynthesis of amino acids, carbon metabolism, and starch and sucrose metabolism pathways were more highly expressed. Our findings also demonstrated that genes involved in defense mechanisms and myeloblastosis (MYB) transcription factor families were mostly upregulated, and the RNA-seq results were validated by quantitative real-time polymerase chain reaction (qRT-PCR). This is the first report on transcriptomics analysis of wheat tassels in response to T. laevis, which will contribute to understanding the interaction of T. laevis and wheat, and may provide higher efficiency control strategies, including developing new methods to increase the resistance of wheat crops to T. laevis-caused wheat common bunt.

Leaf rust of wheat caused by Puccinia triticina (Pt) is one of the most common fungal diseases in the southwest and northwest of China, the middle and lower reaches of the Yangtze River, and the southern part of the Huang-Huai-Hai river basin. Using 13 simple sequence repeat (SSR) markers, we systematically revealed the genotypic diversities, population differentiation and reproduction of Pt isolates in 15 wheat-producing areas in China. A total of 622 isolates were divided into 3 predominant populations, including the eastern Pt populations, consisting of Pt samples from 8 eastern provinces of Beijing, Hebei, Shanxi, Shaanxi, Anhui, Shandong, Henan, and Heilongjiang; the 4 western Pt populations from Gansu, Qinghai, Sichuan, and Inner Mongolia provinces; and the bridge Pt populations including Jiangsu, Hubei, and Yunnan, which communicated the other 2 populations as a “bridge”. The pathogen transmission of eastern Pt populations was more frequent than western Pt populations. The linkage disequilibrium test indicated that the whole Pt population was in a state of linkage disequilibrium. However, populations of Beijing, Hebei, Shaanxi, Jiangsu, Henan, and Heilongjiang provinces showed obvious linkage equilibrium, while the five provinces of Qinghai, Hubei, Anhui, Shandong, and Inner Mongolia supported clonal modes of reproduction.

Tilletia laevis causes common bunt disease in wheat, with severe losses of production yield and seed quality. Metabolomics studies provide detailed information about the biochemical changes at the cell and tissue level of the plants. Ultrahigh-performance liquid chromatography–Q-exactive mass spectrometry (UPLC-QE-MS) was used to examine the changes in wheat grains after T. laevis infection. PCA analysis suggested that T. laevis-infected and non-infected samples were scattered separately during the interaction. In total, 224 organic acids and their derivatives, 170 organoheterocyclic compounds, 128 lipids and lipid-like molecules, 85 organic nitrogen compounds, 64 benzenoids, 31 phenylpropanoids and polyketides, 21 nucleosides, nucleotides, their analogues, and 10 alkaloids and derivatives were altered in hyphal-infected grains. According to The Kyoto Encyclopedia of Genes and genomes analysis, the protein digestion and absorption, biosynthesis of amino acids, arginine and proline metabolism, vitamin digestion and absorption, and glycine, serine, and threonine metabolism pathways were activated in wheat crops after T. laevis infection.

Rust fungi in Pucciniales have caused destructive plant epidemics, have become more aggressive with new virulence, rapidly adapt to new environments, and continually threaten global agriculture. With the rapid advancement of genome sequencing technologies and data analysis tools, genomics research on many of the devastating rust fungi has generated unprecedented insights into various aspects of rust biology. In this review, we first present a summary of the main findings in the genomics of rust fungi related to variations in genome size and gene composition between and within species. Then we show how the genomics of rust fungi has promoted our understanding of the pathogen virulence and population dynamics. Even with great progress, many questions still need to be answered. Therefore, we introduce important perspectives with emphasis on the genome evolution and host adaptation of rust fungi. We believe that the comparative genomics and population genomics of rust fungi will provide a further understanding of the rapid evolution of virulence and will contribute to monitoring the population dynamics for disease management.

Wheat leaf rust (caused by Puccinia triticina Erikss.) is among the major diseases of common wheat. The lack of resistance genes to leaf rust has limited the development of wheat cultivars. Wheat–Agropyron cristatum (A. cristatum) 2P addition line II-9-3 has been shown to provide broad-spectrum immunity to leaf rust. To identify the specific A. cristatum resistance genes and related regulatory pathways in II-9-3, we conducted a comparative transcriptome analysis of inoculated and uninoculated leaves of the resistant addition line II-9-3 and the susceptible cultivar Fukuhokomugi (Fukuho). The results showed that there were 66 A. cristatum differentially expressed genes (DEGs) and 1389 wheat DEGs in II-9-3 during P. triticina infection. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment and gene set enrichment analysis (GSEA) revealed that the DEGs of II-9-3 were associated with plant–pathogen interaction, MAPK signaling pathway–plant, plant hormone signal transduction, glutathione metabolism, and phenylpropanoid biosynthesis. Furthermore, many defense-related A. cristatum genes, such as two NLR genes, seven receptor kinase-encoding genes, and four transcription factor-encoding genes, were identified. Our results indicated that the key step of resistance to leaf rust involves, firstly, the gene expression of chromosome 2P upstream of the immune pathway and, secondly, the effect of chromosome 2P on the co-expression of wheat genes in II-9-3. The disease resistance regulatory pathways and related genes in the addition line II-9-3 thus could play a critical role in the effective utilization of innovative resources for leaf rust resistance in wheat breeding.

Dwarf bunt and common bunt diseases of wheat are caused by Tilletia controversa Kühn and Tilletia foetida Kühn, respectively, and losses caused by these diseases can reach 70–80% in favourable conditions. T. controversa and T. foetida are fungal pathogens belonging to the Exobasidiomycetes within the basidiomycetous smut fungi (Ustilaginomycotina). In order to illuminate the proteomics differences of wheat spikes after the infection of T. controversa and T. foetida, the isobaric tags for relative and absolute quantification (iTRAQ) technique was used for better clarification. A total of 4553 proteins were differentially detected after T. controversa infection; 4100 were upregulated, and 453 were downregulated. After T. foetida infection, 804 differentially expressed proteins were detected; 447 were upregulated and 357 were downregulated. In-depth data analysis revealed that 44, 50 and 82 proteins after T. controversa and 9, 6 and 16 proteins after T. foetida were differentially expressed, which are antioxidant, plant-pathogen interaction and glutathione proteins, respectively, and 9 proteins showed results consistent with PRM. The top 20 KEGG enrichment pathways were identified after pathogen infection. On the basis of gene ontology, the upregulated proteins were linked with metabolic process, catalytic activity, transferase activity, photosynthetic membrane, extracellular region and oxidoreductase activity. The results expanded our understanding of the proteome in wheat spikes in response to T. controversa and T. foetida infection and provide a basis for further investigation for improving the defense mechanism of the wheat crops.

The dynamic of soil-borne disease is closely related to the rhizosphere microbial communities. Maize–soybean relay strip intercropping has been shown to significantly control the type of soybean root rot that tends to occur in monoculture. However, it is still unknown whether the rhizosphere microbial community participates in the regulation of intercropped soybean root rot. In this study, rhizosphere Fusarium and Trichoderma communities were compared in either healthy or root-rotted rhizosphere soil from monocultured and intercropped soybean, and our results showed the abundance of rhizosphere Fusarium in intercropping was remarkably different from monoculture. Of four species identified, F. oxysporum was the most aggressive and more frequently isolated in diseased soil of monoculture. In contrast, Trichoderma was largely accumulated in healthy rhizosphere soil of intercropping rather than monoculture. T. harzianum dramatically increased in the rhizosphere of intercropping, while T. virens and T. afroharzianum also exhibited distinct isolation frequency. For the antagonism test in vitro, Trichoderma strains had antagonistic effects on F. oxysporum with the percentage of mycelial inhibition ranging from 50.59–92.94%, and they displayed good mycoparasitic abilities against F. oxysporum through coiling around and entering into the hyphae, expanding along the cell–cell lumen and even dissolving cell walls of the target fungus. These results indicate maize–soybean relay strip intercropping significantly increases the density and composition proportion of beneficial Trichoderma to antagonize the pathogenic Fusarium species in rhizosphere, thus potentially contributing to the suppression of soybean root rot under the intercropping.

Background and AimsThe dwarf bunt disease of wheat is caused by Tilletia controversa Kühn. This pathogen is primarily involved in the stunted growth of wheat and affects seed quality. Many countries in the world have therefore imposed quarantine bans to prevent the spread of T. controversa. Morphological observations are the main method of detecting teliospores in soil. However, this is a lengthy and laborious process; this method is thus unable to quickly meet the demand for detection of teliospores in the soil.MethodsWe compared PCR, real-time PCR and droplet digital PCR (ddPCR) for the qualitative and quantitative measurement of the teliospores of T. controversa in soil.ResultsWe suggest the use of ddPCR for detection of the soil samples, which was demonstrated to have the most sensitive detection at 2.1 copies/μL. In contract, SYBR Green I real-time PCR could detect 7.97 copies/μL of T. controversa in soil, and this sensitivity was 100 times more sensitive than that of simple PCR.ConclusionThis study was the first report using ddPCR techniques to detect T. controversa teliospores in soil with greatly enhanced sensitivity.

Maize/soybean relay strip intercropping has been widely practiced in Southwest China due to its high productivity and effective application of agricultural resources; however, several seedborne diseases such as seedling blight, pod and seed decay are frequently observed causing severe yield loss and low seed quality. So far, the population and pathogenicity of the seedborne fungi associated with intercropped soybean remain unexplored. In this study, seeds of 12 soybean cultivars screened for intercropping were collected from three growing regions in Sichuan Province of Southwest China, and the seedborne fungi were isolated from the surface-sterilized seeds. Based on sequence analysis of ribosomal DNA internal transcribed spacer (rDNA ITS), 148 isolates were identified into 13 fungal genera, among which Fusarium covered 55.0% as the biggest population followed by Colletotrichum. Furthermore, Fusarium isolates were classified into five distinct species comprising F. fujikuroi, F. proliferatum, F. verticillioides, F. asiaticum and F. incarnatum through sequence analysis of translation elongation factor 1 alpha (EF-1α) and DNA-directed RNA ploymerase II second largest subunit (RPB2). Among them, F. fujikuroi accounted for 51.22% (42/82) and was isolated from 91.7% (11/12) soybean varieties. Pathogenicity assay showed that five Fusarium species were able to infect the seeds of soybean cultivar “Nandou12” and caused water-soaked or rot symptoms, while F. fujikuroi and F. asiaticum had much higher aggressiveness than other species with significant reductions of seed fresh weight and germination percentage. Accordingly, this study indicates that Fusarium species are the dominant seedborne fungi in the intercropped soybean in Sichuan, China, and this provides some useful references for the effective management of seedborne fungal diseases as well as soybean resistance breeding in maize/soybean relay strip intercropping.

Fusarium species are the most detrimental pathogens of soybean root rot worldwide, causing large loss in soybean production. Maize/soybean relay strip intercropping has significant advantages on the increase of crop yields and efficient use of agricultural resources, but its effects on the occurrence and pathogen population of soybean root rot are rarely known. In this study, root rot was investigated in the fields of the continuous maize/soybean strip relay intercropping and soybean monoculture. Fusarium species were isolated from diseased soybean roots and identified based on sequence analysis of translation elongation factor 1α (EF-1α) and RNA polymerase II second largest subunit (RPB2), and the diversity and pathogenicity of these species were also analyzed. Our results showed that intercropping significantly decreased soybean root rot over monoculture. A more diverse Fusarium population including Fusarium solani species complex (FSSC), F. incarnatum-equiseti species complex (FIESC), F. oxysporum, F. fujikuroi, F. proliferatum and F. verticillioides, F. graminearum and F. asiaticum was identified from intercropping while FSSC, FIESC, F. oxysporum, F. commune, F. asiaticum and F. meridionale were found from monoculture. All Fusarium species caused soybean root infection but exhibited distinct aggressiveness. The most aggressive F. oxysporum was more frequently isolated in monoculture than intercropping. FSSC and FIESC were the dominant species complex and differed in their aggressiveness. Additionally, F. fujikuroi, F. proliferatum and F. verticillioides were specifically identified from intercropping with weak or middle aggressiveness. Except for F. graminearum, F. meridionale and F. asiaticum were firstly reported to cause soybean root rot in China. This study indicates maize/soybean relay strip intercropping can reduce soybean root rot, change the diversity and aggressiveness of Fusarium species, which provides an important reference for effective management of this disease.

Wheat leaf rust is one of the most common wheat diseases worldwide and can cause up to 40% wheat yield loss. To combat the growth and spread of leaf rust disease, continual exploration and identification of new and effective resistance genes are needed. Here, we report for the first time a locus conferring leaf rust resistance located on the long arm of Agropyron cristatum chromosome 2P in Triticum aestivum–A. cristatum 2P translocation lines. This study used 50 leaf rust races, including two Chinese major dominant leaf rust races, named by THT and PHT, and other 48 different leaf rust races collected from 11 provinces, 1autonomous region and 1 municipality of China to test the resistance to T. aestivum–A. cristatum 2P chromosome translocation lines and their backcross populations, the results indicated that the novel leaf rust resistance locus was immune or nearly immune to all tested leaf rust races. Four long arm translocation lines with different breakpoints of A. cristatum chromosome 2PL and their backcross populations were tested with leaf rust race THT at the seedling and adult stages and genotyped with 2P-specific STS markers. The results showed that the novel leaf rust resistance locus of the T. aestivum–A. cristatum 2P translocation lines was located in the chromosomal bin FL 0.66–0.86 of 2PL. Therefore, T. aestivum–A. cristatum 2P chromosome translocation lines conferring leaf rust resistance locus could provide a novel disease-resistance resource for future wheat breeding programs.

Intercropping has been considered as a kind of a sustainable agricultural cropping system. In southwest China, maize/soybean strip intercropping has commonly been practised under local limited agricultural land resources. However, heavy rainfall in combination with high humidity and low temperatures cause severe pod and seed deterioration in the maturity and pre-harvesting stages of intercropped soybean. Numerous Fusarium species have been reported as the dominant pathogens of soybean root rot, seedling blight, as well as pod field mold in this area. However, the diversity and pathogenicity of Fusarium species on soybean pods remain unclear. In the current study, diseased soybean pods were collected during the cropping season of 2018 from five different intercropped soybean producing areas. A total of 83 Fusarium isolates were isolated and identified as F. fujikuroi, F. graminearum, F. proliferatum, and F. incarnatum-equiseti species complex based on morphological characteristics and phylogenetic analysis of the nucleotide sequence of EF1-α and RPB2 genes. Pathogenicity tests demonstrated that all Fusarium species were pathogenic to seeds of the intercropped soybean cultivar Nandou12. Fusarium fujikuroi had the maximum disease severity, with a significant reduction of seed germination rate, root length, and seed weight, followed by F. equiseti, F. graminearum, F. proliferatum, and F. incarnatum. Additionally, the diversity of Fusarium species on soybean pods was also considerably distinct according to the geographical origin and soybean varieties. Thus, the findings of the current study will be helpful for the management and resistance breeding of soybean pod decay in the maize/soybean intercropping system.

The wheat powdery mildew resistance gene Pm40, which is located on chromosomal arm 7BS, is effective against nearly all prevalent races of Blumeria graminis f. sp tritici (Bgt) in China and is carried by the common wheat germplasm PI 672538. A set of the F1, F2 and F2:3 populations from the cross of the resistant PI 672538 with the susceptible line L1034 were used to conduct genetic analysis of powdery mildew resistance and construct a high-density linkage map of the Pm40 gene. We constructed a high-density linkage genetic map with a total length of 6.18 cM and average spacing between markers of 0.48 cM.Pm40 is flanked by Xwmc335 and BF291338 at genetic distances of 0.58 cM and 0.26 cM, respectively, in deletion bin C-7BS-1-0.27. Comparative genomic analysis based on EST-STS markers established a high level of collinearity of the Pm40 genomic region with a 1.09-Mbp genomic region on Brachypodium chromosome 3, a 1.16-Mbp genomic region on rice chromosome 8, and a 1.62-Mbp genomic region on sorghum chromosome 7. We further anchored the Pm40 target intervals to the wheat genome sequence. A putative linear index of 85 wheat contigs containing 97 genes on 7BS was constructed. In total, 9 genes could be considered as candidates for the resistances to powdery mildew in the target genomic regions, which encoded proteins that were involved in the plant defense and response to pathogen attack. These results will facilitate the development of new markers for map-based cloning and marker-assisted selection of Pm40 in wheat breeding programs.

Take-all, which is caused by the fungal pathogen, Gaeumannomyces graminis var. tritici (Ggt), is an important soil-borne root rot disease of wheat occurring worldwide. However, the genetic basis of Ggt pathogenicity remains unclear. In this study, transcriptome sequencing for Ggt in axenic culture and Ggt-infected wheat roots was performed using Illumina paired-end sequencing. Approximately 2.62 and 7.76 Gb of clean reads were obtained, and 87% and 63% of the total reads were mapped to the Ggt genome for RNA extracted from Ggt in culture and infected roots, respectively. A total of 3,258 differentially expressed genes (DEGs) were identified with 2,107 (65%) being 2-fold up-regulated and 1,151 (35%) being 2-fold down-regulated between Ggt in culture and Ggt in infected wheat roots. Annotation of these DEGs revealed that many were associated with possible Ggt pathogenicity factors, such as genes for guanine nucleotide-binding protein alpha-2 subunit, cellulase, pectinase, xylanase, glucosidase, aspartic protease and gentisate 1, 2-dioxygenase. Twelve DEGs were analyzed for expression by qRT-PCR, and could be generally divided into those with high expression only early in infection, only late in infection and those that gradually increasing expression over time as root rot developed. This indicates that these possible pathogenicity factors may play roles during different stages of the interaction, such as signaling, plant cell wall degradation and responses to plant defense compounds. This is the first study to compare the transcriptomes of Ggt growing saprophytically in axenic cultures to it growing parasitically in infected wheat roots. As a result, new candidate pathogenicity factors have been identified, which can be further examined by gene knock-outs and other methods to assess their true role in the ability of Ggt to infect roots.

Wheat (Triticum aestivum L.) diseases are major factors responsible for substantial yield losses worldwide, which affect global food security. For a long time, plant breeders have been struggling to improve wheat resistance against major diseases by selection and conventional breeding techniques. Therefore, this review was conducted to shed light on various gaps in the available literature and to reveal the most promising criteria for disease resistance in wheat. However, novel techniques for molecular breeding in the past few decades have been very fruitful for developing broad-spectrum disease resistance and other important traits in wheat. Many types of molecular markers such as SCAR, RAPD, SSR, SSLP, RFLP, SNP, and DArT, etc., have been reported for resistance against wheat pathogens. This article summarizes various insightful molecular markers involved in wheat improvement for resistance to major diseases through diverse breeding programs. Moreover, this review highlights the applications of marker assisted selection (MAS), quantitative trait loci (QTL), genome wide association studies (GWAS) and the CRISPR/Cas-9 system for developing disease resistance against most important wheat diseases. We also reviewed all reported mapped QTLs for bunts, rusts, smuts, and nematode diseases of wheat. Furthermore, we have also proposed how the CRISPR/Cas-9 system and GWAS can assist breeders in the future for the genetic improvement of wheat. If these molecular approaches are used successfully in the future, they can be a significant step toward expanding food production in wheat crops. [ABSTRACT FROM AUTHOR]