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Verticillium wilt of pistachio (Pistacia vera), caused by Verticillium dahliae, is one of the most important diseases in this crop. The most effective and economical control measure is the use of resistant rootstocks. The main objective of this work was to evaluate the resistance of five rootstocks commonly used in pistachio against the disease under field conditions. Pistachio plants (cv. Kerman) grafted on the rootstocks Pistacia atlantica, Pistacia integerrima, Pistacia terebinthus and cv. Platinum and UCBI hybrids, were planted in a field with high inoculum density of V. dahliae in the soil. Plants grafted on P. terebinthus, followed by P. integerrima, showed significantly higher disease severity values than the rest of the rootstocks almost 53 months after planting; also, plants grafted on P. atlantica were more resistant than those grafted on P. integerrima. These results highlighted the intraspecific variability in resistance to the disease. In addition, the disease was monitored in pistachios grafted onto P. terebinthus or UCBI in an established commercial field with three different zones of V. dahliae inoculum density in the soil (none, low or high). In the high‐density zone, the percentage of diseased plants was significantly lower for plants grafted on UCBI than for those grafted on P. terebinthus. The results indicate the importance of rootstock species and specific genotype or seed population used, concentration of V. dahliae inoculum in the soil, and previous history of crops on the plot, on the development of the disease under natural conditions. [ABSTRACT FROM AUTHOR]

SUMMARY Ascorbic acid (AsA) serves as a key antioxidant involved in the various physiological processes and against diverse stresses in plants. Due to the insufficiency of AsA de novo biosynthesis, the AsA regeneration is essential to supplement low AsA synthesis rates. Redox reactions play a crucial role in response to biotic stress in plants; however, how AsA regeneration participates in hydrogen peroxide (H2O2) homeostasis and plant defense remains largely unknown. Here, we identified a Golgi vesicle‐membrane‐localized cytochrome B561 (CytB561) encoding gene, GhB561‐11, involved in AsA regeneration and plant resistance to Verticillium dahliae in cotton. GhB561‐11 was significantly downregulated upon V. dahliae attack. Knocking down GhB561‐11 greatly enhanced cotton resistance to V. dahliae. We found that suppressing GhB561‐11 inhibited the AsA regeneration, elevated the basal level of H2O2, and enhanced the plant defense against V. dahliae. Further investigation revealed that GhB561‐11 interacted with the lipid droplet‐associated protein GhLDAP3 to collectively regulate the AsA regeneration. Simultaneously silencing GhB561‐11 and GhLDAP3 significantly elevated the H2O2 contents and dramatically improved the Verticillium wilt resistance in cotton. The study broadens our insights into the functional roles of CytB561 in regulating AsA regeneration and H2O2 homeostasis. It also provides a strategy by downregulating GhB561‐11 to enhance Verticillium wilt resistance in cotton breeding programs. [ABSTRACT FROM AUTHOR]

Background: As one of the world's most important vegetable crops, eggplant production is often severely affected by verticillium wilt, leading to significant declines in yield and quality. Traditional multispectral disease-imaging equipment is expensive and complicated to operate. Low-cost multispectral devices cannot capture images and cover less information. The traditional approach to early disease diagnosis involves using multispectral disease-imaging equipment in conjunction with machine learning technology. However, this approach has significant limitations in early disease diagnosis, including challenges such as high costs, complex operation, and low model performance. Results: The aim of this study was to combine low-cost multispectral cameras with deep learning technology to detect early stage Verticillium wilt in eggplant effectively. Using the Manual FS-3200T-10GE-NNC multispectral camera to perform multispectral imaging of the leaves of eggplant seedlings at the early infection stage, information fusion was performed on the collected multispectral images, and a five-channel image information fusion model was established. Image information fusion technology was combined with deep learning technology, among which the VGG16-triplet attention model performed the best, achieving a precision of 86.73% on the test set. Model validation on 48- and 72-hour data reached a precision of 75% and 82%, respectively, achieving an early diagnosis of Verticillium wilt. This highlighted the potential of multispectral cameras for early disease detection. Conclusions: In this study, we successfully developed a method for the non-destructive detection of the early stages of eggplant wilt disease by combining multispectral imaging technology with deep learning algorithms. While ensuring high accuracy, this method significantly reduces the cost of experimental equipment. The application of this method can reduce the cost of agricultural equipment and provide a scientific basis for agricultural production practices, helping to reduce losses caused by diseases. [ABSTRACT FROM AUTHOR]

Tomato leaf diseases significantly impact the yield and quality of tomatoes during cultivation, the main of which are septoria leaf spot, leaf curl virus, verticillium wilt, and early blight. These diseases necessitate prompt detection and management strategies to mitigate their deleterious effects on crop productivity. Due to the considerable scale variations in diseased tomato leaves, accurate and rapid detection and diagnosis remain challenging. To address the detection of tomato leaf diseases at different scales, we propose a real-time detection model incorporating a Multi-kernel Inception Aggregation Diffusion Network. In this paper, (1) We present a Multi-kernel Inception Aggregation Diffusion Network (MIADN) for the feature processing stage, which facilitates the aggregation and diffusion of multi-scale features across hierarchical levels, benefiting the detection of targets at various scales. (2) We present the Multi-kernel Inception Module (MKIM), designed to extract multi-scale object features using diverse convolution kernels, thereby enhancing the model's feature fusion and representation capabilities. (3) We incorporate the efficient FasterNet network at the feature extraction stage to preserve feature diversity and improve the model's ability to extract complex target features. (4) Extensive comparative and ablation experiments demonstrate that our method achieves the mean average precision (mAP50) of 96.6%, surpassing the baseline model by 4.1% and the advanced YOLOv9s model by 2.0%. This method provides an effective solution for high-quality tomato cultivation. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Introduction: Verticillium dahliae causes a serious decline in cotton yield and quality, posing a serious threat to the cotton industry. However, the mechanism of resistance to V. dahliae in cotton is still unclear, which limits the breeding of resistant cultivars. Methods: To analyze the defense mechanisms of cotton in response to V. dahliae infection, we compared the defense responses of two upland cotton cultivars from Xinjiang (JK1775, resistant; Z8,susceptible) using transcriptome sequencing at different infection stages. Results: The results revealed a significant differential expression of genes in the two cotton cultivars post V. dahliae infection, with the number of DEGs in JK1775 being higher than that in Z8 at different infection stages of V. dahliae. Interestingly, the DEGs of both JK1775 and Z8 were enriched in the MAPK signaling pathway in the early and late stages of infection. Importantly, the upregulated DEGs in both cultivars were significantly enriched in all stages of the phenylpropanoid metabolic pathway. Some of these DEGs were involved in the regulation of lignin and coumarin biosynthesis, which may be one of the key factors contributing to the resistance of upland cotton cultivars to V. dahliae in Xinjiang. Lignin staining experiments further showed that the lignin content increased in both resistant and susceptible varieties after inoculation with V. dahliae. Discussion: This study not only provides insights into the molecular mechanisms of resistance to Verticillium wilt in Xinjiang upland cotton but also offers important candidate gene resources for molecular breeding of resistance to Verticillium wilt in cotton. [ABSTRACT FROM AUTHOR]

Black root rot (BRR) and Verticillium wilt of cotton are caused by soilborne Berkeleyomyces rouxiae and Verticillium dahliae, respectively, and can individually cause yield loss of a 10–50% in New South Wales (NSW). Both diseases were first detected in a northern valley of NSW but are now present across the state. 'Come Clean Go Clean' is a widely practiced biosecurity measure used to minimize the risk of further introducing the pathogens from one field to another by restricting the movement of soil-contaminated farm equipment and machinery. We rely on cleaning agents to effectively wash down and decontaminate the equipment and machinery. In this study, we examined 12 locally available, commercial disinfectants for their efficacy against B. rouxiae and V. dahliae reproductive structures with and without soil contamination of 10% (w/v). We found a significant interaction between pathogens, disinfectants, and soil amendment (p < 0.01). The germination of B. rouxiae chlamydospores and endoconidia in both the presence and absence of soil contamination was completely suppressed even after a short 10 sec exposure to 70% ethanol and 25% bleach. Both 70% ethanol and bleach were highly lethal at 10 sec exposures in similar assays against V. dahliae microsclerotia and conidia. Some other commercial products were able to reduce the germination rate significantly but did not completely kill microsclerotia and conidia even after 30 min of exposure. The lethal effect against B. rouxiae and V. dahliae warrants further exploration of both 70% ethanol and bleach to improve their field applications. Additionally, the efficacy of most tested commercial disinfectants was time-dependent; however, this is not recommended on the labels for their effectiveness. This study provides an additional integrated disease management option aiming to limit the spread of the pathogens. [ABSTRACT FROM AUTHOR]

Syntaxin of plant (SYP) plays a crucial role in SNARE‐mediated membrane trafficking during endocytic and secretory pathways, contributing to the regulation and execution of plant immunity against pathogens. Verticillium wilt is among the most destructive fungal diseases affecting cotton worldwide. However, information regarding SYP family genes in cotton is scarce. Through genome‐wide identification and transcriptome profiling, we identified GhSYP121, a Qa SNARE gene in Gossypium hirsutum. GhSYP121 is notably induced by Verticillium dahliae, the causal agent of Verticillium wilt in cotton, and acts as a negative regulator of defense against V. dahliae. This is evidenced by the reduced resistance of GhSYP121‐deficient cotton and the increased susceptibility of GhSYP121‐overexpressing lines. Furthermore, the activation of the salicylic acid (SA) pathway by V. dahliae is inversely correlated with the expression level of GhSYP121. GhSYP121 interacts with its cognate SNARE component, GhSNAP33, which is required for the penetration resistance against V. dahliae in cotton. Collectively, GhSYP121, as a member of the cotton SNARE complex, is involved in regulating the SA pathway during plant defense against V. dahliae. This finding enhances our understanding of the potential role of GhSYP121 in these distinct pathways that contribute to plant defense against V. dahliae infection. [ABSTRACT FROM AUTHOR]

Carbon catabolite repression (CCR) is a common transcriptional regulatory mechanism that microorganisms use to efficiently utilize carbon nutrients, which is critical for the fitness of microorganisms and for pathogenic species to cause infection. Here, we characterized two CCR genes, VdCreA and VdCreC, in Verticillium dahliae that cause cotton Verticillium wilt disease. The VdCreA and VdCreC knockout mutants displayed slow growth with decreased conidiation and microsclerotium production and reduced virulence to cotton, suggesting that VdCreA and VdCreC are involved in growth and pathogenicity in V. dahliae. We further generated 36 highly reliable and stable ΔVdCreA and ΔVdCreC libraries to comprehensively explore the dynamic expression of genes and metabolites when grown under different carbon sources and CCR conditions. Based on the weighted gene co-expression network analysis (WGCNA) and correlation networks, VdCreA is co-expressed with a multitude of downregulated genes. These gene networks span multiple functional pathways, among which seven genes, including PYCR (pyrroline-5-carboxylate reductase), are potential target genes of VdCreA. Different carbon source conditions triggered entirely distinct gene regulatory networks, yet they exhibited similar changes in metabolic pathways. Six genes, including 6-phosphogluconolactonase and 2-ODGH (2-oxoglutarate dehydrogenase E1), may serve as hub genes in this process. Both VdCreA and VdCreC could comprehensively influence the expression of plant cell wall-degrading enzyme (PCWDE) genes, suggesting that they have a role in pathogenicity in V. dahliae. The integrated expression profiles of the genes and metabolites involved in the glycolysis/gluconeogenesis and pentose phosphate pathways showed that the two major sugar metabolism-related pathways were completely changed, and GADP (glyceraldehyde-3-phosphate) may be a pivotal factor for CCR under different carbon sources. All these results provide a more comprehensive perspective for further analyzing the role of Cre in CCR. [ABSTRACT FROM AUTHOR]

Verticillium wilt, caused by the soilborne fungus Verticillium dahliae, poses a serious threat to the health of more than 200 plant species worldwide. Although plant rhizosphere‐associated microbiota can influence plant resistance to V. dahliae, empirical evidence underlying Verticillium wilt resistance of perennial trees is scarce. In this study, we systemically investigated the effect of the soil microbiota on the resistance of smoke trees (Cotinus coggygria) to Verticillium wilt using field, greenhouse and laboratory experiments. Comparative analysis of the soil microbiota in the two stands of smoke trees suggested that Bacillus represented the most abundant and key microbial genus related to potential disease suppression. Smoke tree seedlings were inoculated with isolated Bacillus strains, which exhibited disease suppressiveness and plant growth‐promoting properties. Furthermore, repletion of Bacillus agents to disease conducive soil significantly resulted in reduced incidence of smoke tree wilt and increased resistance of the soil microbiota to V. dahliae. Finally, we explored a more effective combination of Bacillus agents with the fungicide propiconazole to combat Verticillium wilt. The results establish a foundation for the development of an effective control for this disease. Overall, this work provides a direct link between Bacillus enrichment and disease resistance of smoke trees, facilitating the development of green control strategies and measurements of soil‐borne diseases. Summary Statement: Bacillus was the key microbial genus responsible for Verticillium wilt disease suppression and plant growth‐promoting and the combination of B. subtilis with a fungicide represented a promising effective control to combat Verticillium wilt. [ABSTRACT FROM AUTHOR]

Cotton verticillium wilt significantly impacts both cotton quality and yield. Selecting disease-resistant varieties and using their resistance genes in breeding is an effective and economical control measure. Accurate severity estimation of this disease is crucial for breeding resistant cotton varieties. However, current methods fall short, slowing the breeding process. To address these challenges, this paper introduces CVW-Etr, a high-precision method for estimating the severity of cotton verticillium wilt. CVW-Etr classifies severity into six levels (L0 to L5) based on the proportion of segmented diseased leaves to lesions. Upon integrating YOLOv8-Seg with MobileSAM, CVW-Etr demonstrates excellent performance and efficiency with limited samples in complex field conditions. It incorporates the RFCBAMConv, C2f-RFCBAMConv, AWDownSample-Lite, and GSegment modules to handle blurry transitions between healthy and diseased regions and variations in angle and distance during image collection, and to optimize the model's parameter size and computational complexity. Our experimental results show that CVW-Etr effectively segments diseased leaves and lesions, achieving a mean average precision (mAP) of 92.90% and an average severity estimation accuracy of 92.92% with only 2.6M parameters and 10.1G FLOPS. Through experiments, CVW-Etr proves robust in estimating cotton verticillium wilt severity, offering valuable insights for disease-resistant cotton breeding applications. [ABSTRACT FROM AUTHOR]

Introduction: Verticillium dahliae causes a serious decline in cotton yield and quality, posing a serious threat to the cotton industry. However, the mechanism of resistance to V. dahliae in cotton is still unclear, which limits the breeding of resistant cultivars. Methods: To analyze the defense mechanisms of cotton in response to V. dahliae infection, we compared the defense responses of two upland cotton cultivars from Xinjiang (JK1775, resistant; Z8,susceptible) using transcriptome sequencing at different infection stages. Results: The results revealed a significant differential expression of genes in the two cotton cultivars post V. dahliae infection, with the number of DEGs in JK1775 being higher than that in Z8 at different infection stages of V. dahliae. Interestingly, the DEGs of both JK1775 and Z8 were enriched in the MAPK signaling pathway in the early and late stages of infection. Importantly, the upregulated DEGs in both cultivars were significantly enriched in all stages of the phenylpropanoid metabolic pathway. Some of these DEGs were involved in the regulation of lignin and coumarin biosynthesis, which may be one of the key factors contributing to the resistance of upland cotton cultivars to V. dahliae in Xinjiang. Lignin staining experiments further showed that the lignin content increased in both resistant and susceptible varieties after inoculation with V. dahliae. Discussion: This study not only provides insights into the molecular mechanisms of resistance to Verticillium wilt in Xinjiang upland cotton but also offers important candidate gene resources for molecular breeding of resistance to Verticillium wilt in cotton. [ABSTRACT FROM AUTHOR]

Sunflower (Helianthus annuus) is a globally significant field crop, and disease resistance is crucial for ensuring yield stability and crop quality. Verticillium dahliae is a notorious soilborne pathogen that causes Verticillium Wilt (VW) and threatens sunflower production worldwide. In this study, we conducted a comprehensive assessment of sunflower resistance to V. dahliae across 231 sunflower cultivar lines, from the Sunflower Association Mapping (SAM) population. We employed EMMAX and ridge regression best linear unbiased prediction (rrBLUP) and identified 148 quantitative trait loci (QTLs) and 23 putative genes associated with V. dahliae resistance, including receptor like kinases, cell wall modification, transcriptional regulation, plant stress signalling and defense regulation genes. Our enrichment and quantitative real-time PCR validation results highlight the importance of membrane vesicle trafficking in the sunflower immune system for efficient signaling and defense upon activation by V. dahliae. This study also reveals the polygenic architecture of V. dahliae resistance in sunflowers and provides insights for breeding sunflower cultivars resistant to VW. This research contributes to ongoing efforts to enhance crop resilience and reduce yield losses due to VW, ultimately benefiting sunflower growers and the agricultural sector. [ABSTRACT FROM AUTHOR]

Potato Verticillium wilt (PVW) caused by Verticillium dahliae is a vascular disease, that seriously affects potato (Solanum tuberosum L.) yield and quality worldwide. V. dahliae occupies the vascular bundle and therefore it cannot efficiently be treated with fungicides. Further, the application of these pesticides causes serious environmental problems. Therefore, it is of great importance to find environmentally friendly biological control methods. In this study, bacterial strains were isolated from agricultural lands on which potato had been cultured for 5 years. Five strains with a broad-spectrum antagonistic activity were selected. Among these five strains, Bacillus velezensis XS142 showed the highest antagonistic activity. To study the mechanism of XS142, by which this strain might confer tolerance to V. dahliae in potato, the genome of strain XS142 was sequenced. This showed that its genome has a high level of sequence identity with the model strain B. velezensis FZB42 as the OrthoANI (Average Nucleotide Identity by Orthology) value is 98%. The fungal suppressing mechanisms of this model strain are well studied. Based on the genome comparison it can be predicted that XS142 has the potential to suppress the growth of V. dahliae by production of bacillomycin D, fengycin, and chitinase. Further, the transcriptomes of potatoes treated with XS142 were analyzed and this showed that XS142 does not induce ISR, but the expression of genes encoding peptides with antifungal activity. Here we showed that XS142 is an endophyte. Further, it is isolated from a field where potato had been cultured for several years. These properties give it a high potential to be used, in the future, as a biocontrol agent of PVW in agriculture. [ABSTRACT FROM AUTHOR]

Compared to traditional manual methods for assessing the cotton verticillium wilt (CVW) hazard level, utilizing deep learning models for foliage segmentation can significantly improve the evaluation accuracy. However, instance segmentation methods for images with complex backgrounds often suffer from low accuracy and delayed segmentation. To address this issue, an improved model, YOLO-VW, with high accuracy, high efficiency, and a light weight, was proposed for CVW hazard level assessment based on the YOLOv10n model. (1) It replaced conventional convolutions with the lightweight GhostConv, reducing the computational time. (2) The STC module based on the Swin Transformer enhanced the expression of foliage and disease spot boundary features, further reducing the model size. (3) It integrated a squeeze-and-excitation (SE) attention mechanism to suppress irrelevant background information. (4) It employed the stochastic gradient descent (SGD) optimizer to enhance the performance and shorten the detection time. The improved CVW severity assessment model was then deployed on a server, and a real-time detection application (APP) for CVW severity assessment was developed based on this model. The results indicated the following. (1) The YOLO-VW model achieved a mean average precision (mAP) of 89.2% and a frame per second (FPS) rate of 157.98 f/s in assessing CVW, representing improvements of 2.4% and 21.37 f/s over the original model, respectively. (2) The YOLO-VW model's parameters and floating point operations per second (FLOPs) were 1.59 M and 7.8 G, respectively, compressed by 44% and 33.9% compared to the original YOLOv10n model. (3) After deploying the YOLO-VW model on a smartphone, the processing time for each image was 2.42 s, and the evaluation accuracy under various environmental conditions reached 85.5%, representing a 15% improvement compared to the original YOLOv10n model. Based on these findings, YOLO-VW meets the requirements for real-time detection, offering greater robustness, efficiency, and portability in practical applications. This model provides technical support for controlling CVW and developing cotton varieties resistant to verticillium wilt. [ABSTRACT FROM AUTHOR]

Fusarium wilt (FW) is widespread in global cotton production, but the mechanism underlying FW resistance in superior‐fiber‐quality Sea Island cotton is unclear. This study reveals that FW resistance has been the target of genetic improvement of Sea Island cotton in China since the 2010s. The key nonsynonymous single nucleotide polymorphism (SNP, T/C) of gene Gbar_D03G001670 encoding protein phosphatase 2C 80 (PP2C80) results in an amino acid shift (L/S), which is significantly associated with FW resistance of Sea Island cotton. Silencing GbPP2C80 increases FW resistance in Sea Island cotton, whereas overexpressing GbPP2C80 reduces FW resistance in Arabidopsis. GbPP2C80 and GbWAKL14 exist synergistically in Sea Island cotton accessions with haplotype forms "susceptible–susceptible" (TA) and "resistant–resistant" (CC), and interact with each other. CRISPR/Cas9‐mediated knockout of GbWAKL14 enhances FW and Verticillium wilt (VW) resistance in upland cotton and overexpression of GbWAKL14 and GbPP2C80 weakens FW and VW resistance in Arabidopsis. GbPP2C80 and GbWAKL14 respond to FW and VW by modulating reactive oxygen species (ROS) content via affecting MPK3 expression. In summary, two tandem genes on chromosome D03, GbPP2C80, and GbWAKL14, functions as cooperative negative regulators in cotton wilt disease defense, providing novel genetic resources and molecular markers for the development of resistant cotton cultivars. [ABSTRACT FROM AUTHOR]

Mitogen‐activated protein kinase (MAPK) cascade is the center of plant signal transduction system that amplify immune signals into cellular responses by phosphorylating diverse substrates. The MAPK cascade consisting of MAPK kinase kinases (MAPKKKs), MAPK kinases (MAPKKs), and MAPKs is well characterized in plants, in which Raf‐like kinases are generally regarded as MAPKKKs. However, it is rarely reported that Raf‐like MAPKKKs function as middle regulators to link MAPK and its downstream transcription factors in plant immunity. Verticillium wilt, caused by the soil‐borne vascular fungus Verticillium dahliae, is a serious disease in many plants, including cotton. The previous studies showed that GhMPK9 (a MAPK) is involved in the response to Verticillium wilt. Here, the Raf‐like kinase GhRAF39_1 is reported as helper regulates the phosphorylation of WRKY transcription factor GhWRKY40a by GhMPK9. The phosphorylated GhWRKY40a can further activate the transcription of GhERF1b to up‐regulate defense‐related genes while inhibit the transcription of GhABF2 to regulate the stomatal opening, thus improving the resistance to Verticillium wilt in cotton. This study reveals a new signaling module of GhMPK9‐GhRAF39_1‐GhWRKY40a to regulate GhERF1b‐ and GhABF2‐mediated defense responses, which triggers plant defense against Verticillium wilt. [ABSTRACT FROM AUTHOR]

Introduction: 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. Methods: 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. Results: 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. Discussion: 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. [ABSTRACT FROM AUTHOR]

BACKGROUND: As a result of the ineffectiveness of existing control methods against Verticillium dahliae, the causal agent of verticillium wilt of olive (Olea europaea; VWO), it is necessary to search for sustainable and environmentally friendly alternatives, such as bioprotection by products based on plant extracts and other naturally synthesized compounds. Therefore, present study aimed to evaluate the effects of seven natural‐based commercial products on the inhibition of mycelial growth, the germination of V. dahliae conidia and microsclerotia, and disease progression in olive plants (cv. Picual). Aluminium lignosulfonate and a copper phosphonate salt (copper phosphite) were included for comparative purposes. RESULTS: The seaweed and willow extracts and copper phosphite inhibited V. dahliae mycelial growth by more than 50% at the high doses tested. Most of the products inhibited conidial germination by up to 90% compared to the control at the high doses tested. However, none of the products showed efficacy above 50% in inhibiting microsclerotia germination. The willow extract was the most effective at reducing disease severity and progression in olive plants, with no significant differences compared to the non‐inoculated negative control. CONCLUSION: The results of the present study suggest that the use of natural‐based products (i.e. seaweed and willow extracts) is a potential sustainable alternative in an integrated VWO control strategy. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Introduction: Verticillium dahliae causes a devastating Verticillium wilt disease on hundreds of plant species worldwide, including cotton. Understanding the interaction mechanism between V. dahliae and its hosts is the prerequisite for developing effective strategies for disease prevention. Methods: Here, based on the previous observation of an xylosidase-encoding gene (VdxyL3) in V. dahliae being obviously up-regulated after sensing root exudates from a cotton variety susceptible to this pathogen, we investigated the function of VdxyL3 in the growth and pathogenesis of V. dahliae by generating its deletion-mutant strains (1VdxyL3). Results: Deleting VdxyL3 led to increased colony expansion rate, conidial production, mycelial growth, carbon and nitrogen utilization capacities, and enhanced stress tolerance and pathogenicity of V. dahliae. VdxyL3 is a secretory protein; however, VdxyL3 failed to induce cell death in N. benthamiana based on transient expression experiment. Transcriptomic analysis identified 1300 genes differentially expressed (DEGs) between wild-type (Vd952) and 1VdxyL3 during infection, including 348 DEGs encoding secretory proteins, among which contained 122 classical secreted proteins and 226 non-classical secreted proteins. It was notable that of the 122 classical secretory proteins, 50 were carbohydrate-active enzymes (CAZymes) and 58 were small cysteine rich proteins (SCRPs), which were required for the pathogenicity of V. dahliae. Conclusion: The RNA-seq data thus potentially connected the genes encoding these proteins to the pathogenesis of V. dahliae. This study provides an experimental basis for further studies on the interaction between V. dahliae and cotton and the pathogenic mechanism of the fungus. [ABSTRACT FROM AUTHOR]

The study explores anaerobic soil disinfection as an alternative to soil fumigants for controlling Verticillium wilt in strawberry crops. For this purpose, two agrowastes close to the strawberry-growing areas of Huelva province were tested as potential amendments for the control of Verticillium wilt: rice bran and residual strawberry extrudate. Furthermore, two application rates were evaluated: 13.50 and 20.00 t/ha for the rice bran and 16.89 and 25.02 t/ha for residual strawberry extrudate. Amended and anaerobically disinfested soils were compared with a non-amended soil under anaerobic conditions, a soil treated with the chemical fungicide metam sodium and an untreated soil. One week before the start of disinfection treatment, these soils were artificially inoculated with 250 microsclerotia/g dry soil of Verticillium dahliae. After disinfestation treatments, pathogens were quantified, and strawberry plants were cropped in a growth chamber to further evaluate Verticillium wilt severity, which was measured with a symptom scale in the same potting soils. Measurements of the anaerobic condition, pH and microbial population densities were performed, and the results showed significant differences between the different amendments. In addition, the treatment with rice bran at 20 t/ha recorded the lowest population density of V. dahliae. Likewise, it was possible to achieve a reduction in foliar disease severity in all amended treatments in similar percentage to those obtained by chemical treatment. These results suggest potential application of this technique for the control of Verticillium wilt in the strawberry-growing area of Huelva, reducing the use of chemical fumigants. [ABSTRACT FROM AUTHOR]

Monitoring of eight pepper genotypes is conducted at Maritsa Vegetable Crop Research Institute, Plovdiv, Bulgaria, during two successive years, under different diseases and pests'management systems. The surveys is carried out, using the standard field inspection methods to detect virus diseases (Tobacco mosaic virus -- TMV, Cucumber mosaic virus -- CMV and Tomato spotted wilt virus -- TSWV, stolbur (Phytoplasma solani), verticillium wilt (Verticillium dahlia Kleb.), brown leaf spots (Alternaria solani Ellis & Martin) the pests: cotton bollworm (Helicoverpa armigera Hubn.), aphids (Homoptera:Aphididae) and thrips (Thysanoptera:Thripidae). Among observed pathogens Alternaria solani Ellis & Martin and Phytoplasma solani were most important for all varieties and breeding lines in all studied production systems as their infestation were on average 27.37% and 31.79%, respectively. During the survey, no plants with symptoms of verticillium wilting were reported in all studied genotypesunder all growing systems. The mean score of virus attack was below 12%. The average rate of brown leaf spots ranged between 29.05% for breeding line K992 to 36.26% for line K995. The established highest Phytoplasma solani attack averaged from 24.10% for Stryama to 32.15% for Kapia UV. Single plants with thrips damage were observed during seedling production and immediately after transplanting in the field. At maturity stage, an infestation of cotton bollworm (Helicoverpa armigera Hubn.) was observed, while aphids' population was low with no colonies established. The lowest infestation rates by H. armigera were reported in pepper genotypes K992 (0.92%) and K995 (0.49%) grown under organic production conditions. [ABSTRACT FROM AUTHOR]

Background and aims: The diversity and stability of belowground microbiota communities influence plant health. However, changes over time in the microbiome composition associated with healthy and diseased trees remains poorly understood. The goal of this study is to understand the changes in belowground microbial community associated with Verticillium wilt of smoke trees to provide a basis for the combination of biological agents and chemical measures to combat Verticillium wilt of smoke trees. Methods: In this study, we used an amplicon-based approach to reveal distinct differences in the diversity, structure and composition of the belowground microbial communities of healthy and diseased smoke trees from June to October. Results: The structure, composition and interactions of the belowground microbial communities are different between healthy and diseased smoke trees from June to October. The co-occurrence networks of the belowground microbial community changed with the severity of the disease over time. Additionally, the structural community of healthy and diseased trees showed a shift in keystone OTUs. Conclusions: This study improves our understanding of the belowground microbial community composition associated with smoke trees in healthy and diseased conditions, offering the potential to both manipulate the plant microbiome to promote plant health and to provide an integrated management strategy of smoke tree. [ABSTRACT FROM AUTHOR]

Verticillium wilt, caused by the pathogenic fungus Verticillium dahliae, has emerged as a severe threat to cotton globally. However, little is known about the genetic diversity of this pathogen in an infected single cotton plant. In this study, we isolated three new V. dahliae strains from the disease stems of Gossypium hirsutum from the cotton field in Western China and assessed their pathogenicity to the cotton cultivar Xinnongmian-1 and its two transgenic lines, as well as two laboratory strains, VD592 and VD991. These three new V. dahliae strains were identified using DNA barcodes of tryptophan synthase (TS), actin (ACT), elongation factor 1-α (EF), and glyceraldehyde-3-phosphate dehydrogenase (GPD). Moreover, the haplotype analysis revealed that the three new races had distinct haplotypes at the TS locus. Furthermore, the results of culture features and genetic diversity of ISSR (inter-simple sequence repeat) revealed that there were separate V. dahliae strains, which were strong defoliating pathotypes belonging to race 2 type, as determined by particular DNA marker recognition. The identified strains demonstrated varied levels of pathogenicity by leaf disc and entire plant inoculation methods. Conservatively, these strains showed some pathogenicity on cotton lines, but were less pathogenic than the reference strains. The findings revealed that several strong defoliating V. dahliae pathotypes coexist on the same cotton plant. It indicats the importance of regular monitoring as an early warning system, as well as the detection and reporting of virulent pathogen strains and their effects on crop response. [ABSTRACT FROM AUTHOR]

SUMMARY: Verticillium wilt (VW) is a devasting disease affecting various plants, including upland cotton, a crucial fiber crop. Despite its impact, the genetic basis underlying cotton's susceptibility or defense against VW remains unclear. Here, we conducted a genome‐wide association study on VW phenotyping in upland cotton and identified a locus on A13 that is significantly associated with VW resistance. We then identified a cystathionine β‐synthase domain gene at A13 locus, GhCBSX3A, which was induced by Verticillium dahliae. Functional analysis, including expression silencing in cotton and overexpression in Arabidopsis thaliana, confirmed that GhCBSX3A is a causal gene at the A13 locus, enhancing SAR‐RBOHs‐mediated apoplastic oxidative burst. We found allelic variation on the TATA‐box of GhCBSX3A promoter attenuated its expression in upland cotton, thereby weakening VW resistance. Interestingly, we discovered that altered artificial selection of GhCBSX3A_R (an elite allele for VW) under different VW pressures during domestication and other improved processes allows specific human needs to be met. Our findings underscore the importance of GhCBSX3A in response to VW, and we propose a model for defense‐associated genes being selected depending on the pathogen's pressure. The identified locus and gene serve as promising targets for VW resistance enhancement in cotton through genetic engineering. Significance Statement: Upland cotton cultivars demonstrate limited resistance to Verticillium wilt (VW), with most studies indicating that upland cotton (AD1) possesses an inherent disadvantage concerning VW resistance. We found that the artificial selection of GhCBSX3A_R, an elite allele for VW resistance, achieved through enhanced reactive oxygen species accumulation, resulted in reduced VW resistance in improved accessions. This evidence supports the idea that the susceptibility against VW may be attributed to a significant decline in VW resistance during upland cotton domestication. [ABSTRACT FROM AUTHOR]

Understanding the extent of heritability of a plant-associated microbiome (phytobiome) is critically important for exploitation of phytobiomes in agriculture. Two crosses were made between pairs of cotton cultivars (Z2 and J11, L1 and Z49) with differential resistance to Verticillium wilt. F2 plants were grown in a field, together with the four parents to study the heritability of cotton rhizosphere microbiome. Amplicon sequencing was used to profile bacterial and fungal communities in the rhizosphere. F2 offspring plants of both crosses had higher average alpha diversity indices than the two parents; parents differed significantly from F2 offspring in Bray–Curtis beta diversity indices as well. Two types of data were used to study the heritability of rhizosphere microbiome: principal components (PCs) and individual top microbial operational taxonomic units (OTUs). For the L1 × Z49 cross, the variance among the F2 progeny genotypes (namely, genetic variance, VT) was significantly greater than the random variability (VE) for 12 and 34 out of top 100 fungal and bacterial PCs, respectively. For the Z2 × J11 cross, the corresponding values were 10 and 20 PCs. For 29 fungal OTUs and 10 bacterial OTUs out of the most abundant 100 OTUs, genetic variance (VT) was significantly greater than VE for the L1 × Z49 cross; the corresponding values for the Z2 × J11 cross were 24 and one. The estimated heritability was mostly in the range of 40% to 60%. These results suggested the existence of genetic control of polygenic nature for specific components of rhizosphere microbiome in cotton. Key points: • F2offspring cotton plants differed significantly from parents in rhizosphere microbial diversity. • Specific rhizosphere components are likely to be genetically controlled by plants. • Common PCs and specific microbial groups are significant genetic components between the two crosses. [ABSTRACT FROM AUTHOR]

Verticillium wilt, a soilborne vascular disease caused by Verticillium dahliae , strongly affects cotton yield and quality. In this study, an isolated rhizosphere bacterium, designated Bacillus velezensis BvZ45-1, exhibited >46% biocontrol efficacy against cotton verticillium wilt under greenhouse and field conditions. Moreover, through crude protein extraction and mass spectrometry analyses, we found many antifungal compounds present in the crude protein extract of BvZ45-1. The purified oxalate decarboxylase Odx_S12 from BvZ45-1 inhibited the growth of V. dahliae Vd080 by reducing the spore yield, causing mycelia to rupture, spore morphology changes, cell membrane rupture, and cell death. Subsequently, overexpression of Odx_S12 in Arabidopsis significantly improved plant resistance to V. dahliae. Through studies of the resistance mechanism of Odx_S12 , V. dahliae was shown to produce oxalic acid (OA), which has a toxic effect on Arabidopsis leaves. Odx_S12 overexpression reduced Arabidopsis OA content, enhanced tolerance to OA, and improved resistance to verticillium wilt. Transcriptomics and quantitative real-time PCR analysis revealed that Odx_S12 promoted a reactive oxygen species burst and a salicylic acid- and abscisic acid-mediated defence response in Arabidopsis. In summary, this study not only identified B. velezensis BvZ45-1 as an efficient biological control agent, but also identified the resistance gene Odx_S12 as a candidate for cotton breeding against verticillium wilt. [ABSTRACT FROM AUTHOR]

Verticillium wilt, caused by Verticillium dahliae, is a soil-borne disease affecting eggplant. Wild eggplant, recognized as an excellent disease-resistant resource against verticillium wilt, plays a pivotal role in grafting and breeding for disease resistance. However, the underlying resistance mechanisms of wild eggplant remain poorly understood. This study compared two wild eggplant varieties, LC-2 (high resistance) and LC-7 (sensitive) at the phenotypic, transcriptomic, and metabolomic levels to determine the molecular basis of their resistance to verticillium wilt. These two varieties exhibit substantial phenotypic differences in petal color, leaf spines, and fruit traits. Following inoculation with V. dahliae, LC-2 demonstrated significantly higher activities of polyphenol oxidase, superoxide dismutase, peroxidase, phenylalanine ammonia lyase, b-1,3 glucanase, and chitinase than did LC-7. RNA sequencing revealed 4,017 differentially expressed genes (DEGs), with a significant portion implicated in processes associated with disease resistance and growth. These processes encompassed defense responses, cell wall biogenesis, developmental processes, and biosynthesis of spermidine, cinnamic acid, and cutin. A gene co-expression analysis identified 13 transcription factors as hub genes in modules related to plant defense response. Some genes exhibited distinct expression patterns between LC-2 and LC-7, suggesting their crucial roles in responding to infection. Further, metabolome analysis identified 549 differentially accumulated metabolites (DAMs) between LC-2 and LC-7, primarily consisting of compounds such as flavonoids, phenolic acids, lipids, and other metabolites. Integrated transcriptome and metabolome analyses revealed the association of 35 gene-metabolite pairs in modules related to the plant defense response, highlighting the interconnected processes underlying the plant defense response. These findings characterize the molecular basis of LC-2 resistance to verticillium wilt and thus have potential value for future breeding of wilt-resistant eggplant varieties. [ABSTRACT FROM AUTHOR]

Summary: The cell wall is the major interface for arbuscular mycorrhizal (AM) symbiosis. However, the roles of cell wall proteins and cell wall synthesis in AM symbiosis remain unclear. We reported that a novel wall‐associated kinase 13 (GhWAK13) positively regulates AM symbiosis and negatively regulates Verticillium wilt resistance in cotton.GhWAK13 transcription was induced by AM symbiosis and Verticillium dahliae (VD) infection. GhWAK13 is located in the plasma membrane and expressed in the arbuscule‐containing cortical cells of mycorrhizal cotton roots.GhWAK13 silencing inhibited AM colonization and repressed gene expression of the mycorrhizal pathway. Moreover, GhWAK13 silencing improved Verticillium wilt resistance and triggered the expression of immunity genes. Therefore, GhWAK13 is considered an immune suppressor required for AM symbiosis and disease resistance. GhWAK7A, a positive regulator of Verticillium wilt resistance, was upregulated in GhWAK13‐silenced cotton plants. Silencing GhWAK7A improved AM symbiosis. Oligogalacturonides application also suppressed AM symbiosis. Finally, GhWAK13 negatively affected the cellulose content by regulating the transcription of cellulose synthase genes.The results of this study suggest that immunity suppresses AM symbiosis in cotton. GhWAK13 affects AM symbiosis by suppressing immune responses. See also the Commentary on this article by Giovanetti & Genre, 242: 1851–1853. [ABSTRACT FROM AUTHOR]

As a universal second messenger, cytosolic calcium (Ca2+) functions in multifaceted intracellular processes, including growth, development and responses to biotic/abiotic stresses in plant. The plant‐specific Ca2+ sensors, calmodulin and calmodulin‐like (CML) proteins, function as members of the second‐messenger system to transfer Ca2+ signal into downstream responses. However, the functions of CMLs in the responses of cotton (Gossypium spp.) after Verticillium dahliae infection, which causes the serious vascular disease Verticillium wilt, remain elusive. Here, we discovered that the expression level of GbCML45 was promoted after V. dahliae infection in roots of cotton, suggesting its potential role in Verticillium wilt resistance. We found that knockdown of GbCML45 in cotton plants decreased resistance while overexpression of GbCML45 in Arabidopsis thaliana plants enhanced resistance to V. dahliae infection. Furthermore, there was physiological interaction between GbCML45 and its close homologue GbCML50 by using yeast two‐hybrid and bimolecular fluorescence assays, and both proteins enhanced cotton resistance to V. dahliae infection in a Ca2+‐dependent way in a knockdown study. Detailed investigations indicated that several defence‐related pathways, including salicylic acid, ethylene, reactive oxygen species and nitric oxide signalling pathways, as well as accumulations of lignin and callose, are responsible for GbCML45‐ and GbCML50‐modulated V. dahliae resistance in cotton. These results collectively indicated that GbCML45 and GbCML50 act as positive regulators to improve cotton Verticillium wilt resistance, providing potential targets for exploitation of improved Verticillium wilt‐tolerant cotton cultivars by genetic engineering and molecular breeding. [ABSTRACT FROM AUTHOR]

Verticillium dahliae is a xylem‐invading fungal pathogen that causes vascular wilt in a wide range of angiosperms. The pathogen uses a variety of virulence factors to invade and colonize its hosts. Here, we report that VdNEP, an NLP (Necrosis and ethylene inducing peptide 1‐Like Protein), functions as one such factor in multiple hosts. Eggplant leaves treated with VdNEP developed necrotic symptoms. Overexpression of VdNEP by incorporating extra copies of the VdNEP gene increased virulence to cotton, eggplant and tomato plants, suggesting its role as a virulence factor in diverse plants. Increased expression of VdNEP among the transformants did not correlate with the number of VdNEP inserts, suggesting that its expression was affected by the genomic context of the insertion sites. Interestingly, a transformant derived from a defoliating strain with high VdNEP transcript levels caused disease symptoms in tomato plants, whereas the corresponding wild‐type strain did not cause visible symptoms. The amount of V. dahliae DNA in plants infected with this VdNEP‐overexpressing transformant was 22 times higher than that in plants infected with the wild‐type isolate, further supporting the critical role of VdNEP in infection. A VdNEP‐EGFP fusion was constructed to follow its localization in fungal cells and during infection. [ABSTRACT FROM AUTHOR]

Root exudates play a key role in the life cycle of Verticillium dahliae, the causal agent of Verticillium wilt diseases, because they induce microsclerotia germination to initiate plant infection through the roots. In olive plants, the genotype and the application of biological control agents (BCAs) or phosphonate salts influence the ability of root exudates to decrease V. dahliae viability. Understanding the chemical composition of root exudates could provide new insights into the mechanisms of olive plant defense against V. dahliae. Therefore, the main goal of this study was to analyze the metabolomic profiles of root exudates collected from the olive cultivars Arbequina, Frantoio, and Picual subjected to treatment with BCAs (Aureobasidium pullulans AP08, Bacillus amyloliquefaciens PAB-024) or phosphonate salts (copper phosphite, potassium phosphite). These treatments were selected due to their effectiveness as inducers of resistance against Verticillium wilt in olive plants. Our metabolomic analysis revealed that the olive cultivars exhibited differences in root exudates, which could be related to the different degrees of susceptibility to V. dahliae. The composition of root exudates also changed with the application of BCAs or phosphonate fertilizer, highlighting the complex and dynamic nature of the interactions between olive cultivars and treatments preventing V. dahliae infections. Thus, the identification of genotype-specific metabolic changes and specific metabolites induced by these treatments emphasizes the potential of resistance inducers for enhancing plant defense and promoting the growth of beneficial microorganisms. [ABSTRACT FROM AUTHOR]

Verticillium dahliae (Vd) and Fusarium oxysporum f. sp. lycopersici (Fol) are two major fungal pathogens that infect tomato plants, causing significant challenges in their control since both pathogens can persist in the soil for several years even in the absence of a host plant and no effective fungicides are available at present. This study investigated the efficacy of two biocontrol formulations, Clonotri (containing Trichoderma and Clonostachys microorganisms) and Strepse (comprising Streptomyces and Pseudomonas microorganisms), against Vd and Fol and their impact on tomato fruit quality and yield under greenhouse conditions. The pathogenicity experiment demonstrated that the Clonotri formulation, containing Trichoderma and Clonostachys spores, significantly reduced Fusarium wilt disease by 32% compared to the control group. However, in the Vd pathogenicity experiment, the formulations did not exhibit disease reduction, although treatment with Strepse, containing Streptomyces and Pseudomonas microorganisms, resulted in a preserved total fruit number when compared to uninfected plants. Analysis of fruit quality attributes revealed no significant differences among the various interventions. Furthermore, Fol infection in the first fruit set significantly increased fruit firmness, while Vd infection resulted in elevated levels of total soluble solids in fruits. These findings demonstrate that the evaluated biocontrol formulations provide a degree of protection against fungal wilt pathogens in tomato plants and can increase yield in greenhouse conditions while having minimal impact on overall fruit quality attributes. [ABSTRACT FROM AUTHOR]

Brinjal, eggplant or aubergine (Solanum melongena L.) belongs to the Solanaceae family and is a widely cultivated warm-season vegetable in India and around the world. Brinjal production and productivity are strongly affected by many biotic stresses, viz., fusarium wilt, verticillium wilt and small leaves of brinjal. Among all the diseases, bacterial wilt (BW) is the most destructive disease in tropical, subtropical, temperate, and humid regions worldwide due to the broad host range and prolonged duration of spore survival. BW disease in brinjal is caused by Ralstonia solanacearum, which belongs to the β-proteobacteria family and is a gram-negative, nonsporeforming, rod-shaped, and soilborne bacterium. BW disease management strategies, such as culture, mechanical, biological, and chemical methods, are ineffective due to the prolonged survival period of the bacterium in the soil and its wide host range. The use of resistant varieties and hybrids against BW disease is the safest method for controlling this disease. Information on the genetics of resistance to BW disease in brinjal is vital for the development of an effective breeding method and for identifying bacterial wilt resistance in preferable brinjal cultivars. The use of molecular markers associated with BW disease resistance gene loci helps to characterize traits of interest and develop resistant varieties and hybrids. This review described recent advances in different control measures. We focused on the importance of marker-assisted selection for identifying bacterial wilt diseases. [ABSTRACT FROM AUTHOR]

Verticillium wilt is a soil-borne disease caused by distinct vegetative compatibility groups (VCG) of the fungus Verticillium dahliae. Defoliating (VCG 1A) and non-defoliating (VCG 2A) pathotypes of V. dahliae have contributed to yield losses of cotton production in Australia. To study the virulence and the infection process of V. dahliae on cotton, two isolates, one representing each VCG, have been transformed with fluorescent protein genes. The transformants maintained their ability to infect the host, and both strains were observed to move through the plant vasculature to induce wilt symptoms. Furthermore, virulence testing suggests that the cotton V. dahliae strains can endophytically colonise common weed plant species found in the Australian landscape, and that is contrasted by their ability to infect and colonise native tobacco plants. The fluorescently labelled strains of V. dahliae not only allowed us to gain a thorough understanding of the infection process but also provided a method to rapidly identify recovered isolates from host colonisation studies. [ABSTRACT FROM AUTHOR]

Cotton Verticillium wilt is mainly caused by the fungus Verticillium dahliae, which threatens the production of cotton. Its pathogen can survive in the soil for several years in the form of microsclerotia, making it a destructive soil-borne disease. The accurate, sensitive, and rapid detection of V. dahliae from complex soil samples is of great significance for the early warning and management of cotton Verticillium wilt. In this study, we combined the loop-mediated isothermal amplification (LAMP) with CRISPR/Cas12a technology to develop an accurate, sensitive, and rapid detection method for V. dahliae. Initially, LAMP primers and CRISPR RNA (crRNA) were designed based on a specific DNA sequence of V. dahliae, which was validated using several closely related Verticillium spp. The lower detection limit of the LAMP-CRISPR/Cas12a combined with the fluorescent visualization detection system is approximately ~10 fg/μL genomic DNA per reaction. When combined with crude DNA-extraction methods, it is possible to detect as few as two microsclerotia per gram of soil, with the total detection process taking less than 90 min. Furthermore, to improve the method's user and field friendliness, the field detection results were visualized using lateral flow strips (LFS). The LAMP-CRISPR/Cas12a-LFS system has a lower detection limit of ~1 fg/μL genomic DNA of the V. dahliae, and when combined with the field crude DNA-extraction method, it can detect as few as six microsclerotia per gram of soil, with the total detection process taking less than 2 h. In summary, this study expands the application of LAMP-CRISPR/Cas12a nucleic acid detection in V. dahliae and will contribute to the development of field-deployable diagnostic productions. [ABSTRACT FROM AUTHOR]

Introduction: Bacillus velezensis occurs extensively in the soil environment. It produces a range of antimicrobial compounds that play an important role in the field of biological control. However, during the actual application process it is often affected by factors such as the medium formulation and fermentation conditions, and therefore biocontrol measures often do not achieve their expected outcomes. Methods: In this study, the B. velezensis BHZ-29 strain was used as the research object. The carbon and nitrogen sources, and inorganic salts that affect the number of viable bacteria and antibacterial potency of B. velezensis BHZ-29, were screened by a single factor test. A Plackett--Burman design experiment was conducted to determine the significant factors affecting the number of viable bacteria and antibacterial potency, and a Box--Behnken design experiment was used to obtain the optimal growth of B. velezensis BHZ-29. The medium formula that produced the highest number of viable bacteria and most antibacterial substances was determined. The initial pH, temperature, amount of inoculant, liquid volume, shaking speed, and culture time were determined by a single factor test. The factors that had a significant influence on the number of viable bacteria of B. velezensis BHZ-29 were selected by an orthogonal test. A Box-- Behnken design experiment was conducted to obtain the optimal fermentation conditions, and highest number of viable bacteria and antibacterial titer. Results: Molasses, peptone, and magnesium sulfate had significant effects on the viable count and antibacterial titer of B. velezensis BHZ-29. The viable count of B. velezensis BHZ-29 increased from 7.83 x 109 to 2.17 x 1010 CFU/mL, and the antibacterial titer increased from 111.67 to 153.13 mm/mL when the optimal media were used. The optimal fermentation conditions for B. velezensis BHZ-29 were as follows: temperature 25.57°C, pH 7.23, culture time 95.90 h, rotation speed 160 rpm, amount of inoculant 2%, and liquid volume 100 ml. After the optimization of fermentation conditions, the number of viable bacteria increased to 3.39 x 1010 CFU/mL, and the bacteriostatic titer increased to 158.85 mm/ml. The plant height and leaf number of cotton plants treated with BHZ-29 fermentation broth were higher than those of cotton inoculated with Verticillium dahliae. The number of bacteria was 1.15 x 107 CFU/g, and the number of fungi was 1.60 x 105 spores/g. The disease index of the cotton seedlings treated with the optimized fermentation broth was 2.2, and a control effect of 93.8% was achieved. B. velezensis BHZ-29 could reduce the disease index of cotton Verticillium wilt and had a controlling effect on the disease. The best effect was achieved in the treatment group with an inoculation concentration of 2 x 108 CFU/ml, the disease index was 14.50, and a control effect of 84.18% was achieved. Discussion: The fermentation process parameters of the number of viable bacteria and antibacterial titer by strain B. velezensis BHZ-29 were optimized to lay a foundation for the practical production and application of strain B. velezensis BHZ-29 in agriculture. [ABSTRACT FROM AUTHOR]

Aubergine (Solanum melongena L.) (Solanaceae) is a widespread crop in the Mediterranean basin. Verticillium dahliae is one of the main soil-borne pathogens affecting the aubergine crop. Its control has traditionally been achieved by soil fumigation with chemical disinfectants. Restrictions on the use of chemical fumigants have led to the search for solutions in genetic resistance using rootstocks. In southeastern Spain, aubergines are grafted for the control of V. dahliae. Two Solanum torvum rootstocks (Hugo F1 and Torpedo) and a Solanum melongena hybrid (Javah F1) were tested against five isolates of V. dahliae obtained from grafted (A1 and A2) and ungrafted (Vd8, Vd17 and Vd66) aubergines compared with the susceptible cultivar Larne F1 under controlled conditions. Isolates from grafted plants infected all three rootstocks, with differences observed in the percentage of plants with symptoms and in the disease symptom severity. Three strains isolated from the ungrafted aubergines (Vd8, Vd17 and Vd66) infected Javah F1 rootstock. The Hugo F1 and Torpedo rootstocks showed a high level of resistance to V. dahliae, while Javah F1 was susceptible to the pathogen. The Hugo F1 and Torpedo rootstocks are suitable for mitigating the effects of Verticillium wilt in Mediterranean aubergine crops. Understanding the nature of the resistance from S. torvum could enhance the benefits of grafting or facilitate the introduction of resistance into commercial cultivars. [ABSTRACT FROM AUTHOR]

Verticillium dahliae Kleb., the causal agent of Verticillium wilts, is a devastating plant disease affecting many plant species. Fungus V. dahliae was detected in a partially artificially established Acer pseudoplatanus L. forest stand in central Slovenia. This finding incited further study about the risk of different sources of V. dahliae isolates for maples in forests and the pathogenicity of three V. dahliae isolates of different origins was tested on saplings of A. pseudoplatanus, A. platanoides L., and A. campestre L. The inoculated saplings exhibited disease symptoms, i.e., leaf necrosis and wilting. At the end of the pathogenicity test, typical xylem browning was visible on the cross-sections, and the pathogen was successfully re-isolated. The isolates showed significant differences in their pathogenicity to specific maple hosts, with the agricultural isolate (originated from bell pepper) being the most aggressive on all three maple species. The disease severity index (DSI) and relative area under the disease progress curve (rAUDPC), as well as the success of re-isolation, indicate that A. platanoides is the most susceptible to inoculation with V. dahliae. In addition, significant differences in sapling biomass were observed between treated and control plants. These results suggest that maples in forest stands are threatened by V. dahliae, and biosecurity measures should be considered and implemented in forest management to reduce the transmission and potential spread of the pathogen. [ABSTRACT FROM AUTHOR]

Smoke tree (Cotinus coggygria) is an important ornamental tree that represents the autumnal landscape of red leaves in Northern China, especially in Beijing. However, Verticillium wilt, caused by the fungus (Verticillium dahliae), has resulted in a high mortality rate for smoke trees, posing a serious threat to the highly valued landscape of red leaves in Beijing. To explore an efficient control measure for Verticillium wilt, we systematically analyzed the applicability and efficacy of multiple treatments for three consecutive years in Xiangshan Park and Badaling Forest Park. From 2021 to 2023, diseased smoke trees in Xiangshan Park were subjected to three application methods (agent irrigation, trunk injection, or a combination of the two) and five candidate agents, namely Bacillus subtilis, azoxystrobin, propiconazole, carbendazim, and prochloraz. Analyses of the data for three consecutive years revealed a decreasing trend in the annual disease incidence rate. Specifically, the combined application of agent irrigation and trunk injection exhibited the highest control effect and a significant improvement in the landscape of red leaves in Beijing. Furthermore, the combination of propiconazole via irrigation plus the trunk injection of carbendazim and prochloraz had the greatest control effect. These suppressive measurements were further used and demonstrated to be effective in Badaling Forest Park. Overall, our study provides an effective disease management means for controlling Verticillium wilt in smoke trees. [ABSTRACT FROM AUTHOR]

DUSPs, a diverse group of protein phosphatases, play a pivotal role in orchestrating cellular growth and development through intricate signaling pathways. Notably, they actively participate in the MAPK pathway, which governs crucial aspects of plant physiology, including growth regulation, disease resistance, pest resistance, and stress response. DUSP is a key enzyme, and it is the enzyme that limits the rate of cell metabolism. At present, complete understanding of the DUSP gene family in cotton and its specific roles in resistance to Verticillium wilt (VW) remains elusive. To address this knowledge gap, we conducted a comprehensive identification and analysis of four key cotton species: Gossypium arboreum, Gossypium barbadense, Gossypium hirsutum, and Gossypium raimondii. The results revealed the identification of a total of 120 DUSP genes in the four cotton varieties, which were categorized into six subgroups and randomly distributed at both ends of 26 chromosomes, predominantly localized within the nucleus. Our analysis demonstrated that closely related DUSP genes exhibited similarities in terms of the conserved motif composition and gene structure. A promoter analysis performed on the GhDUSP gene promoter revealed the presence of several cis-acting elements, which are associated with abiotic and biotic stress responses, as well as hormone signaling. A tissue expression pattern analysis demonstrated significant variations in GhDUSP gene expression under different stress conditions, with roots exhibiting the highest levels, followed by stems and leaves. In terms of tissue-specific detection, petals, leaves, stems, stamens, and receptacles exhibited higher expression levels of the GhDUSP gene. The gene expression analysis results for GhDUSPs under stress suggest that DUSP genes may have a crucial role in the cotton response to stress in cotton. Through Virus-Induced Gene Silencing (VIGS) experiments, the silencing of the target gene significantly reduced the resistance efficiency of disease-resistant varieties against Verticillium wilt (VW). Consequently, we conclude that GH_A11G3500-mediated bispecific phosphorylated genes may serve as key regulators in the resistance of G. hirsutum to Verticillium wilt (VW). This study presents a comprehensive structure designed to provide an in-depth understanding of the potential biological functions of cotton, providing a strong foundation for further research into molecular breeding and resistance to plant pathogens. [ABSTRACT FROM AUTHOR]

Background: In agricultural production, fungal diseases significantly impact the yield and quality of cotton (Gossypium spp.) with Verticillium wilt posing a particularly severe threat. Results: This study is focused on investigating the effectiveness of endophytic microbial communities present in the seeds of disease-resistant cotton genotypes in the control of cotton Verticillium wilt. The technique of 16S ribosomal RNA (16S rRNA) amplicon sequencing identified a significant enrichment of the Bacillus genus in the resistant genotype Xinluzao 78, which differed from the endophytic bacterial community structure in the susceptible genotype Xinluzao 63. Specific enriched strains were isolated and screened from the seeds of Xinluzao 78 to further explore the biological functions of seed endophytes. A synthetic microbial community (SynCom) was constructed using the broken-rod model, and seeds of the susceptible genotype Xinluzao 63 in this community that had been soaked with the SynCom were found to significantly control the occurrence of Verticillium wilt and regulate the growth of cotton plants. Antibiotic screening techniques were used to preliminarily identify the colonization of strains in the community. These techniques revealed that the strains can colonize plant tissues and occupy ecological niches in cotton tissues through a priority effect, which prevents infection by pathogens. Conclusion: This study highlights the key role of seed endophytes in driving plant disease defense and provides a theoretical basis for the future application of SynComs in agriculture. [ABSTRACT FROM AUTHOR]

Verticillium dahliae is a broad host-range pathogen that causes vascular wilt in plants. The Verticillium wilt disease severity assay on plants caused by V. dahliae mainly includes two methods, one is a plant disease grade classification based on disease severity statistics [namely the disease index (DI)], and the other is the V. dahliae biomass quantification in plants (namely the fungal biomass). In this study, the relationships of pathogenicity with the DI, pathogenicity with the biomass, and the correlation analysis of the DI and relative fungal biomass were analysed. The results showed that pathogenicity assessment of V. dahliae strains using the DI method was able to give an intuitive reflection of the pathogenic ability for defoliating and non-defoliating strains; moreover, the method of quantitative PCR for fungal biomass also had high repeatability and stability. As a whole, the correlation coefficient between the DI and fungal biomass values of 28 strains was 0.728, indicating that the two data sets were highly correlated; however, the correlation coefficients of the defoliating strains and non-defoliating strains were only 0.5384 and 0.4547, respectively. In conclusion, the correlation coefficient between the DI and the fungal biomass presented high consistency, which could provide some meaningful exploration for the more accurate pathogenicity identification of V. dahliae. [ABSTRACT FROM AUTHOR]

Aims This study aimed to assess the impact of rocket (Eruca sativa) extract on Verticillium wilt in eggplants, explore rhizospheric microorganisms for disease biocontrol, and evaluate selected strains' induced systemic resistance (ISR) potential while characterizing their genomic and biosynthetic profiles. Methods and results Rocket extract application led to a significant reduction in Verticillium wilt symptoms in eggplants compared to controls. Isolated microorganisms from treated soil, including Paraburkholderia oxyphila EP1, Pseudomonas citronellolis EP2, Paraburkholderia eburnea EP3, and P. oxyphila EP4 and EP5, displayed efficacy against Verticillium dahliae , decreasing disease severity and incidence in planta. Notably, strains EP3 and EP4 triggered ISR in eggplants against V. dahliae. Genomic analysis unveiled shared biosynthetic gene clusters, such as ranthipeptide and non-ribosomal peptide synthetase-metallophore types, among the isolated strains. Additionally, metabolomic profiling of EP2 revealed the production of metabolites associated with amino acid metabolism, putative antibiotics, and phytohormones. Conclusions The application of rocket extract resulted in a significant reduction in Verticillium wilt symptoms in eggplants, while the isolated microorganisms displayed efficacy against V. dahliae , inducing systemic resistance and revealing shared biosynthetic gene clusters, with metabolomic profiling highlighting potential disease-suppressing metabolites. [ABSTRACT FROM AUTHOR]

Symptoms of infection okra plants (Abelmoschus esculentus L.) with Verticillium wilt were recorded during the summer of 2021 in Beni Suef Governorate specifically in Nasser, Beba, and El-Wasta Counties. The symptoms appeared on the external parts of plants, specifically through the alteration of color in the leaves of infected shoots, which transition from a normal color to a light green or yellow hue, had V-shaped chlorotic lesions, loss their turgidity and eventually desiccation, plant stunting and vascular discoloration. The presence of V. tricorpus on okra has not been previously documented. The isolated fungus was identified on the basis of morphological characteristics and molecular traits by utilizing sequencing techniques applied to the internal transcribed space (ITS region) of the ribosomal RNA gene. The analysis of the fungal sequencing data revealed a substantial genetic similarity and close ties between the fungus in question and V. tricorpus by 99.24–100% identity and 97–100% coverage with several strains of V. tricorpus, including the type strain CBS447.54 (NR_126128). The obtained sequences were submitted to the GenBank database and assigned the accession number MZ936483. Pathogenicity test confirmed that V. tricorpus had pathogenic properties, as shown by the appearance of disease symptoms that were similar to those previously noted on okra plants in the surveyed regions. This observation represents the initial documentation of V. tricorpus in relation to the occurrence of Verticillium wilt disease in okra. However, there is currently no information of host resistance to this disease among Egyptian okra cultivars, although it is essential for sustainable disease management. Our study indicate that cultivar of Balady green is more resistance to Verticillium wilt while the Iranian red cv. is more sensitive to infection. [ABSTRACT FROM AUTHOR]

Cotton (Gossypium hirsutum L.) is economically important in Xinjiang, China, for many years; Verticillium wilt, arid climate, and soil salinization reduce cotton production, which has inhibited development of the cotton economy. Thus, creating a microbial agent specifically for cotton to resisting environmental stress is crucial. In this study, water dispersible granules strains (Bacillus tequilensis C-9 and B. sphingosporium A1) were screened for antagonistic bacteria of Verticillium dahliae from the rhizosphere soil of Ferula sinkiangensis K.M. Shen, and screened for carrier and auxiliary that could enhance the survival of strains A1+C-9. To test the efficacy and practicality of the water dispersible granules, their growth-promoting effect was examined. Meanwhile, bursts of reactive oxygen species and changes in the physiological indicators related to cotton growth were detected after the application of these granules to cotton that had been subjected to biotic and abiotic stresses to study the ability of the water dispersible granules to induce immunity in cotton. It was found that the treated group could increase the above-ground and below-ground fresh weight of cotton by 20.06% and 22.71% relative to the control group, and the biocontrol effect on cotton was increased by 46.55%. In salt and drought stress, catalase activity could be increased by 14.61% and 15.52%, respectively, relative to the control group. Comprehensive analyses showed that the application of water dispersible granules that contained A1+C-9 helped the cotton to grow and develop, inhibited V. dahliae from infecting cotton plants, reduced the damage from drought and salt stress, and increased the resilience of cotton to stress. [ABSTRACT FROM AUTHOR]

The most common soilborne diseases affecting the strawberry industry in California include Verticillium wilt due to Verticillium dahliae, charcoal root rot due to Macrophomina phaseolina, and Fusarium wilt due to Fusarium oxysporum f. sp. fragariae. Detection of these pathogens in soil is an important facet of disease management and fumigation recommendations. Whereas the soil populations of both M. phaseolina and V. dahliae can be readily quantified with quantitative PCR (qPCR) assays using DNA extractions with 500 mg of soil, the single-cell nature of the F. oxysporum chlamydospore does not provide enough pathogen DNA from 500-mg extractions to be reliably quantified. Here, we describe an improved DNA extraction protocol from 10 to 15 g of soil that allows for the quantification of F. oxysporum f. sp. fragariae populations below 10 CFU/g. The relationship between results from the TaqMan qPCR assay and pathogen population density in soil was determined by using this extraction method in pathogen-free soils artificially infested with a hygromycin-resistant strain of F. oxysporum f. sp. fragariae to facilitate accurate colony counts when plated on a selective medium. Although the protocol was developed for F. oxysporum f. sp. fragariae, it is applicable for detection and quantification of other soilborne pathogens. [ABSTRACT FROM AUTHOR]

In practical applications, the effectiveness of biological control agents such as Bacillus is often unstable due to different soil environments. Herein, we aimed to explore the control effect and intrinsic mechanism of Bacillus in black soil and red soil in combination with tomato Verticillium wilt. Bacillus application effectively controlled the occurrence of Verticillium wilt in red soil, reducing the incidence by 19.83%, but played a limited role in black soil. Bacillus colonized red soil more efficiently. The Verticillium pathogen decreased by 71.13% and 76.09% after the application of Bacillus combinations in the rhizosphere and bulk of the red soil, respectively, while there was no significant difference in the black soil. Additionally, Bacillus application to red soil significantly promoted phosphorus absorption. Furthermore, it significantly altered the bacterial community in red soil and enriched genes related to pathogen antagonism and phosphorus activation, which jointly participated in soil nutrient activation and disease prevention, promoting tomato plant growth in red soil. This study revealed that the shaping of the bacterial community by native soil may be the key factor affecting the colonization and function of exogenous Bacillus. [ABSTRACT FROM AUTHOR]

Cotton is a critical crop with massive economic implications worldwide. Verticillium wilt is a soil-borne ailment caused by Verticillium dahliae, which harms the growth and development of cotton. Therefore, investigating the genes associated with resistance to verticillium wilt is of particular significance. In this study, we identified the GhIQD1 gene through transcriptome analysis and experimentally characterized the role of the GhIQD1 gene in cotton against V. dahliae. The findings indicated that GhIQD1 acts as a calmodulin-binding protein. The expression of GhIQD1 was the highest in stems, and the expression level increased significantly following infection with V. dahliae. The expression in resistant cotton varieties was higher than in susceptible cotton varieties. Through overexpression of the GhIQD1 gene in tobacco, these transgenic plants exhibited improved resistance to V. dahliae. In contrast, by silencing the GhIQD1 gene in cotton through VIGS, the resistance to V. dahliae was reduced. Following inoculation, the leaves yellowed, and the disease index was higher. Transcriptome analysis of transgenic tobacco 72 h after inoculation indicated that overexpression of GhIQD1 increased the enrichment of the calmodulin pathway and stimulated the production of plant hormones alongside secondary metabolites. Consequently, we investigated the relationship between the GhIQD1 gene and plant disease-resistant hormones SA, JA, and ABA. In summary, this study uncovered the mechanism by which GhIQD1 conferred resistance to V. dahliae in cotton through positive regulation of JA and ABA, providing crucial information for further research on the adaptation of plants to pathogen invasion. [ABSTRACT FROM AUTHOR]

BACKGROUND: Verticillium dahliae is a soil‐borne pathogenic fungus that causes Verticillium wilt disease on more than 400 plant species worldwide. Because of its broad host range and its ability to survive long term in the soil, there are few effective control measures for V. dahliae once it has become established. Accurate, sensitive, and rapid detection of V. dahliae is crucial for limiting pathogen entry into new regional environments and early management of Verticillium wilt. RESULTS: In this study, we developed a method to detect V. dahliae based on recombinase polymerase amplification (RPA) and CRISPR/Cas technology and used fluorescence and lateral flow test strips to monitor the outcomes. Through the establishment and optimization of RPA–CRISPR/Cas13a detection, the sensitivity of the fluorescence method was 1 am for genomic DNA (gDNA) within 20 min, whereas the sensitivity of the lateral flow strip method was 100 am for gDNA in 30 min. The field applicability of RPA–CRISPR/Cas13a was also validated by the detection of V. dahliae on smoke trees (Cotinus coggygria) in Xiangshan Park, Beijing, China. Finally, diplex detection for defoliating and nondefoliating pathotypes of V. dahliae was established by combining CRISPR–Cas12a/Cas13a with specific target genes. CONCLUSION: Taken together, this study achieved rapid, sensitive, and accurate detection of V. dahliae and the differentiation of defoliating and nondefoliating pathotypes and provides potential for field‐deployable diagnostic tools for rapid and ultrasensitive detection. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]