Tomato Wilt Disease Research Articles

Immunomodulating melatonin-decorated silica nanoparticles suppress bacterial wilt (Ralstonia solanacearum) in tomato (Solanum lycopersicum L.) through fine-tuning of oxidative signaling and rhizosphere bacterial community

BackgroundTomato (Solanum lycopersicum L.) production is severely threatened by bacterial wilt, caused by the phytopathogenic bacterium Ralstonia solanacearum. Recently, nano-enabled strategies have shown tremendous potential in crop disease management.ObjectivesThis study investigates the efficacy of biogenic nanoformulations (BNFs), comprising biogenic silica nanoparticles (SiNPs) and melatonin (MT), in controlling bacterial wilt in tomato.MethodsSiNPs were synthesized using Zizania latifolia leaves extract. Further, MT containing BNFs were synthesized through the one-pot approach. Nanomaterials were characterized using standard characterization techniques. Greenhouse disease assays were conducted to assess the impact of SiNPs and BNFs on tomato plant immunity and resistance to bacterial wilt.ResultsThe SiNPs and BNFs exhibited a spherical morphology, with particle sizes ranging from 13.02 nm to 22.33 nm for the SiNPs and 17.63 nm to 21.79 nm for the BNFs, indicating a relatively uniform size distribution and consistent shape across both materials. Greenhouse experiments revealed that soil application of BNFs outperformed SiNPs, significantly enhancing plant immunity and reducing bacterial wilt incidence by 78.29% in tomato plants by maintaining oxidative stress homeostasis via increasing the activities of antioxidant enzymes such as superoxide dismutase (31.81%), peroxidase (32.9%), catalase (32.65%), and ascorbate peroxidase (47.37%) compared to untreated infected plants. Additionally, BNFs induced disease resistance by enhancing the production of salicylic acid and activating defense-related genes (e.g., SlPAL1, SlICS1, SlNPR1, SlEDS, SlPD4, and SlSARD1) involved in phytohormones signaling in infected tomato plants. High-throughput 16 S rRNA sequencing revealed that BNFs promoted growth of beneficial rhizosphere bacteria (Gemmatimonadaceae, Ramlibacter, Microscillaceae, Anaerolineaceae, Chloroplast and Phormidium) in both healthy and diseased plants, while suppressing R. solanacearum abundance in infected plants.ConclusionOverall, these findings suggest that BNFs offer a more promising and sustainable approach for managing bacterial wilt disease in tomato plants.Graphical

Novel Fungicide from Waste Oil Palm Fruit Bunch for Sustainable Management of Tomato Wilt Disease Caused by Fusarium oxysporum - experimental and in silico studies

Tomato wilt, a widespread and persistent disease caused by the fungus Fusarium oxysporum, can be effectively controlled through biological approaches, promoting ecosystem sustainability and enhancing agricultural productivity and quality. This study explored the use of empty oil palm fruit bunch (EOPFB) waste as a natural fungicide to control tomato Fusarium wilt disease through in vitro, pilot scale, and molecular docking experiments. EOPFB aqueous extract showed the highest mycelium growth inhibition of 68.3 %, with the commercial fungicide Mancozeb exhibiting a moderate 42.30 % inhibition, and sterile distilled water having a negligible effect of 0.24 %. The inhibition rate varied with concentration, peaking at 72.63 % at 0.15 g/mL and dropping to 54.80 % at 0.01 g/mL. In terms of wilt disease incidence and severity, EOPFB performed best, recording the lowest incidence of 47.20 % and severity of 15.58 %. Mancozeb followed closely, with an incidence of 46.30 % and severity of 26.23 %. Sterile distilled water, on the other hand, showed the highest incidence of 76.28 % and severity of 56.17 %. Molecular docking simulations corroborated the experimental results, validating the potency of EOPFB as an inhibitor of trypsin from F. oxysporum, and identifying 2-amino-5,6-dimethyl-3 H-pyrimidine-4-one as the primary active compound responsible for the fungicidal activity. This phytochemical, abundant in the crude ethanolic extract of EOPFB, was found to have a higher binding affinity (6.1 kcal/mol) than Mancozeb (4.8 kcal/mol), at the trypsin target. These findings demonstrate the potential of EOPFB, an agricultural waste product, as a sustainable and eco-friendly solution for managing tomato Fusarium wilt disease, contributing to increased tomato production. It also highlights the potential of 2-amino-5,6-dimethyl-3 H-pyrimidine-4-one as a lead compound for the development of novel, natural fungicides, and underscores the value of EOPFB as a rich source of bioactive compounds with agricultural and biotechnological applications.

Identification of hub genes and potential networks by centrality network analysis of PCR amplified Fusarium oxysporum f. sp. lycopersici EF1α gene

BackgroundFusarium wilt is a devastating soil-borne fungal disease of tomato across the world. Conventional method of disease prevention including usage of common pesticides and methods like soil solarisation are usually ineffective in the treatment of this disease. Therefore, there is an urgent need to identify virulence related genes in the pathogen which can be targeted for fungicide development.ResultsPathogenicity testing and phylogenetic classification of the pathogen used in this study confirmed it as Fusarium oxysporum f. sp. lycopersici (Fol) strain. A recent discovery indicates that EF1α, a protein with conserved structural similarity across several fungal genera, has a role in the pathogenicity of Magnaporthe oryzae, the rice blast fungus. Therefore, in this study we have done structural and functional classification of EF1α to understand its role in pathogenicity of Fol. The protein model of Fol EF1α was created using the template crystal structure of the yeast elongation factor complex EEF1A:EEF1BA which showed maximum similarity with the target protein. Using the STRING online database, the interactive information among the hub genes of EF1α was identified and the protein–protein interaction network was recognized using the Cytoscape software. On combining the results of functional analysis, MCODE, CytoNCA and CytoHubba 4 hub genes including Fol EF1α were selected for further investigation. The three interactors of Fol EF1α showed maximum similarity with homologous proteins found in Neurospora crassa complexed with the known fungicide, cycloheximide. Through the sequence similarity and PDB database analysis, homologs of Fol EF1α were found: EEF1A:EEF1BA in complex with GDPNP in yeast and EF1α in complex with GDP in Sulfolobus solfataricus. The STITCH database analysis suggested that EF1α and its other interacting partners interact with guanosine diphosphate (GDPNP) and guanosine triphosphate (GTP).ConclusionsOur study offers a framework for recognition of several hub genes network in Fusarium wilt that can be used as novel targets for fungicide development. The involvement of EF1α in nucleocytoplasmic transport pathway suggests that it plays role in GTP binding and thus apart from its use as a biomarker, it may be further exploited as an effective target for fungicide development. Since, the three other proteins that were found to be tightly associated Fol EF1α have shown maximum similarity with homologous proteins of Neurospora crassa that form complex with fungicide- Cycloheximide. Therefore, we suggest that cycloheximide can also be used against Fusarium wilt disease in tomato. The active site cavity of Fol EF1α can also be determined for computational screening of fungicides using the homologous proteins observed in yeast and Sulfolobus solfataricus. On this basis, we also suggest that the other closely associated genes that have been identified through STITCH analysis, they can also be targeted for fungicide development.

Impact of biosynthesized silver nanoparticles (AgNPs) on fusarium wilt disease in tomato: In vitro and In vivo studies

Impact of biosynthesized silver nanoparticles (AgNPs) on fusarium wilt disease in tomato: In vitro and In vivo studies

First report of bacterial wilt caused by Clavibacter michiganensis subsp. michiganensis affecting tomato in Iğdır

Tomato wilt disease caused by Clavibacter mihiganensis subsp. michiganensis (Cmm) is one of the most destructive tomato diseases and causes significant crop loss in both greenhouse and field tomato production areas worldwide. In this study, the presence of the causal agent of bacterial wilt disease in tomato plants was investigated in Aras Valley. Isolation was made from diseased plant samples and it was determined whether the strains were pathogenic by cellulase activity and HR test. The virulence, morphological and biochemical characteristics of the strains were determined. Strains that fatty acid methyl ester extraction, isolation and purification were performed were identified at species and subspecies level with % similarity index using gas chromatography system. The diagnosis was confirmed with the Biolog Gen III System and all strains were identified at the subspecies level with a % similarity index. As a result of this study, 57 strains were obtained in the isolation, and 39 of the strains were determined not to be pathogenic. Strain 18 was determined as the pathogen causing the most damage to tomato plants with 100 % disease severity. Strains were identified as Cmm at subspecies level with a similarity index of 71-87 % using gas chromatography system and 54-75 % similarity index with Biolog Gen III System. According to the heat map created, it was determined that the strains consisted of two main clusters. The presence of pathogen in Aras Valley was proved for the first time by this study.

Impact of biosynthesized silver nanoparticles (AgNPs) on fusarium wilt disease in tomato: In vitro and In vivo studies Hafez, M. S., Abdou, El-S., Shaat, M. M. and Abd ElGawad, T. I. Plant Pathology Department, Faculty of Agriculture, Minia Universit

Impact of biosynthesized silver nanoparticles (AgNPs) on fusarium wilt disease in tomato: In vitro and In vivo studies Hafez, M. S., Abdou, El-S., Shaat, M. M. and Abd ElGawad, T. I. Plant Pathology Department, Faculty of Agriculture, Minia Universit

Application of Endophytic Bacteria from Tomato Stems to Control Bacterial Wilt Disease in Tomato and Enhance Plant Growth

Application of Endophytic Bacteria from Tomato Stems to Control Bacterial Wilt Disease in Tomato and Enhance Plant Growth

Unveiling the efficacy of a bacterial antagonist in the management of tomato wilt disease caused by Ralstonia solanacearum

The long-term application of agrochemicals, including pesticides, in intensive agricultural practices to protect crops from biotic stresses results in the emergence of resistance among phytopathogens and the ineffectiveness of chemical applications. The microbiological strategies, in contrast, minimize the reliance on chemicals and, hence, reduce environmental and human health risks. Bacterial wilt of tomato induced by Ralstonia solanacearum is one of the most destructive diseases worldwide that requires urgent attention to develop a safer and more efficient method to control the phytopathogen. The present work was conducted in the year 2022-2023. In this study, the bacterial wilt was managed by an antagonist bacterium, Pseudomonas fluorescens, exhibiting variable morphological, biochemical, and plant growth-promoting activities. The P. fluorescens inhibited the growth of R. solanacearum in plate assay at different time intervals (0-48 h). The SEM image of R. solanacearum cells cultured with P. fluorescens revealed pores, distortion, and fragmented cell envelope, while the untreated bacterial cells were uniform and smooth. Tomato plants infected with R. solanacearum showed 89% disease incidence compared to uninfected but PGPR-inoculated tomato plants. Application of P. fluorescens reduced disease incidence by 63% compared to R. solanacearum infested plants. Furthermore, plant length enhanced by 21 and 26% significantly following bacterial inoculation after three and four weeks of growth. Conclusively, this study emphasizes the effectiveness of using PGPR as a potent strategy for managing wilt disease in vegetable crops, especially tomato.

Screening of some commonly cultivated tomato varieties against Fusarium wilt in Jama’are, Bauchi State, Nigeria

Fusarium wilt disease of tomato is caused by Fusarium oxysporum f.sp. lycopersici and is a limiting factor to tomato production in Nigeria. This research was conducted to assess the effect of Fusarium oxysporum on some tomato varieties cultivated in Jama’are, Bauchi state. The objectives of this research work were to isolate and identify pathogen (Fusarium oxysporum) from the soil, to determine the incidence and severity of Fusarium wilt on different varieties of Tomato grown in Jama’are, and to determine the most resistance variety among four different varieties. The experiment was laid in a Completely Randomized design (CRD) with four replications. The four tomato varieties (Roma Savanna, Rio Grande, UC82B and Tropimech) were inoculated at the root region by making shallow groove around the base of the plant in the root region and placing 3 g mycelia plug of 7 days old pure culture of F. oxysporum face-down close to the root of the seedling and covered with soil. At two weeks intervals after treatments the seedlings were then observed for symptoms of wilt infection. Data on disease incidence (%), were collected based on observation after which they were analyzed using Analysis of Variance (ANOVA) test. The Disease Incidence Percentage for Roma Savanna, Rio Grande, UC28B and Tropimech were evaluated to be 51.6%, 18.2 %,38.4 % and 43.3 % respectively while the Disease Severity Percentage were 72.1%, 24.8%, 40.9% and 62.3% respectively. The findings of this research reveals that; there is a significant effect of F. oxysporum on the tomato varieties, all the varieties were severely infected to some degree, and that of all species, Roma savanna was moderately resistant to F. oxysporum to some degree. This agrees with other findings in the literature, as Fusarium oxysporum affects different varieties of tomato at different growth stages. Also, it was observed that screening excises involving selecting of non-infected seedlings with Fusarium species and other fungi should be done prior to transplanting.

Host RNAi-mediated silencing of Fusarium oxysporum f. sp. lycopersici specific-fasciclin-like protein genes provides improved resistance to Fusarium wilt in Solanum lycopersicum.

Tomato transgenics expressing dsRNA against FoFLPs act as biofungicides and result in enhanced disease resistance upon Fol infection, by downregulating the endogenous gene expression levels of FoFLPs within Fol. Fusarium oxysporum f. sp. lycopersici (Fol) hijacks plant immunity by colonizing within the host and further instigating secondary infection causing vascular wilt disease in tomato that leads to significant yield loss. Here, RNA interference (RNAi) technology was used to determine its potential in enduring resistance against Fusarium wilt in tomato. To gain resistance against Fol infection, host-induced gene silencing (HIGS) of Fol-specific genes encoding for fasciclin-like proteins (FoFLPs) was done by generating tomato transgenics harbouring FoFLP1, FoFLP4 and FoFLP5 RNAi constructs confirmed by southern hybridizations. These tomato transgenics were screened for stable siRNA production in T0 and T1 lines using northern hybridizations. This confirmed stable dsRNAhp expression in tomato transgenics and suggested durable trait heritability in the subsequent progenies. FoFLP-specific siRNAs producing T1 tomato progenies were further selected to ascertain its disease resistance ability using seedling infection assays. We observed a significant reduction in FoFLP1, FoFLP4 and FoFLP5 transcript levels in Fol, upon infecting their respective RNAi tomato transgenic lines. Moreover, tomato transgenic lines, expressing intended siRNA molecules in the T1 generation, exhibit delayed disease onset with improved resistance. Furthermore, reduced fungal colonization was observed in the roots of Fol-infected T1 tomato progenies, without altering the plant photosynthetic efficiency of transgenic plants. These results substantiate the cross-kingdom dsRNA or siRNA delivery from transgenic tomato to Fol, leading to enhanced resistance against Fusarium wilt disease. The results also demonstrated that HIGS is a successful approach in rendering resistance to Fol infection in tomato plants.

Greenhouse tomato farmers' knowledge, perceptions, and management of tomato bacterial wilt (Ralstonia solanacearum) disease

A major constraint to tomato cultivation is bacterial wilt disease. The use of greenhouses to cultivate tomato is vital to controlling the bacterial wilt disease. Bacterial wilt can be successfully managed when farmers are well-informed with better knowledge of bacterial wilt in tomatoes. This study was conducted to assess farmers’ knowledge and experiences on the cultivation practices, prevalence, detection, spread, and control of bacterial wilt disease in tomato in greenhouses in the Volta, Eastern, Central, and Greater Accra regions of Ghana. Questionnaires were administered for fifty (50) greenhouse farmers, purposefully selected using a database of greenhouse tomato producers in southern Ghana provided by the Ministry of Food and Agriculture (MOFA). Frequency data was analyzed using descriptive statistical analysis. The majority (86%) of respondents had formal education. Most of the greenhouses in operation were in the Greater Accra Region, and none was under cultivation in the Volta region at the time of the study. Most respondents have been involved in greenhouse tomato cultivation for barely three years. The frequency of greenhouse tomatoes production varied from one region to the other. Only 28% of greenhouse farmers knew the test to detect the disease with 64% of greenhouse farmers without any knowledge about how the disease spreads. 62% of respondents used roughing and burying of the infected plants to control the disease. Out of the 54 greenhouses (domes) surveyed, 12 were infected with the bacterial wilt disease. Greenhouse farmers had little knowledge on the spread, detection, and control of the bacterial wilt disease of tomato. The findings of this study would lead to the design of targeted training programs on cultivation practices, detection, spread and management of bacterial wilt of tomato to increase yield and boost income levels of greenhouse tomato farmers in Ghana. Key words: bacterial wilt, tomatoes, spread, detection, control, greenhouse, farmers, constraints

Resistance Risk and Molecular Mechanism of Tomato Wilt Pathogen Fusarium oxysporum f. sp. lycopersici to Pyraclostrobin.

Tomato wilt disease caused by Fusarium oxysporum f. sp. lycopersici (Fol) results in a decrease in tomato yield and quality. Pyraclostrobin, a typical quinone outside inhibitor (QoI), inhibits the cytochrome bc1 complex to block energy transfer. However, there is currently limited research on the effectiveness of pyraclostrobin against Fol. In this study, we determined the activity of pyraclostrobin against Fol and found the EC50 values for pyraclostrobin against 100 Fol strains (which have never been exposed to QoIs before). The average EC50 value is 0.3739 ± 0.2413 μg/mL, indicating a strong antifungal activity of pyraclostrobin against Fol, as shown by unimodal curves of the EC50 values. Furthermore, we generated five resistant mutants through chemical taming and identified four mutants with high-level resistance due to the Cytb-G143S mutation and one mutant with medium-level resistance due to the Cytb-G137R mutation. The molecular docking results indicate that the Cytb-G143S or Cytb-G137R mutations of Fol lead to a change in the binding mode of Cytb to pyraclostrobin, resulting in a decrease in affinity. The resistant mutants exhibit reduced fitness in terms of mycelial growth (25 and 30 °C), virulence, and sporulation. Moreover, the mutants carrying the Cytb-G143S mutation suffer a more severe fitness penalty compared to those carrying the Cytb-G137R mutation. There is a positive correlation observed among azoxystrobin, picoxystrobin, fluoxastrobin, and pyraclostrobin for resistant mutants; however, no cross-resistance was detected between pyraclostrobin and pydiflumetofen, prochloraz, or cyazofamid. Thus, we conclude that the potential risk of resistance development in Fol toward pyraclostrobin can be categorized as ranging from low to moderate.

Differential expression and gene correlation analyses reveal core CAZymes in Fusarium oxysporum f.sp. lycopersici exposed to mutagen and heat stress

AbstractFusarium oxysporum f.sp. lycopersici is the casual organism of tomato wilt disease which is a devastating plant pathogen responsible for annual loss of this fruit crop. An important step in fungal growth and development is germination, that is transformation of dormant spores into a growing hypha. For this, a set of special type of enzymes known as CAZymes are required for construction and remodelling of polysaccharides and penetration into the host. This necessitates to understand the underlying mechanism of hyphal growth and the crucial CAZymes involved in the process. Therefore, a comparative transcriptomic study of F. oxysporum f.sp. lycopersici using publically available RNA‐seq data under two perturbed conditions was taken up to find out the core differentially expressed genes (DEGs) followed by CAZyme annotation and their co‐expression. Among these two perturbations, 514 core DEGs were found, from which 32 CAZymes were annotated, with majority of them being glycosyl hydrolases and glycosyl transferases. Most of these CAzymes were also found to be downregulated in both the stresses. Further, among these annotated DEGs, the interaction network revealed the association of CAZymes with zinc finger transcription factors, enzymes associated with fatty acid biosynthesis, and sulphur and citrate metabolism. Hence, in the present study, the key or core CAZymes and their association with other enzymes were revealed in the impaired fungus which are otherwise crucial for growth in normal conditions.

Pathogenicity of Fusarium oxysporum f. sp. lycopersici (Sacc.) Isolates in Causing Tomato Wilt Disease on Two Tomato (Solanum lycopersicum L) Varieties

Pathogenicity test of Fusarium oxysporum isolates in causing tomato wilt disease (TWD) was carried out on two tomato varieties (UC 82B and Rio-grande) in a screen house located at the Teaching and Research Farm of Federal University of Agriculture, Makurdi during 2015 cropping season. The experiment was a 2 x 11 factorial laid out in Completely Randomized Design (CRD) and replicated three times. F. oxysporum isolates tested were coded as: FoAs1, FoAs2, FoAg, FoNb, FoSb, FoAm, FoAk, FoOr, FoAd and FoUAM together with an uninoculated control. All the isolates of F. oxysporum tested were pathogenic, causing wilt on the plants from 3 weeks after inoculation (WAI), with severity of wilt been significantly higher (P≤0.05) in FoUAM. Isolates of F. oxysporum showed significant difference (P≤0.05) both in incidence and severity compared with the control. Effect of F. oxysporum isolates on some agronomic characteristics such as plant height, number of fruits, fruit weight and number of branches on the two varieties of tomato at 12 weeks after sowing (WAS) was significantly different (P≤0.05) with the control experiment. Highest severity score of 5.00 was calculated in FoUAM while the least of 4.33 was in FoSb compare with uninoculated value of 2.50. It is therefore, concluded that the two tomato varieties are pathogenic to F. oxysporum isolates and illicit disease in tomato hence reduced the yield of the crop.

Crude Saponins from Chenopodium quinoa Willd. Reduce Fusarium Wilt Infection in Tomato Seedlings

Quinoa saponins are pentacyclic triterpene compounds composed of one triterpenoid glycoside and two different sugar chains. Previous studies have showed that natural quinoa saponins showed little or no antifungal activity, and there are few reports about their antifungal effects in recent decades. Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (FOL) is the most serious for tomatoes in the field and under greenhouse conditions. The main objective of this study was to investigate the effectiveness of different concentrations and application modes of crude saponins from quinoa bran against the causal pathogen of tomato wilt under a greenhouse experiment. The results showed that the anti-FOL activity of quinoa saponins was weak in vitro, but significantly enhanced in vivo. Tomato seeds and seedlings treated with solution of quinoa saponins at 0.5 and 1.0 g/L significantly reduced the disease incidence (%) of tomato Fusarium wilt. The treatment types of saponin solution have influence on the preventive effects (%) of tomato seedlings against Fusarium wilt, among them, root soaking > foliar spray > seed soaking. The treatment of seed soaking with quinoa saponins inhibited germination of tomato seeds to some extent. However, the germination rate of tomato seeds after saponin soaking was comparable to the chemical pesticide (thiram carboxin); therefore, it could be used to control tomato wilt disease. This is due to the fact that the antifungal activity of quinoa saponins in vivo was much higher than that in vitro when the saponin concentration was between 0.5–1.0 g/L, indicating that the antifungal activity of quinoa saponins may be achieved mainly by inducing resistance. This investigation supports the potential use of quinoa saponins as a supplier of antifungal compounds, and could be the foundation for a future study examining the use of quinoa bran as a new resource against FOL.

Evaluation of the Humic Acid Efficiency and the Bioagent Chaetomium globosum in Controlling Fusarium Wilt Disease in Tomato

Fusarium wilt disease presents a significant menace to tomato plants, necessitating effective preventative measures. Five isolates of Fusarium oxysporum f.sp. lycopersici from infected tomato plants from different regions in Missan province, each showing varying levels of pathogenicity. Differences in the ability of tomato seedlings to be infected by these isolates were also observed during a pot experiment, ranging from highly pathogenic to non-pathogenic. To address this issue, the efficacy of the bioagent Chaetomium globosum and humic acid in controlling the pathogenic fungus F. oxysporum f.sp. lycopersici and improving the growth indicators of tomato plants in the presence of the causative fungus. Two concentrations of humic acid (8 and 4 mL/l) were tested to determine their efficacy in reducing the growth of pathogenic fungi in vitro. We found that the growth of the pathogenic fungi decreased to 41.06% at a concentration of 8 ml/l, compared to the control, which showed no growth. Moreover, when used alone or in combination with humic acid, C. globosum significantly reduced the percentage and severity of infection. The treatment involving the interaction between the bioagent and humic acid at a concentration of 8 ml/l recorded an infection percentage of 14.4% and an infection severity of 11.87%, compared to the pathogenic fungus treatment that resulted in 53% infection and 43.80% severity, respectively. Moreover, the use of C. globosum and different concentrations of humic acid raised all growth indices in all treatments whether in the presence or absence of pathogenic funus. These improvements included increased fresh and dry weight of the shoot and root, enhanced plant height and leaf area, as well as a significant increase in the content of total phenols and total chlorophyll in the leaves. The findings of this study highlight the potential of C. globosum and humic acid as effective control measures against fusarium wilt disease in tomato plants. The results indicate their ability to suppress the pathogenic fungus and promote plant growth. These findings hold promise for the development of sustainable agricultural practices.

Investigation of the Impact of Mycogenic Titanium and Selenium Nanoparticles on Fusarium Wilt Infection of Tomato Plant

We applied biosynthesized titanium and selenium nanoparticles, prepared using a fungal water extract of Trichoderma harzianum (T. harzianum), to eradicate tomota wilt infection. Transmission electron microscopy (TEM), Scanning electron microscopy (SEM), Energy Dispersive X-Ray analysis, and Transmission electron microscopy/X-ray diffraction (TEM/XRD) techniques were used to characterize the spherical metal nanoparticles, whose diameters were 16.0 nm for selenium nanoparticles (SeNPs) and 50.0 nm for titanium nanoparticles (TiNPs). This confirmed the efficient biosynthesis of the nanoparticles. Under greenhouse conditions, the effectiveness of TiNPs and SeNPs produced by nonpathogenic fungi (T. harzianum) against the pathogen responsible for the tomato wilt disease, Fusarium oxysporum (F. oxysporum), was studied. Based on the results, the most efficient method for combating the pathogen that causes tomato wilt was used in open fields, whereas pot studies were conducted in greenhouse conditions. All tested treatments considerably lowered tomato plant wilt disease in both the greenhouse and the open field. The disease severity was reduced by 20.4% using TiNPs at high concentrations of 150 ppm and by 41.5% using SeNPs. Compared to conventional antibiotics, the antibacterial activity assessment of the biosynthesized TiNPs and SeNPs revealed a significant effect versus pathogenic bacteria and fungi, with a negligible influence on the examined human and animal microflora. The findings showed that biosynthesized TiNPs and SeNPs can be applied to suppress the plant pathogen F. oxysporum in a way that is safe for the microflora of humans and animals. This is the first instance where the nanocidal activity of biological TiNPs and SeNPs has been used against the pathogen that causes tomato wilt.

Response of bacterial biovars to inheritance pattern of bacterial wilt disease in tomato

ABSTRACT Knowledge of the genetic control of disease tolerance is necessary for effective transfer of desirable genes in resultant progenies leading to development of disease tolerant cultivars and hybrids. Bacterial wilt (BW) disease-resistant varieties of tomato (Solanum lycopersicum L.) express differential reactions to bacterial biovars distributed in specific regions. Genetic control of host tolerance to BW disease was studied using the generations P1, P2, F1, F2, BC1, BC2 of three tolerant × susceptible crosses (“Utkal Kumari × CLN- 2460E;” “Utkal Kumari × CLN-2764A;” “Utkal Deepti × CLN-2460E”) involving two tolerant (“Utkal Kumari,” “Utkal Deepti”), and two susceptible (“CLN-2460E” and “CLN-2764A”) genotypes exposed to the virulent bacterial biovars III and VI. The inheritance pattern indicated tolerance to BW disease was conditioned by a single dominant gene in three crosses when exposed to both biovars. Genetic control of bacterial wilt resistance did not depend on the interaction between genotype and bacterial biovars tested. Predominance of non-additive gene action and duplicate epistasis for conditioning of first appearance and incidence of BW disease in crosses indicated hindrance in progress through conventional selection. Selection for improvement of BW disease-related traits should be delayed to later segregating generations. The modified bulk method of selection appeared to be best for improving BW disease-related traits of tomato. Tomato hybrids tolerant to BW disease could be developed with the involvement of one parent tolerant to this disease.

Boosting the Biocontrol Efficacy of Bacillus amyloliquefaciens DSBA-11 through Physical and Chemical Mutagens to Control Bacterial Wilt Disease of Tomato Caused by Ralstonia solanacearum.

Bacterial wilt disease of tomato (Solanum lycopersicum L.), incited by Ralstonia solanacearum (Smith), is a serious agricultural problem in India. In this investigation, chemical mutagenic agents (NTG and HNO2 treatment) and ultraviolet (UV) irradiation have been used to enhance the antagonistic property of Bacillus amyloliquefaciens DSBA-11 against R. solanacearum UTT-25 towards an effective management of tomato wilt disease. The investigation established the fact that maximum inhibition to R. solanacearum UTT-25 was exerted by the derivative strain MHNO2-20 treated with nitrous acid (HNO2) and then by the derivative strain MNTG-21 treated with NTG. The exertion was significantly higher than that of the parent B. amyloliquefaciens DSBA-11. These two potential derivatives viz. MNTG-21, MHNO2-20 along with MUV-19, and a wild derivative strain of B. amyloliquefaciens i.e.,DSBA-11 were selected for GC/MS analysis. Through this analysis 18 major compounds were detected. Among the compounds thus detected, the compound 3-isobutyl hexahydropyrrolo (1,2), pyrazine-1,4-dione (4.67%) was at maximum proportion in the variant MHNO2-20 at higher retention time (RT) of 43.19 s. Bio-efficacy assessment observed a record of minimum intensity (9.28%) in wilt disease and the highest bio-control (88.75%) in derivative strain MHNO2-20-treated plants after 30 days of inoculation. The derivative strain MHNO2-20, developed by treating B. amyloliquefaciens with nitrous acid (HNO2), was therefore found to have a higher bio-efficacy to control bacterial wilt disease of tomato under glasshouse conditions than a wild-type strain.

First Report of Ralstonia pseudosolanacearum Causing Wilt Disease in Tomato (Solanum lycopersicum L.) Plants from Mexico.

Mexico produces more than four million tons of tomato fruits and ranks tenth worldwide. In February 2022, tomato plants in a greenhouse in Culiacan, Sinaloa State, were affected by wilt diseases with an incidence of 20% and irreversible wilt and death of the infected plants (severity up 70%). When cut stems from affected plants, a reddish to brown discoloration of the vascular system was observed and these were disinfected with 1% NaClO for 5 min and then placed in a humid chamber. Characteristic milky-white exudate was obtained. From that exudate, irregular, mucoid, and white colonies with pink centres were obtained on casamino peptone glucose (CPG) plates supplemented with 1% 2,3,5-triphenyl 15 tetrazolium chloride (TZC); these characteristics are typical of the Ralstonia solanacearum species complex (RSSC) (Garcia et al., 2019). Identification of the pathogen was done by PCR using specific primer pairs reported by Paudel et al. (2022), RssC-wF3 (5'-TATATATCCTCGACTTTTCCATGAAGCTGTG-3') - RssCwR3 (5'-CTATATATATACCCCACTTGTTGAGGAACTG-3') and Rpseu-wF5 (5'-TTTTATTTTTTTGGTGTCCGGGCCAAGATAG-3') - Rpseu-wR5 (5'- TTATATTACTCGAACGTGCTGCAAAACCACT-3'), which amplified fragments of 162 and 251 bp for RSSC and Ralstonia pseudosolanacearum, respectively. Additionally, 759 (5'-GTCGCCGTCAACTCACTTTCC-3') - 760 (5'-GTCGCCGTCAGCAATGCGGAATCG-3') (Opina, et al., 1997) and Nmult21:1F (5'-CGTTGATGAGGCGCGCAATTT-3') - Nmult22:RR (5'- TCGCTTGACCCTATAACGAGTA-3') (Fegan and Prior, 2005) were used to generate 282 and 144 bp amplicons for RSSC and phylotype I, respectively. Subsequen to making the specific detection, the representative strain ClnMx was used to generate a sequence for the endoglucanase (egl) gene for separation into sequevars by using the primers Endo-F (5'- ATGCATGCCGCTGGTCGCCGC-3') and Endo-R (5'-GCGTTGCCCGGCACGAACACC-3'), which amplified a fragment of 750 bp (Fegan et al., 1998). The egl sequence (GenBank Access ON542479) showed 100% identity with the well-defined R. pseudosolanacearum sequevar 14, which was isolated from tomato plants from Senegal (UW763, I-14 GenBank Access CP051174) (Steidl et al., 2021), as well as, the strain MAFF 301070 (GenBank Access AB508612) from Japanese tomato. For pathogenicity tests, four 1-month-old tomato plants were infected using an insulin syringe that contained a pure bacterial suspension with approximately 2x108 CFU/mL. For each plant, 20 µL was infiltrated into the axil of the third upper leaf, and for untreated controls, tomato plants were infiltrated with sterile water. All plants were kept at 28°C under greenhouse conditions. Symptoms resembling those observed in the field were observed in inoculated plants six days after inoculation, and the plant pathogen was recovered on TZC medium. To confirm the bacteria identification a PCR using the specific primer pairs mentioned early was carried out. In contrast, water-treated control plants remained healthy. Koch's postulates were carried out twice with similar results. Ralstonia solanacearum species complex (RSSC) causes severe economic losses in many countries of the world because of their capability to infect a wide range of host plants, including potato, tomato, eggplant, tobacco, and, banana, among others. Ralstonia pseudosolanacearum has been reported to cause tomato wilt disease mainly on the Afro-Eurasian continent in areas such as Senegal, Cambodia, and Japan (Klass et al., 2019). To our knowledge, this is the first report of R. pseudosolanacearum causing bacterial wilt diseases in tomato plants from Mexico and because, the control of this bacteria is a challenge by the long survival time in soil, water, and infected plant tissues, the identification of this important pathogen could provide relevant information for developing management strategies.