Soil-borne Fungal Pathogens Research Articles

Screening of microRNAs and target genes involved in Sclerotinia sclerotiorum (Lib.) infection in Brassica napus L.

BackgroundRapeseed (Brassica napus L.) is the third largest source of vegetable oil in the world, and Sclerotinia sclerotiorum (Lib.) is a major soil-borne fungal plant pathogen that infects more than 400 plant species, including B. napus. Sclerotinia stem rot caused an annual loss of 10 − 20% in rapeseed yield. Exploring the molecular mechanisms in response to S. sclerotiorum infection in B. napus is beneficial for breeding and cultivation of resistant varieties. To gain a better understanding of the mechanisms regarding B. napus tolerance to Sclerotinia stem rot, we employed a miRNAome sequencing approach and comprehensively investigated global miRNA expression profile among five relatively resistant lines and five susceptible lines of oilseed at 0, 24, and 48 h post-inoculation.ResultsIn this study, a total of 40 known and 1105 novel miRNAs were differentially expressed after S. sclerotiorum infection, including miR156, miR6028, miR394, miR390, miR395, miR166, miR171, miR167, miR164, and miR172. Furthermore, 8,523 genes were predicted as targets for these differentially expressed miRNAs. These target genes were mainly associated with disease resistance (R) genes, signal transduction, transcription factors, and hormones. Constitutively expressing miR156b (OX156b) plants strengthened Arabidopsis resistance against S. sclerotiorum accompanied by smaller necrotic lesions, whereas blocking miR156 expression in Arabidopsis (MIM156) led to greater susceptibility to S. sclerotiorum disease, associated with extensive cell death of necrotic lesions.ConclusionsThis study reveals the distinct difference in miRNA profiling between the relatively resistant lines and susceptible lines of B. napus in response to S. sclerotiorum. The identified differentially expressed miRNAs related to sclerotinia stem rot resistance are involved in regulating resistance to S. sclerotiorum in rapeseed by targeting genes related to R genes, signal transduction, transcription factors, and hormones. miR156 positively modulates the resistance to S. sclerotiorum infection by restricting colonization of S. sclerotiorum mycelia. This study provides a broad view of miRNA expression changes after S. sclerotiorum infection in oilseed and is the first to elucidate the function and mechanism underlying the miR156 response to S. sclerotiorum infection in oilseed rape.

Molecular interactions between the soilborne pathogenic fungus Macrophomina phaseolina and its host plants.

Mentioned for the first time in an article 1971, the occurrence of the term "Macrophomina phaseolina" has experienced a steep increase in the scientific literature over the past 15 years. Concurrently, incidences of M. phaseolina-caused crop diseases have been getting more frequent. The high levels of diversity and plasticity observed for M. phasolina genomes along with a rich equipment of plant cell wall degrading enzymes, secondary metabolites and putative virulence effectors as well as the unusual longevity of microsclerotia, their asexual reproduction structures, make this pathogen very difficult to control and crop protection against it very challenging. During the past years several studies have emerged reporting on host defense measures against M. phaseolina, as well as mechanisms of pathogenicity employed by this fungal pathogen. While most of these studies have been performed in crop systems, such as soybean or sesame, recently interactions of M. phaseolina with the model plant Arabidopsis thaliana have been described. Collectively, results from various studies are hinting at a complex infection cycle of M. phaseolina, which exhibits an early biotrophic phase and switches to necrotrophy at later time points during the infection process. Consequently, responses of the hosts are complex and seem coordinated by multiple defense-associated phytohormones. However, at this point no robust and strong host defense mechanism against M. phaseolina has been described.

Microbe-induced gene silencing boosts crop protection against soil-borne fungal pathogens.

Small RNA (sRNA)-mediated trans-kingdom RNA interference (RNAi) between host and pathogen has been demonstrated and utilized. However, interspecies RNAi in rhizospheric microorganisms remains elusive. In this study, we developed a microbe-induced gene silencing (MIGS) technology by using a rhizospheric beneficial fungus, Trichoderma harzianum, to exploit an RNAi engineering microbe and two soil-borne pathogenic fungi, Verticillium dahliae and Fusarium oxysporum, as RNAi recipients. We first detected the feasibility of MIGS in inducing GFP silencing in V. dahliae. Then by targeting a fungal essential gene, we further demonstrated the effectiveness of MIGS in inhibiting fungal growth and protecting dicotyledon cotton and monocotyledon rice plants against V. dahliae and F. oxysporum. We also showed steerable MIGS specificity based on a selected target sequence. Our data verify interspecies RNAi in rhizospheric fungi and the potential application of MIGS in crop protection. In addition, the in situ propagation of a rhizospheric beneficial microbe would be optimal in ensuring the stability and sustainability of sRNAs, avoiding the use of nanomaterials to carry chemically synthetic sRNAs. Our finding reveals that exploiting MIGS-based biofungicides would offer straightforward design and implementation, without the need of host genetic modification, in crop protection against phytopathogens.

The soil-borne fungal pathogen Athelia rolfsii: past, present, and future concern in legumes.

Legumes are ubiquitous, low-cost meals that are abundant in protein, vitamins, minerals, and calories. Several biotic constraints are to blame for the global output of legumes not meeting expectations. Fungi, in particular, are substantial restrictions that not only hinder production but also pose a serious health risk to both human and livestock consumption. Athelia rolfsii (Syn. Sclerotium rolfsii) is a dangerous pathogenic fungus that attacks most crops, causing massive yield losses. Legumes are no longer immune to this dreadful fungus, which can potentially result in a 100% yield loss. The initial disease symptoms based on the formation of brown color lesions at the point of infection and further development of mycelia, followed by yellowing and wilting of the whole plant. To tackle such situation, various strategies, i.e., management in cultural practices, disease-free plant growth, genetic changes, crop hybridization and in vitro culture techniques have been undertaken. This present review encapsulates the entire situation, from sclerotial dissemination through infection development and control in legume crops, with the goal of developing a tangible understanding of sustainable legume production improvements. Further study in this area might be led in an integrated manner as a result of this information, which could contribute to a better understanding of the processes of disease incidence, resistance mechanism, and its control, and fostering greater inventiveness in the production of legumes.

Summer Rice–Winter Potato Rotation Suppresses Various Soil-Borne Plant Fungal Pathogens

Growing potatoes (Solanum tuberosum) using the idle rice fields in Southern China and the Indo-Gangetic Plains of India in the winter season through the rice–potato rotation (RC) system could support future food security. However, the modulation capacity of the RC system on soilborne fungal pathogens is still unclear. In the current study, a pot experiment was designed and conducted to monitor the dynamics of soil fungal community composition between the potato monoculture (CC) system and the RC system, where the two systems were set with the same soil conditions: autoclaving with fertilization; autoclaving without fertilization; autoclave-free with fertilization; and autoclave-free without fertilization. Then, the uncultivated soil (CK) and root-zone soil samples of conditions under the two systems were collected, and then soil physiochemical properties and enzymatic activities were determined. Next, the high-variable region (V5–V7) of fungal 18S rRNA genes of the samples were amplified and sequenced through the PCR technique and the Illumina Miseq platform, respectively. Finally, the fungal species diversity and composition, as well as the relative abundance of fungal pathogens annotated against the Fungiuld database in soil samples, were also investigated. The results showed that the RC could significantly (p < 0.05) increase soil fungal species diversity and decrease the relative abundance of soil fungal pathogens, where the RC could suppress 23 soil fungal pathogens through cultivating the rice during the summer season and 93.75% of the remaining pathogens through winter-season cultivation. Seven-eighths of the conditions under RC have lower pathogenic MGIDI indices (6.38 to 7.82) than those of the CC (7.62 to 9.63). Notably, both rice cultivation and winter planting reduced the abundance of the pathogenic strain ASV24 under the Colletotrichum genus. The bipartite fungal network between the pathogens and the non-pathogens showed that the pathogenic members could be restricted through co-occurring with the non-pathogenic species and planting crops in the winter season. Finally, the redundancy analysis (RDA) indicated that soil pH, electronic conductivity, available phosphorus content, and various enzyme activities (cellulase, urease, sucrase, acid phosphatase, catalase, polyphenol oxidase) could be the indicators of soil fungal pathogens. This experiment demonstrated that the rice–potato rotation system outperformed the potato monoculture on suppressing soilborne fungal pathogenic community.

Temperature mediates microbial multitrophic communities assembly and soil-borne fungal pathogens in coastal ecosystems

A better understanding of assembly processes of multitrophic microbial communities and soil-borne fungal pathogens is important to predict changes in ecosystem functions. However, the drivers governing the microbial community assembly processes and soil-borne fungal pathogen distributions, particularly in the coastal ecosystem is unclear. Here, we examined the shifts in soil bacterial, fungal, protistan communities and soil-borne fungal pathogens across 2000 km in the coastal areas of eastern Australia. Determinism predominated the multitrophic microbial community assembly with mean annual temperature (MAT) differentially mediates deterministic and stochastic processes. Contrasting to protistan community, bacterial and fungal community assembly tended to be more deterministic in higher MAT regions. A total of 609 soil-borne fungal pathogens were identified with the total relative abundances ranged from 0.34 % to 52.29 % in a given soil sample. The profile of soil-borne fungal pathogens was significantly correlated with bacterial and protistan communities. Random forest indicated that MAT was the main driver of soil-borne fungal pathogens. Altogether, our work provided evidence that MAT is a critical factor driving the belowground microbial community assembly and soil-borne fungal pathogen patterns and highlighted the potential importance of trophic interactions in predicting dynamics of soil-borne fungal pathogens under the changing environment.

Engineered coumarin accumulation reduces mycotoxin-induced oxidative stress and disease susceptibility.

Coumarins can fight pathogens and are thus promising for crop protection. Their biosynthesis, however, has not yet been engineered in crops. We tailored the constitutive accumulation of coumarins in transgenic Nicotiana benthamiana, Glycine max and Arabidopsis thaliana plants, as well as in Nicotiana tabacum BY-2 suspension cells. We did so by overexpressing A. thaliana feruloyl-CoA 6-hydroxylase 1 (AtF6'H1), encoding the key enzyme of scopoletin biosynthesis. Besides scopoletin and its glucoside scopolin, esculin at low level was the only other coumarin detected in transgenic cells. Mechanical damage of scopolin-accumulating tissue led to a swift release of scopoletin, presumably from the scopolin pool. High scopolin levels in A. thaliana roots coincided with reduced susceptibility to the root-parasitic nematode Heterodera schachtii. In addition, transgenic soybean plants were more tolerant to the soil-borne pathogenic fungus Fusarium virguliforme. Because mycotoxin-induced accumulation of reactive oxygen species and cell death were reduced in the AtF6'H1-overexpressors, the weaker sensitivity to F. virguliforme may be caused by attenuated oxidative damage of coumarin-hyperaccumulating cells. Together, engineered coumarin accumulation is promising for enhanced disease resilience of crops.

Dazomet changes microbial communities and improves morel mushroom yield under continuous cropping.

Morels (Morchella spp.) are highly prized and popular edible mushrooms. The outdoor cultivation of morels in China first developed at the beginning of the 21st century. Several species, such as Morchella sextelata, M. eximia, and M. importuna, have been commercially cultivated in greenhouses. However, the detriments and obstacles associated with continuous cropping have become increasingly serious, reducing yields and even leading to a complete lack of fructification. It has been reported that the obstacles encountered with continuous morel cropping may be related to changes in the soil microbial community. To study the effect of dazomet treatment on the cultivation of morel under continuous cropping, soil was fumigated with dazomet before morel sowing. Alpha diversity and beta diversity analysis results showed that dazomet treatment altered the microbial communities in continuous cropping soil, which decreased the relative abundance of soil-borne fungal pathogens, including Paecilomyces, Trichoderma, Fusarium, Penicillium, and Acremonium, increased the relative abundance of beneficial soil bacteria, including Bacillius and Pseudomonas. In addition, the dazomet treatment significantly increased the relative abundance of morel mycelia in the soil and significantly improved morel yield under continuous cropping. These results verified the relationship between the obstacles associated with continuous cropping in morels and the soil microbial community and elucidated the mechanism by which the obstacle is alleviated when using dazomet treatment.

Soybean red crown rot: Current knowledge and future challenges

AbstractSoybean red crown rot (RCR) is caused by the soilborne fungal pathogen Calonectria ilicicola. It causes root rot, leaf senescence, early defoliation, plant wilting and even death, often resulting in a severe reduction in seed yield and quality. C. ilicicola is an extremely difficult pathogen to control owing to its wide host range and long survival in soil and plant debris. Currently, no practically effective fungicide for this pathogen is available. Breeding for resistance is considered the most effective and environmentally acceptable means of disease management, but no C. ilicicola‐resistant soybean variety has yet been identified. Given this situation, it is necessary to promote our understanding of the mechanism of RCR incidence and the development of a comprehensive and sustainable RCR management strategy urgently.

Impact of the fungal pathogen Fusarium oxysporum on the taxonomic and functional diversity of the common bean root microbiome

BackgroundPlants rely on their root microbiome as the first line of defense against soil-borne fungal pathogens. The abundance and activities of beneficial root microbial taxa at the time prior to and during fungal infection are key to their protective success. If and how invading fungal root pathogens can disrupt microbiome assembly and gene expression is still largely unknown. Here, we investigated the impact of the fungal pathogen Fusarium oxysporum (fox) on the assembly of rhizosphere and endosphere microbiomes of a fox-susceptible and fox-resistant common bean cultivar.ResultsIntegration of 16S-amplicon, shotgun metagenome as well as metatranscriptome sequencing with community ecology analysis showed that fox infections significantly changed the composition and gene expression of the root microbiome in a cultivar-dependent manner. More specifically, fox infection led to increased microbial diversity, network complexity, and a higher proportion of the genera Flavobacterium, Bacillus, and Dyadobacter in the rhizosphere of the fox-resistant cultivar compared to the fox-susceptible cultivar. In the endosphere, root infection also led to changes in community assembly, with a higher abundance of the genera Sinorhizobium and Ensifer in the fox-resistant cultivar. Metagenome and metatranscriptome analyses further revealed the enrichment of terpene biosynthesis genes with a potential role in pathogen suppression in the fox-resistant cultivar upon fungal pathogen invasion.ConclusionCollectively, these results revealed a cultivar-dependent enrichment of specific bacterial genera and the activation of putative disease-suppressive functions in the rhizosphere and endosphere microbiome of common bean under siege.

Application of Hydrothermal Time Models to Predict Sclerotial Germination of Athelia rolfsii.

Athelia rolfsii, causal agent of "southern blight" disease, is a soilborne fungal pathogen with a wide host range of more than 500 species. This study's objectives were to (i) quantify the effects of two environmental factors, temperature and soil moisture, on germination of A. rolfsii inoculum (sclerotia), which is a critical event for the onset of disease epidemics and (ii) predict the timing of sclerotial germination by applying population-based threshold-type hydrothermal time (HTT) models. We conducted in vitro germination experiments with three isolates of A. rolfsii isolated from peanuts, which were tested at five temperatures (T), ranging from 17 to 40°C, four matric potentials (Ψm) between -0.12 and -1.57 MPa, and two soil types (fine sand and loamy fine sand), using a factorial design. When Ψm was maintained between -0.12 and -0.53 MPa, T from 22 to 34°C was found to be conducive to sclerotial germination (>50%). The HTT models were fitted for a range of T (22 to 34°C) and Ψm (-0.12 to -1.57 MPa) that accounted for 84% or more of variation in the timing of sclerotial germination. The estimated base T ranged between 0 and 4.5°C and the estimated base Ψm between -2.96 and -1.52 MPa. The results suggest that the HTT modeling approach is a suitable means of predicting the timing of A. rolfsii sclerotial germination. This HTT methodology can potentially be tested to fine-tune fungicide application timing and in-season A. rolfsii management strategies. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Molecular Characterization of Defense of Brassica napus (Oilseed Rape) to Rhizoctonia solani AG2-1 Confirmed by Functional Analysis in Arabidopsis thaliana.

Rhizoctonia solani is a necrotrophic, soilborne fungal pathogen associated with significant establishment losses in Brassica napus (oilseed rape; OSR). The anastomosis group (AG) 2-1 of R. solani is the most virulent to OSR, causing damping-off, root and hypocotyl rot, and seedling death. Resistance to R. solani AG2-1 in OSR has not been identified, and the regulation of OSR defense to its adapted pathogen, AG2-1, has not been investigated. In this work, we used confocal microscopy to visualize the progress of infection by sclerotia of AG2-1 on B. napus varieties with contrasting disease phenotypes. We defined their defense response using gene expression studies and functional analysis with Arabidopsis thaliana mutants. Our results showed existing variation in susceptibility to AG2-1 and plant growth between OSR varieties, and differential expression of genes of hormonal and defense pathways related to auxin, ethylene, jasmonic acid, abscisic acid, salicylic acid, and reactive oxygen species regulation. Auxin, abscisic acid signaling, and the MYC2 branch of jasmonate signaling contributed to the susceptibility to AG2-1, while induced systemic resistance was enhanced by NAPDH RBOHD, ethylene signaling, and the ERF/PDF branch of jasmonate signaling. These results pave the way for future research, which will lead to the development of Brassica crops that are more resistant to AG2-1 of R. solani and reduce dependence on chemical control options.

Pathogenic and Saprophytic Growth Rates of Fusarium oxysporum f. sp. vasinfectum Interact to Affect Variation in Inoculum Density and Interannual Infection Risk.

Fusarium wilt of cotton caused by the soilborne fungal pathogen Fusarium oxysporum f. sp. vasinfectum race 4 (FOV4) is a contemporary epidemic affecting cotton production in Far West Texas. The spatial distribution of soilborne FOV4 can be heterogeneous at small scales, and the factors that lead to this heterogeneity require investigation. Hypothetical causes include dissemination of spores through soils and variable saprophytic growth of the fungus. In the field, FOV4 DNA was quantified from soil during and after the cotton-growing season, and though the average amounts of DNA were not different between these time points, the variances of DNA across space were significantly different. Variability was higher when pathogenic growth of the fungus was expected owing to the presence of live cotton plants and lower when saprophytic growth was expected after cropping. In sterile-environment growth chamber experiments, the abundance of organic matter influenced the fungal vegetative growth rate and maximum amount as measured through quantitative PCR and the timing of the fungus' increasing its rate of spore production as measured through dilution plating. To investigate movement of spores in soils, spore mobility in experimental columns was quantified. Soil composition and organic matter abundance affected spore mobility, indicating that the timing of spore production relative to the availability of growth resources will affect the spatial spread of FOV4 and suggesting that soil properties affect the retention of conidia. The spatial spread of FOV4 through soil varies temporally and is affected by the shift between pathogenic and saprophytic growth modes of the fungus.

Enhancement of resistance against fungal pathogens in peanut (Arachis hypogaea L.) cultivar L14 by heterologous expression of gene encoding chitinase 42 kDa from Trichoderma asperellum SH16

This study aims to extracellularly express the 42 kDa chitinase from Trichoderma asperellum SH16 in peanut (Arachis hypogaea) roots under the regulation of a tissue-specific Asy promoter and the driving of the signal peptide of the rice amylase 3D gene. The 42 kDa chitinase genes were employed in this study, which included one wild-type sequence (Chi42) and two synthetic sequences (syncodChi42–1 and syncodChi42–2) optimized for codon usage for plant expression. From 102 42 kDa chitinase transgenic peanut lines, seven elite lines were discovered, among them two for syncodChi42–1, three for syncodChi42–2, and two for Chi42. The study also revealed chitinase from two synthetic genes had higher expression levels than chitinase from the wild-type gene, with the highest specific activities of approximately 901 U/mg and 1124 U/mg, respectively, compared to 739 U/mg. All seven elite transgenic peanut lines exhibited strong extracellular chitinase activity, and some of them were able to fight the soil-borne pathogenic fungus Sclerotium rolfsii in both in vitro and in vivo assays. The significant chitinase activity of seven elite transgenic peanut lines, particularly five lines carrying one of two synthetic genes, suggests that transgenic crops might be used to prevent phytopathogenic fungi.

Contrasting interactions of the aggressive Verticillium longisporum lineage A1/D1 and the non‐aggressive A1/D2 lineage with roots of oilseed rape

AbstractVerticillium longisporum is a soilborne fungal vascular pathogen of Brassica crops. Three hybrid lineages originating from three independent hybridization events of four haploid Verticillium parents are known. V. longisporum lineage A1/D2 has only been found in horseradish and previous pathogenicity tests have shown that it is non‐aggressive on oilseed rape (Brassica napus). The aim of this study was to investigate the interaction of the non‐aggressive lineage A1/D2 with oilseed rape in comparison with the aggressive A1/D1 lineage. Although A1/D2 did not cause severe disease symptoms, the study revealed restricted root infection and systemic colonization by this lineage, as assessed by quantitative PCR of fungal DNA. Assessment of root surface colonization by confocal laser microscopy indicated early sporulation, disorganized loosely attached hyphal growth and hyphal coiling of A1/D2. By contrast, A1/D1 showed organized hyphal growth tight to the root surface, as well as hyphal swellings at points of penetration. Vascular colonization was studied with transmission electron microscopy, which revealed less vessel colonization, higher intensity of vascular responses and lower plant cell wall degradation in A1/D2‐treated plants in comparison with A1/D1. Vascular responses consisted of strong coating of vessel walls and fungal hyphae with an electron‐dense material, indicating higher intensity of plant defence in comparison with A1/D1. The results, for the first time, demonstrate stem colonization and thus a basic level of pathogenic interaction of non‐aggressive strains of A1/D2 with oilseed rape associated with a lack of severe disease symptoms, which indicate tolerance in oilseed rape to lineage A1/D2.

Biomolecules from Chaetomium globosum Possessing Antimicrobial Compounds Potentially Inhibits Fusarium Wilt of Tomato.

Wilt disease caused by Fusarium solani, a soil-borne plant pathogenic fungus, is a serious disease in tomato causing economic losses. In the present study, among the nine isolates of Chaetomium globosum Kunze (Chg1-Chg9), screened against F. solani No. Fs-1, Chg2 exhibited the maximum inhibition (49.2 %), followed by the isolates Chg6 (47.4%) and Chg1 (46.3%) in dual culture. Further, the crude secondary metabolites from these three isolates showed maximum reduction of the mycelial growth of Fs-1 compared to control. Gas Chromatography-Mass Spectrometry (GC-MS) analysis of the metabolites from Chg2 revealed the presence of major compounds, viz., benzothiazole, 2-(2-hydroxyethylthio) (7.51%); 9,12,15- octadecatrienoic acid, 2,3-bis[(trimethylsilyl)oxy] propyl ester (3.13%); and hexadecanoic acid, 1- (hydroxymethyl)-1,2-ethanediyl ester (2.69%). Fourier transform infrared (FTIR) spectroscopy revealed the presence of alcohol, secondary amine, aliphatic primary amine, carboxylic acid, allene, conjugated ketone, imine/oxime, sulphate, and halo compound groups with a weak to strong range of intensity. Pure compound of benzothiazole @ 5000 ppm exhibited higher antagonistic activity of Fs-1which decreased at lower concentration. In pot culture experiment, dipping tomato seedlings cv. PKM1 in 2% ethyl acetate-fractionated biomolecules of Chg2 recorded a minimum disease incidence of 20.0%, whereas seedlings dipped in the culture filtrate of Chg2 showed a disease incidence of 26.6% compared to control (86.6%). Besides, these seedlings treated with the culture filtrate showed the highest plant height of 37.68 cm and the maximum number of leaves and flowers (39.6 and 5.48, respectively), followed by those treated with the biomolecules of Chg2 (C. globosum) and the culture filtrate of T. asperellum. The results revealed that the antifungal efficacy of C. globosum Chg2 is due to the presence of antimicrobial metabolites including benzothiazole. Exploring the use of this novel antifungal compound in the management of plant diseases is highly warranted.

The presence of the plant pathogen Fusarium graminearum as a soil inoculum enhances the rhizosphere survival of bacterial biocontrol strains aimed at the pathogen

AbstractThe serious wheat pathogen Fusarium graminearum causes both root rot and head blight. Some classical biocontrol tests were first used to explore the biocontrol ability of 39 Pseudomonas fluorescens strains. The five most antifungal strains B4, P13, UTPf127, UTPf125 and UTPf105 were selected to screen known antifungal antibiotic genes and greenhouse experiments. The ability of bacteria to colonize wheat rhizosphere and their effect on plant growth in the presence and absence of soil F. graminearum inoculum was studied under greenhouse conditions. Overall, biocontrol bacteria populations were significantly higher in both wheat endo‐and ectorhizosphere of pathogen‐inoculated soil than in healthy soil. The population of all strains differently decreased with time. On day 28, endorhizosphere populations of strain B4 could be detected in inoculated but not healthy soil, while UTPf127 populations remained high in endorhizospheres at all tested times. Isolate B4 and UTPf105 showed the most substantial plant growth in pathogen‐inoculated soil compared to pathogen‐inoculated soil without added bacteria. UTPf127‐treated plants grew better in control soil than when the pathogen was present. In contrast, UTPf125 and P13 showed little effect on plant growth. These results point to complex interactions between pathogen and biocontrol bacteria and suggest that a fungal pathogen in the soil can affect the survival of potential bacterial biological control agents. Additionally, they highlight the importance of screening and evaluating potential biocontrol bacteria against soilborne fungal pathogens by in vivo tests rather than relying on plate screenings.

Assessment of Genetic Variability and Evolutionary Relationships of Rhizoctonia solani Inherent in Legume Crops.

Rhizoctonia solani is one of the most common soil-borne fungal pathogens of legume crops worldwide. We collected rDNA-ITS sequences from NCBI GenBank, and the aim of this study was to examine the genetic diversity and phylogenetic relationships of various R. solani anastomosis groups (AGs) that are commonly associated with grain legumes (such as soybean, common bean, pea, peanut, cowpea, and chickpea) and forage legumes (including alfalfa and clover). Soybean is recognized as a host for multiple AGs, with AG-1 and AG-2 being extensively investigated. This is evidenced by the higher representation of sequences associated with these AGs in the NCBI GenBank. Other AGs documented in soybean include AG-4, AG-7, AG-11, AG-5, AG-6, and AG-9. Moreover, AG-4 has been extensively studied concerning its occurrence in chickpea, pea, peanut, and alfalfa. Research on the common bean has been primarily focused on AG-2, AG-4, and AG-1. Similarly, AG-1 has been the subject of extensive investigation in clover and cowpea. Collectively, AG-1, AG-2, and AG-4 have consistently been identified and studied across these diverse legume crops. The phylogenetic analysis of R. solani isolates across different legumes indicates that the distinct clades or subclades formed by the isolates correspond to their specific anastomosis groups (AGs) and subgroups, rather than being determined by their host legume crop. Additionally, there is a high degree of sequence similarity among isolates within the same clade or subclade. Principal coordinate analysis (PCoA) further supports this finding, as isolates belonging to the same AGs and/or subgroups cluster together, irrespective of their host legume. Therefore, the observed clustering of R. solani AGs and subgroups without a direct association with the host legume crop provides additional support for the concept of AGs in understanding the genetic relationships and evolution of R. solani.

Functional Characterization of Verticillium dahliae Race 3-Specific Gene VdR3e in Virulence and Elicitation of Plant Immune Responses.

Verticillium dahliae is a soilborne fungal pathogen that causes disease on many economically important crops. Based on the resistance or susceptibility of differential cultivars in tomato, isolates of V. dahliae are divided into three races. Avirulence (avr) genes within the genomes of the three races have also been identified. However, the functional role of the avr gene in race 3 isolates of V. dahliae has not been characterized. In this study, bioinformatics analysis showed that VdR3e, a cysteine-rich secreted protein encoded by the gene characterizing race 3 in V. dahliae, was likely obtained by horizontal gene transfer from the fungal genus Bipolaris. We demonstrate that VdR3e causes cell death by triggering multiple defense responses. In addition, VdR3e localized at the periphery of the plant cell and triggered immunity depending on its subcellular localization and the cell membrane receptor BAK1. Furthermore, VdR3e is a virulence factor and shows differential pathogenicity in race 3-resistant and -susceptible hosts. These results suggest that VdR3e is a virulence factor that can also interact with BAK1 as a pathogen-associated molecular pattern (PAMP) to trigger immune responses. IMPORTANCE Based on the gene-for-gene model, research on the function of avirulence genes and resistance genes has had an unparalleled impact on breeding for resistance in most crops against individual pathogens. The soilborne fungal pathogen, Verticillium dahliae, is a major pathogen on many economically important crops. Currently, avr genes of the three races in V. dahliae have been identified, but the function of avr gene representing race 3 has not been described. We investigated the characteristics of VdR3e-mediated immunity and demonstrated that VdR3e acts as a PAMP to activate a variety of plant defense responses and induce plant cell death. We also demonstrated that the role of VdR3e in pathogenicity was host dependent. This is the first study to describe the immune and virulence functions of the avr gene from race 3 in V. dahliae, and we provide support for the identification of genes mediating resistance against race 3.

Plant Physiological Assessments on Promising New HLB-Tolerant Citrus Rootstocks after Inoculation with the Phytopathogenic Ascomycete Rosellinia necatrix

Citriculture is considered the most important fruit industry and involves the cultivation of several fruit varieties, which are susceptible to many plant pathogens. In this sense, soil-borne pathogenic fungi, such Rosellinia necatrix, threaten citrus fruit production worldwide because they can cause fruit loss. Therefore, we assayed the physiological reaction of novel citrus rootstocks against white root rot disease during long-term management. Data from above-ground symptoms and chlorophyll content were periodically obtained during the experimental process. In addition, plant leaf area and percentage of biomass reduction were determined for each rootstock when the experiment finished. The behavior of the inoculated tolerant rootstocks was as follows: the lowest symptom rate of SAUDPC was achieved by B11R5T25 and N40R3T25; AMB+CZO manifested the highest disease incidence; B11R5T25 and A+VOLK × Orange 19-11-8 displayed the highest and the lowest chlorophyll content, respectively; AMB+CZO and A+VOLK × Orange 19-11-8 showed the highest biomass reduction, and the lowest was detected in B11R5T25 and N40R2T19; concerning the leaf area, N40R1T18, N40R3T25 and N40R2T19 showed the lowest response, and 2247 × 2075-01-2 achieved the highest rate. In summary, B11R5T25 and N40R3T25 displayed the lowest disease rate.