The zinc finger protein StMR1 affects the pathogenicity and melanin synthesis of Setosphaeria turcica and directly regulates the expression of DHN melanin synthesis pathway genes.

The infection and colonization of pathogenic fungi are often regulated by transcription factors. In our previous study, the zinc finger protein-encoding gene StMR1 was found to be highly expressed during the infection process of Setosphaeria turcica, the pathogen causing northern corn leaf blight. Evolutionary tree analysis showed that this gene was associated with regulatory factors of melanin synthesis. However, the regulatory mechanism of melanin synthesis and its effect on pathogenicity remain unclear. In this study, the function of StMR1 was analyzed by gene knockout. When the expression level of StMR1 in the mutants was significantly reduced, the colony color became lighter, the mycelia were curved and transparent, and the mutant showed a significant loss of pathogenicity. In addition, compared with wild-type, the accumulation of melanin decreased significantly in ΔStmr1. RNA-seq analysis revealed 1,981 differentially expressed genes between the wild-type and knockout mutant, among which 39 genes were involved in melanin metabolism. qPCR revealed that the expression levels of six key genes in the melanin synthesis pathway were significantly reduced. ChIP-PCR and yeast one-hybrid assays confirmed that StMR1 directly binds to the promoters of St3HNR, St4HNR, StPKS, and StLAC2 in the DHN melanin synthesis pathway and regulates gene expression. The C2H2-type zinc fingers and Zn(Ⅱ)2Cys6 binuclear cluster in StMR1 were important for the binding to targets.

Omnivorous Notoxus trinotatus Pic (Coleoptera: Anthicidae) is a newly recognized vector of northern leaf blight in maize.

The adaptations of omnivorous insects to food are manifested in a multifaceted manner, and the availability of food resources directly determines insect feeding tendencies, which contribute to a complex insect-food relationship and impact insect functionality in the environment. Stable isotope analysis was applied to test the feeding preference and further define the functional role of omnivorous beetles in cropland. Our results confirmed that as an omnivorous beetle, the fungivorous nature of Notoxus trinotatus accounted for a prominent proportion food selection at the adult stage, and more importantly, this dietary feature contributed to the dispersal of the northern corn leaf blight in maize (NLB) during the feeding trials. In addition to the preference for fungi, water supplementation was an essential element extending adult longevity, which directly prolonged the contact time of adults with pathogenic fungi in agricultural fields. Consistent with the herbivorous characteristics of beetles, before the emergence of NLB fungal pathogens, corn tissues served as the main food, which provided the beetles with more opportunities to transmit fungal pathogen propagules. We conclude that the role of N. trinotatus in carrying NLB pathogen is due to its feeding on this plant mycopathogen, and an increased abundance of beetles carrying the pathogen may increase the rate of NLB disease infestation. More focus should be concentrated on the functions of fungivorous beetles, not only as pathogen-transmitting pests, but also as an element among the balanced biotic factors in farmland.

Occurrence and Distribution of Physiological Races of Exserohilum turcicum in Maize-Growing Regions of Mexico.

Turcicum leaf blight (TLB) is a common foliar disease of maize in Mexico that is caused by the fungal pathogen Exserohilum turcicum. The most effective management strategy against TLB is monogenic race-specific resistance. Among the 140 E. turcicum isolates from symptomatic leaves collected from maize fields in Mexico, 100 were obtained from tropical (Veracruz) and temperate areas (Estado de México) between 2010 and 2019, and 40 isolates were obtained from tropical (Sinaloa, Tamaulipas, Veracruz, and Chiapas), subtropical (Nayarit, Jalisco, and Guanajuato), and temperate areas (Estado de Mexico, Hidalgo, and Puebla) collected in 2019. All the isolates caused TLB symptoms on the positive control (ht4), showing that they were all pathogenic. Six physiological races of E. turcicum (2, 3, 23, 3N, 23N, and 123N) were identified based on resistant or susceptible responses displayed by five maize differential genotypes (A619Ht1, A619Ht2, A619Ht3, B68HtN, and A619ht4). The most common was race 23, accounting for 68% of the isolates, followed by races 23N, 123N, 3, 2, and 3N at 15, 8, 6, 2, and 1%, respectively. Race 123N was able to infect the greatest number of maize differential genotypes used in the study. Race 123N was detected in Sinaloa and Estado de México. Race 3 was detected in Nayarit and Jalisco. Race 2 was detected in Jalisco, Estado de México, and Veracruz, and race 3N was detected in Tamaulipas. Race 23 was equally dominant in the tropical, subtropical, and temperate regions, while race 123N was more common in the tropical environment, and race 23N was more common in the tropical and temperate environments. There was no evidence for shifts in the races between 2010 and 2019.

Genomic regions associated with virulence in Setosphaeria turcica identified by linkage mapping in a biparental population.

Northern corn leaf blight (NCLB) and sorghum leaf blight (SLB) are significant diseases of maize and sorghum, respectively, caused by the filamentous fungus Setosphaeria turcica. Strains of S. turcica are typically host-specific and infect either maize or sorghum. Host specificity in this pathogen is attributed to a single locus for maize and a second distinct locus for sorghum. To identify the genetic basis of host specificity in S. turcica, we generated a biparental population of S. turcica by crossing strains specific to maize and sorghum, phenotyped the population for leaf blight on sorghum and maize, genotyped the population to create a linkage map of S. turcica, and located candidate virulence regions. A total of 190 ascospores from 35 pseudothecia were isolated from the cross of maize and sorghum-specific strains. Greenhouse phenotyping of the biparental population (n = 144) showed independent inheritance of virulence, as indicated by a 1:1:1:1 segregation for virulence to maize, sorghum, both maize and sorghum, and avirulence to both crops. The population and host-specific parent strains were genotyped using genome skim sequencing on an Illumina NovaSeq 6000 platform resulting in over 780 million reads. A total of 32,635 variants including single nucleotide polymorphisms and indels were scored. There was evidence for a large deletion in the sorghum-specific strain of S. turcica. A genetic map consisting of 17 linkage groups spanning 3,069 centimorgans was constructed. Virulence to sorghum and maize mapped on distinct linkage groups with a significant QTL detected for virulence to maize. Furthermore, a single locus each for the in vitro traits hyphal growth rate and conidiation were identified and mapped onto two other linkage groups. In vitro traits did not correlate with in planta virulence complexity, suggesting that virulence on both hosts does not incur a fitness cost. Hyphal growth rate and conidiation were negatively correlated, indicating differences in hyphal growth versus dispersal ability for this pathogen. Identification of genetic regions underlying virulence specificity and saprotrophic growth traits in S. turcica provides a better understanding of the S. turcica- Andropogoneae pathosystem.

Evaluation of Maize Hybrids for Identifying Resistance to Northern Corn Leaf Blight in Northeast China.

Northern corn leaf blight (NCLB) caused by Setosphaeria turcica is one of the most devastating foliar diseases in maize (Zea mays L.), resulting in great economic losses worldwide. The mutation of the pathogen exacerbates the occurrence and harmfulness of NCLB in China. Therefore, there is an urgent need for evaluating and cultivating resistant hybrids. Here, the response of 239 maize hybrids approved in Northeast China to NCLB was evaluated during 2019 and 2020. The results showed that 92 (38.49%) and 75 (31.38%) hybrids were rated as moderately resistant and resistant, respectively, which together constituted the predominant resistant categories. We observed that maize hybrids from different certified sources had different levels of resistance to NCLB, whose disease parameter values varied significantly (P < 0.05) among 52 main cultivated hybrids. In 2019 and 2020, the average size of the lesions increased from 21.02 to 21.06 cm2, the average lesion density decreased from 1.36 to 1.33 lesions/100 cm2, and more than 30% of the hybrids registered final disease severity scores between 10 and 30%. The area under the disease progress curve of the main cultivated hybrids ranged from 57.96 to 986.86 cm2 in 2019 and from 50.75 to 1,028.65 cm2 in 2020. Correlation analysis revealed a significant relationship (P < 0.0001) among four disease parameters. Current research has shown that many maize hybrids in Northeast China are resistant to NCLB, suggesting that the large-scale cultivation of susceptible hybrids has led to the occurrence and prevalence of the disease. This study should assist growers in purposefully selecting resistant commercial hybrids to contribute to the management of NCLB.

Conserved defense responses between maize and sorghum to Exserohilum turcicum.

BACKGROUND: Exserohilum turcicum is an important pathogen of both sorghum and maize, causing sorghum leaf blight and northern corn leaf blight. Because the same pathogen can infect and cause major losses for two of the most important grain crops, it is an ideal pathosystem to study plant-pathogen evolution and investigate shared resistance mechanisms between the two plant species. To identify sorghum genes involved in the E. turcicum response, we conducted a genome-wide association study (GWAS). RESULTS: Using the sorghum conversion panel evaluated across three environments, we identified a total of 216 significant markers. Based on physical linkage with the significant markers, we detected a total of 113 unique candidate genes, some with known roles in plant defense. Also, we compared maize genes known to play a role in resistance to E. turcicum with the association mapping results and found evidence of genes conferring resistance in both crops, providing evidence of shared resistance between maize and sorghum. CONCLUSIONS: Using a genetics approach, we identified shared genetic regions conferring resistance to E. turcicum in both maize and sorghum. We identified several promising candidate genes for resistance to leaf blight in sorghum, including genes related to R-gene mediated resistance. We present significant advancements in the understanding of host resistance to E. turcicum, which is crucial to reduce losses due to this important pathogen.

A Genome Resource of Setosphaeria turcica, Causal Agent of Northern Leaf Blight of Maize.

The heterothallic ascomycete Setosphaeria turcica (anamorph Exserohilum turcicum) causes northern corn leaf blight, which results in devastating yield losses and a reduction in feed value. Although genome sequences of two model strains of the pathogen are available (https://mycocosm.jgi.doe.gov/mycocosm/home), previous drafts were assembled using short read technologies, making evolutionary and genetic linkage inferences difficult. Here, race 23N of S. turcica strain Et28A was sequenced again using Illumina HiSeq and PacBio Sequel technologies, and assembled to approximately 43,480,261 bp on 30 scaffolds. In all, 13,183 protein-coding genes were predicted, 13,142 of them were well annotated. This S. turcica genome resource is important for understanding the genetics behind pathogen evolution and infection mechanisms.

Plant resistance and leaf chemical characteristic jointly shape phyllosphere bacterial community.

Phyllosphere bacteria have an important role in plant growth and resistance to pathogen infection and are partially influenced by plant genotype and leaf environment. How plant resistance to pathogens and leaf chemical characteristics shape the phyllosphere bacterial communities is unclear. In this study, the phyllosphere bacterial communities of maize hybrids with various resistance to Setosphaeria turcica were compared using the high-throughput sequencing and large-scale culturing methods. The results showed that Shannon and Simpson indices of phyllosphere bacterial communities were markedly higher in the highly resistant hybrid (HR) compared with the susceptible one. Hierarchical clustering analysis, unweighted UniFrac principal component analysis (PCoA) and the analysis of similarities (ANOSIM) demonstrated that the phyllosphere bacterial communities were significantly distinct between resistant and susceptible hybrids. The redundancy analysis (RDA) demonstrated that leaf chemical characteristics, including nitrogen and phosphorus concentration, and disease resistance play an important role in shaping the phyllosphere bacterial community. Linear discriminant effect size (LEfSe) analysis indicated that Bacillus, Pseudomonas and Tumebacillus were the biomarker species in the phyllosphere of HR. Biocontrol bacteria against S. turcica (such as Pseudomonas and Bacillus) were isolated from the phyllosphere of HR by large-scale culturing. The work contributes to understanding of the phyllosphere bacterial community assembly and provides a new clue to screening for strong biocontrol bacteria from HR and to facilitating future breeding efforts for enhancing disease resistance.

Population structure and genetic diversity of Setosphaeria turcica from corn in Heilongjiang province, China.

AIM: The aims of this study were to identify races and mating types of Setosphaeria turcica causing northern corn leaf blight in Heilongjiang province of China and analyse the genetic diversity of S. turcica isolates using SSR markers. METHODS AND RESULTS: Based on gene-for-gene interactions, 13 races of S. turcica (races 0, 1, 2, 3, 12, 13, 23, 123, N, 1N, 12N, 3N and 23N) were isolated from infected corn plants in Heilongjiang province. Races 0 and 1 were the predominant races, and race 23N was identified for the first time in the region. Using two pairs of specific primers, three mating types, 'a', 'Aa' and 'A', were identified, with 'a' being the predominant mating type. SSR markers were used to analyse genetic diversity of 60 S. turcica isolates. Five SSR primers were polymorphic, which resulted in 45 reproducible bands with 2-15 bands for each primer. Cluster analysis separated the isolates into five groups at a similarity coefficient of 0·84. Analysis of molecular variance showed that there was significant correlation between SSR groups and mating type of the isolates. No significant correlation was found between SSR groups and physiological races or geographical location of the isolates. CONCLUSIONS: The work reported that races 0 and 1 were the predominant races, and race 23N was identified for the first time in Heilongjiang province with 'a' being the predominant mating type. There was significant correlation between SSR groups and mating type of S. turcica isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results provide information on population structure and genetic diversity of S. turcica causing Northern corn leaf blight, which will facilitate the development of effective disease management programs.

Expression, purification, and characterization of a novel laccase from Setosphaeria turcica in Eschericha coli.

Laccases are multicopper oxidases (E.C. 1.10.3.2) that catalyze the oxidation of many phenolic compounds. In this study, a novel laccase, Stlac4, from Setosphaeria turcica was cloned and expressed in Escherichia coli by insertion into the pET-30a expression plasmid. The recombinant laccase was purified and visualized on SDS-PAGE as a single band with an apparent molecular weight of 71.5 KDa, and confirmed by Western blot. The maximum activity of the purified laccase was 127.78 U · mg-1 , the optimum temperature and pH value were 60 °C and 4.0 respectively, measured by oxidation of 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS). Purified laccase activity under different metal ions and an inhibitor were tested, revealing that laccase activity increased by approximately 434.8% with Fe3+ , and 217.4% with Cu2+ at 10 mmol · L-1 concentrations, Mn2+ increased the laccase activity only at 5 mmol · L-1 , while Na+ increased activity at 1 mmol · L-1 but inhibited activity at 5 and 10 mmol · L-1 . SDS increased laccase activity at 1 mmol · L-1 , and inhibited activity at 5 and 10 mmol · L-1 .

The catalytic subunit of type 2A protein phosphatase negatively regulates conidiation and melanin biosynthesis in Setosphaeria turcica.

Northern corn leaf blight caused by Setosphaeria turcica is a major fungal disease responsible for significant reductions in maize yield worldwide. Eukaryotic type 2A protein phosphatase (PP2A) influences growth and virulence in a number of pathogenic fungi, but little is known about its roles in S. turcica. Here, we functionally characterized S. turcica StPP2A-C, which encodes the catalytic C subunit of StPP2A. StPP2A-C deletion slowed colony growth, conidial germination, and appressorium formation but increased conidiation, melanin biosynthesis, glycerol content, and disease lesion size on maize. These effects were associated with expression changes in genes related to calcium signaling, conidiation, laccase activity, and melanin and glycerol biosynthesis, as well as changes in intra- and extracellular laccase activity. A pull-down screen for candidate StPP2A-c interactors revealed an interaction between StPP2A-c and StLac1. Theoretical modeling and yeast two-hybrid experiments confirmed that StPP2A-c interacted specifically with the copper ion binding domain of StLac1 and that Cys267 of StPP2A-c was required for this interaction. StPP2A-C expression thus appears to promote hyphal growth and reduce pathogenicity in S. turcica, at least in part by altering melanin synthesis and laccase activity; these insights may ultimately support the development of novel strategies for biological management of S. turcica.

Recent advances in the population biology and management of maize foliar fungal pathogens Exserohilum turcicum, Cercospora zeina and Bipolaris maydis in Africa.

Maize is the most widely cultivated and major security crop in sub-Saharan Africa. Three foliar diseases threaten maize production on the continent, namely northern leaf blight, gray leaf spot, and southern corn leaf blight. These are caused by the fungi Exserohilum turcicum, Cercospora zeina, and Bipolaris maydis, respectively. Yield losses of more than 10% can occur if these pathogens are diagnosed inaccurately or managed ineffectively. Here, we review recent advances in understanding the population biology and management of the three pathogens, which are present in Africa and thrive under similar environmental conditions during a single growing season. To effectively manage these pathogens, there is an increasing adoption of breeding for resistance at the small-scale level combined with cultural practices. Fungicide usage in African cropping systems is limited due to high costs and avoidance of chemical control. Currently, there is limited knowledge available on the population biology and genetics of these pathogens in Africa. The evolutionary potential of these pathogens to overcome host resistance has not been fully established. There is a need to conduct large-scale sampling of isolates to study their diversity and trace their migration patterns across the continent.

Molecular and biological characteristics of a novel chrysovirus infecting the fungus phytopathogenic Setosphaeria turcica f.sp. sorghi.

A new double-stranded RNA (dsRNA) virus has been identified in the filamentous fungus Setosphaeria turcica f.sp. sorghi, whose genome consists of four segments (dsRNA1-4). Each dsRNA carries single open reading frame (ORF) flanked by 5' and 3' untranslated regions (UTRs) containing strictly conserved termini. The putative protein encoded by dsRNA1 showed 80.50% identity to the RNA-dependent RNA polymerase (RdRp) of the most closely related virus, Alternaria alternata chrysovirus 1 (AaCV1), belonging to the Chrysoviridae. dsRNA2 encodes the putative coat protein, while dsRNA3 and dsRNA4 respectively encode the hypothetical proteins of unknown functions. Phylogenetic analysis based on the RdRp protein indicated the virus clustered with members of the genus Betachrysovirus in the family Chrysoviridae. Based on the dsRNA profile, amino acid sequence comparisons, and phylogenetic analyses, the mycovirus is thought to be a new member of the family Chrysoviridae and designated as Setosphaeria turcica chrysovirus 1 (StCV1). Moreover, obvious differences were observed in the colony, mycelial and spore morphology between StCV1-infected and virus-cured strains of S. turcica f.sp. sorghi. StCV1 infection strongly reduced colony growth rate, spore production ability and virulence on host fungus. To our knowledge, this is the first report about mycovirus infecting S. turcica f.sp. sorghi and also the first chrysovirus infecting S. turcica.

The Septin Gene StSep4 Contributes to the Pathogenicity of Setosphaeria turcica by Regulating the Morphology, Cell Wall Integrity, and Pathogenic Factor Biosynthesis.

Septins are a conserved group of GTP-binding proteins found in all eukaryotes and are the fourth-most abundant cytoskeletal proteins. Septins of some pathogenic fungi are involved in morphological changes related to infection. Our previous studies have identified four core septins (StSep1-4) in Setosphaeria turcica, the causal agent of northern corn leaf blight, while only StSep4 is significantly upregulated during the invasive process. We therefore used forchlorfenuron (FCF), the specific inhibitor of septin, and ΔStSep4 knockout mutants to further clarify the role of septins in S. turcica pathogenicity. FCF treatment caused a dose-dependent reduction in S. turcica colony growth, delayed the formation of infection structures, and reduced the penetration ability. ΔStSep4 knockout mutants displayed abnormal mycelium morphology, slow mycelial growth, conidiation deficiency, delayed appressorium development, and weakened pathogenicity. StSep4 deletion also broke cell wall integrity, altered chitin distribution, decreased the melanin content, and disrupted normal nuclear localization. A transcriptomic comparison revealed that genes differentially expressed between ΔStSep4 and WT were enriched in terms of ribosomes, protein translation, membrane components, and transmembrane transport activities. Our results demonstrate that StSep4 is required for morphology and pathogenicity in S. turcica, making it a promising target for the development of novel fungicides.

Population Genomic Evidence for a Repeated Introduction and Rapid Expansion of the Fungal Maize Pathogen Setosphaeria turcica in Europe.

Modern agricultural practices, climate change, and globalization foster the rapid spread of plant pathogens, such as the maize fungal pathogen Setosphaeria turcica, which causes Northern corn leaf blight and expanded into Central Europe during the twentieth century. To investigate the rapid expansion of S. turcica, we sequenced 121 isolates from Europe and Kenya. Population genomic inference revealed a single genetically diverse cluster in Kenya and three clonal lineages with low diversity, as well as one cluster of multiple clonal sublineages in Europe. Phylogenetic dating suggests that all European lineages originated through sexual reproduction outside Europe and were subsequently introgressed multiple times. Unlike isolates from Kenya, European isolates did not show sexual recombination, despite the presence of both MAT1-1 and MAT1-2 mating types. For the clonal lineages, coalescent model selection supported a selectively neutral model with strong exponential population growth, rather than models with pervasive positive selection caused by host defense resistance or environmental adaptation. Within clonal lineages, phenotypic variation in virulence to different monogenic resistances, which defines the pathogen races, suggests that these races may originate from repeated mutations in virulence genes. Association testing based on k-mers did not identify genomic regions linked to pathogen races, but it did uncover strongly differentiated genomic regions between clonal lineages, which harbor genes with putative roles in pathogenicity. In conclusion, the expansion and population growth of S. turcica in Europe are mainly driven by an expansion of the maize cultivation area and not by rapid adaptation.

StRAB4 gene is required for filamentous growth, conidial development, and pathogenicity in Setosphaeria turcica.

Setosphaeria turcica, the fungal pathogen responsible for northern corn leaf blight in maize, forms specialized infectious structures called appressoria that are critical for fungal penetration of maize epidermal cells. The Rab family of proteins play a crucial role in the growth, development, and pathogenesis of many eukaryotic species. Rab4, in particular, is a key regulator of endocytosis and vesicle trafficking, essential for filamentous growth and successful infection by other fungal pathogens. In this study, we silenced StRAB4 in S. turcica to gain a better understanding the function of Rab4 in this plant pathogen. Phenotypically, the mutants exhibited a reduced growth rate, a significant decline in conidia production, and an abnormal conidial morphology. These phenotypes indicate that StRab4 plays an instrumental role in regulating mycelial growth and conidial development in S. turcica. Further investigations revealed that StRab4 is a positive regulator of cell wall integrity and melanin secretion. Functional enrichment analysis of differentially expressed genes highlighted primary enrichments in peroxisome pathways, oxidoreductase and catalytic activities, membrane components, and cell wall organization processes. Collectively, our findings emphasize the significant role of StRab4 in S. turcica infection and pathogenicity in maize and provide valuable insights into fungal behavior and disease mechanisms.

Novel factors contributing to fungal pathogenicity at early stages of Setosphaeria turcica infection.

The fungal pathogen Setosphaeria turcica causes leaf blight on maize, which leads to considerable crop losses. However, how S. turcica establishes sustained systemic infection is largely unknown. Here, we report several novel factors contributing to S. turcica pathogenicity, identified using a genomic and transcriptional screen at different stages of S. turcica appressorium development. We identified two cytoskeleton regulators, SLM1 and SLM2, that are crucial for hypha and appressorium development. The SLM1 and SLM2 transcripts accumulated during germling stage but their levels were notably reduced at the appressorium stage. Deletion of SLM2 dramatically affected cell morphology, penetration ability, and pathogenicity. We also identified three different types of S. turcica glycosyl hydrolases that are critical for plant cell wall degradation. Their transcripts accumulated during the appressorium infection stage induced by cellophane and maize leaf. Most importantly, we characterized a novel and specific S. turcica effector, appressorium-coupled effector 1 (StACE1), whose expression is coupled to appressorium formation in S. turcica. This protein is required for maize infection and induces cell death on expression in Nicotiana benthamiana. These observations suggest that the phytopathogen S. turcica is primed in advance with multiple strategies for maize infection, which are coupled to appressorium formation at the early infection stages.

The Secreted Ribonuclease SRE1 Contributes to Setosphaeria turcica Virulence and Activates Plant Immunity.

During the plant infection process, pathogens can secrete several effectors. Some of the effectors are well-known for their roles in regulating plant immunity and promoting successful pathogen colonization. However, there are few studies on the ribonuclease (RNase) effectors secreted by fungi. In the present study, we discovered a secretable RNase (SRE1) in the secretome of Setosphaeria turcica that was significantly upregulated during the early stages of S. turcica infection in maize. Knockdown of SRE1 significantly reduced the virulence of S. turcica. SRE1 can induce cell death in maize and Nicotiana benthamiana. However, unlike the conventional hypersensitive response (HR) caused by other effectors, SRE1 is not dependent on its signal peptide (SP) or plant receptor kinases (such as BAK1 and SOBIR1). SRE1-induced cell death depends upon its enzymatic activity and the N-terminal ß-hairpin structure. SRE1 relies on its N-terminal ß-hairpin structure to enter cells, and then degrades plant's RNA through its catalytic activity causing cytotoxic effects. Additionally, SRE1 enhances N. benthamiana's resistance to pathogenic fungi and oomycetes. In summary, SRE1 promotes the virulence of S. turcica, inducing plant cell death and activating plant immune responses.

Molecular and Morphological Characterization of Exserohilum turcicum (Passerini) Leonard and Suggs Causing Northern Corn Leaf Blight of Maize in Bihar.

Maize is considered the third most important cereal crop in Asia after rice and wheat. Many diseases affect this crop due to the cultivation of various hybrids. This research aimed to characterize the causative agent of northern corn leaf blight disease in Bihar, India, caused by Exserohilum turcicum (Passerini) Leonard and Suggs. Leaf samples were collected from infected fields in five maize growing districts of Bihar in 2020-2022. A total of 45 fungal isolates from 135 samples were examined for cultural, morphological, and molecular characteristics and were identified as E. turcicum. The isolates were grouped into four groups based on colony color, i.e., olivaceous brown, blackish brown, whitish black, and grayish, and into two groups based on regular and irregular margins. The conidial shapes were observed to be elongated and spindle-shaped with protruding hilum, with conidial septa ranging from 2-12. Similarly, conidial length varied from 52.94 µm to 144.12 µm. ß-tubulin gene sequences analysis made it possible to verify the identities of fungal strains and the phylogenetic relationships of all isolates, which were clustered in the same clade. The ß-tubulin gene sequences of all the isolates showed a high level of similarity (100%) with reference isolates from GenBank accession numbers KU670342.1, KU670344.1, KU670343.1, KU670341.1, and KU670340.1. The findings of this study will serve as a baseline for future studies and will help to minimize yield losses.

Protein kinase A participates in hyphal and appressorial development by targeting Efg1-mediated transcription of a Rab GTPase in Setosphaeria turcica.

The cyclic adenosine monophosphate (cAMP) signalling pathway plays an important role in the regulation of the development and pathogenicity of filamentous fungi. cAMP-dependent protein kinase A (PKA) is the conserved element downstream of cAMP, and its diverse mechanisms in multiple filamentous fungi are not well known yet. In the present study, gene knockout mutants of two catalytic subunits of PKA (PKA-C) in Setosphaeria turcica were created to illustrate the regulatory mechanisms of PKA-Cs on the development and pathogenicity of S. turcica. As a result, StPkaC2 was proved to be the main contributor of PKA activity in S. turcica. In addition, it was found that both StPkaC1 and StPkaC2 were necessary for conidiation and invasive growth, while only StPkaC2 played a negative role in the regulation of filamentous growth. We reveal that only StPkaC2 could interact with the transcription factor StEfg1, and it inhibited the transcription of StRAB1, a Rab GTPase homologue coding gene in S. turcica, whereas StPkaC1 could specifically interact with a transcriptional regulator StFlo8, which could rescue the transcriptional inhibition of StEfg1 on StRAB1. We also demonstrated that StRAB1 could positively influence the biosynthesis of chitin in hyphae, thus changing the filamentous growth. Our findings clarify that StPkaC2 participates in chitin biosynthesis to modulate mycelium development by targeting the Efg1-mediated transcription of StRAB1, while StFlo8, interacting with StPkaC1, acts as a negative regulator during this process.