Soil-borne Fungal Pathogens Research Articles

CAP superfamily proteins (VdPRYs) manipulate plant immunity and contribute to the virulence of Verticillium dahliae

ABSTRACT CAP (cysteine-rich secretory proteins, antigen5, pathogenesis-related proteins) superfamily proteins are widely distributed, can be subdivided into 11 subfamilies, and form a unique branch in fungi, named PRY proteins. Verticillium dahliae is a soil-borne fungal pathogen of vascular plants that causes plant Verticillium wilt. However, the roles of CAP superfamily proteins in this fungus is unclear. Here, four CAP superfamily members with a conserved domain were identified in V. dahliae: VdPRY1, VdPRY2, VdPRY3, and VdPRY4. VdPRY1 and VdPRY3 were found to be key in suppressing plant immune responses. Moreover, these four members are highly expressed during early infection of cotton by V. dahliae. Deleting VdPRY1, VdPRY2, or VdPRY3 reduced the fungus’s ability to cause disease, but VdPRY4 deletion did not affect virulence. Deletion of any of four members did not impact fungal growth or carbon source use. Yeast two-hybrid experiments suggest that these proteins may function through interactions with each other. This investigation has, for the initial time, elucidated the pivotal roles of V. dahliae CAP superfamily proteins in inhibiting plant immunity and exerting virulence during interaction with the host plant.

Morphological, Biochemical and Molecular Characterization of Plant Growth Promoting Rhizobacteria with Synergistic Effect against Fusarium oxysporum

Currently, plant growth-promoting rhizosphere bacteria (PGPR) are heavily exploited as microbial inoculants in agricultural production among which Pseudomonas sp. and Bacillus sp. are widely used in plant growth promotion and disease control as excellent inoculum strains. Plant growth-promoting rhizobacteria has the potential to be used in agriculture to promote plant growth and health through various mechanisms. Therefore, identifying novel strains tailored to specific agricultural requirements continues to be a key area of research. In the present research work, the strains of PGPR isolated from soybean rhizosphere at Department of Plant Pathology and characterized at morphological, biochemical and molecular level. These strains are now used in ‘Biomix’ formulation a microbial consortium product developed at the Department of Plant Pathology VNMKV, Parbhani to benefit the farmers. Studying the molecular, biochemical, and morphological traits of these microbial strains which are now involved in VNMKV produced ‘Biomix formulation is crucial to maintain its consistency, purity and optimize its formulation. Molecular characterization and phylogenetic analysis will ensure accurate identification and classification of the isolates, enhancing the biofertilizer's effectiveness in agriculture. All three isolates have been identified as per Bergey’s Manual for the determination of bacteriology. All isolates tested positive for starch hydrolysis, citrate utilization, catalase activity, indole production, gelatine hydrolysis, and tetrazolium reduction. Additionally, the 16S rDNA gene sequences of the various potential isolates were validated as P. fluorescens VNMKV1, P. putida VNMKV1, and B. subtilis VNMKV1. The potential of these novel strains of plant growth-promoting rhizobacteria as biocontrol agents was explored by screening against soil-borne pathogenic fungi highlighting their potential to be used as bioinoculant agents.

Detection of novel pathogenic variants of Fusarium oxysporum f. sp. lactucae in California

AbstractFusarium wilt, caused by the soilborne fungal pathogen Fusarium oxysporum f. sp. lactucae (FOL), is an economically important disease of lettuce (Lactuca sativa) throughout the world. Four pathogenic races of FOL are reported, though only race 1 is known to exist in the United States. Recently, Californian lettuce growers have expressed increasing concern about Fusarium wilt, as some race 1‐resistant cultivars have exhibited susceptibility, and some susceptible cultivars have displayed reduced disease severity. To determine whether such changes in disease patterns are responses to potentially novel variants, we collected infected plants from commercial fields in the Salinas Valley and Santa Maria regions, isolated the fungus, and conducted a series of pathogenicity tests using a set of FOL race differential cultivars over 2 years (2022 and 2023) in controlled conditions. Pathogenicity tests revealed two new FOL variants that elicited novel disease reaction patterns on the set of differential cultivars that have not been previously described. Isolates Fol621 and Fol621s were less aggressive on race 1‐susceptible Banchu Red Fire, designated Variant‐1. Isolate VSP‐0916 incited severe Fusarium wilt on race 1‐resistant Costa Rica No. 4, designated Variant‐2. Moreover, VSP‐0916 exhibited high aggressiveness and the ability to induce disease in other race 1‐resistant cultivars. This study is the first documentation, to our knowledge, of the occurrence of FOL‐inciting Fusarium wilt on race 1‐resistant lettuce in the United States. Our work provides valuable information critical for the development of Fusarium wilt management strategies, including broad‐spectrum resistance breeding efforts against multiple FOL races.

Inoculation of Bacillus velezensis Bv-116 and its bio-organic fertilizer serve as an environmental friendly biocontrol strategy against cucumber Fusarium wilt.

The aim of this study was to evaluate the effects of Bacillus velezensis Bv-116 and its bio-organic fertilizer on the control of cucumber Fusarium wilt caused by Fusarium oxysporum f. sp. Cucumerinum (FOC), the promotion of growth of cucumber seedlings, and the soil microbial community. B. velezensis Bv-116 exhibited an inhibition rate of 84.93% against FOC, as well as broad-spectrum inhibitory activities against other soil-borne plant pathogenic fungi. Fermentation products of B. velezensis Bv-116 destroyed the cell structure of FOC and inhibited the growth of FOC mycelium. These products were identified as volatile antimicrobial gases, proteases and cellulases. In the greenhouse pot experiment, both B. velezensis Bv-116 and its bio-organic fertilizer exhibited significant promoting effects on cucumber growth, and a significant reduction in the incidence and disease severity index of cucumber wilt (p < 0.05). Analysis of the microbial community structure of cucumber rhizosphere soil revealed that inoculation of B. velezensis Bv-116 and its bio-organic fertilizer increased the abundance of genera with biocontrol capabilities against plant pathogens. In addition, inoculation of the bio-organic fertilizer reversed the excessive proliferation of Fusarium and Acidobacteria. Our results suggest the potential of inoculating B. velezensis Bv-116 and its bio-organic fertilizer as an environmentally friendly biocontrol strategy against cucumber wilt.

A Novel Secreted Protein of Fusarium oxysporum Promotes Infection by Inhibiting PR-5 Protein in Plant.

Fusarium oxysporum, an important soilborne fungal pathogen that causes serious Fusarium wilt disease, secretes diverse effectors during the infection. In this study, we identified a novel secreted cysteine-rich protein, FolSCP1, which contains unknown protein functional domain. Here, we characterized FolSCP1 as a secreted virulence factor that promotes the pathogen infection of host plants by inhibiting diverse plant defence responses. FolSCP1 interacted with the pathogenesis-related 5 (PR-5) protein SlPR5, a positive regulator of tomato plant immunity against multiple tomato pathogens, and effectively attenuated the antifungal activity of the tomato PR-5 protein. FoSCP1, a homologue of FolSCP1, was secreted by a F. oxysporum isolate from infected tobacco and targeted the tobacco PR-5 protein NtPR5 to suppress plant defence for further infection. In summary, our study revealed a fungal virulence strategy in which F. oxysporum secrete effectors that interfere with plant immunity by binding to the PR-5 protein of the host plant and inhibiting its biological activity, thereby promoting fungal infection.

Enhancing Date Palm (Phoenix dactylifera L.) Resilience: Strategies Against Bayoud Spread in Moroccan Oases

Abstract “Bayoud”, a vascular disease caused by the soilborne fungal pathogen Fusarium oxysporum f. sp. albedinis (Foa), is the principal enemy of palm trees that puts at stake the future of the date palm industry in Morocco. The parasite attacks the palms through the roots and colonizes the entire vascular system, leading to wilt and ultimately palm death. Effective disease management is a major step towards the success of all sustainable development projects in the oasis. This review delineates diverse strategies for managing Bayoud disease, including prophylactic measures, chemical and cultural controls, genetic selection for resistant varieties, and exploration of biochemical and molecular markers. An integrated management approach was proposed that underscores early detection, sustainable agricultural practices, genetic resistance, biological control, community involvement, and continuous research and development. To contain the disease in already-infested areas and prevent its spread to new date palm plantations in oases.

Exogenous Application of dsRNA—Inducing Silencing of the Fusarium oxysporum Tup1 Gene and Reducing Its Virulence

Fusarium oxysporum is a widespread soil-borne fungal pathogen that can infect various plants, causing wilt and root rot diseases. The root rot disease of Atractylodes macrocephala caused by F. oxysporum is among the most serious diseases associated with continuous cropping, significantly hindering its sustainable development. In this study, we aimed to investigate the effect of exogenous application of double-stranded RNA (dsRNA) on silencing the F. oxysporum Tup1 gene to reduce its virulence and to evaluate its potential application in controlling root rot disease in A. macrocephala. The Tup1 gene was amplified from the F. oxysporum genome, and different lengths of Tup1-dsRNA were designed and synthesized. The uptake of dsRNA by the fungus was verified using Tup1-dsRNA labeled with fluorescein, and in vitro dsRNA treatment experiments were conducted to assess its impact on the growth and virulence of F. oxysporum. Additionally, Tup1-dsRNA was applied to the roots of A. macrocephala to evaluate its effectiveness in controlling root rot disease. The experimental results showed that F. oxysporum could effectively uptake exogenously applied Tup1-dsRNA, significantly reducing Tup1 gene expression. All lengths of Tup1-dsRNA inhibited fungal growth and caused morphological changes in the fungal hyphae. Further plant experiments and Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) analysis indicated that Tup1-dsRNA treatment significantly reduced the incidence of root rot disease in A. macrocephala, which was supported by the reduction in peroxidase (POD) and catalase (CAT) enzyme activities, malondialdehyde (MDA) content, and proline (Pro) levels in treated root tissues. This study demonstrated that exogenous dsRNA could reduce the virulence of F. oxysporum by silencing the Tup1 gene and effectively mitigate the root rot disease it causes in A. macrocephala. The successful application of Tup1-dsRNA provided strong evidence for the potential of RNA interference (RNAi) technology in plant disease control. Future research could further optimize the design and application of dsRNA to enhance its practical value in agriculture.

Biosynthesis and Signaling of Strigolactones Act Synergistically With That of ABA and JA to Enhance Verticillium dahliae Resistance in Cotton (Gossypium hirsutum L.).

Verticillium wilt (VW) caused by the soil-borne fungal pathogen Verticillium dahliae reduces cotton productivity and quality. Numerous studies have explored the genetic and molecular mechanisms regulating VW resistance in cotton, but the role and mechanism of strigolactone (SL) is still elusive. We investigated the function of SL in cotton's immune response to V. dahliae infection by exogenously applying SL analog, blocking or enhancing biosynthesis of endogenous SLs in combination with comparative transcriptome analysis and by exploring cross-talk between SL and other phytohormones. Silencing GhDWARF27 and applying the SL analog GR24 or overexpressing GhDWARF27 decreased and enhanced V. dahliae resistance, respectively. Transcriptome analysis revealed SL-mediated activation of abscisic acid (ABA) and jasmonic acid (JA) biosynthesis and signaling pathways. Enhanced ABA biosynthesis and signaling led to increased activity of antioxidant enzymes and reduced buildup of excess reactive oxygen species. Enhanced JA biosynthesis and signaling facilitated transcription of JA-dependent disease resistance genes. One of the components of the SL signal transduction pathway, GhD53, was found to interact with GhNCED5 and GhLOX2, the key enzymes of ABA and JA biosynthesis, respectively. We revealed the molecular mechanism underlying SL-enabled V. dahliae resistance and provided potential solutions for improving VW resistance in cotton.

Tailor-made green composts with suppressive properties against tracheofusariosis of wild rocket (Diplotaxis tenuifolia): Useful option for sustainable circular horticulture

Intensive cultivation systems of wild rocket (Diplotaxis tenuifolia) are threatened by the outbreak of tracheofusariosis caused by the soil-borne fungal pathogen Fusarium oxysporum f. sp. raphani. The disease is difficult to eradicate by curative methods due to the endophytic progression of the pathogen. Suppressive composts can help to prevent this adversity by providing organic matter carrying chemical and biological components that can directly and/or indirectly interfere with the plant-pathogen system. In this study, a collection of seven promising composts from different selected green feedstocks were investigated for their suppressive properties towards wild rocket Fusarium wilting. In planta assays identified two composts, obtained from olive tree pruning + plant and tomato residues, and from olive waste + straw + wool residues, with the highest levels of suppressiveness, influenced by both application dose and plant development stage. These active composts showed overlapping 13C NMR profiles as in vivo suppressiveness, being positively correlated with the organic aliphatic alkyl, O-Aryl and carboxyl C fractions and subsequently with the hydrophobic index and alkyl ratio, and negatively with the lignin ratio. On the contrary, although biotic compost components showed antifungal effects, biological properties did not correlate with in planta wilting suppression. Consistently, reflectance spectroscopy in the Vis-NIR-SWIR highlighted the clustering of samples depending on their feedstock and chemical composition. The results allow the hypothesis of a plant-mediated suppression mechanism. The composting of crop residues improves the circularity of horticulture and makes it possible to produce, from selected matrices, tailored compost towards the sustainable cultivation of wild rocket.

The Kelch Repeat Protein VdKeR1 Is Essential for Development, Ergosterol Metabolism, and Virulence in Verticillium dahliae.

Verticillium dahliae is a soil-borne fungal pathogen that can cause severe vascular wilt in many plant species. Kelch repeat proteins are essential for fungal growth, resistance, and virulence. However, the function of the Kelch repeat protein family in V. dahliae is unclear. In this study, a Kelch repeat domain-containing protein DK185_4252 (VdLs.17 VDAG_08647) included in the conserved VdPKS9 gene cluster was identified and named VdKeR1. Phylogenetic analysis demonstrated a high degree of evolutionary conservation of VdKeR1 and its homologs among fungi. The experimental results showed that the absence of VdKeR1 impaired vegetative growth, microsclerotia development, and pathogenicity of V. dahliae. Osmotic and cell wall stress analyses suggested that VdKeR1-deleted mutants were more tolerant to NaCl, sorbitol, CR, and CFW, while more sensitive to H2O2 and SDS. In addition, analyses of the relative expression level of sqe and the content of squalene and ergosterol showed that VdKeR1 mediates the synthesis of squalene and ergosterol by positively regulating the activity of squalene epoxidase. In conclusion, these results indicated that VdKeR1 was involved in the growth, stress resistance, pathogenicity, and ergosterol metabolism of V. dahliae. Investigating VdKeR1 provided theoretical and experimental foundations for subsequent control of Verticillium wilt.

Phylogenetic relatedness and habitat affect seedling abundance of a mid-montane humid evergreen broad-leaved forest in the Gaoligong Mountains, Southwestern China

The neighborhood effect has become an important framework with which to study the mechanisms that maintain the coexistence of tree species. Phylogenetic relatedness among neighboring plants directly affects species coexistence and the maintenance of tree diversity. And some studies have reported that seedling performance is negatively correlated with phylogenetic relatedness, which termed phylogenetic negative density dependence. Soil-borne fungal pathogens affected seedling performance of phylogenetically related host species, i.e., phylogenetic Janzen–Connell effect. Seedlings may be particularly vulnerable to habitat and neighbor characteristics. Although previous studies have demonstrated the influence of neighborhood effects, phylogenetic relatedness, and habitat filtering on seedling survival, growth, and mortality, the effect of variation in these factors on seedling abundance remains unclear. To address this question, we used a 4-ha (200 m × 200 m) and monitored four-year (2020–2023) seedling dataset from a mid-montane humid evergreen broad-leaved subtropical forest in the Gaoligong Mountains, Yunnan, Southwestern China, and which consisted of 916 seedlings belonging to 56 species. The results of generalized linear mixed models showed no significant effect of conspecific adult neighbors on seedling abundance at any of the intervals evaluated. In contrast, we found evidence of phylogenetic distance density dependence in the forests of the Gaoligong Mountains. Specifically, there was a significant positive effect of the relative average phylogenetic distance between heterospecific adult neighbors and focal seedlings on focal seedling abundance in 2020; however, the relative average phylogenetic distance between heterospecific seedling neighbors and focal seedlings had a significant negative effect on seedling abundance over the four-year period (2020–2023). Among the habitat factors, only light (canopy opening) had a negative effect on seedling abundance in all four years. Light resources may be a limiting factor for seedlings, and determine seedling dynamics in subtropical forests. Overall, our results demonstrated that phylogenetic density dependence and habitat filtering affected subtropical seedling abundance. Our findings provide new evidence of the impact of phylogenetic density dependence on seedling abundance in a subtropical mid-montane humid evergreen broad-leaved forest and highlight the need to incorporate the neighborhood effect, phylogenetic relatedness, and habitat factors in models assessing seedling abundance.

Polymorphic microsatellite markers for genetic analysis of Berkeleyomyces rouxiae, a causal agent of black root rot

AbstractBerkeleyomyces rouxiae, a soil-borne fungal pathogen, causes black root rot on various plant species including carrot, okra, tobacco, lettuce, and other important crops. Here we analyzed the entire genome sequence of a Japanese isolate of B. rouxiae and discovered 25 microsatellite sequences. These sequences were amplified from the genomic DNA of several Japanese isolates by PCR and sequenced, revealing that the microsatellite sequences were polymorphic among isolates. These markers are useful for phylogenetic analysis, epidemiological population analysis, identification of specific isolates, and other genetic investigation of B. rouxiae.

Unraveling the ecological niche signals of Verticillium dahliae: Insights from Mediterranean landscapes.

Verticillium wilt (VW), caused by the soil-borne plant pathogenic fungus Verticillium dahliae, is a major disease impacting olive crops globally. In view of the lack of effective post-infection treatments, exclusion and avoidance strategies are essential in disease management. Assessing the risks posed by this pathogen is essential to prevent the spread and to ensure selection of suitable sites for new plantations. This study aimed to elucidate the environmental factors driving V. dahliae establishment in the Andalusia region, in southern Spain, an emblematic Mediterranean landscape for olive cultivation. To this end, we explored ecological niche signals for this fungal pathogen by analyzing 62 environmental variables across 1.6 million hectares dedicated to olive and cotton cultivation, using a 15-yr survey data on VW incidence on presence-absence from both olive and cotton fields. To ensure robust identification of ecological niche signals, we employed randomization-based, non-parametric univariate tests to compare presence records with the broader sampling universe (including absence records). Our findings identified key environmental variables that are associated significantly with V. dahliae presence, including temperature range seasonality (including mean diurnal and annual ranges), summer temperature (maximum of the warmest month, mean of the warmest quarter), and moisture and water availability (near-surface humidity, potential evapotranspiration, vapor pressure) as core niche variables for V. dahliae. Our results replicated the pathogen's known distribution, identifying the Guadalquivir Valley as a particularly high-risk area in view of its mild winters and distinct rainy seasons, providing new insights into the specific environmental conditions that facilitate the pathogen's survival and spread. Furthermore, this study introduces a novel approach to niche modeling that prioritizes variables with consistent effects and significant impact on the presence and distribution of V. dahliae and identifies potential data artifacts. This approach enhances our understanding of ecological requirements in V. dahliae and informs targeted management strategies.

Evaluation of Sheath Blight Resistance in Diverse Rice Varieties Under Artificial Inoculation and Field Conditions

Rice (Oryza sativa L.) a vital food crop for nearly half of the global population, facing significant threats from various biotic and abiotic stresses. Among biotic stresses, diseases pose the most common challenge. Sheath blight, caused by soil-borne fungal pathogen Rhizoctonia Solani Kühn (R. solani), is a major concern, leading to substantial damage to rice and cereal crops worldwide. This study evaluated sheath blight resistance across diverse rice varieties, including short grain, long grain, aromatic, hybrid, and wild types, under both artificial inoculation and field conditions. Ten host plant species were assessed for resistance to sheath blight caused by Rhizoctonia Solani, using physiological screening one-week post-inoculation. The highly virulent R. solani isolate (AG-1 IA anastomosis group) was cultured on PDA media, covering the entire plate 3 to 5 days. Screening of cultivated rice varieties, was conducted using field conditions and humidified chamber methods, with results confirmed through molecular characterization. Among the ten cultivated rice varieties, Arize 6444 and Dhanya 748 exhibited the lowest disease index, classifying them as moderately resistant. In contrast, Kalanamak and Pusa Basmati were highly susceptible, displaying high disease indices. Among wild rice accessions, Oryza rufipogon showed a disease index of five or less. Visual ratings indicated the highest susceptibility in Kalanamak, followed by Pusa Basmati and Swarna sub-1, while the lowest visual ratings were observed in wild rice accessions Oryza australiensis and Oryza rufipogon across all tested hosts. The findings underscore the importance of identifying resistant rice varieties to manage sheath blight effectively. The moderately resistant cultivars and wild rice accessions identified in this study offer valuable genetic resources for breeding programs aimed at enhancing sheath blight resistance in rice.

Analysis of Ginkgo biloba Root Exudates and Inhibition of Soil Fungi by Flavonoids and Terpene Lactones.

Ginkgo biloba is abundant in secondary metabolites, including flavonoids and terpenoids. While the majority of research has focused on the role of these compounds in disease resistance, their specific contribution to pathogen defense has been rarely explored. In this study, we collected root exudates from hydroponically cultivated ginkgo seedlings and conducted a metabolomic analysis. We identified several primary metabolites mainly comprising amino acids and nucleotides, while secondary metabolites consisted of various compounds, including bioactive compounds such as flavonoids and terpenoids. Focusing on the secondary metabolites with relatively higher abundance in the exudates, we selected a mixture of flavonoids and terpenoids for in vitro inhibition experiments against two soil-borne fungal pathogens, Fusarium oxysporum f. sp. cucumerinum that causes cucumber wilt and Rhizoctonia solani AG-8 that causes wheat root rot. The results indicated that the growth rate of both fungus cells was significantly reduced with the increasing concentration of the flavonoid and terpenoid mixture extracted from ginkgo and was completely inhibited at a concentration of 5 mg/mL. Further experiments revealed that this mixture of flavonoids and terpenoids had a destructive effect on the cellular structure of both fungi, thereby reducing cell viability and achieving an antifungal effect. These findings provide a foundation for further research into the use of ginkgo extracts in biological control.

GC-MS analysis and antifungal potential of flower extract of Acacia nilotica subsp. Indica against Macrophomina phaseolina

Macrophomina phaseolina is a wide host ranged soil-borne fungal plant pathogen. It infects more than 500 host plant species belonging to 100 families. Many important oil-seed and leguminous crops are known to be attacked by this devastating plant pathogen. In the present study, antifungal potential of flowers of a leguminous tree Acacia nilotica subsp. indica, was assessed against this pathogen through bioassays guided fractionation. Initially, methanolic extracts of 1 %–5 % of leaf, flower, root-bark and stem-bark of the plant species under consideration were evaluated for their antifungal potential against the target pathogen. Among these, the best antifungal activity was shown by flower extract. The reduction in growth of the test fungal strain was 27–49 %, 4–40 % and 2–27 % due to flower, root-bark and leaf extracts, respectivey, over control. Flower extract was partitioned using n-hexane, chloroform, ethyl acetate and n-butanol as the solvents. Bioassays guided study of these fractions of methanolic extract of flower revealed that high antifungal potential was shown by n-hexane and chloroform fractions against M. phaseolina causing 26–53 % and 28–50 % decline in fungal biomass, respectively, as compared to that of control. GC-MS analysis of chloroform fraction revealed the presence of 27 compounds in this fraction. Among these cyclopentanol,-1-methyl (10.93 %) was the predominant compound followed by methyl, 4,4-dimethyl butanoate (7.04 %), 1-pentanol (6.80 %), 2-propanol, 1-cyclopropyl (6.11 %), 1H,imidazole-4-5-dihydro-2-methyl (5.93 %), trichloroethane (5.91 %), carbonic acid-ethyl hexyl ester (4.59 %), 1,4-butandiol,2,3-bis(methylene)- (4.54 %) and [S]-3,4-dimethyl pentanol (4.48 %).

Detection of fusarium wilt-induced physiological impairment in strawberry plants using hyperspectral imaging and machine learning

AbstractStrawberry (Fragraria x ananassa) is a crop affected by various soil-borne fungal pathogens with mostly non-specific foliar symptoms and often requiring laboratory isolation for correct diagnosis. Moreover, these nonspecific foliar symptoms, appreciated by the human eye, appear after some time following infection by the pathogen. Early detection of plant diseases is one of the primary objectives in agriculture because it may contribute to identifying more tolerant cultivars in breeding programs and optimise pesticide use in agricultural production with earlier applications in emerging disease foci. New technologies, such as remote sensing and machine learning (ML) algorithms, have arisen as potential tools to improve the ability to detect and classify different crop diseases. The combined use of hyperspectral imagery and ML algorithms were investigated to detect and classify the physiological stress caused by early infections of Fusarium wilt in strawberry plants. Six ML models, namely artificial neural network, decision tree, K-nearest neighbour, support vector machine, multinomial logistic regression and Naïve Bayes were developed to estimate physiological stress associated with Fusarium wilt disease. The results showed that stomatal conductance (gs) and photosynthesis (A) declined even without visual symptoms of the disease. Among the six ML models evaluated, the artificial neural network model showed the highest classification performance with an overall accuracy of 81%, regardless of the physiological parameter utilized for model training. Moreover, the artificial neural network accurately predicted the absolute values of both physiological parameters (gs and A) based on the complete spectral signature from visually healthy foliar tissue, achieving coefficients of determination of 84% and 81%, respectively. Consequently, ML models utilizing physiological response data and hyperspectral imaging exhibited remarkable robustness, facilitating the estimation of Fusarium wilt severity in strawberry plants even without visual symptoms.

VdP5CDH is involved in melanin formation, stress resistance and play a regulatory role in virulence of Verticillium dahliae.

Verticillium dahliae, a soil-borne fungal pathogen, can cause cotton Verticillium wilt. In this study, VdP5CDH, the member of the ALDH_F4-17 family of carboxylate dehydrogenases, was identified in the genome of V. dahliae and investigated function in regulating virulence by generating gene deletion mutants and complementary mutants. Homologous recombination method was used to construct mutants, transcriptome sequencing revealed gene-related metabolic pathways, and disease degree of cotton was observed through pathogen infection experiments. The conidial surface of VdP5CDH deletion strains was dented and shriveled, and the number of conidial spores increased. Compared with the wild-type (WT), the mycelial diameter of deletion mutants increased by 10.59%-11.16%, the mycelial growth showed irregular branching patterns, and misaligned arrangement. Although capable of penetrating cellophane, deletion mutants were unable to produce melanin. VdP5CDH was mainly associated with glucose metabolism, nitrogen metabolism, ABC transporter activity as well as various amino acid metabolic processes. After gene knockout, raffinose and pectin were used as the main carbon sources to promote the growth of strains and the growth rate of deletion strains in the medium containing raffinose was higher than that of WT. Consequently, the deletion mutant strains decreased utilization efficiency with which they utilized various nitrogen sources. The deletion mutants maintain responsiveness to osmotic stress and oxidative stress stimuli. Additionally, compared to WT strains, the deletion mutant strains exhibited differences in culture temperature tolerance, UV exposure response, and fungicide sensitivity. After cotton was infected with deletion strains conidial suspension, its disease index increased dramatically, while it gradually decreased after spraying with 2 mM glutamate in batches. With the increase of spraying times, the effect was more significant, and the disease index decreased by 18.95%-19.66% at 26 dpi. These results indicated that VdP5CDH regulates the pathogenicity of fungi and controls mycelia growth, melanin formation, conidia morphology, abiotic stress resistance, and the expression of infecting structure-related genes.

A pooled mycoviral resource in a strain of Rhizoctonia solani are regulators of fungal virulence

Rhizoctonia solani is a widespread and devastating soil-borne plant fungal pathogen that causes diseases, including rice sheath blight, which are difficult to control. Some mycoviruses are potential biocontrol agents for the control of fungal diseases. In order to investigate the factors that influence the virulence of R. solani and search for mycoviruses with the potential for biocontrol of R. solani, a rice-infecting R. solani strain, ZJXD1–1, was isolated and confirmed to contain eight mycoviruses via dsRNA extraction and high-throughput sequencing. The identified mycoviruses belong to families of Endornaviridae (RsEV11 and RsEV12) and Mitoviridae (RsMV125 to RsMV129), and an unclassified Toti-like clade (RsTLV1). The C39 domain in RsEV12, which shares a close evolutionary relationship with bacteria, is observed for the first time in a mycovirus. Strains with different virus combinations were obtained through viral horizontal transfer, and pathogenicity test deduced that the Endornaviruses RsEV11 and RsEV12, and Mitovirus RsMV129 might potentially enhance the pathogenicity of R. solani, while RsMV125 might reduce the virulence or interfere with the function of other Mitoviruses. Furthermore, virus curing via protoplast regeneration and viral horizontal transfer demonstrated that RsMV129 is the causal agent of R. solani hypervirulence. Overall, our study provided the resource pool of viruses that may contribute to the discovery of new biocontrol agents against R. solani and enhance our understanding of the pathogenesis of R. solani regulated by mycoviruses.

Analyzing the defense response mechanism of Atractylodes macrocephala to Fusarium oxysporum through small RNA and degradome sequencing.

Fusarium oxysporum is a significant soil-borne fungal pathogen that affects over 100 plant species, including crucial crops like tomatoes, bananas, cotton, cucumbers, and watermelons, leading to wilting, yellowing, growth inhibition, and ultimately plant death. The root rot disease of A. macrocephala, caused by F. oxysporum, is one of the most serious diseases in continuous cropping, which seriously affects its sustainable development. In this study, we explored the interaction between A. macrocephala and F. oxysporum through integrated small RNA (sRNA) and degradome sequencing to uncover the microRNA (miRNA)-mediated defense mechanisms. We identified colonization of F. oxysporum in A. macrocephala roots on day 6. Nine sRNA samples were sequenced to examine the dynamic changes in miRNA expression in A. macrocephala infected by F. oxysporum at 0, 6, and 12 days after inoculation. Furthermore, we using degradome sequencing and quantitative real-time PCR (qRT-PCR), validated four miRNA/target regulatory units involved in A. macrocephala-F. oxysporum interactions. This study provides new insights into the molecular mechanisms underlying A. macrocephala's early defense against F. oxysporum infection, suggesting directions for enhancing resistance against this pathogen.