Important Plant Pathogenic Fungi Research Articles

Insights into the cumulative effect of Colletotrichum gloeosporioides and Fusarium acutatum causing anthracnose-twister disease complex of onion

Colletotrichum is an important plant pathogenic fungi that causes anthracnose/-twister disease in onion. This disease was prevalent in the monsoon season from August to November months and the symptoms were observed in most of the fields. This study aimed to investigate the pathogenicity and cumulative effect, if any of Colletotrichum gloeosporioides and Fusarium acutatum. The pot experiment was laid out to identify the cause responsible for inciting anthracnose-twister disease, whether the Colletotrichum or Fusarium or both, or the interaction of pathogens and GA3. The results of the pathogenicity test confirmed that C. gloeosporioides and F. acutatum are both pathogenic. C. gloeosporioides caused twisting symptoms independently, while F.acutatum independently caused only neck elongation. The independent application of GA3 did not produce any symptoms, however, increased the plant height. The combined treatment of C. gloeosporioides and F. acutatum caused twisting, which enhanced upon interaction with GA3 application giving synergistic effect. The acervuli were found in lesions infected with C. gloeosporioides after 8 days of inoculation on the neck and leaf blades. Symptoms were not observed in untreated control plants. Koch's postulates were confirmed by reisolating the same pathogens from the infected plants.

The APSES Transcription Factor SsStuA Regulating Cell Wall Integrity Is Essential for Sclerotia Formation and Pathogenicity in Sclerotinia sclerotiorum.

APSES (Asm1p, Phd1p, Sok2p, Efg1p, and StuAp) family transcription factors play crucial roles in various biological processes of fungi, however, their functional characterization in phytopathogenic fungi is limited. In this study, we explored the role of SsStuA, a typical APSES transcription factor, in the regulation of cell wall integrity (CWI), sclerotia formation and pathogenicity of Sclerotinia sclerotiorum, which is a globally important plant pathogenic fungus. A deficiency of SsStuA led to abnormal phosphorylation level of SsSmk3, the key gene SsAGM1 for UDP-GlcNAc synthesis was unable to respond to cell wall stress, and decreased tolerance to tebuconazole. In addition, ΔSsStuA was unable to form sclerotia but produced more compound appressoria. Nevertheless, the virulence of ΔSsStuA was significantly reduced due to the deficiency of the invasive hyphal growth and increased susceptibility to hydrogen peroxide. We also revealed that SsStuA could bind to the promoter of catalase family genes which regulate the expression of catalase genes. Furthermore, the level of reactive oxygen species (ROS) accumulation was found to be increased in ΔSsStuA. In summary, SsStuA, as a core transcription factor involved in the CWI pathway and ROS response, is required for vegetative growth, sclerotia formation, fungicide tolerance and the full virulence of S. sclerotiorum.

A New Biocontrol Agent Bacillus velezensis SF334 against Rubber Tree Fungal Leaf Anthracnose and Its Genome Analysis of Versatile Plant Probiotic Traits.

Natural rubber is an important national strategic and industrial raw material. The leaf anthracnose of rubber trees caused by the Colletotrichum species is one of the important factors restricting the yields of natural rubber. In this study, we isolated and identified strain Bacillus velezensis SF334, which exhibited significant antagonistic activity against both C. australisinense and C. siamense, the dominant species of Colletotrichum causing rubber tree leaf anthracnose in the Hainan province of China, from a pool of 223 bacterial strains. The cell suspensions of SF334 had a significant prevention effect for the leaf anthracnose of rubber trees, with an efficacy of 79.67% against C. siamense and 71.8% against C. australisinense. We demonstrated that SF334 can lead to the lysis of C. australisinense and C. siamense mycelia by causing mycelial expansion, resulting in mycelial rupture and subsequent death. B. velezensis SF334 also harbors some plant probiotic traits, such as secreting siderophore, protease, cellulase, pectinase, and the auxin of indole-3-acetic acid (IAA), and it has broad-spectrum antifungal activity against some important plant pathogenic fungi. The genome combined with comparative genomic analyses indicated that SF334 possesses most genes of the central metabolic and gene clusters of secondary metabolites in B. velezensis strains. To our knowledge, this is the first time a Bacillus velezensis strain has been reported as a promising biocontrol agent against the leaf anthracnose of rubber trees caused by C. siamense and C. australisinense. The results suggest that B. velezensis could be a potential candidate agent for the leaf anthracnose of rubber trees.

Genomics of the expanding pine pathogen Lecanosticta acicola reveals patterns of ongoing genetic admixture.

Lecanosticta acicola is a fungal pathogen causing severe defoliation, growth reduction, and even death in more than 70 conifer species. Despite the increasing incidence of this species, little is known about its population dynamics. Two divergent lineages have been described that have now been found together in regions of France and Spain, but it is unknown how these mixed populations evolve. Here we present the first reference genome for this important plant pathogenic fungi and use it to study the population genomics of 70 isolates from an affected forest in the north of Spain. We find signs of introgression between the two main lineages, indicating that active mating is occurring in this region which could propitiate the appearance of novel traits in this species. We also study the phenotypic differences across this population based on enzymatic activities on 20 compounds.

First Report of Cucurbita Blossom Blight and Fruit Rot Caused by Choanephora cucurbitarum in Mexico.

In December 2022, blossom blight, abortion, and soft rot of fruits were observed on Cucurbita pepo L. var. Zucchini in Mexico under greenhouse conditions (temperatures of 10 to 32°C and relative humidity up to 90%). The disease incidence in about 50 plants analyzed was around 70% with a severity of nearly 90%. Mycelial growth on flower petals and fruit rot with brown sporangiophores was observed. Ten disinfested fruit tissues in 1% NaClO for 5 min and then rinsed twice in distilled water from the lesion edges were placed on potato dextrose agar culture medium (PDA) supplemented with acid lactic and then, the morphological characterization was carried out in V8 agar medium. After 48 h of growth at 27°C, the colonies were pale yellow with diffuse cottony mycelia that were non-septate and hyaline and produced both sporangiophores bearing sporangiola and sporangia. The sporangiola were brown, ranged from ellipsoid to ovoid, and had longitudinal striations that measured 22.7 to 40.5 (29.8) μm x 16.08 to 21.9 (14.5) μm long and wide, respectively (n=100). The sporangia were subglobose, had a diameter of 127.2 to 281.09 (201.7) μm (n=50), and contained ovoid sporangiospores that measured 26.5 to 63.1 (46.7) μm x 20.07 to 34.7 (26.3) μm long and wide, respectively (n=100) which had hyaline appendages at the ends. Based on these characteristics, the fungus was identified as Choanephora cucurbitarum (Ji-Hyun et al. 2016). For molecular identification, DNA fragments for the internal transcribed spacer (ITS) and the large subunit rRNA 28S (LSU) regions for two representative strains (CCCFMx01 and CCCFMx02) were amplified and sequenced with the primer pairs ITS1-ITS4 and NL1-LR3 (White et al. 1990; Vilgalys and Hester 1990). The ITS and LSU sequences were deposited in GenBank database (Accession numbers OQ269823-24 and OQ269827-28, respectively) for both strains. The Blast alignment showed from 99.84 to 100% identity with Choanephora cucurbitarum strains JPC1 (MH041502, MH041504), CCUB1293 (MN897836), PLR2 (OL790293), and CBS 178.76 (JN206235, MT523842). To confirm the specie identification, the evolutionary analyses were conducted from the concatenated sequences of the ITS and LSU of C. cucurbitarum and other mucoralean species with the Maximum Likelihood method and Tamura-Nei model included in the software MEGA11. The pathogenicity test was demonstrated using five surface-sterilized zucchini fruits inoculated with a sporangiospores suspension containing a concentration of 1 x 105 esp/mL on two sites per fruit (20 µL each) that previously were wounded with a sterile needle. For fruit control, 20 µL of sterile water was used. Three days after inoculation under humidity conditions at 27°C, white mycelia and sporangiola growth with a soaked lesion were observed. That fruit damage was not observed on the control fruits. C.cucurbitarum was reisolated from lesions on PDA and V8 medium which was confirmed by morphological characterization fulfilling Koch's postulates. Blossom blight, abortion, and soft rot of fruits caused by C. cucurbitarum were observed on Cucurbita pepo and C. moschata in Slovenia and Sri Lanka (Žerjav and Schroers 2019; Emmanuel et al. 2021). This pathogen has the capability to infect a wide variety of plants worldwide (Kumar et al. 2022; Ryu et al. 2022). There are no reports of C. cucurbitarum causing agricultural losses in Mexico, and this is the first report causing the disease symptoms in Cucurbita pepo in this country; however, this fungus was found in the soil of papaya-producing areas and it is considered an important plant pathogenic fungus. Therefore, strategies for their control are highly recommended to avoid spreading the disease (Cruz-Lachica et al. 2018).

Ubiquitination of acetyltransferase Gcn5 contributes to fungal virulence in Fusarium graminearum.

The histone acetyltransferase general control non-depressible 5 (Gcn5) plays a critical role in the epigenetic landscape and chromatin modification for regulating a wide variety of biological events. However, the post-translational regulation of Gcn5 itself is poorly understood. Here, we found that Gcn5 was ubiquitinated and deubiquitinated by E3 ligase Tom1 and deubiquitinating enzyme Ubp14, respectively, in the important plant pathogenic fungus Fusarium graminearum. Tom1 interacted with Gcn5 in the nucleus and subsequently ubiquitinated Gcn5 mainly at K252 to accelerate protein degradation. Conversely, Ubp14 deubiquitinated Gcn5 and enhanced its stability. In the deletion mutant Δubp14, protein level of Gcn5 was significantly reduced and resulted in attenuated virulence in the fungus by affecting the mycotoxin production, autophagy process, and the penetration ability. Our findings indicate that Tom1 and Ubp14 show antagonistic functions in the control of the protein stability of Gcn5 via post-translational modification and highlight the importance of Tom1-Gcn5-Ubp14 circuit in the fungal virulence. IMPORTANCE Post-translational modification (PTM) enzymes have been reported to be involved in regulating numerous cellular processes. However, the modification of these PTM enzymes themselves is largely unknown. In this study, we found that the E3 ligase Tom1 and deubiquitinating enzyme Ubp14 contributed to the regulation of ubiquitination and deubiquitination of acetyltransferase Gcn5, respectively, in Fusarium graminearum, the causal agent of Fusarium head blight of cereals. Our findings provide deep insights into the modification of acetyltransferase Gcn5 and its dynamic regulation via ubiquitination and deubiquitination. To our knowledge, this work is the most comprehensive analysis of a regulatory network of ubiquitination that impinges on acetyltransferase in filamentous pathogens. Moreover, our findings are important because we present the novel roles of the Tom1-Gcn5-Ubp14 circuit in fungal virulence, providing novel possibilities and targets to control fungal diseases.

Sclerotinia sclerotiorum Agglutinin Modulates Sclerotial Development, Pathogenicity and Response to Abiotic and Biotic Stresses in Different Manners.

Sclerotinia sclerotiorum is an important plant pathogenic fungus of many crops. Our previous study identified the S. sclerotiorum agglutinin (SSA) that can be partially degraded by the serine protease CmSp1 from the mycoparasite Coniothyrium minitans. However, the biological functions of SSA in the pathogenicity of S. sclerotiorum and in its response to infection by C. minitans, as well as to environmental stresses, remain unknown. In this study, SSA disruption and complementary mutants were generated for characterization of its biological functions. Both the wild-type (WT) of S. sclerotiorum and the mutants were compared for growth and sclerotial formation on potato dextrose agar (PDA) and autoclaved carrot slices (ACS), for pathogenicity on oilseed rape, as well as for susceptibility to chemical stresses (NaCl, KCl, CaCl2, sorbitol, mannitol, sucrose, sodium dodecyl sulfate, H2O2) and to the mycoparasitism of C. minitans. The disruption mutants (ΔSSA-175, ΔSSA-178, ΔSSA-225) did not differ from the WT and the complementary mutant ΔSSA-178C in mycelial growth. However, compared to the WT and ΔSSA-178C, the disruption mutants formed immature sclerotia on PDA, and produced less but larger sclerotia on ACS; they became less sensitive to the eight investigated chemical stresses, but more aggressive in infecting leaves of oilseed rape, and more susceptible to mycoparasitism by C. minitans. These results suggest that SSA positively regulates sclerotial development and resistance to C. minitans mycoparasitism, but negatively regulates pathogenicity and resistance to chemical stresses.

The STRIPAK complex orchestrates cell wall integrity signalling to govern the fungal development and virulence of Fusarium graminearum.

Striatin-interacting phosphatases and kinases (STRIPAKs) are evolutionarily conserved supramolecular complexes that control various important cellular processes such as signal transduction and development. However, the role of the STRIPAK complex in pathogenic fungi remains elusive. In this study, the components and function of the STRIPAK complex were investigated in Fusarium graminearum, an important plant-pathogenic fungus. The results obtained from bioinformatic analyses and the protein-protein interactome suggested that the fungal STRIPAK complex consisted of six proteins: Ham2, Ham3, Ham4, PP2Aa, Ppg1, and Mob3. Deletion mutations of individual components of the STRIPAK complex were created, and observed to causea significant reduction in fungal vegetative growth and sexual development, and dramatically attenuae virulence, excluding the essential gene PP2Aa. Further results revealed that the STRIPAK complex interacted with the mitogen-activated protein kinase Mgv1, a key component in the cell wall integrity pathway, subsequently regulating the phosphorylation level and nuclear accumulation of Mgv1 to control the fungal stress response and virulence. Our results also suggested that the STRIPAK complex was interconnected with the target of rapamycin pathway through Tap42-PP2A cascade. Taken together, our findings revealed that the STRIPAK complex orchestrates cell wall integrity signalling to govern the fungal development and virulence of F. graminearum and highlighted the importance of the STRIPAK complex in fungal virulence.

Antagonistic transcriptome profile reveals potential mechanisms of action on Xanthomonas oryzae pv. oryzicola by the cell-free supernatants of Bacillus velezensis 504, a versatile plant probiotic bacterium.

Bacterial leaf streak (BLS) of rice is a severe disease caused by the bacterial pathogen Xanthomonas oryzae pv. oryzicola (Xoc) that has gradually become the fourth major disease on rice in some rice-growing regions in southern China. Previously, we isolated a Bacillus velezensis strain 504 that exhibited apparent antagonistic activity against the Xoc wild-type strain RS105, and found that B. velezensis 504 was a potential biocontrol agent for BLS. However, the underlying mechanisms of antagonism and biocontrol are not completely understood. Here we mine the genomic data of B. velezensis 504, and the comparative transcriptomic data of Xoc RS105 treated by the cell-free supernatants (CFSs) of B. velezensis 504 to define differentially expressed genes (DEGs). We show that B. velezensis 504 shares over 89% conserved genes with FZB42 and SQR9, two representative model strains of B. velezensis, but 504 is more closely related to FZB42 than SQR9, as well as B. velezensis 504 possesses the secondary metabolite gene clusters encoding the essential anti-Xoc agents difficidin and bacilysin. We conclude that approximately 77% of Xoc RS105 coding sequences are differentially expressed by the CFSs of B. velezensis 504, which significantly downregulates genes involved in signal transduction, oxidative phosphorylation, transmembrane transport, cell motility, cell division, DNA translation, and five physiological metabolisms, as well as depresses an additional set of virulence-associated genes encoding the type III secretion, type II secretion system, type VI secretion system, type IV pilus, lipopolysaccharides and exopolysaccharides. We also show that B. velezensis 504 is a potential biocontrol agent for bacterial blight of rice exhibiting relative control efficiencies over 70% on two susceptible cultivars, and can efficiently antagonize against some important plant pathogenic fungi including Colletotrichum siamense and C. australisinense that are thought to be the two dominant pathogenic species causing leaf anthracnose of rubber tree in Hainan province of China. B. velezensis 504 also harbors some characteristics of plant growth-promoting rhizobacterium such as secreting protease and siderophore, and stimulating plant growth. This study reveals the potential biocontrol mechanisms of B. velezensis against BLS, and also suggests that B. velezensis 504 is a versatile plant probiotic bacterium.

Genetic and Transcriptional Regulatory Mechanisms of Lipase Activity in the Plant Pathogenic Fungus Fusarium graminearum.

Lipases, which catalyze the hydrolysis of long-chain triglycerides, diglycerides, and monoglycerides into free fatty acids and glycerol, participate in various biological pathways in fungi. In this study, we examined the biological functions and regulatory mechanisms of fungal lipases via two approaches. First, we performed a systemic functional characterization of 86 putative lipase-encoding genes in the plant-pathogenic fungus Fusarium graminearum. The phenotypes were assayed for vegetative growth, asexual and sexual reproduction, stress responses, pathogenicity, mycotoxin production, and lipase activity. Most mutants were normal in the assessed phenotypes, implying overlapping roles for lipases in F. graminearum. In particular, FgLip1 and Fgl1 were revealed as core extracellular lipases in F. graminearum. Second, we examined the lipase activity of previously constructed transcription factor (TF) mutants of F. graminearum and identified three TFs and one histone acetyltransferase that significantly affect lipase activity. The relative transcript levels of FgLIP1 and FGL1 were markedly reduced or enhanced in these TF mutants. Among them, Gzzc258 was identified as a key lipase regulator that is also involved in the induction of lipase activity during sexual reproduction. To our knowledge, this study is the first comprehensive functional analysis of fungal lipases and provides significant insights into the genetic and regulatory mechanisms underlying lipases in fungi. IMPORTANCE Fusarium graminearum is an economically important plant-pathogenic fungus that causes Fusarium head blight (FHB) on wheat and barley. Here, we constructed a gene knockout mutant library of 86 putative lipase-encoding genes and established a comprehensive phenotypic database of the mutants. Among them, we found that FgLip1 and Fgl1 act as core extracellular lipases in this pathogen. Moreover, several putative transcription factors (TFs) that regulate the lipase activities in F. graminearum were identified. The disruption mutants of F. graminearum-lipase regulatory TFs all showed defects in sexual reproduction, which implies a strong relationship between sexual development and lipase activity in this fungus. These findings provide valuable insights into the genetic mechanisms regulating lipase activity as well as its importance to the developmental stages of this plant-pathogenic fungus.

FgGyp8 as a putative FgRab1 GAP is required for growth and pathogenesis by regulating FgSnc1-mediated secretory vesicles fusion in Fusarium graminearum

Fusarium graminearum is an important plant pathogenic fungus that causes disease and yield reduction in many cereal crops, such as wheat and barley. Gyp8 stimulates GTP hydrolysis on Ypt1 in yeast. However, the functions of Gyp8 in plant pathogenic fungi are still unknown. In this study, we investigated the roles of FgGyp8 in F. graminearum by genetic and pathological analyses. Through gene knockout and phenotypic analyses, we found that FgGyp8 is required for vegetative growth in F. graminearum. The conidiation, conidial size and number of septa per conidium of ΔFggyp8 mutant are significantly reduced when compared to the wild type PH-1. Furthermore, FgGyp8 is crucial for pathogenicity on wheat coleoptiles and wheat heads. FgGyp8 contains a conserved TBC domain. Domain deletion analysis showed that the TBC domain, C- and N-terminal regions of FgGyp8 are all important for its biological functions in F. graminearum. Moreover, we showed that FgGyp8 catalyzes the hydrolysis of the GTP on FgRab1 to GDP in vitro, indicating that FgGyp8 is a GTPase-activating protein (GAP) for FgRab1. In addition, we demonstrated that FgGyp8 is required for FgSnc1-mediated fusion of secretory vesicles with the plasma membrane in F. graminearum. Finally, we showed that FgGyp8 has functional redundancy with another FgRab1 GAP, FgGyp1, in F. graminearum. Taken together, we conclude that FgGyp8 is required for vegetative growth, conidiogenesis, pathogenicity and acts as a GAP for FgRab1 in F. graminearum.

Components of TOR and MAP kinase signaling control chemotropism and pathogenicity in the fungal pathogen Verticillium dahliae

Filamentous fungi can sense useful resources and hazards in their environment and direct growth of their hyphae accordingly. Chemotropism ensures access to nutrients, contact with other individuals (e.g., for mating), and interaction with hosts in the case of pathogens. Previous studies have revealed a complex chemotropic sensing landscape during host-pathogen interactions, but the underlying molecular machinery remains poorly characterized. Here we studied mechanisms controlling directed hyphal growth of the important plant-pathogenic fungus Verticillium dahliae towards different chemoattractants. We found that the homologs of the Rag GTPase Gtr1 and the GTPase-activating protein Tsc2, an activator and a repressor of the TOR kinase respectively, play important roles in hyphal chemotropism towards nutrients, plant-derived signals, and heterologous α-pheromone of Fusarium oxysporum. Furthermore, important roles of these regulators were identified in fungal development and pathogenicity. We also found that the mitogen-activated protein kinase (MAPK) Fus3 is required for chemotropism towards nutrients, while the G protein-coupled receptor (GPCR) Ste2 and the MAPK Slt2 control chemosensing of plant-derived signals and α-pheromone. Our study establishes V. dahliae as a suitable model system for the analysis of fungal chemotropism and discovers new components of chemotropic signaling during growth and host-pathogen interactions of V. dahliae.

Design, synthesis, antifungal activity and molecular docking of ring-opened pimprinine derivative containing (thio)amide structure.

To obtain new environmentally friendly fungicides, we used the natural product pimprinine as the lead compound, and designed and synthesized two series of ring-opening derivatives of pimprinine containing amide/thioamide. We then studied their antifungal activity against six common plant pathogenic fungi in vitro. Most of the target compounds have good antifungal activity against six important plant pathogenic fungi in vitro. At a concentration of 50 μg ml-1 , compound 3o showed prominent antifungal effects on Alternaria solani and Rhioctornia solani, with inhibition rates of 91.8% and 97.4%, and a 50% effective concentration (EC50 ) of 6.2255 and 0.6969 μg ml-1 respectively. The EC50 of compound 3o against Alternaria solani was significantly lower than that of boscalid (13.0380 μg ml-1 ) and flutriafol (11.9057 μg ml-1 ). In addition, compound 3o had good antifungal activity against Sclerotinia sclerotiorum, cucumber powdery mildew, cucumber Botrytis cinerea and Phytophthora capsici in vivo; the antifungal activity of compound 3o against cucumber Botrytis cinerea is 91.7%. At the same time, docking results for highly active compound 3o with the presumed target succinate dehydrogenase and the molecular docking prediction scores of all compounds further indicate its possible antifungal activity mechanism. The designed and optimized derivative 3o of ring-opening pimprinine has good antifungal activity and can be used as a new antifungal drug for further research. © 2023 Society of Chemical Industry.

An Accurate, Affordable, and Precise Resazurin-Based Digital Imaging Colorimetric Assay for the Assessment of Fungicide Sensitivity Status of Fungal Populations

This study aimed at the development and validation of an accurate, more affordable, and precise digital imaging resazurin-based fungicide sensitivity colorimetric assay (COL-assay) for fungal plant pathogens from the genera Mycosphaerella and Pyricularia. This proposed digital imaging assay was based on colorimetric estimates of resazurin reduction, which was used as a metabolic indicator of fungal respiration activity on microplate cultures. As fungal model systems, we used the yellow and black Sigatoka pathogens [Mycosphaerella musicola (Mm) and M. fijiensis (Mf), respectively] and the wheat blast pathogen, Pyricularia oryzae Triticum lineage (PoTl), which were previously characterized for QoI, DMI, and SDHI fungicide sensitivity. We then compared the classical spectrophotometry detection assay (SPEC-assay) with the proposed COL-assay based on the analyses of digital images of the microplates’ cultures captured with mobile phone cameras on a handmade trans-illuminator built for poorly equipped labs. Qualitatively, in terms of accuracy, there was full correspondence between the SPEC-assay and the COL-assay according to the fungal EC50 or the relative growth classes on QoI, SDHI, and DMI fungicides for both Mycosphaerella and Pyricularia pathogens. We also observed a strong to very strong correlation coefficient between the COL-assay and the SPEC-assay fungicide sensitivity values for the QoI azoxystrobin, the SDHI fluxapyroxad, and the DMI tebuconazole. Our conclusion was that the COL-assay had a similar accuracy as the SPEC-assay (i.e., resulted in similar fungicide-sensitivity categories for both resistant or sensitive fungal isolates) and high precision. By openly sharing here the COL-assay’s full methodology, and the blueprints of the handmade trans-illuminator, we foresee its adoption by poorly equipped labs throughout the country as an affordable venue for monitoring the fungicide resistance status of populations of important fungal plant pathogens such as M. fijiensis, M. musicola, and P. oryzae Triticum and Oryza lineages.

Rigidoporus microporus and the white root rot disease of rubber

AbstractRigidoporus microporus is an economically important plant pathogenic fungus causing particularly severe losses to the rubber industry worldwide. The pathogen is responsible for white root rot (WRR) disease, infecting the host roots via white fibrous mycelia, causing vascular disfunction, and visible symptoms including leaf discolouration and dieback in severely infected trees. The final stage of the disease is characterized by the appearance of basidiocarps at the tree collar. The development of WRR in rubber plantations is dependent on fungal diversity in the soil, pH, temperature, and cation levels. Several ‘‐omics’ approaches have been undertaken to understand how R. microporus functions with the objective, ultimately, to control WRR. Unfortunately, no resistant rubber clone has been identified to date. The disease is managed through physical and chemical methods that are laborious and negatively impact the environment, respectively. Recent developments in research on R. microporus shed light on potential sustainable routes to WRR disease control using beneficial microorganisms and natural compounds. This review discusses the characteristics of R. microporus isolates from different geographical origins, the pathogenicity and virulence mechanisms of the necrotrophic fungal pathogen, factors that influence the development of WRR, recent findings from the multi‐omics studies, and control methods that are available to combat this economically important pathogen.

Diversity of Antimicrobial Peptide Genes in Bacillus from the Andaman and Nicobar Islands: Untapped Island Microbial Diversity for Disease Management in Crop Plants.

Taxonomic and functional characterization of a total of 90 bacterial isolates representing bulk and rhizosphere soils of diverse niches of Andaman and Nicobar Islands, India were carried out. Twelve bacterial isolates were found promising for the biological suppression of agriculturally important fungal and bacterial plant pathogens such as Ralstonia solanacearum, Xanthomonas oryzae pv. oryzae, and Colletotrichum gloeosporioides. The 16S rRNA gene sequence analysis revealed their identity as belonging to Bacillus subtilis, Bacillus amyloliquefaciens, and Lysinibacillus sphaericus. The isolates were positive for plant growth promotion (PGP) traits including siderophore production, and nutrient solubilization especially phosphorous, zinc, and potassium. Interestingly, the PCR test confirmed the presence of 62 antimicrobial peptides (AMP) biosynthesis genes specific to the genus Bacillus. Whilst all tested species of Bacillus harboured the bacD biosynthesis gene, the B. subtilis (Ba_Abi), and B. amyloliquefaciens (Ba_Abi) harboured the maximum AMP biosynthesis genes analysed in the study. Upon in planta evaluation, the biocontrol potential of the bacterial isolates against leaf spot disease of chilli was observed. The study culminated in the isolation and identification of diverse Bacillus species for exploitation as bioinoculants for plant health management programmes.

A novel victorivirus isolated from the tobacco spot blight fungus Stagonosporopsis cucurbitacearum in China.

Stagonosporopsis cucurbitacearum is an important plant-pathogenic fungus that causes stem and leaf blight diseases in a variety of crops. Here, we report the characterization of a novel victorivirus, tentatively named "Stagonosporopsis cucurbitacearum victorivirus 1" (ScVV-1), isolated from the S. cucurbitacearum isolate M-7. The ScVV-1 genome is 5,165bp in length with a predicted GC content of 60.1% and contains two large open reading frames (ORF 1 and ORF2) encoding putative proteins that share significant sequence similarity with coat proteins (CPs) and RNA-dependent RNA polymerases (RdRps) of mycoviruses of the family Totiviridae. The ScVV-1 RdRp appears to be translated using a stop-initiation pentanucleotide UAAUG sequence. Phylogenetic analysis based on CP and RdRp amino acid (aa) sequences both indicated that ScVV-1 belongs to the genus Victorivirus in the family Totiviridae. To our knowledge, this is the first full-length genome sequence of a victorivirus infecting S. cucurbitacearum.

The effects of different ambient pH on the pathogenicity of Fusarium sulphureum and reactive oxygen species metabolism in F. sulphureum inoculation muskmelon fruits

Fusarium sulphureum is an important plant pathogenic fungus, which can lead to fusarium rot of muskmelon. During fungal infection host plant, environmental pH plays crucial role by secreting acid or base to regulates the arsenals of enzymes which contribute to fungal pathogenicity. In this study, in order to define the acid-alkali property of F. sulphureum and determinate the effect of ambient pH values on reactive oxygen species (ROS) metabolism and pathogenicity during F. sulphureum infection muskmelon fruits. F. sulphureum spore suspension was modified with different pH values buffer solution and then was inoculated muskmelon fruits. The change of pH value in lesion tissue, the pathogenicity, as well as ROS metabolism in inoculated muskmelon fruits were investigated and analyzed. The results indicated that the pH value of lesion tissue increased from 5.92 on 0 day to 9.24 on 7 day after inoculation, the lesion diameter in fruit inoculated with spore suspension at pH 6 was significantly higher than those at pH 3, 5, 7 and 9. Meanwhile, the inoculated fruits with pH 6 spore suspension showed higher NOX activity, O2−. production and H2O2 content, cell membrane permeability, as well as Malondialdehyde （MDA）content. However, the activities of antioxidant enzyme (such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT)) and the activities of key enzymes and the contents of non-enzymatic antioxidants involved in ascorbate-glutathione cycle (AsA-GSH) were remarkedly increased in fruits inoculated with spore suspension at pH 3, comparing with pH 5, 6, 7, 9. Based on the present results, F. sulphureum inoculation led to pH increasing of the tissue around the inoculation site, and weak alkaline environment increased the pathogenicity of F. sulphureum infection muskmelon by stimulating ROS metabolism.

Xenorhabdus szentirmaii Bakteri Metabolitlerinin Bazı Önemli Bitki Patojeni Funguslara Karşı Antifungal Etkilerinin Belirlenmesi

This study was carried out to determine the antifungal effects of the supernatant produced by the bacterium Xenorhabdus szentirmaii, which is associated with soil-inhabiting entomopathogenic nematodes, on important plant pathogenic fungi, Fusarium verticilliodes, Fusarium oxysporum f.sp lycopersici, Fusarium oxysporum f.sp radicis lycopersici, Botrytis cinerea, Sclerotinia sclerotiorum and Phytophthora nicotianae. The effects of 1, 3, 5 and 7% concentrations of the supernatant produced by X. szentirmaii bacteria on mycelium growth of the fungal pathogens was determined at 3, 7 and 14 days after application (dap). The effects of the supernatant on the formation of sporangia and the emergence of zoospores from the sporangium of Phytophthora nicotianae was also determined. According to the results of the study, the highest dose of the supernatant (7% concentration) prevented mycelium development of all tested pathogens. The effect of the supernatant decreased at 14 dap against F. verticilliodes, F. oxysporum f.sp lycopersici, F. oxysporum f.sp radicis lycopersici. The antifungal effect on S. sclerotiorum completely disappeared in 14 days whereas the effect continued even in 14 days against B. cinerea and P. nicotianae. Xenorhabdus szentirmaii supernatant at a concentration of 7% was highly efficacious on the formation of sporangia and the emergence of zoospores within the sporangium of P. nicotianae at a concentration of 7%.

New molecular markers for distinguishing the main phylogenetic lineages within Alternaria section Alternaria

Several members of Alternaria section Alternaria are economically important plant-pathogenic fungi that cause disease on a wide range of host types and plant tissues. The production of Alternaria-derived mycotoxins can lead to significant post-harvest losses due to contamination of agricultural products. Multiple Alternaria species are listed as regulated organisms, which are monitored by international plant protection programmes. These taxa often share high levels of both morphological and phylogenetic similarity, and the establishment of molecular markers that are able to distinguish among them unambiguously has proven to be quite challenging. Previously, we examined fine-scale genome-wide phylogenetic patterns and proposed a list of candidate genes for development into informative markers that are diagnostic for the main section Alternaria lineages. Here, we design primer sets and sequence three new markers (ASA-05, ASA-10, ASA-19) in order to evaluate their diagnostic performance. Sequence data for 49 new independent test taxa were combined with existing data for 38 taxa, and phylogenetic analyses revealed that, in general, the new molecular markers consistently classified section Alternaria strains into one of the main lineages. A discrepancy among the three markers for lineage assignment was observed only for one strain. We also sequenced a commonly used marker in molecular phylogenetics of fungi (rpb2, RNA polymerase II second largest subunit) and found it was outperformed by all three of the new markers. We suggest that an ASA marker presented here could form the basis of a convenient one-locus test for rapid and routine diagnostic screening of unknown section Alternaria strains.