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PAMP-triggered immunity (PTI) is the first layer of plant defense response that occurs on the plant plasma membrane. Recently, the application of a rhizobacterium, Bacillus amyloliquefaciens strain PMB05, has been demonstrated to enhance flg22Pst- or harpin-triggered PTI response such as callose deposition. This PTI intensification by PMB05 further contributes to plant disease resistance to different bacterial diseases. Under the demand for rapid and large-scale screening, it has become critical to establish a non-staining technology to identify microbial strains that can enhance PTI responses. Firstly, we confirmed that the expression of the GSL5 gene, which is required for callose synthesis, can be enhanced by PMB05 during PTI activation triggered by flg22 or PopW (a harpin from Ralstonia solanacearum). The promoter region of the GSL5 gene was further cloned and fused to the coding sequence of gfp. The constructed fragments were used to generate transgenic Arabidopsis plants through a plant transformation vector. The transgenic lines of AtGSL5-GFP were obtained. The analysis was performed by infiltrating flg22Pst or PopW in one homozygous line, and the results exhibited that the green fluorescent signals were observed until after 8 h. In addition, the PopW-induced fluorescent signal was significantly enhanced in the co-treatment with PMB05 at 4 h after inoculation. Furthermore, by using AtGSL5-GFP to analyze 13 Bacillus spp. strains, the regulation of PopW-induced fluorescent signal was observed. And, the regulation of these fluorescent signals was similar to that performed by callose staining. More importantly, the Bacillus strains that enhance PopW-induced fluorescent signals would be more effective in reducing the occurrence of bacterial wilt. Taken together, the technique by using AtGSL5-GFP would be a promising platform to screen plant immunity-intensifying microbes to control bacterial wilt.

The bacterial wilt disease caused by Ralstonia solanacearum is a significant threat to eggplant production. Breeding and promoting resistant varieties is one of the most effective methods to manage bacterial wilt. Conducting QTL (quantitative trait locus) mapping of resistant genes can substantially enhance the breeding of plant resistance to bacterial wilt. In this study, a population of 2200 F2 individuals derived from resistant and susceptible materials was utilized to establish extreme resistance and susceptibility pools. Following resequencing analysis of the parents and extreme pools, the QTL were examined using the DEEP-BSA software and QTLseqr R package (version 0.7.5.2). The results revealed that the detection of 10 QTL sites on chromosomes 5, 8, 9, and 11 by the five algorithms of the DEEP-BSA software. Additionally, the candidate region of 62 Mb–72 Mb on chromosome 5 was identified in all five algorithms of the DEEP-BSA software, as well as by the QTLseqr R package. Subsequent gene annotation uncovered 276 genes in the candidate region of 62 Mb–72 Mb on chromosome 5. Additionally, RNA-seq results indicated that only 13 genes had altered expression levels following inoculation with R. solanacearum in the resistant materials. Based on the expression levels, SMEL4_05g015980.1 and SMEL4_05g016110.1 were identified as candidate genes. Notably, SNP annotation identified a non-synonymous mutation in the exonic region of SMEL4_05g015980.1 and a variant in the promoter region of SMEL4_05g016110.1. The research findings have practical significance for the isolation of bacterial wilt resistance genes in eggplant and the development of resistance to bacterial wilt varieties in eggplant.

Bacterial wilt (BW) of tomatoes, caused by Ralstonia solanacearum, is a devastating disease that results in large annual yield losses worldwide. Management of BW of tomatoes is difficult due to the soil-borne nature of the pathogen. One of the best ways to mitigate the losses is through breeding for disease resistance. Moreover, plant height (PH) is a crucial element related to plant architecture, which determines nutrient management and mechanical harvesting in tomatoes. An intraspecific F2 segregating population (NC 11212) of tomatoes was developed by crossing NC 84173 (tall, BW susceptible) × CLN1466EA (short, BW resistant). We performed quantitative trait loci (QTL) mapping using single nucleotide polymorphic (SNP) markers and the NC 11212 F2 segregating population. The QTL analysis for BW resistance revealed a total of three QTLs on chromosomes 1, 2, and 3, explaining phenotypic variation (R2) ranging from 3.6% to 14.9%, whereas the QTL analysis for PH also detected three QTLs on chromosomes 1, 8, and 11, explaining R2 ranging from 7.1% to 11%. This work thus provides information to improve BW resistance and plant architecture-related traits in tomatoes.

Biochar is increasingly being recognized as an effective soil amendment to enhance plant health and improve soil quality, but the complex relationships among biochar, plant resistance, and the soil microbial community are not clear. In this study, biochar derived from an invasive plant (Solidago canadensis L.) was used to investigate its impacts on bacterial wilt control, soil quality, and microbial regulation. The results reveal that the invasive plant biochar application significantly reduced the abundance of Ralstonia solanacearum in the soil (16.8–32.9%) and wilt disease index (14.0–49.2%) and promoted tomato growth. The biochar treatment increased the soil organic carbon, nutrient availability, soil chitinase, and sucrase activities under pathogen inoculation. The biochar did not influence the soil bacterial community diversity, but significantly increased the relative abundance of beneficial organisms, such as Bacillus and Sphingomonas. Biochar application increased the number of nodes, edges, and the average degree of soil microbial symbiotic network, thereby enhancing the stability and complexity of the bacterial community. These findings suggest that the invasive plant biochar produces win–win effects on plant–soil systems by suppressing soilborne wilt disease, enhancing the stability of the soil microbial community network, and promoting resource utilization, indicating its good potential in sustainable soil management.

Many bacterial plant pathogens have a broad host range important for their life cycle. Alternate hosts from plant families other than the main (primary) host support the survival and dissemination of the pathogen population even in absence of main host plants. Metabolic peculiarities of main and alternative host plants can affect genetic diversity within and between the pathogen populations isolated from those plants. Strains of Gram-positive bacterium Curtobacterium flaccumfaciens were identified as being causal agents of bacterial spot and wilt diseases on leguminous plants, and other crop and weed plants, collected in different regions of Russia. Their biochemical properties and susceptibility to copper compounds have been found to be relatively uniform. According to conventional PCR assays, all of the isolates studied were categorised as pathovar Curtobacterim flaccumfaciens pv. flaccumfaciens, a pathogen of legumes. However, the strains demonstrated a substantial diversity in terms of virulence on several tested host plants and different phylogenetic relationships were revealed by BOX-PCR and alanine synthase gene (alaS) sequencing.

Bacterial wilt is a significant soil-borne disease that poses a threat to mulberry production yield and quality of agricultural production worldwide. However, the disease resistance mechanisms dependent on root exudates are not well understood. In this present study, we investigated the antibacterial mechanisms of the main active substances (erucamide, oleamide, and camphor bromide) present in mulberry root exudates (MRE) against Ralstonia pseudosolanacearum (Rp), the causal agent of bacterial wilt. Our findings revealed that these three active substances inhibited the growth activity of Rp by affecting the cell morphology and extracellular polysaccharide content, as well as triggering a burst of reactive oxygen species. The active substances induced oxidative stress, leading to a decrease in Rp growth. Additionally, the expression levels of key genes in the hrp gene cluster (hrpB, hrpX, and hrpF) and other virulence-related genes (such as ripAW, ripAE, Rs5-4819, Rs5-4374, ace, egl3, and pehB) were significantly reduced upon treatment with the active substances. Further pathogenicity experiments demonstrated that root exudates (at a concentration of 1.5 mg·mL−1) delayed or slowed down the occurrence of bacterial wilt in mulberry. These findings provide valuable insight into the antimicrobial mechanisms of MRE against Rp and lay a theoretical foundation for the development and application of biocontrol agents to control mulberry bacterial wilt.

Bacterial wilt (BW), caused by Ralstonia solanacearum, is one of the devastating diseases in tomatoes (Solanum lycopersicum L.). The use of resistant cultivars and breeding for genetic resistance is the most effective, economical, and environmentally friendly management strategy for this disease. It is necessary to screen diverse germplasm and cultivated genotypes to identify resistant resources and to develop resistant cultivars in tomatoes to combat the changing pathogen isolates. This study evaluated 40 United States Department of Agriculture (USDA) tomato accessions for their BW resistance to the R. solanacearum isolate P822 under greenhouse conditions. The tomato plants were inoculated and visually assessed to observe their symptoms, and the disease severity was scored on a scale of 0 to 4 (0 = no leaf wilted, 1 = 25% of leaves wilted, 2 = 50% leaves wilted, 3 = 75% of leaves wilted, and 4 = 100% leaves wilted). Five accessions (PI 645370, PI 647306, PI 600993, PI 355110, and PI 270210) were observed as BW resistance, with PI 645370 showing the greatest resistance. The broad-sense heritability for BW resistance was estimated as 59.9% and 42.8% based on a 0–4 scale of disease incidence and the disease severity index, respectively. Two distinct clusters (sub-populations) were detected among 39 of the 40 accessions. The five identified BW-resistant accessions were distributed in both clusters, suggesting a likely difference in the genetic base among the five resistance accessions. The resistant accessions will contribute significantly to the tomato breeding program to develop new cultivars with BW resistance.

Bacterial wilt, caused by the Ralstonia solanacearum species complex, is one of the most damaging bacterial diseases in tobacco and other Solanaceae crops. In this study, we conducted an analysis and comparison of transcriptome landscape changes in seedling roots of three tobacco BC4F5 lines, C244, C010, and C035, with different resistance to bacterial wilt at 3, 9, 24, and 48 h after R. solanacearum infection. A number of biological processes were highlighted for their differential enrichment between C244, C010, and C035, especially those associated with cell wall development, protein quality control, and stress response. Hence, we performed a genome-wide identification of seven cell wall development-related gene families and six heat shock protein (Hsp) families and proposed that genes induced by R. solanacearum and showing distinct expression patterns in C244, C010, and C035 could serve as a potential gene resource for enhancing bacterial wilt resistance. Additionally, a comparative transcriptome analysis of R. solanacearum-inoculated root samples from C244 and C035, as well as C010 and C035, resulted in the identification of a further 33 candidate genes, of which Nitab4.5_0007488g0040, a member of the pathogenesis-related protein 1 (PR-1) family, was found to positively regulate bacterial wilt resistance, supported by real-time quantitative PCR (qRT-PCR) and virus-induced gene silencing (VIGS) assays. Our results contribute to a better understanding of molecular mechanisms underlying bacterial wilt resistance and provide novel alternative genes for resistance improvement.

Pentatricopeptide repeat (PPR) proteins, with tandem 30–40 amino acids, were characterized as one kind of nucleus coding protein. They have been demonstrated to play important roles in RNA editing, plant growth and development, and plant immunity. Although the PPR gene family has been characterized in some plant species, less is known about this family in peanut, especially their functions in response to Ralstonia solanacearum. In this study, we performed a genome-wide analysis to identify PPR genes and their functions in resistance to R. solanacearum. Here, 389, 481, and 1079 PPR genes were identified from Arachis duranensis, Arachis ipaensis, and Arachis hypogaea, respectively. Allopolyploidization was the main reason for the increased number of the AhPPR members. Gene duplication brought about 367 pairs of homologous genes of PPRs in A. hypogaea. Whole-genome replication, tandem repeats, scattered repeats, and unconnected repeats constituted the replication types. The substitution rates of nonsynonymous (Ka) versus synonymous (Ks) of all homologous pairs were less than 1.0, suggesting that the homologous AhPPRs underwent intense purifying selection pressure and remained conserved in both structure and function. RNA-seq and RT-qPCR analyses showed that AhPPR598 gene was highly expressed in the aerial part of peanut and involved in response to R. solanacearum. The transient expression of AhPPR598 in Nicotiana benthamiana induced the HR-mediated cell death, up-regulated expression of resistant marker genes, and enhanced the resistance to R. solanacearum, suggesting AhPPR598 was a positive regulator of immunity by regulating the JA and SA pathways. These results provide a new understanding of the origin, distribution, and evolution of the AhPPR gene family and potential gene resources for peanut-resistant breeding.

Potato is an important crop, used not only for food production but also for various industrial applications. With the introduction of the potato as a staple food strategy, the potato industry in China has grown rapidly. However, issues related to bacterial wilt, exacerbated by factors such as seed potato transportation and continuous cropping, have become increasingly severe in the primary potato cultivation regions of China, leading to significant economic losses. The extensive genetic diversity of Ralstonia solanacearum (R. solanacearum), which is the pathogen of bacterial wilt, has led to a lack of highly resistant potato genetic resources. There is a need to identify and cultivate potato varieties with enhanced resistance to reduce the adverse impact of this disease on the industry. We screened 55 accessions of nine different wild potato species against the bacterial wilt pathogen R. solanacearum PO2-1, which was isolated from native potato plants and belongs to phylotype II. Three accessions of two species (ACL24-2, PNT880-3, and PNT204-23) were identified with high resistance phenotypes to the tested strains. We found these accessions also showed high resistance to different phylotype strains. Among them, only PNT880-3 was capable of flowering and possessed viable pollen, and it was diploid. Consistent with the high resistance, decreased growth of R. solanacearum was detected in PNT880-3. All these findings in our study reveal that the wild potato PNT880-3 was a valuable resistance source to bacterial wilt with breeding potential.

Continuous cropping is the primary cultivation method in Chinese facility agriculture, and the challenge of it stands as a global issue in soil remediation. Growing tomatoes continuously on the same plot for an extended period can result in outbreaks of tomato bacterial wilt. It is caused by the soil-borne bacterium Ralstonia solanacearum, a widespread plant pathogen that inflicts considerable damage on economically significant crops worldwide. Simultaneously, this plant pathogen proves extremely resilient, as it can adhere to plant residues and persist through the winter, continuing to infect plants in subsequent years. Scientists have dedicated considerable efforts towards finding effective methods to manage this disease. This article delineates the characteristics of tomato bacterial wilt and the various types of pathogenic bacteria involved. It systematically reviews the progress in research aimed at controlling tomato bacterial wilt, encompassing both physical and biological aspects concerning soil and plants. Emphasis is placed on the principles and current applications of these control measures, alongside proposed improvements to address their limitations. It is anticipated that the future of tomato bacterial wilt control will revolve around the development of a novel environmental protection system and efficient control strategies, focusing on microecological management and enhancing tomato resistance against bacterial wilt through breeding.

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.

Ralstonia solanacearum is the causal agent of bacterial wilt, one of the most destructive diseases of solanaceous plants, affecting staple crops worldwide. The bacterium survives in water, soil, and other reservoirs, and is difficult to control. In this sense, the use of three specific lytic R. solanacearum bacteriophages was recently patented for bacterial wilt biocontrol in environmental water and in plants. To optimize their applications, the phages and the bacterium need to be accurately monitored and quantified, which is laborious and time-consuming with biological methods. In this work, primers and TaqMan probes were designed, and duplex and multiplex real-time quantitative PCR (qPCR) protocols were developed and optimized for the simultaneous quantification of R. solanacearum and their phages. The quantification range was established from 108 to 10 PFU/mL for the phages and from 108 to 102 CFU/mL for R. solanacearum. Additionally, the multiplex qPCR protocol was validated for the detection and quantification of the phages with a limit ranging from 102 targets/mL in water and plant extracts to 103 targets/g in soil, and the target bacterium with a limit ranging from 103 targets/mL in water and plant extracts to 104 targets/g in soil, using direct methods of sample preparation.

The foundation of effectively predicting plant disease in the early stage using deep learning algorithms is ideal for addressing food insecurity, inevitably drawing researchers and agricultural specialists to contribute to its effectiveness. The input preprocessor, abnormalities of the data (i.e., incomplete and nonexistent features, class imbalance), classifier, and decision explanation are typical components of a plant disease detection pipeline based on deep learning that accepts an image as input and outputs a diagnosis. Data sets related to plant diseases frequently display a magnitude imbalance due to the scarcity of disease outbreaks in real field conditions. This study examines the effects of several preprocessing methods and class imbalance approaches and deep learning classifiers steps in the pipeline for detecting plant diseases on our data set. We notably want to evaluate if additional preprocessing and effective handling of data inconsistencies in the plant disease pipeline may considerably assist deep learning classifiers. The evaluation’s findings indicate that contrast limited adaptive histogram equalization (CLAHE) combined with image sharpening and generative adversarial networks (GANs)-based approach for resampling performed the best among the preprocessing and resampling techniques, with an average classification accuracy of 97.69% and an average F1-score of 97.62% when fed through a ResNet-50 as the deep learning classifier. Lastly, this study provides a general workflow of a disease detection system that allows each component to be individually focused on depending on necessity.

PAMP-triggered immunity (PTI) is the first layer of plant defense response that occurs on the plant plasma membrane. Recently, the application of a rhizobacterium, Bacillus amyloliquefaciens strain PMB05, has been demonstrated to enhance flg22 Pst - or harpin-triggered PTI response such as callose deposition. This PTI intensification by PMB05 further contributes to plant disease resistance to different bacterial diseases. Under the demand for rapid and large-scale screening, it has become critical to establish a non-staining technology to identify microbial strains that can enhance PTI responses. Firstly, we confirmed that the expression of the GSL5 gene, which is required for callose synthesis, can be enhanced by PMB05 during PTI activation triggered by flg22 or PopW (a harpin from Ralstonia solanacearum ). The promoter region of the GSL5 gene was further cloned and fused to the coding sequence of gfp . The constructed fragments were used to generate transgenic Arabidopsis plants through a plant transformation vector. The transgenic lines of At GSL5 -GFP were obtained. The analysis was performed by infiltrating flg22 Pst or PopW in one homozygous line, and the results exhibited that the green fluorescent signals were observed until after 8 h. In addition, the PopW-induced fluorescent signal was significantly enhanced in the co-treatment with PMB05 at 4 h after inoculation. Furthermore, by using At GSL5 -GFP to analyze 13 Bacillus spp. strains, the regulation of PopW-induced fluorescent signal was observed. And, the regulation of these fluorescent signals was similar to that performed by callose staining. More importantly, the Bacillus strains that enhance PopW-induced fluorescent signals would be more effective in reducing the occurrence of bacterial wilt. Taken together, the technique by using At GSL5 -GFP would be a promising platform to screen plant immunity-intensifying microbes to control bacterial wilt.

Ralstonia solanacearum is a bacterial phytopathogen affecting staple crops, originally from tropical and subtropical areas, whose ability to survive in temperate environments is of concern under global warming. In this study, two R. solanacearum strains from either cold or warm habitats were stressed by simultaneous exposure to natural oligotrophy at low (4 °C), temperate (14 °C), or warm (24 °C) temperatures in environmental water. At 4 °C, the effect of temperature was higher than that of oligotrophy, since R. solanacearum went into a viable but non-culturable (VBNC) state, which proved to be dependent on water nutrient contents. Resuscitation was demonstrated in vitro and in planta. At 14 °C and 24 °C, the effect of oligotrophy was higher than that of temperature on R. solanacearum populations, displaying starvation-survival responses and morphological changes which were stronger at 24 °C. In tomato plants, starved, cold-induced VBNC, and/or resuscitated cells maintained virulence. The strains behaved similarly regardless of their cold or warm areas of origin. This work firstly describes the natural nutrient availability of environmental water favoring R. solanacearum survival, adaptations, and resuscitation in conditions that can be found in natural settings. These findings will contribute to anticipate the ability of R. solanacearum to spread, establish, and induce disease in new geographical and climatic areas.

Bacillus spp. has the potential to control bacterial and fungal diseases of crops. In vitro study, Bacillus amyloliquefaciens DSBA-11 showed best to inhibit the growth of Ralstonia pseudosolanacearum as compared to Bacillus cereus JHTBS-7, B. pumilus MTCC-7092, B. subtilis DTBS-5 and B. licheniformis DTBL-6.Three primers sets from nucleotide sequences of polyketide antibiotic synthase genes viz., macrolactin, difficidin and bacillaene of B. amyloliquefaciens FZB42 were designed and standardized protocol for simultaneous detection of polyketide antibiotics-producing strains of Bacillus spp. by multiplex—PCR with products size of 792 bp, 705 bp and 616 bp respectively. All the strains of B. amyloliquefaciens contained three polyketide antibiotic synthase genes, and B. subtilis possessed difficidin and macrolactin, whereas B. cereus JHTBS-7, B. pumilus MTCC-7092 and B. licheniformis DTBL-6 did not contain any polyketide antibiotic genes. By using this technique, polyketide-producing strains of Bacillus spp. were screened within a short period with high accuracy. These polyketide synthase genes were cloned by using a T&A vector to study the role of these genes in producing antibiotics that suppressed the growth of R. pseudosolanacearum under both in vitro and in vivo conditions. Bio-efficacy of cloned products of these genes macrolactin, bacillaene, and difficidin along with parent strain B. amyloliquefaciens DSBA-11 inhibited the growth of R. pseudosolanacearum and formed 1.9 cm2, 1.9 cm2, 1.7 cm2 and 3.3 cm2 inhibition area under in vitro conditions respectively. Minimum bacterial wilt disease intensity (29.3%) with the highest biocontrol efficacy (57.72%) was found in tomato cv. Pusa Ruby (susceptible to wilt disease) was treated with B. amyloliquefaciens DSBA-11 followed by cloned products of difficidin and macrolactin under glasshouse conditions. Hence, the developed multiplex protocol might be helpful for screening polyketide antibiotics producing potential strains of Bacillus spp. from soil which can apply for managing the wilt disease of tomatoes. The polyketide antibiotics produced by bacteria might have a significant role suppression of R. pseudosolanacearum due to the disintegration of cells.

Beneficial host-associated bacteria can assist plant protection against pathogens. In particular, specific microbes are able to induce plant systemic resistance. However, it remains largely elusive which specific microbial taxa and functions trigger plant immune responses associated with disease suppression. Here, we experimentally studied this by setting up two independent microcosm experiments that differed in the time at which plants were exposed to the pathogen and the soil legacy (i.e., soils with historically suppressive or conducive). Overall, we found soil legacy effects to have a major influence on disease suppression irrespective of the time prior to pathogen exposure. Rhizosphere bacterial communities of tomato plants were significantly different between the two soils, with potential beneficial strains occurring at higher relative abundances in the suppressive soil. Root transcriptome analysis revealed the soil legacy to induce differences in gene expression, most importantly, genes involved in the pathway of phenylpropanoid biosynthesis. Last, we found genes in the phenylpropanoid biosynthesis pathway to correlate with specific microbial taxa, including Gp6, Actinomarinicola, Niastella, Phaeodactylibacter, Longimicrobium, Bythopirellula, Brevundimonas, Ferruginivarius, Kushneria, Methylomarinovum, Pseudolabrys, Sphingobium, Sphingomonas, and Alterococcus. Taken together, our study points to the potential regulation of plant systemic resistance by specific microbial taxa, and the importance of soil legacy on disease incidence and eliciting plant-defense mechanisms.

The environmental bacterium Pseudomonas mosselii produces antagonistic secondary metabolites with inhibitory effects on multiple plant pathogens, including Ralstonia solanacearum, the causal agent of bacterial wilt. In this study, an engineered P. mosselii strain was generated to express R. solanacearum ripAA, which determines the incompatible interactions with tobacco plants. The ripAA gene, together with its native promoter, was integrated into the P. mosselii chromosome. The resulting strain showed no difference in antimicrobial activity against R. solanacearum. Promoter-LacZ fusion and RT-PCR experiments demonstrated that the ripAA gene was transcribed in culture media. Compared with that of the wild type, the engineered strain reduced the disease index by 9.1% for bacterial wilt on tobacco plants. A transcriptome analysis was performed to identify differentially expressed genes in tobacco plants, and the results revealed that ethylene- and jasmonate-dependent defense signaling pathways were induced. These data demonstrates that the engineered P. mosselii expressing ripAA can improve biological control against tobacco bacterial wilt by the activation of host defense responses.

Microbial bio-products are becoming an appealing and viable alternative to chemical pesticides for effective management of crop diseases. These bio-products are known to have potential to minimize agrochemical applications without losing crop yield and also restore soil fertility and productivity. In this study, the inhibitory efficacy of 2,4-diacetylphloroglucinol (DAPG) produced by Pseudomonas fluorescens VSMKU3054 against Ralstonia solanacearum was assessed. Biochemical and functional characterization study revealed that P. fluorescens produced hydrogen cyanide (HCN), siderophore, indole acetic acid (IAA) and hydrolytic enzymes such as amylase, protease, cellulase and chitinase, and had the ability to solubilize phosphate. The presence of the key antimicrobial encoding gene in the biosynthesis of 2,4-diacetylphloroglucinol (DAPG) was identified by PCR. The maximum growth and antimicrobial activity of P. fluorescens was observed in king’s B medium at pH 7, 37 °C and 36 h of growth. Glucose and tryptone were found to be the most suitable carbon and nitrogen sources, respectively. DAPG was separated by silica column chromatography and identified by various methods such as UV-Vis, FT-IR, GC-MS and NMR spectroscopy. When R. solanacearum cells were exposed to DAPG at 90 µg/mL, the cell viability was decreased, reactive oxygen species (ROS) were increased and chromosomal DNA was damaged. Application of P. fluorescens and DAPG significantly reduced the bacterial wilt incidence. In addition, P. fluorescens was also found effective in promoting the growth of tomato seedlings. It is concluded that the indigenous isolate P. fluorescens VSMKU3054 could be used as a suitable biocontrol agent against bacterial wilt disease of tomato.

Bacterial wilt caused by the β-proteobacterium Ralstonia solanacearum is one of the most destructive soil-borne pathogens in peppers (Capsicum annuum L.) worldwide. Cultivated pepper fields in Korea face a continuous spread of this pathogen due to global warming. The most efficient and sustainable strategy for controlling bacterial wilt is to develop resistant pepper varieties. Resistance, which is quantitatively inherited, occurs differentially depending on R. solanacearum isolates. Therefore, in this study, we aimed to identify resistance quantitative trait loci (QTLs) in two F2 populations derived from self-pollination of a highly resistant pepper cultivar ‘Konesian hot’ using a moderately pathogenic ‘HS’ isolate and a highly pathogenic ‘HWA’ isolate of R. solanacearum for inoculation, via genotyping-by-sequencing analysis. QTL analysis revealed five QTLs, Bwr6w-7.2, Bwr6w-8.1, Bwr6w-9.1, Bwr6w-9.2, and Bwr6w-10.1, conferring resistance to the ‘HS’ isolate with R2 values of 13.05, 12.67, 15.07, 10.46, and 9.69%, respectively, and three QTLs, Bwr6w-5.1, Bwr6w-6.1, and Bwr6w-7.1, resistant to the ‘HWA’ isolate with phenotypic variances of 19.67, 16.50, and 12.56%, respectively. Additionally, six high-resolution melting (HRM) markers closely linked to the QTLs were developed. In all the markers, the mean disease index of the paternal genotype was significantly lower than that of the maternal genotype. The QTLs and HRM markers are expected to be useful for the development of pepper varieties with high resistance to bacterial wilt.

Ralstonia solanacearum is a causative agent of bacterial wilt and therefore poses a serious threat to cultivated peppers (Capsicum annuum L.). Although attempts have been made to control bacterial wilt by grafting, the disease resistance mechanisms that protect grafted peppers are poorly understood. Here, we grew grafted peppers composed of the rootstock Buyeding or Weishi and the scion Xinfeng 2. Following infection by R. solanacearum, we assessed the differences in lipid peroxidation, cellular structure, root secondary metabolism, and biomass, between grafted plants and controls. The grafted plants exhibited a greater root biomass than the control plants after infection. The root cell ultrastructure of the grafted plants showed only slight injury relative to that in the controls, and the roots of the grafted peppers were partially resistant to R. solanacearum. Grafted pepper plants showed lower levels of lipid peroxidation. Lignin content, salicylic acid levels, and the activities of phenylalanine ammonia lyase (PAL), peroxidase (POD), catalase (CAT), and polyphenol oxidase (PPO), were also higher in grafted plants. All of these effects occurred concomitantly with increased R. solanacearum resistance. Taken together, our findings demonstrate that grafting can significantly improve the disease resistance of pepper. Moreover, our results suggest that the Weishi rootstock may be very useful for the prevention and control of bacterial wilt in cultivated peppers.

Ralstonia solanacaerum is one of the most devastating bacteria causing bacterial wilt disease in more than 200 species of plants, especially those belonging to the family Solanaceae. To cope with this pathogen, plants have evolved different resistance mechanisms depending on signal transduction after perception. Phosphorylation is the central regulatory component of the signal transduction pathway. We investigated a comparative phosphoproteomics analysis of the stems of resistant and susceptible tomatoes at 15 min and 30 min after inoculation with Ralstonia solanacearum to determine the phosphorylated proteins involved in induced resistance. Phosphoprotein profiling analyses led to the identification of 969 phosphoproteins classified into 10 functional categories. Among these, six phosphoproteins were uniquely identified in resistant plants including cinnamyl alcohol dehydrogenase 1 (CAD1), mitogen-activated protein kinase kinase kinase 18 (MAPKKK18), phospholipase D delta (PLDDELTA), nicotinamide adenine dinucleotide transporter 1 (NDT1), B3 domain-containing transcription factor VRN1, and disease resistance protein RPM1 (RPM1). These proteins are typically involved in defense mechanisms across different plant species. qRT-PCR analyses were performed to evaluate the level of expression of these genes in resistant and susceptible tomatoes. This study provides useful data, leading to an understanding of the early defense mechanisms of tomatoes against R. solanacearum.

Ralstonia solanacearum is a pathogen that causes bacterial wilt producing severe damage in staple solanaceous crops. Traditional control has low efficacy and/or environmental impact. Recently, the bases of a new biotechnological method by lytic bacteriophages vRsoP-WF2, vRsoP-WM2 and vRsoP-WR2 with specific activity against R. solanacearum were established. However, some aspects remain unknown, such as the survival and maintenance of the lytic activity after submission to a preservation method as the lyophilization. To this end, viability and stability of lyophilized vRsoP-WF2, vRsoP-WM2 and vRsoP-WR2 and their capacity for bacterial wilt biocontrol have been determined against one pathogenic Spanish reference strain of R. solanacearum in susceptible tomato plants in different conditions and making use of various cryoprotectants. The assays carried out have shown satisfactory results with respect to the viability and stability of the bacteriophages after the lyophilization process, maintaining high titers throughout the experimental period, and with respect to the capacity of the bacteriophages for the biological control of bacterial wilt, controlling this disease in more than 50% of the plants. The results offer good prospects for the use of lyophilization as a conservation method for the lytic bacteriophages of R. solanacearum in view of their commercialization as biocontrol agents.

Ralstonia solanacearum is the causative agent of bacterial wilt, one of the most destructive plant diseases. While chemical control has an environmental impact, biological control strategies can allow sustainable agrosystems. Three lytic bacteriophages (phages) of R. solanacearum with biocontrol capacity in environmental water and plants were isolated from river water in Europe but not fully analysed, their genomic characterization being fundamental to understand their biology. In this work, the phage genomes were sequenced and subjected to bioinformatic analysis. The morphology was also observed by electron microscopy. Phylogenetic analyses were performed with a selection of phages able to infect R. solanacearum and the closely related phytopathogenic species R. pseudosolanacearum. The results indicated that the genomes of vRsoP-WF2, vRsoP-WM2 and vRsoP-WR2 range from 40,688 to 41,158 bp with almost 59% GC-contents, 52 ORFs in vRsoP-WF2 and vRsoP-WM2, and 53 in vRsoP-WR2 but, with only 22 or 23 predicted proteins with functional homologs in databases. Among them, two lysins and one exopolysaccharide (EPS) depolymerase, this type of depolymerase being identified in R. solanacearum phages for the first time. These three European phages belong to the same novel species within the Gyeongsanvirus, Autographiviridae family (formerly Podoviridae). These genomic data will contribute to a better understanding of the abilities of these phages to damage host cells and, consequently, to an improvement in the biological control of R. solanacearum.

Bacterial wilt, caused by Ralstonia solanacearum, is an emerging constraint in amaranth production in Benin. Host resistance is the most sustainable disease control measure. Ten amaranth varieties including A2002, Bresil (B) -Sel, Madiira 2, AC-NL, GARE ES13-7, Madiira 1, UG-AMES13-2, AM-NKGN, IP-5-Sel and a local variety from Benin were screened for resistance to bacterial wilt. The study was conducted in a screen house and in the naturally contaminated open field during a consecutive rainy and dry season using a randomized complete block design with four and three replications, respectively. In the screen house, plants were inoculated by drenching a 40 mL of bacterial suspension containing 108 CFU/mL of R. solanacearum strain NCBI 5 GenBank N° MH397250 at the collar region. The bacterial wilt incidence (BWI) and the area under the disease progress curve (AUDPC) suggested differential reactions of amaranth varieties to the pathogen. BWI and AUDPC were low for UG-AMES13-2, moderate for Madiira 2, AM-NKGN and the local variety and very high for A2002, Bresil (B) -Sel, AC-NL, GARE ES13-7, Madiira 1 and IP-5-Sel. The World Vegetable Center’s UG-AMES13-2 can be considered as first choice, which is resistant to R. solanacearum, and should be scaled up for seed production towards supporting farmers.

Finding sources of resistance to bacterial wilt (BW) caused by Ralstonia solanacearum species complex is a crucial step toward the development of improved bacterial wilt-resistant tomato varieties. Here, we evaluated new sources of bacterial wilt-tolerant/resistant tomato lines and identified associated phylotype/sequevar of R. solanacearum strains in Benin. Eighteen F5 lines and five checks were evaluated in two hotspots: the experimental site of the World Vegetable Center, Cotonou Benin, and the Laboratory of Genetics, Biotechnology and Seed Science of the University of Abomey-Calavi. Experiments were laid out in a randomized complete block design with four replicates. Data were collected on bacterial wilt incidence, horticultural and fruit traits and yield components. Across the two experiments, the F5 lines showed no wilting, while the local variety ‘Tounvi’ used as susceptible check showed 57.64% wilting. The wilting was due to BW and was associated with sequevars I-14, I-18 and I-31 of phylotype I. AVTO1803, AVTO1955-6 and H7996 were the highest yielding lines with 20.29 t·ha−1, 17.66 t·ha−1 and 17.07 t/ha, respectively. The sources of resistance to BW can be recommended to national agricultural system for dissemination or used in tomato breeding programs.

Soil acidity is one of the main constraints to crop production worldwide. In Ethiopia, the problem of soil acidity has been increasing. Currently, more than 40% of cultivated land in the country has a soil pH < 5.5. Recently, bacterial wilt (caused by Ralstonia solanacearum) has become a serious problem, reaching epidemic levels in some of the major potato growing districts in the country. However, it is currently unknown if the current outbreak of bacterial wilt in potato production is associated with soil acidification or not. To examine the association between bacterial wilt and soil acidification, we conducted a field survey and field experiments and detected and characterised R. solanacearum strains. The study showed that 50% of potato fields were very strongly acidic (pH 4.5–5.0) and bacterial wilt incidence was higher in potato fields with low soil pH. The field experiments indicated that lime application significantly increased soil pH (p < 0.001) and reduced bacterial wilt incidence (p < 0.001). The more lime was applied, the stronger the positive effect on soil pH and the stronger the reduction in bacterial wilt incidence. Bacterial wilt incidence was on average 10.8% under 12 t/ha lime application, while it was about 40% in control plots (without lime) after 90 days. All R. solanacearum strains isolated from the symptomatic potato plants were Phylotype II. Our findings show that the current outbreak of bacterial wilt in Ethiopia is associated with soil acidification. They add to the understanding of the risk factors for bacterial wilt in potato. Aside from farm hygiene, sanitation and cultural practices, addressing soil acidification using lime needs to be considered as an additional component of an integrated package to deal with bacterial wilt in potato under acidic soil conditions.

Bacterial wilt (BW) of tomatoes, caused by Ralstonia solanacearum , is a devastating disease that results in large annual yield losses worldwide. Management of BW of tomatoes is difficult due to the soil-borne nature of the pathogen. One of the best ways to mitigate the losses is through breeding for disease resistance. Moreover, plant height (PH) is a crucial element related to plant architecture, which determines nutrient management and mechanical harvesting in tomatoes. An intraspecific F 2 segregating population (NC 11212) of tomatoes was developed by crossing NC 84173 (tall, BW susceptible) × CLN1466EA (short, BW resistant). We performed quantitative trait loci (QTL) mapping using single nucleotide polymorphic (SNP) markers and the NC 11212 F 2 segregating population. The QTL analysis for BW resistance revealed a total of three QTLs on chromosomes 1, 2, and 3, explaining phenotypic variation (R 2 ) ranging from 3.6% to 14.9%, whereas the QTL analysis for PH also detected three QTLs on chromosomes 1, 8, and 11, explaining R 2 ranging from 7.1% to 11%. This work thus provides information to improve BW resistance and plant architecture-related traits in tomatoes.

Many bacterial plant pathogens have a broad host range important for their life cycle. Alternate hosts from plant families other than the main (primary) host support the survival and dissemination of the pathogen population even in absence of main host plants. Metabolic peculiarities of main and alternative host plants can affect genetic diversity within and between the pathogen populations isolated from those plants. Strains of Gram-positive bacterium Curtobacterium flaccumfaciens were identified as being causal agents of bacterial spot and wilt diseases on leguminous plants, and other crop and weed plants, collected in different regions of Russia. Their biochemical properties and susceptibility to copper compounds have been found to be relatively uniform. According to conventional PCR assays, all of the isolates studied were categorised as pathovar Curtobacterim flaccumfaciens pv. flaccumfaciens , a pathogen of legumes. However, the strains demonstrated a substantial diversity in terms of virulence on several tested host plants and different phylogenetic relationships were revealed by BOX-PCR and alanine synthase gene ( alaS ) sequencing.

Biochar is increasingly being recognized as an effective soil amendment to enhance plant health and improve soil quality, but the complex relationships among biochar, plant resistance, and the soil microbial community are not clear. In this study, biochar derived from an invasive plant ( Solidago canadensis L.) was used to investigate its impacts on bacterial wilt control, soil quality, and microbial regulation. The results reveal that the invasive plant biochar application significantly reduced the abundance of Ralstonia solanacearum in the soil (16.8-32.9%) and wilt disease index (14.0-49.2%) and promoted tomato growth. The biochar treatment increased the soil organic carbon, nutrient availability, soil chitinase, and sucrase activities under pathogen inoculation. The biochar did not influence the soil bacterial community diversity, but significantly increased the relative abundance of beneficial organisms, such as Bacillus and Sphingomonas . Biochar application increased the number of nodes, edges, and the average degree of soil microbial symbiotic network, thereby enhancing the stability and complexity of the bacterial community. These findings suggest that the invasive plant biochar produces win-win effects on plant-soil systems by suppressing soilborne wilt disease, enhancing the stability of the soil microbial community network, and promoting resource utilization, indicating its good potential in sustainable soil management.

Bacterial wilt is a significant soil-borne disease that poses a threat to mulberry production yield and quality of agricultural production worldwide. However, the disease resistance mechanisms dependent on root exudates are not well understood. In this present study, we investigated the antibacterial mechanisms of the main active substances (erucamide, oleamide, and camphor bromide) present in mulberry root exudates (MRE) against Ralstonia pseudosolanacearum ( Rp ), the causal agent of bacterial wilt. Our findings revealed that these three active substances inhibited the growth activity of Rp by affecting the cell morphology and extracellular polysaccharide content, as well as triggering a burst of reactive oxygen species. The active substances induced oxidative stress, leading to a decrease in Rp growth. Additionally, the expression levels of key genes in the hrp gene cluster ( hrpB , hrpX , and hrpF ) and other virulence-related genes (such as ripAW , ripAE , Rs5-4819 , Rs5-4374 , ace , egl3 , and pehB ) were significantly reduced upon treatment with the active substances. Further pathogenicity experiments demonstrated that root exudates (at a concentration of 1.5 mg·mL -1 ) delayed or slowed down the occurrence of bacterial wilt in mulberry. These findings provide valuable insight into the antimicrobial mechanisms of MRE against Rp and lay a theoretical foundation for the development and application of biocontrol agents to control mulberry bacterial wilt.

Potato is an important crop, used not only for food production but also for various industrial applications. With the introduction of the potato as a staple food strategy, the potato industry in China has grown rapidly. However, issues related to bacterial wilt, exacerbated by factors such as seed potato transportation and continuous cropping, have become increasingly severe in the primary potato cultivation regions of China, leading to significant economic losses. The extensive genetic diversity of Ralstonia solanacearum ( R. solanacearum ), which is the pathogen of bacterial wilt, has led to a lack of highly resistant potato genetic resources. There is a need to identify and cultivate potato varieties with enhanced resistance to reduce the adverse impact of this disease on the industry. We screened 55 accessions of nine different wild potato species against the bacterial wilt pathogen R. solanacearum PO2-1, which was isolated from native potato plants and belongs to phylotype II. Three accessions of two species (ACL24-2, PNT880-3, and PNT204-23) were identified with high resistance phenotypes to the tested strains. We found these accessions also showed high resistance to different phylotype strains. Among them, only PNT880-3 was capable of flowering and possessed viable pollen, and it was diploid. Consistent with the high resistance, decreased growth of R. solanacearum was detected in PNT880-3. All these findings in our study reveal that the wild potato PNT880-3 was a valuable resistance source to bacterial wilt with breeding potential.

Grafting is increasingly becoming an indispensable tool that minimizes the risks associated with intensive vegetable production systems, including soil-borne diseases. This study assesses the performance of two cherry tomato hybrids (‘Cheramy’ and ‘Sheeja’) grafted onto three tomato and five eggplant local rootstock genotypes (cultivated/wild) under Ralstonia solanacearum (bacterial wilt)-infested greenhouse soil. The impact of grafting on growth, yield and fruit physical quality was mainly influenced by the response of rootstocks to disease resistance. The non-grafted plants of both the cultivars were severely affected by bacterial wilt, thus presenting high susceptibility to disease. Eggplant rootstocks imparted moderate to high resistance against bacterial wilt in both the scions, while tomato (cultivated or wild) rootstocks did not improve disease resistance, except ‘Anagha’, which provided resistance to scion cv. ‘Cheramy’. In general, scion cv. ‘Cheramy’, grafted or non-grafted, showed superior growth, yield and fruit quality compared to ‘Sheeja’. The most productive graft combinations for both the cultivars involved resistant rootstocks, i.e., ‘Sheeja’ onto eggplant rootstock ‘Surya’, and ‘Cheramy’ onto tomato rootstock ‘Anagha’. Fruit quality attributes such as ascorbic acid and lycopene contents were considerably higher, and the total soluble solids (TSS) content was considerably lower in scion cv. ‘Cheramy’, whether grafted or non-grafted, than those involving scion cv. ‘Sheeja’. The grafting effect on fruit chemical quality attributes was not promising, except grafting ‘Sheeja’ onto ‘Sopim’ for TSS, ‘Sheeja’ onto ‘Sotor’ for lycopene and ‘Cheramy’ onto ‘Ponny’ for total phenols, though no clear connection with disease incidence was in these grafts. Conclusively, eggplant rootstock imparted wilt resistance, while both eggplant and tomato rootstock grafting was beneficial to both scion cultivars in boosting the overall production and economic gains, especially for ‘Cheramy’ grafted onto ‘Anagha’ rootstock under bacterial wilt infested soil of greenhouse.

Ralstonia solanacearum is the pathogen responsible for wilting, yield losses, and death in tomato plants. The use of resistant cultivars has been proven as the most appropriate solution to controlling this pathogen. Therefore, further study of host-plant resistance mechanisms in tomatoes is urgently needed. 1H-NMR (nuclear magnetic resonance) spectroscopy combined with multivariate data analysis has been used to identify the biochemical compounds that play a crucial role in the defense mechanisms of tomato against bacterial wilt. Eleven metabolites consisting of amino acids, sugars and organic acids were identified and presented at different concentrations in each cultivar. Leucine and valine were determined as distinguishable metabolites of resistant and susceptible cultivars. Permata and Hawaii 7996 as resistant cultivars had a significant decrease of valine after inoculation about 1.5–2 times compared to the susceptible cultivar (GM2). Meanwhile, the resistant cultivars had a higher level of leucine, about 1.3–1.5 times compared to the susceptible ones. Synthesis of leucine and valine are linked as a member of the pyruvate family. Therefore, the decrease in valine may be related to the higher need for leucine to form the leucine-rich receptor, which plays a role in the plant’s immune system against the bacterial wilt.

The bacterial wilt pathogen, first known as Bacillus solanacearum, has undergone numerous taxonomic changes since its first description in 1896. The history and significance of this pathogen is covered in this review with an emphasis on the advances in technology that were used to support each reclassification that finally led to the current separation of Ralstonia solanacearum into three genomic species. Frequent name changes occurred as methodology transitioned from phenotypic, biochemical, and molecular studies, to genomics and functional genomics. The diversity, wide host range, and geographical distribution of the bacterial wilt pathogen resulted in its division into three species as genomic analyses elucidated phylogenetic relationships among strains. Current advances in phylogenetics and functional genomics now open new avenues for research into epidemiology and control of the devastating bacterial wilt disease.

Bacterial wilt disease caused by Ralstonia solanacearum is a devastating plant disease that inflicts heavy losses to the large number of economic host plants it infects. The potential of dried powder of the Conyza canadensis to control bacterial wilt (BW) of tomato was explored in vitro and in planta. Three application times (16 days before transplanting (DBT), 8 DBT and 0 DBT), three plastic-mulch durations (10 days plastic mulching (DPM), 5DPM and 0DPM) and four doses viz. 0 g, 8 g, 16 g and 24 g of the plant powder were evaluated. SEM analysis was also conducted to observe the change in bacterial cell morphology. Ethanol extract of dried C. canadensis in different concentrations inhibited the in vitro growth of R. solanacearum by as much as 98% of that produced by ampicillin. As evident from the scanning electron micrograph, the highest concentration produced severe morphologic changes, such as rupture of the bacterial cell walls and cell contents leaked out. Results from application time and dose experiment demonstrated that the highest powder dose viz. 24 g kg−1 mixed with infested soil 16 DBT gave maximum root length (34.0 ± 2.5 cm), plant height (74.3 ± 4.7 cm), fresh biomass (58.3 ± 4.3 g), reduction in bacterial population (1.52 log10) and resulted in lowest AUDPC value (1156.6). In case of mulching duration and dose experiment the maximum root length (39.6 ± 3.2 cm), plant height (78.3 ± 5.8 cm), fresh biomass (65.6 ± 4.9 g) reduction in bacterial population (1.59 log10) and lowest AUDPC value (1251.6) was achieved through the application of highest powder dose viz. 24 g kg−1 and longest plastic mulching duration of 10 DPM. The better results of highest dose and longer application time can be explained on the basis of higher amounts of anti-microbial plant bio-active compounds in highest dose and the longer exposure time of the pathogen to these chemicals. The better results of longer mulching duration are due to faster and more complete decomposition (because of 10-days-long plastic-mulch-provided increased solar heat) of the dried powder which produced more amounts of volatile and non-volatile bactericidal compounds. Our results clearly suggest that the use of 24 g kg−1 dried plant powder of C. canadensis plastic-mulched for two weeks could be used as a reliable component of the integrated disease management program against BW.

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 HNO 2 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 MHNO 2 -20 treated with nitrous acid (HNO 2 ) 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, MHNO 2 -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 MHNO 2 -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 MHNO 2 -20-treated plants after 30 days of inoculation. The derivative strain MHNO 2 -20, developed by treating B. amyloliquefaciens with nitrous acid (HNO 2 ), was therefore found to have a higher bio-efficacy to control bacterial wilt disease of tomato under glasshouse conditions than a wild-type strain.

Ralstonia solanacearum is a causative agent of bacterial wilt in many important crops throughout the world. How to control bacterial wilt caused by R. solanacearum is a major problem in agriculture. In this study, we aim to isolate the biocontrol agents that have high efficacy in the control of bacterial wilt. Three new bacterial strains with high antimicrobial activity against R. solanacearum GMI1000 were isolated and identified. Our results demonstrated that these bacteria could remarkably inhibit the disease index of host plant infected by R. solanacearum. It was indicated that strain GZ-34 (CCTCC No. M 2016353) showed an excellent protective effect to tomato under greenhouse conditions. Strain GZ-34 was characterized as Escherichia coli based on morphology, biochemistry, and 16S rRNA analysis. We identified that the main antimicrobial compounds produced by E. coli GZ-34 were cyclo(l-Pro-d-Ile) and cyclo(l-Pro-l-Phe) using electrospray ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance (NMR) analysis. The two active compounds also interfered with the expression levels of some pathogenicity-contributors of R. solanacearum. Furthermore, cyclo(l-Pro-l-Phe) effectively inhibited spore formation of Magnaporthe grisea, which is a vital pathogenesis process of the fungal pathogen, suggesting cyclic dipeptides from E. coli are promising potential antimicrobial agents with broad-spectrum activity to kill pathogens or interfere with their pathogenesis.

Bacterial wilt is a destructive disease caused by the phytopathogen Ralstonia solanacearum (R. solanacearum), which is widely found in various tobacco-growing areas all over the world. Botanical bactericidal substances have gradually emerged as a hot topic in modern pesticide research. In this study, the antibacterial activities of two phytochemicals (resveratrol and coumarin) against R. solanacearum and their in vivo and in vitro efficacy for controlling tobacco bacterial wilt were evaluated. We rule out significant biological effects of both phytochemicals using transmission electron microscope (TEM) and fluorescence microscope, which suppressed the growth of R. solanacearum. Furthermore, we demonstrated that the toxicity mechanisms mainly involved damaging bacterial cell membrane and preventing swarming motility and biofilm formation. A further pot experiment demonstrated that coumarin and resveratrol significantly inhibited early adhesion and colonization of R. solanacearum in tobacco plants and the corresponding control efficacies were 68% and 85% after incubation for 13 days, respectively. The findings of this study suggest that both resveratrol and coumarin have potential as non-toxic antimicrobial strategies for controlling tobacco bacterial wilt disease.

Many cereals accumulate hydroxamic acids involved in defense of plant against various fungi, bacteria, and insects. 2,4-dihydroxy-7-methoxy-1,4-benzoxazine-3-one, commonly known as DIMBOA, is one of the principal cyclic hydroxamic acids in aqueous extracts of maize. The aim of this study was to evaluate the antibacterial activity of the isolated DIMBOA and its derivatives 2-benzoxazolinone (BOA), 6-chloro-2-benzoxazolinone (CDHB), and 2-mercaptobenzothiazole (MBT) against Ralstonia solanacearum. MBT showed the strongest antibacterial activity, followed by CDHB and DIMBOA, with minimum inhibitory concentrations (MICs) of 50, 100 and 200 mg/L, respectively, better than the BOA with 300 mg/L. These compounds also significantly affect bacterial growth, reduce biofilm formation, and inhibit swarming motility within 24 h. This paper is the first to report the anti-R. solanacearum activity of DIMBOA from Z. mays. The bioassay and pot experiment results suggested that DIMBOA and its derivatives exhibit potential as a new matrix structure of designing target bactericide or elicitor for controlling tobacco bacterial wilt. Further studies must evaluate the efficacy of DIMBOA and its derivatives in controlling bacterial wilt under natural field conditions where low inoculum concentrations exist.

Protocatechualdehyde (PCA) is an important plant-derived natural product that has been associated with a wide variety of biological activities and has been widely used in medicine as an antioxidant, anti-aging and an anti-inflammatory agent. However, fewer reports concerning its antibacterial effects on plant-pathogenic bacteria exist. Therefore, in this study, protocatechualdehyde was evaluated for its antibacterial activity against plant pathogens along with the mechanism of its antibacterial action. PCA at 40 μg/mL was highly active against R. solanacearum and significantly inhibited its growth. The minimum bactericidal concentration and minimum inhibitory concentration values for PCA were 40 μg/mL and 20 μg/mL, respectively. Further investigation of the mechanism of action of PCA via transmission electron microscopy and biological assays indicated that the destruction of the cell structure, the shapes and the inhibition of biofilm formation were important. In addition, the application of PCA effectively reduced the incidence of bacterial wilt on tobacco under greenhouse conditions, and the control efficiency was as high as 92.01% at nine days after inoculation. Taken together, these findings suggest that PCA exhibits strong antibacterial activity against R. solanacearum and has the potential to be applied as an effective antibacterial agent for controlling bacterial wilt caused by R. solanacearum.

Coumarins are important plant-derived natural products with wide-ranging bioactivities and extensive applications. In this study, we evaluated for the first time the antibacterial activity and mechanisms of action of coumarins against the phytopathogen Ralstonia solanacearum, and investigated the effect of functional group substitution. We first tested the antibacterial activity of 18 plant-derived coumarins with different substitution patterns, and found that daphnetin, esculetin, xanthotol, and umbelliferone significantly inhibited the growth of R. solanacearum. Daphnetin showed the strongest antibacterial activity, followed by esculetin and umbelliferone, with MICs of 64, 192, and 256 mg/L, respectively, better than the archetypal coumarin with 384 mg/L. We further demonstrated that the hydroxylation of coumarins at the C-6, C-7 or C-8 position significantly enhanced the antibacterial activity against R. solanacearum. Transmission electron microscope (TEM) and fluorescence microscopy images showed that hydroxycoumarins may interact with the pathogen by mechanically destroying the cell membrane and inhibiting biofilm formation. The antibiofilm effect of hydroxycoumarins may relate to the repression of flagellar genes fliA and flhC. These physiological changes in R. solanacearum caused by hydroxycoumarins can provide information for integral pathogen control. The present findings demonstrated that hydroxycoumarins have superior antibacterial activity against the phytopathogen R. solanacearum, and thus have the potential to be applied for controlling plant bacterial wilt.