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Fusarium pseudograminearum is a soilborne, hemibiotrophic phytopathogenic fungus that causes Fusarium crown rot and Fusarium head blight in wheat. The basic leucine zipper proteins (bZIPs) are evolutionarily conserved transcription factors that play crucial roles in a range of growth and developmental processes and the responses to biotic and abiotic stresses. However, the roles of bZIP transcription factors remains unknown in F. pseudograminearum. In this study, a bZIP transcription factor Fpkapc was identified to localize to the nucleus in F. pseudograminearum. A mutant strain (Δfpkapc) was constructed to determine the role of Fpkapc in growth and pathogenicity of F. pseudograminearum. Transcriptomic analyses revealed that many genes involved in basic metabolism and oxidation-reduction processes were downregulated, whereas many genes involved in metal iron binding were upregulated in the Δfpkapc strain, compared with the wild type (WT). Correspondingly, the mutant had severe growth defects and displayed abnormal hyphal tips. Conidiation in the Fpkapc mutant was reduced, with more conidia in smaller size and fewer septa than in the WT. Also, relative to WT, the Δfpkapc strain showed greater tolerance to ion stress, but decreased tolerance to H2O2. The mutant caused smaller disease lesions on wheat and barley plants, but significantly increased TRI gene expression, compared with the WT. In summary, Fpkapc plays multiple roles in governing growth, development, stress responses, and virulence in F. pseudograminearum. [ABSTRACT FROM AUTHOR]

Simple Summary: Biological control of plant diseases caused by fungal pathogens using antagonistic microorganisms including Bacilli has been considered to be an effective and safe alternative to chemical fungicides. Fusarium crown rot of wheat is a serious fungal disease affecting yield and grain quality. In this study, a newly isolated strain of Bacillus subtilis YB-15 from soil of wheat rhizosphere significantly inhibited Fusarium crown rot as well as improved growth of wheat seedlings. Multiple potential biocontrol and growth-promoting attributes of Bacillus subtilis YB-15 were determined in vitro and according to the whole genome sequencing analysis. Overall, the results demonstrated that Bacillus subtilis YB-15 has great potential for practical application in controlling plant fungal diseases and improving plant growth. Fusarium crown rot caused by Fusarium pseudograminearum is one of the most devastating diseases of wheat worldwide causing major yield and economic losses. In this study, strain YB-15 was isolated from soil of wheat rhizosphere and classified as Bacillus subtilis by average nucleotide identity analysis. It significantly reduced Fusarium crown rot with a control efficacy of 81.50% and significantly improved the growth of wheat seedlings by increasing root and shoot fresh weight by 11.4% and 4.2%, respectively. Reduced Fusarium crown rot may have been due to direct antagonism by the production of β-1, 3-glucanase, amylase, protease and cellulase, or by the ability of B. subtilis YB-15 to induce defense-related enzyme activities of wheat seedlings, both alone and in seedlings infected with F. pseudograminearum. Improved plant growth may be related to the ability of B. subtilis YB-15 to secrete indole acetic acid and siderophores, as well as to solubilize phosphorus. In addition, the genome of strain YB-15 was determined, resulting in a complete assembled circular genome of 4,233,040 bp with GC content of 43.52% consisting of 4207 protein-encoding genes. Sequencing the B. subtilis YB-15 genome further revealed genes for encoding carbohydrate-active enzymes, biosynthesis of various secondary metabolites, nutrient acquisition, phytohormone production, chemotaxis and motility, which could explain the potential of strain YB-15 to be plant growth-promoting bacteria and biological control agent. B. subtilis YB-15 appears to be a promising biocontrol agent against Fusarium crown rot as well as for wheat growth promotion. [ABSTRACT FROM AUTHOR]

Fusarium pseudograminearum megabirnavirus 1 (FpgMBV1) was isolated from the hypovirulent strain FC136-2A of F. pseudograminearum as a novel double-stranded RNA mycovirus belonging to the family Megabirnaviridae. Here we examined the effects of FpgMBV1 on colony morphology and pathogenicity of F. pseudograminearum. Through hyphal tip culture, we obtained virus-free progeny of strain FC136-2A, referred to as FC136-2A-V–. FpgMBV1 was transferred horizontally to another virus-free strain, WZ-8A-HygR-V–. The progeny obtained through horizontal transfer was referred to as WZ-8A-HygR-V+. Colony morphology was similar between the FpgMBV1-positive and -negative strains. The ability to penetrate cellophane in vitro was lost, and pathogenicity on wheat plants was reduced significantly in the FpgMBV1-positive strains relative to the FpgMBV1-negative strains. Microscopic observations showed a 6-h delay in the formation of appressoria-like structures in FC136-2A relative to FC136-2A-V–. Mycelium extension was significantly longer in wheat coleoptiles infected by WZ-8A-HygR-V– than in that infected by WZ-8A-HygR-V+ at 12 and 20 h after inoculation (hai). In addition, expression of five genes that encode cell wall-degrading enzymes differed significantly between FpgMBV1-positive and -negative strains at 12 and 20 hai during early infection of wheat cells by conidia. This study provides evidence for the hypovirulence effect of FpgMBV1 on F. pseudograminearum and suggests that the underlying mechanism involves unsuccessful early infection and perhaps cell wall degradation. [ABSTRACT FROM AUTHOR]

Fusarium pseudograminearum causes crown rot in wheat. This study aimed to assess the effects of the bacterial strain QTH8 isolated from Cotinus coggygria rhizosphere soil against F. pseudograminearum. Bacterial strain QTH8 was identified as Bacillus halotolerans in accordance with the phenotypic traits and the phylogenetic analysis of 16S rDNA and gyrB gene sequence. Culture filtrates of bacterial strain QTH8 inhibited the mycelial growth of F. pseudograminearum and resulted in mycelial malformation such as tumor formation, protoplast condensation, and mycelial fracture. In addition, bacterial strain QTH8 also inhibited the mycelial growth of Hainesia lythri, Pestalotiopsis sp., Botrytis cinerea, Curvularia lunata, Phyllosticta theaefolia, Fusarium graminearum, Phytophthora nicotianae, and Sclerotinia sclerotiorum. The active compounds produced by bacterial strain QTH8 were resistant to pH, ultraviolet irradiation, and low temperature, and were relatively sensitive to high temperature. After 4 h exposure, culture filtrates of bacterial strain QTH8—when applied at 5%, 10%, 15%, 20%, 25%, and 30%—significantly reduced conidial germination of F. pseudograminearum. The coleoptile infection assay proved that bacterial strain QTH8 reduced the disease index of wheat crown rot. In vivo application of QTH8 to wheat seedlings decreased the disease index of wheat crown rot and increased root length, plant height, and fresh weight. Iturin, surfactin, and fengycin were detected in the culture extract of bacterial strain QTH8 by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Bacterial strain QTH8 was identified for the presence of the ituC, bacA, bmyB, spaS, srfAB, fend, and srfAA genes using the specific polymerase chain reaction primers. B. halotolerans QTH8 has a vital potential for the sustainable biocontrol of wheat crown rot. [ABSTRACT FROM AUTHOR]

Abstract Background Fusarium crown rot is major disease in wheat. However, the wheat defense mechanisms against this disease remain poorly understood. Results Using tandem mass tag (TMT) quantitative proteomics, we evaluated a disease-susceptible (UC1110) and a disease-tolerant (PI610750) wheat cultivar inoculated with Fusarium pseudograminearum WZ-8A. The morphological and physiological results showed that the average root diameter and malondialdehyde content in the roots of PI610750 decreased 3 days post-inoculation (dpi), while the average number of root tips increased. Root vigor was significantly increased in both cultivars, indicating that the morphological, physiological, and biochemical responses of the roots to disease differed between the two cultivars. TMT analysis showed that 366 differentially expressed proteins (DEPs) were identified by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment in the two comparison groups, UC1110_3dpi/UC1110_0dpi (163) and PI610750_3dpi/PI610750_0dpi (203). It may be concluded that phenylpropanoid biosynthesis (8), secondary metabolite biosynthesis (12), linolenic acid metabolites (5), glutathione metabolism (8), plant hormone signal transduction (3), MAPK signaling pathway-plant (4), and photosynthesis (12) contributed to the defense mechanisms in wheat. Protein-protein interaction network analysis showed that the DEPs interacted in both sugar metabolism and photosynthesis pathways. Sixteen genes were validated by real-time quantitative polymerase chain reaction and were found to be consistent with the proteomics data. Conclusion The results provided insight into the molecular mechanisms of the interaction between wheat and F. pseudograminearum.

Fusarium crown rot, caused by Fusarium pseudograminearum , is a devastating disease affecting the yield and quality of cereal crops. Peroxisomes are single-membrane organelles that play a critical role in various biological processes in eukaryotic cells. To functionally characterise peroxisome biosynthetic receptor proteins FpPEX5 and FpPEX7 in F. pseudograminearum , we constructed deletion mutants, Δ FpPEX5 and Δ FpPEX7 , and complementary strains, Δ FpPEX5-C and Δ FpPEX7-C , and analysed the functions of FpPEX5 and FpPEX7 proteins using various phenotypic observations. The deletion of FpPEX5 and FpPEX7 resulted in a significant deficiency in mycelial growth and conidiation and blocked the peroxisomal targeting signal 1 and peroxisomal targeting signal 2 pathways, which are involved in peroxisomal matrix protein transport, increasing the accumulation of lipid droplets and reactive oxygen species. The deletion of FpPEX5 and FpPEX7 may reduce the formation of toxigenic bodies and decrease the pathogenicity of F. pseudograminearum. These results indicate that FpPEX5 and FpPEX7 play vital roles in the growth, asexual reproduction, virulence, and fatty acid utilisation of F. pseudograminearum. This study provides a theoretical basis for controlling stem rot in wheat. [ABSTRACT FROM AUTHOR]

Sharp eyespot and Fusarium crown rot, mainly caused by soil-borne fungi Rhizoctonia cerealis and Fusarium pseudograminearum, are destructive diseases of major cereal crops including wheat (Triticum aestivum). However, the mechanisms underlying wheat-resistant responses to the two pathogens are largely elusive. In this study, we performed a genome-wide analysis of wall-associated kinase (WAK) family in wheat. As a result, a total of 140 TaWAK (not TaWAKL) candidate genes were identified from the wheat genome, each of which contains an N-terminal signal peptide, a galacturonan binding domain, an EGF-like domain, a calcium binding EGF domain (EGF-Ca), a transmembrane domain, and an intracellular Serine/Threonine protein kinase domain. By analyzing the RNA-sequencing data of wheat inoculated with R. cerealis and F. pseudograminearum, we found that transcript abundance of TaWAK-5D600 (TraesCS5D02G268600) on chromosome 5D was significantly upregulated, and that its upregulated transcript levels in response to both pathogens were higher compared with other TaWAK genes. Importantly, knock-down of TaWAK-5D600 transcript impaired wheat resistance against the fungal pathogens R. cerealis and F. pseudograminearum, and significantly repressed expression of defense-related genes in wheat, TaSERK1, TaMPK3, TaPR1, TaChitinase3, and TaChitinase4. Thus, this study proposes TaWAK-5D600 as a promising gene for improving wheat broad resistance to sharp eyespot and Fusarium crown rot (FCR) in wheat.

Fusarium pseudograminearum is a major pathogen for the destructive disease Fusarium crown rot (FCR) of wheat (Triticum aestivum). The cytosolic Acetoacetyl-CoA thiolase II (AACT) is the first catalytic enzyme in the mevalonate pathway that biosynthesizes isoprenoids in plants. However, there has been no investigation of wheat cytosolic AACT genes in defense against pathogens including Fusarium pseudograminearum. Herein, we identified a cytosolic AACT-encoding gene from wheat, named TaAACT1, and demonstrated its positively regulatory role in the wheat defense response to F. pseudograminearum. One haplotype of TaAACT1 in analyzed wheat genotypes was associated with wheat resistance to FCR. The TaAACT1 transcript level was elevated after F. pseudograminearum infection, and was higher in FCR-resistant wheat genotypes than in susceptible wheat genotypes. Functional analysis indicated that knock down of TaAACT1 impaired resistance against F. pseudograminearum and reduced the expression of downstream defense genes in wheat. TaAACT1 protein was verified to localize in the cytosol of wheat cells. TaAACT1 and its modulated defense genes were rapidly responsive to exogenous jasmonate treatment. Collectively, TaAACT1 contributes to resistance to F. pseudograminearum through upregulating the expression of defense genes in wheat. This study sheds new light on the molecular mechanisms underlying wheat defense against FCR.

Fusarium pseudograminearum is one of the most damaging Fusarium species that causes root, crown, and foot rots in wheat. Identification of resistant germplasm is one of the most efficient and environmentally sound control methods. However, up to date, limited wheat genotypes with partial resistance are available. Therefore, in this study, the seedling resistance reaction of 200 bread wheat genotypes plus 6 control genotypes obtained from CIMMYT to Fusarium pseudograminearum was determined under growth room conditions. Out of the 200 tested genotypes; 1 (0.5%), 35 (17.5%), 112 (56%), 45 (22.5%), and 7 (3.5%) were resistant, moderately resistant, moderately susceptible, susceptible and very susceptible to Fusarium pseudograminearum, respectively. Resistant and moderately resistant genotypes could be used in breeding studies for developing crown rot-resistant cultivars. [ABSTRACT FROM AUTHOR]

Wheat crown rot caused by Fusarium pseudograminearum has become an important soilborne disease and affects the quality and yield of wheat. In order to detect the sensitivity of F. pseudograminearum to fludioxonil in Henan Province, China, 105 isolates of F. pseudograminearum were collected from six cities in 2019. Sensitivity was determined by the mycelial growth rate method, then the methods of least-significant difference (LSD) and SPSS cluster were used for result analysis. The sensitivity of F. pseudograminearum to carbendazim and tebuconazole was determined and the correlation coefficient which existed between fludioxonil and the two fungicides, carbendazim and tebuconazole was analyzed. The results showed that the minimum inhibitory concentration (MIC) of fludioxonil was 0.240 0 μg/mL. The sensitivity frequency distribution was a continuous single peak curve, and the EC50 value ranged from 0.002 7 to 0.047 0 μg/mL. The average EC50 value of (0.026 3 ± 0.010 1) μg/mL, could be used as the sensitivity baseline of the pathogen to fludioxonil. The variance analysis results showed that the sensitivity of the different cities to fludioxonil was different, and the EC50 value ranged from 0.015 0 to 0.033 5 μg/mL. The maximum EC50 value of the isolate from Zhongmu County Zhengzhou City was 16.78 times bigger than the minimum value. Cluster analysis showed that there was no significant correlation between the sensitivity of isolates to fludioxonil and their geographical origin. The mean EC50 values of carbendazimand tebuconazole against the pathogens were (0.788 1 ± 0.315 3) μg/mL and (0.088 6 ± 0.145 3) μg/mL, respectively. There was no significant correlation between the sensitivity of the isolates to fludioxonil, carbendazim and tebuconazole. The results of greenhouse trials showed that the control efficacy of fludioxonil suspension seed coat agent could reach 58.00% (75.0 μg a.i/g) in 2020 and 63.69% (50.0 μg a.i/g) in 2021 when used to treat wheat as a seed dressing agent. The results of this study provide the basis for the rational use of fludioxonil in the control of wheat crown rot and provide information for monitoring the sensitivity of pathogenic fungi to fungicides. [ABSTRACT FROM AUTHOR]

Wheat stem rot caused by Fusarium pseudograminearum has become the main disease in Huang-Huai wheat area, which poses a great threat to the stable and high yield of wheat. In order to detect the sensitivity to carbendazim of F. pseudograminearum causing wheat stembase rot in Henan Province, China, 90 isolates of were collected from 8 counties in 2019, the inhibitory activities of these isolates were determined by mycelial growth rate method. The results were analyzed by variance analysis and cluster analysis, and the sensitivity of tebuconazole and fludioxonil was also tested, to analyze the correlation efficient existed between carbendazim and the two fungicides, tebuconazole and fludioxonil. The results showed that the minimum inhibitory concentration of carbendazim was 2.4 μg/mL, the EC50 values ranged from 0.436 to 1.73 μg/mL, the maximum value was 3.98 times the minimum value, and the average EC50 value was (0.750 ± 0.291) μg/mL. The sensitivity frequency distribution map showed that although there were subpopulations with low sensitivity to multiple bacteria, there were still 61 strains tested within the corresponding main peak range, and the sensitivity frequency distribution was a continuous single peak curve, which could be used as the sensitivity baseline of the pathogen to carbendazim. The results of variance analysis showed that the sensitivity of the different county to carbendazim was less different, and the average EC50 value in different cities ranged from 0.604 μg/mL to 1.04 μg/mL, and the lowest and highest strains were Hongqi and Huixian from Xinxiang, respectively, with a difference of 1.72 times. The susceptibility of the strains to carbendazim was significantly different in the same city, and the difference of EC50 value was 3.98 times in Nanyang Neixiang. Cluster analysis showed that there was no significant correlation between the sensitivity of F. pseudograminearum to carbendazem in Henan Province and their geographical origin. There was no significant correlation between the sensitivity of wheat stem rot pathogen to carbendazim and its sensitivity to tebuconazole and fludioxonil. The results of greenhouse control showed that 50% carbendazim wettable powder can be used for wheat seed dressing treatment and the highest control efficacy on wheat stem rot up to 76.66% when the treatment dosage was 3.90 mg/g. The results of this study laid a theoretical foundation for the chemical control of wheat stem rot with carbendazim and provide important information for the sensitivity monitoring of pathogenic bacteria to fungicides. [ABSTRACT FROM AUTHOR]

Abstract C2H2 zinc finger transcription factors such as FlbC and Msn2, have broad regulatory roles in fungal growth and conidiation. In the present study, we cloned and characterized a C2H2 zinc finger transcription factor gene, FpCzf14, in the wheat pathogen Fusarium pseudograminearum. FpCzf14 was localized to the nuclei. The expression of FpCzf14 was significantly upregulated in conidia, suggesting that FpCzf14 might contribute to conidiation. Further analysis of the fpczf14-deleted mutant (Δfpczf14) demonstrated that it exhibited defect in conidiation, and this defect was restored in the complemented strain Δfpczf14-C expressing FpCzf14, demonstrating that FpCzf14 was essential for conidiation. Moreover, FpCzf14 was required for mycelial growth and pathogenicity of F. pseudograminearum. Microscopic observation results showed that Δfpczf14 produced only very few penetration pegs and invasive hyphae inside host tissues compared with WT and Δfpczf14-C. Additionally, results of reverse transcription quantitative PCR (RT-qPCR) showed that FpCzf14 regulated expression of several conidiation-related genes in F. pseudograminearum. In conclusion, FpCzf14, as a core regulatory gene in conidiation, provides new insights into the mechanism of conidiation in F. pseudograminearum.

The cereal disease Fusarium crown rot (FCR), caused by the fungal pathogen Fusarium pseudograminearum (Fp), is a major constraint to cereal production worldwide. Nitrogen (N) fertilizer is estimated to be approximately 30% of the input costs for grain growers in Australia and is the primary driver of yield and grain protein levels. When targeting high yield and protein, generous nitrogen fertilizer applications are thought to result in large biomass production, which exacerbates FCR severity, reducing grain yield and quality. This research was undertaken to investigate the effect of temporal N availability in high-protein bread and durum wheat varieties on FCR severity. Laboratory and controlled environment experiments assessed the relationship between FCR and N at a mechanistic and plant level. An in vitro study demonstrated an increase in Fp mycelial growth under increased N availability, especially when N was supplied as urea compared with ammonium nitrate. Similarly, under controlled environmental conditions, increased soil N availability promoted FCR severity within infected plants. Stem N transfer efficiency was significantly decreased under FCR infection in both bread and durum wheat varieties by 4.5% and 10.2%, respectively. This new research demonstrates that FCR not only decreases yield and grain quality but appears to have previously unrecognised detrimental impacts on nitrogen-use efficiency in wheat. This indicates that the current impact of losses from FCR may also decrease N-use inefficiencies, as well as yield and quality penalties. An improved understanding of the interactions and restrictions of FCR infection may allow growers to better manage the disease through manipulation of the soil’s temporal N availability.

Fusarium crown rot (FCR) is an important and devastating disease of wheat ( Triticum aestivum ) caused by the fungus Fusarium pseudograminearum and related pathogens. Using two distinct susceptible cultivars, we investigated the isolation frequencies of F. pseudograminearum and quantified its biomass accumulation and the levels of the associated toxins deoxynivalenol (DON) and DON-3-glucoside (D3G) in inoculated field-grown wheat plants. We detected F. pseudograminearum in stem, peduncle, rachis, and husk tissues, but not in grains, whereas DON and D3G accumulated in stem, rachis, husk, and grain tissues. Disease severity was positively correlated with the frequency of pathogen isolation, F. pseudograminearum biomass, and mycotoxin levels. The amount of F. pseudograminearum biomass and mycotoxin contents in asymptomatic tissue of diseased plants were associated with the distance of the tissue from the diseased internode and the disease severity of the plant. Thus, apparently healthy tissue may harbor F. pseudograminearum and contain associated mycotoxins. This research helps clarify the relationship between F. pseudograminearum occurrence, F. pseudograminearum biomass, and mycotoxin accumulation in tissues of susceptible wheat cultivars with or without disease symptoms, providing information that can lead to more effective control measures., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Xu, Shi, Liu, Li, Wang, Han, Liu, Wang, Liu, Fan, Wang, Feng, Song, Zhou and Xu.)

Fusarium pseudograminearum is an important plant pathogen that invades many crops (Zhang et al. 2018). Since it was first discovered in Australia in 1951, F. pseudograminearum has been reported in many countries and regions and caused huge economic losses (Burgess et al. 2001). In 2012, crown rot of wheat caused by F. pseudograminearum was discovered for the first time in Henan Province, China (Li et al. 2012). Wheat (Triticum aestivum L.) is one of the most important food crops in Xinjiang Uygur Autonomous Region (XUAR), with 1.07 million hectares cultivated in 2020. In June 2023, a survey of crown rot disease was carried out in winter wheat cv. Xindong 20 in Hotan area, XUAR, China (80.148907°E, 37.051474°N). About 5% of wheat plants showed symptoms of crown rot such as browning of the stem base and white head. The disease was observed in 85% of wheat fields. In order to identify the pathogens, 36 pieces of diseased stem basal tissue, 0.5 cm in length, were collected and sterilized with 75% alcohol for 30s and 5% NaOCl solution for 2 min, then rinsed three times with sterile water and placed on potato dextrose agar (PDA) medium at 25°C. A total of 27 isolates with consistent morphological characteristics were obtained using single-spore technique (Leslie and Summerell. 2006), and the isolation rate was 75%. The isolates grew rapidly on PDA, produced large numbers of fluffy white hyphae, and pink pigment accumulated in the medium. The isolates were grown on 2% mung bean flour medium and identified by morphological and molecular methods. Macroconidia were abundant, relatively slender, curved to almost straight, commonly two to seven septate, and averaged 22 to 72 × 1.8 to 4.9 μm. Microconidia were not observed. The morphological characters are consistent with Fusarium (Aoki and O'Donnell. 1999). Two isolates (LP-1 and LP-3) were selected for molecular identification. Primers EF1/EF2 (5'-ATGGGTAAGGARGACAAGAC-3'/5'-GGARGTACCAGTSATCATG-3') were used to amplify a portion of the EF-1α gene (O'Donnell et al. 1998). The two 696 bp PCR products were sequenced and submitted to GenBank. The EF-1α gene sequences (GenBank Accession No: PP062794 and PP062795) shared 99.9% identity (695/696) with published F.pseudograminearum sequences (e.g., OP105187, OP105184, OP105179, OP105173). The identification was further confirmed by F. pseudograminearum species-specific PCR primers Fp1-1/Fp1-2 (Aoki and O'Donnell. 1999). The expected PCR products of 518 bp were produced only in F. pseudograminearum. Pathogenicity tests of LP-1 and LP-3 isolates were performed on 7-day-old seedlings of winter wheat cv. Xindong 20 using the drip inoculation method with a 10-μl of a 106 macroconidia ml-1 suspension near the stem base (Xu et al. 2017). The experiment was repeated five times in a 20 to 25°C greenhouse. Control seedlings were treated with sterile water. After 4 weeks, wheat seedling death and crown browning occurred in the inoculated plants with over 90% incidence. No symptoms were observed in the control plants. The pathogen was reisolated from the inoculated plants by the method described above and identified by morphological and PCR amplification using F. pseudograminearum species-specific primers Fp1-1/Fp1-2. No F. pseudograminearum was isolated from the control plants, fulfilling Koch's postulates. To our knowledge, this is the first report of F. pseudograminearum causing crown rot of winter wheat in XUAR of China. Since F. pseudograminearum can cause great damage to wheat, one of the most important food crops in China, necessary measures should be taken to prevent the spread of F. pseudograminearum to other regions.

The soil-borne fungi Fusarium pseudograminearum and Rhizoctonia cerealis are the major pathogens for the economically important diseases Fusarium crown rot (FCR) and sharp eyespot of common wheat (Triticum aestivum), respectively. However, there has been no report on the broad resistance of wheat genes against both F. pseudograminearum and R. cerealis. In the current study, we identified TaWAK-6D , a wall-associated kinase (WAK) which is an encoding gene located on chromosome 6D, and demonstrated its broad resistance role in the wheat responses to both F. pseudograminearum and R. cerealis infection. TaWAK-6D transcript induction by F. pseudograminearum and R. cerealis was related to the resistance degree of wheat and the gene expression was significantly induced by exogenous pectin treatment. Silencing of TaWAK-6D compromised wheat resistance to F. pseudograminearum and R. cerealis , and repressed the expression of a serial of wheat defense-related genes. Ectopic expression of TaWAK-6D in Nicotiana benthamiana positively modulated the expression of several defense-related genes. TaWAK-6D protein was determined to localize to the plasma membrane in wheat and N. benthamiana. Collectively, the TaWAK-6D at the plasma membrane mediated the broad resistance responses to both F. pseudograminearum and R. cerealis in wheat at the seedling stage. This study, therefore, concludes that TaWAK-6D is a promising gene for improving wheat broad resistance to FCR and sharp eyespot. [ABSTRACT FROM AUTHOR]

The soil-borne fungi Fusarium pseudograminearum and Rhizoctonia cerealis are the major pathogens for the economically important diseases Fusarium crown rot (FCR) and sharp eyespot of common wheat (Triticum aestivum), respectively. However, there has been no report on the broad resistance of wheat genes against both F. pseudograminearum and R. cerealis. In the current study, we identified TaWAK-6D, a wall-associated kinase (WAK) which is an encoding gene located on chromosome 6D, and demonstrated its broad resistance role in the wheat responses to both F. pseudograminearum and R. cerealis infection. TaWAK-6D transcript induction by F. pseudograminearum and R. cerealis was related to the resistance degree of wheat and the gene expression was significantly induced by exogenous pectin treatment. Silencing of TaWAK-6D compromised wheat resistance to F. pseudograminearum and R. cerealis, and repressed the expression of a serial of wheat defense-related genes. Ectopic expression of TaWAK-6D in Nicotiana benthamiana positively modulated the expression of several defense-related genes. TaWAK-6D protein was determined to localize to the plasma membrane in wheat and N. benthamiana. Collectively, the TaWAK-6D at the plasma membrane mediated the broad resistance responses to both F. pseudograminearum and R. cerealis in wheat at the seedling stage. This study, therefore, concludes that TaWAK-6D is a promising gene for improving wheat broad resistance to FCR and sharp eyespot.

Fusarium crown rot (FCR) of wheat, an important soil-borne disease, presents a worsening trend year by year, posing a significant threat to wheat production. Fusarium pseudograminearum cv. b was reported to be the dominant pathogen of FCR in China. Peroxisomes are single-membrane organelles in eukaryotes that are involved in many important biochemical metabolic processes, including fatty acid β-oxidation. PEX11 is important proteins in peroxisome proliferation, while less is known in the fungus F. pseudograminearum. The functions of FpPEX11a, FpPEX11b, and FpPEX11c in F. pseudograminearum were studied using reverse genetics, and the results showed that FpPEX11a and FpPEX11b are involved in the regulation of vegetative growth and asexual reproduction. After deleting FpPEX11a and FpPEX11b, cell wall integrity was impaired, cellular metabolism processes including active oxygen metabolism and fatty acid β-oxidation were significantly blocked, and the production ability of deoxynivalenol (DON) decreased. In addition, the deletion of genes of FpPEX11a and FpPEX11b revealed a strongly decreased expression level of peroxisome-proliferation-associated genes and DON-synthesis-related genes. However, deletion of FpPEX11c did not significantly affect these metabolic processes. Deletion of the three protein-coding genes resulted in reduced pathogenicity of F. pseudograminearum. In summary, FpPEX11a and FpPEX11b play crucial roles in the growth and development, asexual reproduction, pathogenicity, active oxygen accumulation, and fatty acid utilization in F. pseudograminearum.

Heat shock protein Hsp104, a homolog of the bacterial chaperone ClpB and plant Hsp100, plays an essential part in the response to heat and various chemical agents in Saccharomyces cerevisiae. However, their functions remain largely unknown in plant fungal pathogens. Here, we report the identification and functional characterization of a plausible ortholog of yeast Hsp104 in Fusarium pseudograminearum, which we termed FpHsp104. Deletion mutant of FpHsp104 displayed severe defects in the resistance of heat shock during F. pseudograminearum mycelia and conidia when exposed to extreme heat. We also found that the protein showed dynamic localization to small particles under high temperature. However, no significant differences were detected in osmotic, oxidative, or cell wall stress responses between the wild-type and Δfphsp104 strains. Quantitative real-time PCR analysis showed that FpHsp104 was upregulated in the conidia, and disruption of FpHsp104 gene resulted in defects in conidia production, morphology, and germination. The transcript levels of conidiation-related genes of FpFluG, FpVosA, FpWetA, and FpAbaA were reduced in the Δfphsp104 mutant vs. the wild-type strain, but heat-shocked mRNA splicing repair was not affected in Δfphsp104. Moreover, Δfphsp104 mutant also showed attenuated virulence, but its DON synthesis was normal. These data from the first study of Hsp104 in F. pseudograminearum strongly suggest that FpHsp104 gene is an important element in the heat tolerance, development, and pathogenicity processes of F. pseudograminearum.

Heat shock protein Hsp104, a homolog of the bacterial chaperone ClpB and plant Hsp100, plays an essential part in the response to heat and various chemical agents in Saccharomyces cerevisiae. However, their functions remain largely unknown in plant fungal pathogens. Here, we report the identification and functional characterization of a plausible ortholog of yeast Hsp104 in Fusarium pseudograminearum , which we termed FpHsp104. Deletion mutant of FpHsp104 displayed severe defects in the resistance of heat shock during F. pseudograminearum mycelia and conidia when exposed to extreme heat. We also found that the protein showed dynamic localization to small particles under high temperature. However, no significant differences were detected in osmotic, oxidative, or cell wall stress responses between the wild-type and Δ fphsp104 strains. Quantitative real-time PCR analysis showed that FpHsp104 was upregulated in the conidia, and disruption of FpHsp104 gene resulted in defects in conidia production, morphology, and germination. The transcript levels of conidiation-related genes of FpFluG , FpVosA , FpWetA , and FpAbaA were reduced in the Δ fphsp104 mutant vs. the wild-type strain, but heat-shocked mRNA splicing repair was not affected in Δ fphsp104. Moreover, Δ fphsp104 mutant also showed attenuated virulence, but its DON synthesis was normal. These data from the first study of Hsp104 in F. pseudograminearum strongly suggest that FpHsp104 gene is an important element in the heat tolerance, development, and pathogenicity processes of F. pseudograminearum. [ABSTRACT FROM AUTHOR]

Ipconazole is a broad-spectrum triazole fungicide that is highly effective against Fusarium pseudograminearum. However, its risk of developing resistance and mechanism are not well understood in F. pseudograminearum. Here, the sensitivities of 101 F. pseudograminearum isolates to ipconazole were investigated, and the average EC 50 value was 0.1072 μg/mL. Seven mutants resistant to ipconazole were obtained by fungicide adaption, with all but one showing reduced fitness relative to the parental isolates. Cross-resistance was found between ipconazole and mefentrifluconazole and tebuconazole, but none between ipconazole and pydiflumetofen, carbendazim, fludioxonil, or phenamacril. In summary, these findings suggest that there is a low risk of F. pseudograminearum developing resistance to ipconazole. Additionally, a point mutation, G464S, was seen in FpCYP51B and overexpression of FpCYP51A , FpCYP51B and FpCYP51C was observed in ipconazole-resistant mutants. Assays, including transformation and molecular docking, indicated that G464S conferred ipconazole resistance in F. pseudograminearum. [Display omitted] • The baseline sensitivity of F. pseudograminearum to ipconazole was established. • F. pseudograminearum demonstrated a low ipconazole resistance risk. • The mutation G464S in FpCYP51B and overexpression of FpCYP51s could confer ipconazole resistance in F. pseudograminearum. [ABSTRACT FROM AUTHOR]

Fusarium crown rot caused by Fusarium pseudograminearum is one of the most devastating diseases of wheat worldwide causing major yield and economic losses. In this study, strain YB-15 was isolated from soil of wheat rhizosphere and classified as Bacillus subtilis by average nucleotide identity analysis. It significantly reduced Fusarium crown rot with a control efficacy of 81.50% and significantly improved the growth of wheat seedlings by increasing root and shoot fresh weight by 11.4% and 4.2%, respectively. Reduced Fusarium crown rot may have been due to direct antagonism by the production of β-1, 3-glucanase, amylase, protease and cellulase, or by the ability of B. subtilis YB-15 to induce defense-related enzyme activities of wheat seedlings, both alone and in seedlings infected with F. pseudograminearum. Improved plant growth may be related to the ability of B. subtilis YB-15 to secrete indole acetic acid and siderophores, as well as to solubilize phosphorus. In addition, the genome of strain YB-15 was determined, resulting in a complete assembled circular genome of 4,233,040 bp with GC content of 43.52% consisting of 4207 protein-encoding genes. Sequencing the B. subtilis YB-15 genome further revealed genes for encoding carbohydrate-active enzymes, biosynthesis of various secondary metabolites, nutrient acquisition, phytohormone production, chemotaxis and motility, which could explain the potential of strain YB-15 to be plant growth-promoting bacteria and biological control agent. B. subtilis YB-15 appears to be a promising biocontrol agent against Fusarium crown rot as well as for wheat growth promotion.

Fusarium pseudograminearum causes crown rot in wheat. This study aimed to assess the effects of the bacterial strain QTH8 isolated from Cotinus coggygria rhizosphere soil against F. pseudograminearum. Bacterial strain QTH8 was identified as Bacillus halotolerans in accordance with the phenotypic traits and the phylogenetic analysis of 16S rDNA and gyrB gene sequence. Culture filtrates of bacterial strain QTH8 inhibited the mycelial growth of F. pseudograminearum and resulted in mycelial malformation such as tumor formation, protoplast condensation, and mycelial fracture. In addition, bacterial strain QTH8 also inhibited the mycelial growth of Hainesia lythri, Pestalotiopsis sp., Botrytis cinerea, Curvularia lunata, Phyllosticta theaefolia, Fusarium graminearum, Phytophthora nicotianae, and Sclerotinia sclerotiorum. The active compounds produced by bacterial strain QTH8 were resistant to pH, ultraviolet irradiation, and low temperature, and were relatively sensitive to high temperature. After 4 h exposure, culture filtrates of bacterial strain QTH8—when applied at 5%, 10%, 15%, 20%, 25%, and 30%—significantly reduced conidial germination of F. pseudograminearum. The coleoptile infection assay proved that bacterial strain QTH8 reduced the disease index of wheat crown rot. In vivo application of QTH8 to wheat seedlings decreased the disease index of wheat crown rot and increased root length, plant height, and fresh weight. Iturin, surfactin, and fengycin were detected in the culture extract of bacterial strain QTH8 by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Bacterial strain QTH8 was identified for the presence of the ituC, bacA, bmyB, spaS, srfAB, fend, and srfAA genes using the specific polymerase chain reaction primers. B. halotolerans QTH8 has a vital potential for the sustainable biocontrol of wheat crown rot.

Fusarium crown rot (FCR) caused by the fungus Fusarium pseudograminearum (Fp) is an important disease of wheat that reduces yield and grain quality in many countries, including Australia. In this study, we investigated mating type idiomorph composition, putative chemotype and population genotypic structure of 98 Fp isolates from Western Australia (WA) and the eastern Australian states of New South Wales, Victoria and South Australia. Our results revealed the expected 1:1 mating type ratio for isolates from the eastern states while there was significant variation to a 1:1 mating type composition with isolates from WA. A chemotype-specific PCR assay indicated that all Fp isolates from eastern states and WA segregated for the 3-ADON trichothecene chemotype. Genetic diversity assessed using 21 cleaved amplified polymorphic sequence markers revealed a high level of genotypic variation within and between Fp populations from eastern Australian states and WA. Analysis of molecular variance (AMOVA) showed significant difference between populations from eastern states and WA. The genetic diversity measured by Shannon index ranged from 0.95 to 2.30 with the lowest and highest values detected in Hart and Rowena populations, respectively in the eastern states. Index of association analysis showed no significant linkage of markers among isolates within 60% of the populations, suggesting that sexual reproduction may be occurring in the pathogen from those locations. These results improve understanding of Fp population dynamics across Australia and highlight the importance of monitoring for shifts in the population which could have implications for disease management. [ABSTRACT FROM AUTHOR]

Summary: An emerging experimental framework suggests that plants under biotic stress may actively seek help from soil microbes, but empirical evidence underlying such a 'cry for help' strategy is limited.We used integrated microbial community profiling, pathogen and plant transcriptive gene quantification and culture‐based methods to systematically investigate a three‐way interaction between the wheat plant, wheat‐associated microbiomes and Fusarium pseudograminearum (Fp).A clear enrichment of a dominant bacterium, Stenotrophomonas rhizophila (SR80), was observed in both the rhizosphere and root endosphere of Fp‐infected wheat. SR80 reached 3.7 × 107 cells g−1 in the rhizosphere and accounted for up to 11.4% of the microbes in the root endosphere. Its abundance had a positive linear correlation with the pathogen load at base stems and expression of multiple defence genes in top leaves. Upon re‐introduction in soils, SR80 enhanced plant growth, both the below‐ground and above‐ground, and induced strong disease resistance by boosting plant defence in the above‐ground plant parts, but only when the pathogen was present.Together, the bacterium SR80 seems to have acted as an early warning system for plant defence. This work provides novel evidence for the potential protection of plants against pathogens by an enriched beneficial microbe via modulation of the plant immune system. [ABSTRACT FROM AUTHOR]

Background: Fusarium crown rot is major disease in wheat. However, the wheat defense mechanisms against this disease remain poorly understood. Results: Using tandem mass tag (TMT) quantitative proteomics, we evaluated a disease-susceptible (UC1110) and a disease-tolerant (PI610750) wheat cultivar inoculated with Fusarium pseudograminearum WZ-8A. The morphological and physiological results showed that the average root diameter and malondialdehyde content in the roots of PI610750 decreased 3 days post-inoculation (dpi), while the average number of root tips increased. Root vigor was significantly increased in both cultivars, indicating that the morphological, physiological, and biochemical responses of the roots to disease differed between the two cultivars. TMT analysis showed that 366 differentially expressed proteins (DEPs) were identified by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment in the two comparison groups, UC1110_3dpi/UC1110_0dpi (163) and PI610750_3dpi/PI610750_0dpi (203). It may be concluded that phenylpropanoid biosynthesis (8), secondary metabolite biosynthesis (12), linolenic acid metabolites (5), glutathione metabolism (8), plant hormone signal transduction (3), MAPK signaling pathway-plant (4), and photosynthesis (12) contributed to the defense mechanisms in wheat. Protein-protein interaction network analysis showed that the DEPs interacted in both sugar metabolism and photosynthesis pathways. Sixteen genes were validated by real-time quantitative polymerase chain reaction and were found to be consistent with the proteomics data. Conclusion: The results provided insight into the molecular mechanisms of the interaction between wheat and F. pseudograminearum. [ABSTRACT FROM AUTHOR]

Fusarium crown rot (FCR) and Fusarium head blight (FHB) are caused by Fusarium pseudograminearum and are newly emerging diseases of wheat in China. In this study, we characterized FpPPR1, a gene that encodes a protein with 12 pentatricopeptide repeat (PPR) motifs. The radial growth rate of the ΔFpppr1 deletion mutant was significantly slower than the wild type strain WZ-8A on potato dextrose agar plates and exhibited significantly smaller colonies with sector mutations. The aerial mycelium of the mutant was almost absent in culture tubes. The ΔFpppr1 mutant was able to produce spores, but spores of abnormal size and altered conidium septum shape were produced with a significant reduction in sporulation compared to wild type. ΔFpppr1 failed to cause disease on wheat coleoptiles and barley leaves using mycelia plugs or spore suspensions. The mutant phenotypes were successfully restored to the wild type levels in complemented strains. FpPpr1-GFP signals in spores and mycelia predominantly overlapped with Mito-tracker signals, which substantiated the mitochondria targeting signal prediction of FpPpr1. RNAseq revealed significant transcriptional changes in the ΔFpppr1 mutant with 1,367 genes down-regulated and 1,333 genes up-regulated. NAD-binding proteins, thioredoxin, 2Fe-2S iron-sulfur cluster binding domain proteins, and cytochrome P450 genes were significantly down-regulated in ΔFpppr1, implying the dysfunction of mitochondria-mediated reductase redox stress in the mutant. The mating type idiomorphic alleles MAT1-1-1, MAT1-1-2, and MAT1-1-3 in F. pseudograminearum were also down-regulated after deletion of FpPPR1 and validated by real-time quantitative PCR. Additionally, 21 genes encoding putative heterokaryon incompatibility proteins were down-regulated. The yellow pigmentation of the mutant was correlated with reduced expression of PKS12 cluster genes. Taken together, our findings on FpPpr1 indicate that this PPR protein has multiple functions in fungal asexual development, regulation of heterokaryon formation, mating-type, and pathogenesis in F. pseudograminearum.

Guanosine 3′,5′-cyclic monophosphate (cGMP) is an important signaling molecule in plants. cGMP and guanylyl cyclases (GCs), enzymes that catalyze the synthesis of cGMP from GTP, are involved in several physiological processes and responses to environmental factors, including pathogen infections. Using in vitro analysis, we demonstrated that recombinant BdGUCD1 is a protein with high guanylyl cyclase activity and lower adenylyl cyclase activity. In Brachypodium distachyon, infection by Fusarium pseudograminearum leads to changes in BdGUCD1 mRNA levels, as well as differences in endogenous cGMP levels. These observed changes may be related to alarm reactions induced by pathogen infection. As fluctuations in stress phytohormones after infection have been previously described, we performed experiments to determine the relationship between cyclic nucleotides and phytohormones. The results revealed that inhibition of cellular cGMP changes disrupts stress phytohormone content and responses to pathogen. The observations made here allow us to conclude that cGMP is an important element involved in the processes triggered as a result of infection and changes in its levels affect jasmonic acid. Therefore, stimuli-induced transient elevation of cGMP in plants may play beneficial roles in priming an optimized response, likely by triggering the mechanisms of feedback control.

Fusarium crown rot (FCR) and Fusarium head blight (FHB) are caused by Fusarium pseudograminearum and are newly emerging diseases of wheat in China. In this study, we characterized FpPPR1 , a gene that encodes a protein with 12 pentatricopeptide repeat (PPR) motifs. The radial growth rate of the Δ Fpppr1 deletion mutant was significantly slower than the wild type strain WZ-8A on potato dextrose agar plates and exhibited significantly smaller colonies with sector mutations. The aerial mycelium of the mutant was almost absent in culture tubes. The Δ Fpppr1 mutant was able to produce spores, but spores of abnormal size and altered conidium septum shape were produced with a significant reduction in sporulation compared to wild type. Δ Fpppr1 failed to cause disease on wheat coleoptiles and barley leaves using mycelia plugs or spore suspensions. The mutant phenotypes were successfully restored to the wild type levels in complemented strains. FpPpr1-GFP signals in spores and mycelia predominantly overlapped with Mito-tracker signals, which substantiated the mitochondria targeting signal prediction of FpPpr1. RNAseq revealed significant transcriptional changes in the Δ Fpppr1 mutant with 1,367 genes down-regulated and 1,333 genes up-regulated. NAD-binding proteins, thioredoxin, 2Fe-2S iron-sulfur cluster binding domain proteins, and cytochrome P450 genes were significantly down-regulated in Δ Fpppr1 , implying the dysfunction of mitochondria-mediated reductase redox stress in the mutant. The mating type idiomorphic alleles MAT1-1-1, MAT1-1-2 , and MAT1-1-3 in F. pseudograminearum were also down-regulated after deletion of FpPPR1 and validated by real-time quantitative PCR. Additionally, 21 genes encoding putative heterokaryon incompatibility proteins were down-regulated. The yellow pigmentation of the mutant was correlated with reduced expression of PKS12 cluster genes. Taken together, our findings on FpPpr1 indicate that this PPR protein has multiple functions in fungal asexual development, regulation of heterokaryon formation, mating-type, and pathogenesis in F. pseudograminearum. [ABSTRACT FROM AUTHOR]

The fungus F. pseudograminearum can cause the destructive disease Fusarium crown rot (FCR) of wheat, an important staple crop. Functional roles of FCR resistance genes in wheat are largely unknown. In the current research, we characterized the antifungal activity and positive-regulatory function of the cysteine-rich repeat receptor-like kinase TaCRK-7A in the defense against F. pseudograminearum in wheat. Antifungal assays showed that the purified TaCRK-7A protein inhibited the growth of F. pseudograminearum. TaCRK-7A transcript abundance was elevated after F. pseudograminearum attack and was positively related to FCR-resistance levels of wheat cultivars. Intriguingly, knocking down of TaCRK-7A transcript increased susceptibility of wheat to FCR and decreased transcript levels of defense-marker genes in wheat. Furthermore, the transcript abundances of TaCRK-7A and its modulated-defense genes were responsive to exogenous jasmonate treatment. Taken together, these results suggest that TaCRK-7A can directly inhibit F. pseudograminearum growth and mediates FCR-resistance by promoting the expression of wheat defense genes in the jasmonate pathway. Thus, TaCRK-7A is a potential gene resource in FCR-resistant wheat breeding program.

Fusarium pseudograminearum is an important pathogen of Fusarium crown rot and Fusarium head blight, which is able to infect wheat and barley worldwide, causing great economic losses. Transcription factors (TFs) of the basic leucine zipper (bZIP) protein family control important processes in all eukaryotes. In this study, we identified a gene, designated FpAda1, encoding a bZIP TF in F. pseudograminearum. The homolog of FpAda1 is also known to affect hyphal growth in Neurospora crassa. Deletion of FpAda1 in F. pseudograminearum resulted in defects in hyphal growth, mycelial branching and conidia formation. Pathogenicity assays showed that virulence of the Δfpada1 mutant was dramatically decreased on wheat coleoptiles and barley leaves. However, wheat coleoptile inoculation assay showed that Δfpada1 could penetrate and proliferate in wheat cells. Moreover, the FpAda1 was required for abnormal nuclear morphology in conidia and transcription of FpCdc2 and FpCdc42. Taken together, these results indicate that FpAda1 is an important transcription factor involved in growth and development in F. pseudograminearum. [ABSTRACT FROM AUTHOR]

Fusarium pseudograminearum-induced crown rot causes significant reduction to wheat production worldwide. To date, efforts to develop effective resistance to this disease have been hampered by the quantitative nature of resistance trait and a lack of understanding of the molecular pathogenesis. Non-ribosomal peptides have important roles in development, pathogenicity, and toxins in many plant pathogens, while less is known in F. pseudograminearum. In this work, we studied the expression and function of a nonribosomal peptide gene FpNPS9 in F. pseudograminearum. We determined the expression of FpNPS9 which was significantly up regulated during the infection of wheat. A deletion mutant Δfpnps9 produced in this study displayed a normal growth and conidiation phenotype, however, hyphae polar growth was obviously affected. Deoxynivalenol production in this mutant was significantly reduced and the infection of wheat coleoptiles and wheat spikelet was attenuated. The Δfpnps9 showed serious defects on the extension of infectious hyphae in plant and inhibition of roots elongation compared with the wild type. The complementation assay using a FpNPS9-GFP fusion construct fully restored the defects of the mutant. GFP signal was detected in the germinating conidia and infectious hyphae in coleoptiles of the infected plants. Interestingly, the signal was not observed when it was grown on culture medium, suggesting that the expression of FpNPS9 was regulated by an unknown host factor. This observation was supported by the result of qRT-PCR. In summary, we provided new knowledge on FpNPS9 expression in F. pseudograminearum and its function in F. pseudograminearum pathogenicity in wheat. [ABSTRACT FROM AUTHOR]

Fusarium crown rot of wheat has been spreading in the Huanghuai wheat‐growing area in China since 2010, leading to a potential yield loss. To investigate the pathogens associated with this disease in Jiangsu and Shandong provinces in recent years, 617 Fusarium isolates were isolated from nine sites in these two provinces between 2014 and 2016. Of these isolates, 372 were identified as Fusarium pseudograminearum, and the remaining isolates were identified as F. asiaticum and F. graminearum, suggesting that F. pseudograminearum is becoming a predominant causative pathogen of crown rot of wheat in eastern China. Trichothecene gene detection and chemical analyses of trichothecenes indicated that the F. pseudograminearum isolates belonged to the 3‐ADON or 15‐ADON chemotype, and one isolate had the NIV genotype but produced no detectable NIV. 3‐ADON isolates were predominant in Jiangsu, whereas 15‐ADON isolates were prevalent in Shandong. The mating type of the F. pseudograminearum isolates were identified. MAT‐1 and MAT‐2 existed, but in most collections, particularly those in Jiangsu, the ratios of the two mating types deviated significantly from an expected 1:1 ratio. The reason for the occurrence of F. pseudograminearum is hypothesized, and the chemotype and mating type distribution of this species in these two provinces are analysed. [ABSTRACT FROM AUTHOR]

Wheat crown rot caused by Fusarium spp. is a common disease worldwide. Both Fusarium pseudograminearum and Fusarium graminearum infect wheat crown and produce mycotoxin leading to grain loss due to white head. F. pseudograminearum (Fp) was reported in wheat from Henan Province of China a couple of years ago. The wheat crown rot (CR) caused by this new pathogen is as an emerging severe disease of wheat, which has recently expanded to several provinces in China and is, therefore, under rapid investigation. Colonization of wheat tissue by Fp is accomplished though the formation of a septated foot-shaped appressoria and generation of a penetration peg to break through the internal cells of leaf sheath. The molecular mechanism by which Fp regulates the pathogenesis on wheat host is unclear. Here, we report FpPDE1, a P-type ATPase-encoding predicted PDE1 orthologue gene of Magnaporthe oryzae, belonging to the DRS2 subfamily of aminophospholipid translocases. The gene deletion of FpPDE1 with the split-marker approach did not obviously affect hyphae growth and conidiation, but led to an attenuated virulence on wheat base stem and root. Our finding indicates that the putative aminophospholipid translocases is not essential for the infectious hyphae development in Fp.

Heat shock protein 70s (Hsp70s) are a class of molecular chaperones that are highly conserved and ubiquitous in organisms ranging from microorganisms to plants and humans. Hsp70s play key roles in cellular development and protecting living organisms from environmental stresses such as heat, drought, salinity, acidity, and cold. However, their functions in pathogenic fungi are largely unknown. Here, a total of 14 FpHsp70 genes were identified in Fusarium pseudograminearum, including 3 in the mitochondria, 7 in the cytoplasm, 2 in the endoplasmic reticulum (ER), 1 in the nucleus, and 1 in the plastid. However, the exon–intron boundaries and protein motifs of the FpHsp70 have no consistency in the same subfamily. Expression analysis revealed that most FpHsp70 genes were up-regulated during infection, implying that FpHsp70 genes may play important roles in F. pseudograminearum pathogenicity. Furthermore, knockout of an ER lumenal Hsp70 homolog FpLhs1 gene reduced growth, conidiation, and pathogenicity in F. pseudograminearum. These mutants also showed a defect in secretion of some proteins. Together, FpHsp70s might play essential roles in F. pseudograminearum and FpLhs1 is likely to act on the development and virulence by regulating protein secretion.

Mefentrifluconazole, a triazole fungicide, exhibits remarkable efficacy in combating Fusarium spp. The mean EC 50 value of mefentrifluconazole against 124 isolates of Fusarium pseudograminearum was determined to be 1.06 μg/mL in this study. Fungicide taming produced five mefentrifluconazole-resistant mutants with resistance factors ranging from 19.21 to 111.34. Compared to the original parental isolates, the fitness of three resistant mutants was much lower, while the remaining two mutants displayed enhanced survival fitness. There was evidence of positive cross-resistance between tebuconazole and mefentrifluconazole. Mefentrifluconazole resistance in F. pseudograminearum can be conferred by FpCYP51B L144F , which was identified in four mutants according to molecular docking and site-directed transformation experiments. Overexpression of FpCYP51s was also detected in the resistant mutants. In conclusion, mefentrifluconazole has a low-to-medium resistance risk in F. pseudograminearum , and the L144F mutation in FpCYP51B and the increased expression level of FpCYP51s may be responsible for mefentrifluconazole resistance in F. pseudograminearum .

Fusarium crown rot (FCR) caused by Fusarium pseudograminearum is a serious threat to wheat production worldwide. This study aimed to assess the effects of Talaromyces muroii strain TM28 isolated from root of Panax quinquefolius against F. pseudograminearum . The strain of TM28 inhibited mycelial growth of F. pseudograminearum by 87.8% at 72 h, its cell free fermentation filtrate had a strong antagonistic effect on mycelial growth and conidial germination of F. pseudograminearum by destroying the integrity of the cell membrane. In the greenhouse, TM28 significantly increased wheat fresh weight and height in the presence of pathogen Fp , it enhanced the antioxidant defense activity and ameliorated the negative effects of F. pseudograminearum , including disease severity and pathogen abundance in the rhizosphere soil, root and stem base of wheat. RNA-seq of F. pseudograminearum under TM28 antagonistic revealed 2,823 differentially expressed genes (DEGs). Most DEGs related to cell wall and cell membrane synthesis were significantly downregulated, the culture filtrate of TM28 affected the pathways of fatty acid synthesis, steroid synthesis, glycolysis, and the citrate acid cycle. T. muroii TM28 appears to have significant potential in controlling wheat Fusarium crown rot caused by F. pseudograminearum., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Yang, Cui, Wei, Li, Hu, Yang, Wu, Zhao, Li, Wang and Yang.)

Fusarium pseudograminearum is the dominant pathogen causing Fusarium crown rot (FCR) of wheat. Phenamacril is a 2-cyanoacrylate fungicide, having a control effect on diseases caused by Fusarium spp. The objective of this study was to investigate the inhibitory effect of phenamacril on F. pseudograminearum and its control efficacy against FCR. The results showed that phenamacril had a strong inhibitory effect on the mycelial growth of F. pseudograminearum , EC 50 values of phenamacril to 63 tested strains were in the range of 0.0998 to 0.5672 μg/ml, and the average EC 50 value was 0.3403 ± 0.0872 μg/ml and could be used as the baseline sensitivity of F. pseudograminearum to phenamacril. Phenamacril reduced the germination rate of conidia of F. pseudograminearum , and the EC 50 value was 5.0273 to 26.4814 μg/ml. In addition, we found that phenamacril had a teratogenic effect on conidia and blastotubules, which increased the ratio of conidial germination from the middle cells and showed high efficacy on the sporulation quantity of F. pseudograminearum with an EC 50 value in the range of 0.0770 to 0.1064 μg/ml. There was no significant correlation between the sensitivity of F. pseudograminearum to phenamacril and its sensitivity to fludioxonil, carbendazim, tebuconazole, and kresoxim-methyl. In vitro and greenhouse assays showed that the treatment with 0.125 μl of active ingredient per gram recorded the best control effect on wheat crown rot, reaching 87.8 and 77.3%, respectively. In two experimental sites in Luoyang, phenamacril also had great control effect against FCR, reaching 83.9%. It was proven that phenamacril has a superior control effect against FCR. This study has laid a foundation for the study of the mechanism of action of phenamacril against F. pseudograminearum and provided a theoretical basis for the application of phenamacril to control FCR., Competing Interests: The author(s) declare no conflict of interest.

Background: Fusarium pseudograminearum is one of the dominant pathogens of Fusarium crown rot (FCR) worldwide. Unfortunately, no fungicides have yet been registered for the control of FCR in wheat in China. Pydiflumetofen, a new-generation succinate dehydrogenase inhibitor, exhibits excellent inhibitory activity to Fusarium spp. A resistance risk assessment of F. pseudograminearum to pydiflumetofen and the resistance mechanism involved have not yet been investigated., Results: The median effective concentration (EC 50 ) value of 103 F. pseudograminearum isolates to pydiflumetofen was 0.0162 μg mL -1 , and the sensitivity exhibited a unimodal distribution. Four resistant mutants were generated by fungicide adaption, which possessed similar or impaired fitness compared to corresponding parental isolates based on the results of mycelial growth, conidiation, conidium germination rate, and virulence determination. Pydiflumetofen showed strong positive cross-resistance with cyclobutrifluram and fluopyram but no cross-resistance with carbendazim, phenamacril, tebuconazole, fludioxonil, or pyraclostrobin. Sequence alignment revealed that pydiflumetofen-resistant F. pseudograminearum mutants had two single-point mutations of A83V or R86K in FpSdhC 1 . Molecular docking further confirmed that point mutation of A83V or R86K in FpSdhC 1 could confer resistance of F. pseudograminearum to pydiflumetofen., Conclusion: Fusarium pseudograminearum shows an overall moderate risk of developing resistance to pydiflumetofen, and point mutation FpSdhC 1 A83V or FpSdhC 1 R86K could confer pydiflumetofen resistance in F. pseudograminearum. This study provided vital data for monitoring the emergence of resistance and developing resistance management strategies for pydiflumetofen. © 2023 Society of Chemical Industry., (© 2023 Society of Chemical Industry.)

In two experiments on a farm practicing conservation agriculture, the grain yield of a range of wheat cultivars was significantly (p < 0.001) negatively related to the post-harvest population densities of Pratylenchus thornei in the soil profile to 45 cm depth. In a third and fourth experiment with different rotations, methyl bromide fumigation significantly (p < 0.05) decreased (a) a low initial population density of P. thornei in the soil profile to 90 cm depth and (b) a high initial population of P. thornei to 45 cm depth, and a medium level of the crown rot fungus, Fusarium pseudograminearum, at 0–15 cm depth to a low level. For a range of wheat and durum cultivars, grain yield and response to fumigation were highly significantly (p < 0.001) related to (a) the P. thornei tolerance index of the cultivars in the third experiment, and (b) to both the P. thornei tolerance index and the crown rot resistance index in the fourth experiment. In the latter, grain yield was significantly (p < 0.001) positively related to biomass at anthesis and negatively related to percentage whiteheads at grain fill growth stage. One barley cultivar was more tolerant to both diseases than the wheat and durum cultivars. Crop rotation, utilizing crop cultivars resistant and tolerant to both P. thornei and F. pseudograminearum, is key to success for conservation farming in this region.

Fusarium crown rot (FCR), caused predominantly by Fusarium pseudograminearum (Fp) in Australia, is an important fungal disease of wheat and barley. FCR causes significant yield losses and reduced grain quality worldwide. This study investigated the population dynamics of FCR‐causing F. pseudograminearum isolates from Western Australia (WA), a major wheat‐growing region. Wheat samples were collected from a total of seven different sites in 2008 and 2015. Two sites, Tammin and Karlgarin, with moderate to high FCR incidence, were intensively sampled in both years. The results revealed significant increase in Fp isolation frequency between 2008 and 2015. Over 86% of 1100 Fusarium isolates were Fp in 2015 compared with 59% of 639 isolates from 2008. Mating type idiomorphs, toxin chemotypes and population genetic structures were determined for a subset of 279 Fp isolates (132 isolates from 2008 and 165 from 2015). Mating type analysis revealed differences in MAT1‐1 and MAT1‐2 distributions between Tammin and Karlgarin for both years. Results also showed that 97.6% of Fp isolates analysed had the 3‐ADON trichothecene chemotype. Additionally, for the first time in Australia, the 15‐ADON chemotype was identified in 2.3% and 2.4% of Fp isolates from 2008 and 2015, respectively. The genetic structure of Fp population determined using 21 cleaved amplified polymorphic sequence (CAPS) markers revealed a high level of genetic variation within and between populations. In addition, 2015 isolates from Tammin and Karlgarin were significantly more aggressive (P < 0.0001) than 2008 isolates. This finding may have implications in managing this significant fungal disease. [ABSTRACT FROM AUTHOR]

Heat shock protein 70s (Hsp70s) are a class of molecular chaperones that are highly conserved and ubiquitous in organisms ranging from microorganisms to plants and humans. Hsp70s play key roles in cellular development and protecting living organisms from environmental stresses such as heat, drought, salinity, acidity, and cold. However, their functions in pathogenic fungi are largely unknown. Here, a total of 14 FpHsp70 genes were identified in Fusarium pseudograminearum , including 3 in the mitochondria, 7 in the cytoplasm, 2 in the endoplasmic reticulum (ER), 1 in the nucleus, and 1 in the plastid. However, the exon–intron boundaries and protein motifs of the FpHsp70 have no consistency in the same subfamily. Expression analysis revealed that most FpHsp70 genes were up-regulated during infection, implying that FpHsp70 genes may play important roles in F. pseudograminearum pathogenicity. Furthermore, knockout of an ER lumenal Hsp70 homolog FpLhs1 gene reduced growth, conidiation, and pathogenicity in F. pseudograminearum. These mutants also showed a defect in secretion of some proteins. Together, FpHsp70s might play essential roles in F. pseudograminearum and FpLhs1 is likely to act on the development and virulence by regulating protein secretion. [ABSTRACT FROM AUTHOR]

The disease crown rot, caused predominantly by the fungal pathogen Fusarium pseudograminearum, is a major disease of winter cereals in many regions of the world, including Australia. A methodology is proposed, using response curves, to robustly estimate the relationship between grain yield and increasing crown rot pathogen burdens. Using data from a field experiment conducted in northern New South Wales, Australia in 2016, response curves were derived for five commercial wheat cultivars exposed to six increasing rates of crown rot inoculum, where the rates served to establish a range of crown rot pathogen burdens. In this way, the response curve methodology is fundamentally different from alternate approaches that rely on genetic or environmental variation to establish a range in pathogen burdens over which yield loss relationships are estimated. By manipulating only the rates of crown rot inoculum and, thus, pathogen burden directly, the number of additional confounding factors and interactions are minimized, enabling the robust estimation of the rate of change in yield due to increasing crown rot pathogen burdens for each cultivar. The methodology revealed variation in the rate of change in yield between cultivars, along with the extent of crown rot symptoms expressed by the cultivars. Variation in the rate of change in yield between cultivars provides definitive evidence of differences in the tolerance of commercial Australian wheat cultivars to crown rot caused by F. pseudograminearum, while variation in the extent of crown rot symptoms signifies differences in the resistance of the cultivars to this disease. The response curve methodology also revealed variation in how the different mechanisms of tolerance and resistance act to limit yield losses due to crown rot for different cultivars. [ABSTRACT FROM AUTHOR]

The basic helix-loop-helix (bHLH) transcriptional regulatory proteins are key players in a wide array of developmental processes and abiotic stress responses. The bHLH domain has been identified and functionally characterized from fungi to humans. However, no systematic characterization of a bHLH family has been reported in Fusarium pseudograminearum. Fusarium pseudograminearum is an important plant pathogen that causes crown rot of wheat and barley. In this study, 17 FpbHLH genes with conserved residues in the bHLH domain were identified using a whole-genome searching approach. In addition, transcription profiling results showed that 11 FpbHLH genes were responsive to infection – eight of them (FpbHLH2, FpbHLH6, FpbHLH7, FpbHLH8, FpbHLH11, FpbHLH13, FpbHLH14, FpbHLH15) showed down-regulated expression, while FpbHLH5, FpbHLH9 and FpbHLH10 were induced both in the compatible interaction and incompatible interaction. Furthermore, knockout of FpbHLH9 gene reduced the virulence in a susceptible wheat cultivar. Together, these results suggest that FpbHLHs may play different roles in virulence of F. pseudograminearum, and provide a good basis for further investigation. [ABSTRACT FROM AUTHOR]

The distribution frequency of pathogenic fungi associated with root and crown rot of winter wheat (Triticum aestivum) from 104 fields in the North China Plain was determined during the period from 2013 to 2016. The four most important species identified were Bipolaris sorokiniana (24.0% from roots; 33.7% from stems), Fusarium pseudograminearum (14.9% from roots; 27.8% from stems), Rhizoctonia cerealis (1.7% from roots; 4.4% from stems), and Gaeumannomyces graminis var. tritici (9.8% from roots; 4.4% from stems). We observed that the recovered species varied with the agronomic zone. Fusarium pseudograminearum was predominant in regions 1 and 3, whereas F. graminearum, F. acuminatum, and R. cerealis were predominant in regions 2 and 4. The incidence of F. pseudograminearum and R. cerealis was significantly different between regions 1 and 4, while no significant association was found in the distribution of the other species and the agronomic zones. A negative correlation between the frequency of occurrence of F. pseudograminearum and mean annual precipitation during 2013–2016 (r = −0.71; P < 0.01) in the North China Plain and a positive correlation between the mean annual precipitation during 2013–2016 and the frequency of occurrence of F. asiaticum (r = 0.74; P < 0.01) were observed. Several Fusarium species were also found with low frequencies of ~2.1%−3.4 % (F. graminearum, F. acuminatum, and F. sinensis) and ~0.1%−1.3% (F. equiseti, F. oxysporum, F. proliferatum, F. culmorum, F. avenaceum, and F. asiaticum). In more than 93% of the fields, from the root and crown tissues of wheat, two or more root and crown rot species were isolated. The coexistence of Fusarium spp. and B. sorokiniana in one field (65.4%) or in individual plants (11.6%) was more common than for the other species combinations. Moreover, this is the first report on the association between F. sinensis and root and crown rot of wheat. Our results would be useful in the framing guidelines for the management of root and crown rot fungi in wheat in different agronomic zones of the North China Plain.

Fusarium crown rot (FCR), caused by Fusarium pseudograminearum , is one of the most important diseases impacting wheat production in the Huanghuai region, the most important wheat-growing region of China. The current study found that the SDHI fungicide pydiflumetofen, which was recently developed by Syngenta Crop Protection, provided effective control of 67 wild-type F. pseudograminearum isolates in potato dextrose agar, with an average EC 50 value of 0.060 ± 0.0098 μg/ml (SE). Further investigation revealed that the risk of fungicide resistance in pydiflumetofen was medium to high. Four F. pseudograminearum mutants generated by repeated exposure to pydiflumetofen under laboratory conditions indicated that pydiflumetofen resistance was associated with fitness penalties. Mutants exhibited significantly ( P < 0.05) reduced sporulation in mung bean broth and significantly ( P < 0.05) reduced pathogenicity in wheat seedlings. Sequence analysis indicated that the observed pydiflumetofen resistance of the mutants was likely associated with amino acid changes in the different subunits of the succinate dehydrogenase target protein, including R18L and V160M substitutions in the FpSdhA sequence; D69V, D147G, and C257R in FpSdhB; and W78R in FpSdhC. This study found no evidence of cross-resistance between pydiflumetofen and the alternative fungicides tebuconazole, fludioxonil, carbendazim, or fluazinam, which all have distinct modes of action and could therefore be used in combination or rotation with pydiflumetofen to reduce the risk of resistance emerging in the field. Taken together, these results indicate that pydiflumetofen has potential as a novel fungicide for the control of FCR caused by F. pseudograminearum and could therefore be of great significance in ensuring high and stable wheat yields in China., Competing Interests: The author(s) declare no conflict of interest.