Pathogen Ralstonia Solanacearum Research Articles

Fermented chrysanthemum stem as a source of natural phenolic compounds to alleviate tomato bacterial wilt disease

Abstract Natural antimicrobial compounds (NACs) in the plant stem are crucial for replacing conventional synthetic pesticides in the control of soil-borne diseases, and microbial fermentation can enhance their concentration and bioactivity. In this study, the stems of ten plant species were collected for fermentation by probiotic bacteria Bacillus amyloliquefaciens T-5 to identify the most effective plant resource for controlling tomato bacterial wilt disease and discover key NACs. Chrysanthemum stem was identified as an optimal fermentation substrate, as its water-soluble extracts (WSEs) significantly inhibited the growth of pathogenic Ralstonia solanacearum and effectively alleviated tomato wilt under greenhouse conditions. Key metabolites, primarily phenolic acids including 2-hydroxy-3-phenylpropanoic acid (PLA), 3-(4-hydroxyphenyl)-propionic acid (HPPA), and mandelic acid (MA), were determined by metabolomics, all of which significantly inhibited the growth of R. solanacearum at a concentration of 0.2 mM, with only HPPA effectively controlling tomato wilt. Thus, fermented chrysanthemum stem contains NACs that are effective against bacterial wilt, providing a green option for controlling soil-borne diseases.

Bioprospecting of Selected Trichoderma spp. for Its Plant Growth Promoting and Biocontrol Properties

Bioprospecting for Trichoderma strains from peat soil collected at MARDI Sessang Station, Saratok, Sarawak, revealed promising candidates as plant growth-promoting microbes (PGPM) and producers of phytohormones such as indole-3-acetic acid (IAA) and gibberellic acid (GA3). The study aimed to identify Trichoderma strains with dual functions in biocontrol and plant growth enhancement. A total of seventeen strains were isolated and screened for their biocontrol activity against significant plant pathogens—Ralstonia solanacearum and Colletotrichum gloeosporioides and also their potential to produce IAA and GA3. Among the isolates, one strain exhibited strong antagonistic activity against R. solanacearum, a pathogen responsible for bacterial wilt, suggesting its potential for biocontrol applications. Additionally, two strains showed significant inhibitory effects against C. gloeosporioides, the causative agent of anthracnose. Four strains demonstrated the ability to produce IAA, an essential auxin for root development and overall plant growth promotion and two strains were identified as GA3 producers, contributing to plant growth by promoting cell elongation and germination. This study highlights the potential of Trichoderma strains from peat soils in Sarawak as valuable resources for sustainable agriculture. The dual functionality of these strains as biocontrol agents and phytohormone producers makes them promising candidates for integrated pest management (IPM) and biofertilizer development. Future research should focus on field trials and further molecular characterization to fully explore the potential of these Trichoderma isolates in enhancing crop productivity.

A Ralstonia solanacearum type III effector alters the actin and microtubule cytoskeleton to promote bacterial virulence in plants.

Cellular responses to biotic stress frequently involve signaling pathways that are conserved across eukaryotes. These pathways include the cytoskeleton, a proteinaceous network that senses external cues at the cell surface and signals to interior cellular components. During biotic stress, dynamic cytoskeletal rearrangements serve as a platform from which early immune-associated processes are organized and activated. Bacterial pathogens of plants and animals use proteins called type III effectors (T3Es) to interfere with host immune signaling, thereby promoting virulence. We previously found that RipU, a T3E from the soilborne phytobacterial pathogen Ralstonia solanacearum, co-localizes with the plant cytoskeleton. Here, we show that RipU from R. solanacearum K60 (RipUK60) associated with and altered the organization of both the actin and microtubule cytoskeleton. We found that pharmacological disruption of the tomato (Solanum lycopersicum) cytoskeleton promoted R. solanacearum K60 colonization. Importantly, tomato plants inoculated with R. solanacearum K60 lacking RipUK60 (ΔripUK60) had reduced wilting symptoms and significantly reduced root colonization when compared to plants inoculated with wild-type R. solanacearum K60. Collectively, our data suggest that R. solanacearum K60 uses the type III effector RipUK60 to remodel cytoskeletal organization, thereby promoting pathogen virulence.

Lotus resistance against Ralstonia is enhanced by Mesorhizobium and does not impair mutualism.

Legumes establish nitrogen-fixing symbioses with rhizobia. On the contrary, they can be attacked concomitantly by pathogens, raising the question of potential trade-offs between mutualism and immunity. In order to study such trade-offs, we used a tripartite system involving the model legume Lotus japonicus, its rhizobial symbiont Mesorhizobium loti and the soilborne pathogen Ralstonia solanacearum. We investigated the impact of mutualism on plant defense and the reciprocal influence of plant defense on mutualism. We found that Lotus age-related resistance against Ralstonia was improved by the interaction with rhizobia especially when nodulation is triggered. Conversely, age-related resistance did not compromise nodule organogenesis or functioning under pathogen attack. Proteomic characterization indicates that this resistance is associated with distinct proteome modifications in roots and nodules. This resistance questions the concept of interference between efficient defense reactions and mutualistic interactions and is of great interest for agricultural purposes as it not only restricts pathogen colonization, but would also preserve nitrogen fixation and yield.

Isolation and identification of antifungal, antibacterial and nematocide agents from marine bacillus gottheilii MSB1

Pathogenic fungi employ numerous strategies to colonize plants, infect them, reduce crop yield and quality, and cause significant losses in agricultural production. The increasing use of chemical pesticides has led to various ecological and environmental issues, including the emergence of resistant weeds, soil compaction, and water pollution, all negatively impacting agricultural sustainability. Additionally, the extensive development of synthetic fungicides has adverse effects on animal and human health, prompting the exploration of alternative approaches and green strategies for phytopathogen control. Microorganisms living in sponges represent a promising source of novel bioactive secondary metabolites, potentially useful in developing new nematicidal and antimicrobial agents. This study focuses on extracting bioactive compounds from endosymbiotic bacteria associated with the marine sponge Hyrtios erect sp. (collected from NIOF Station, Hurghada, Red Sea, Egypt) using various organic solvents. Bacillus sp. was isolated and identified through 16 S rRNA gene sequencing. The biocidal activity of Bacillus gotheilii MSB1 extracts was screened against plant pathogenic bacteria, fungi, and nematodes. The n-butanol extract showed significant potential as a biological fungicide against Alternaria alternata and Fusarium oxysporum. Both n-hexane and ethyl acetate extracts exhibited negative impacts against the plant pathogenic bacteria Erwinia carotovora and Ralstonia solanacearum, whereas the n-butanol extract had a positive effect. Regarding nematicidal activity, ethyl acetate and n-butanol extracts demonstrated in-vitro activity against the root-knot nematode Meloidogyne incognita, which causes serious vegetable crop diseases, but the n-hexane extract showed no positive effects. The findings suggest that bioactive compounds from endosymbiotic bacteria associated with marine sponges, particularly B. gotheilii MSB1, hold significant potential as alternative biological control agents against plant pathogens. The n-butanol extract, in particular, displayed promising biocidal activities against various plant pathogenic fungi, bacteria, and nematodes. These results support further exploration and development of such bioactive compounds as sustainable, environmentally friendly alternatives to synthetic pesticides and fungicides in agricultural practices.

The FERONIA-RESPONSIVE TO DESICCATION 26 module regulates vascular immunity to Ralstonia solanacearum.

Some pathogens colonize plant leaves, but others invade the roots, including the vasculature, causing severe disease symptoms. Plant innate immunity has been extensively studied in leaf pathosystems; however, the precise regulation of immunity against vascular pathogens remains largely unexplored. We previously demonstrated that loss of function of the receptor kinase FERONIA (FER) increases plant resistance to the typical vascular bacterial pathogen Ralstonia solanacearum. Here, we show that upon infection with R. solanacearum, root xylem cell walls in Arabidopsis thaliana become highly lignified. FER is specifically upregulated in the root xylem in response to R. solanacearum infection, and inhibits lignin biosynthesis and resistance to this pathogen. We determined that FER interacts with and phosphorylates the transcription factor RESPONSIVE TO DESICCATION 26 (RD26), leading to its degradation. Overexpression and knockout of RD26 demonstrated that it positively regulates plant resistance to R. solanacearum by directly activating the expression of lignin-related genes. Tissue-specific expression of RD26 in the root xylem confirmed its role in vascular immunity. We confirmed that the FER-RD26 module regulates lignin biosynthesis and resistance against R. solanacearum in tomato (Solanum lycopersicum). Taken together, our findings unveil that the FER-RD26 cascade governs plant immunity against R. solanacearum in vascular tissues by regulating lignin deposition. This cascade may represent a key defense mechanism against vascular pathogens in plants.

Antibacterial, Herbicidal, and Plant Growth-Promoting Properties of Streptomyces sp. STD57 from the Rhizosphere of Adenophora stricta.

Bacterial wilt triggered by the soil-borne pathogenic bacterium Ralstonia solanacearum is one of the most serious diseases in tomato plants, leading to huge economic losses worldwide. Biological control is considered an environmentally friendly and sustainable way to manage soil-borne diseases. In this study, Streptomyces sp. STD57 isolated from the rhizosphere of Adenophora stricta showed strong antibacterial activity against R. solanacearum. Pot experiments showed that strain STD57 exhibited a significant biocontrol effect (81.7%) on tomato bacterial wilt in the greenhouse environment. Furthermore, strain STD57 could inhibit the growth of weeds (Amaranthus retroflexus, Portulaca oleracea, and Echinochloa crusgalli) but promote the growth of crops (wheat, rice, and tomato). The plant growth-promoting substance was identified as indoleacetic acid (IAA) by high-pressure liquid chromatography-mass spectrometry and genome analysis. Coarse separation of the fermented extracts revealed that the antibacterial and herbicidal substances were mainly in the fermentation supernatant and belonged to different products. These findings suggested that strain STD57 may be a potential biocontrol and bioherbicide agent useful in agriculture.

A Nonessential Sfp-Type Phosphopantetheinyl Transferase Contributes Significantly to the Pathogenicity of Ralstonia solanacearum.

4'-Phosphopantetheinyl transferases (PPTases) play important roles in the posttranslational modifications of bacterial carrier proteins, which are involved in various metabolic pathways. Here, we found that RsacpS and RspcpS encoded a functional AcpS-type and Sfp-type PPTase, respectively, in Ralstonia solanacearum GMI1000, and both are capable of modifying R. solanacearum AcpP1, AcpP2, AcpP3, and AcpP5 proteins. RspcpS is located on the megaplasmid, which does not affect strain growth and fatty acid synthesis but significantly contributes to the virulence of R. solanacearum and preferentially participates in secondary metabolism. We found that deletion of RspcpS did not affect the abilities of cellulose degradation, biofilm formation, and resistance to NaCl, sodium dodecyl sulfate, and H2O2 and attenuated R. solanacearum pathogenicity only in the assay of soil-drenching infection but not stem injection of tomato. It is hypothesized that RsPcpS plays a role in cell viability in complex environments and in the process during which the strain recognizes and approaches plants. These results suggest that both RsAcpS and RsPcpS may be potential targets for controlling diseases caused by R. solanacearum.

Siderophore interactions drive the ability of Pseudomonas spp. consortia to protect tomato against Ralstonia solanacearum.

The soil-borne bacterial pathogen Ralstonia solanacearum causes significant losses in Solanaceae crop production worldwide, including tomato, potato, and eggplant. To efficiently prevent outbreaks, it is essential to understand the complex interactions between pathogens and the microbiome. One promising mechanism for enhancing microbiome functionality is siderophore-mediated competition, which is shaped by the low iron availability in the rhizosphere. This study explores the critical role of iron competition in determining microbiome functionality and its potential for designing high-performance microbiome engineering strategies. We investigated the impact of siderophore-mediated interactions on the efficacy of Pseudomonas spp. consortia in suppressing R. solanacearum , both in vitro and in vivo. Our findings show that siderophore production significantly enhances the inhibitory effects of Pseudomonas strains on pathogen growth, while other metabolites are less effective under iron-limited conditions. Moreover, siderophores play a crucial role in shaping interactions within the consortia, ultimately determining the level of protection against bacterial wilt disease. This study highlights the key role of siderophores in mediating consortium interactions and their impact on tomato health. Our results also emphasize the limited efficacy of other secondary metabolites in iron-limited environments, underscoring the importance of siderophore-mediated competition in maintaining tomato health and suppressing disease.

A bacterial effector manipulates plant metabolism, cell death, and immune responses via independent mechanisms.

Bacterial pathogens inject effector proteins inside plant cells to manipulate cellular functions and achieve a successful infection. The soil-borne pathogen Ralstonia solanacearum (Smith), the causal agent of bacterial wilt disease, secretes > 70 different effectors inside plant cells, although only a handful of them have been thoroughly characterized. One of these effectors, named RipI, is required for full R. solanacearum pathogenicity. RipI associates with plant glutamate decarboxylases (GADs) to promote the accumulation of gamma-aminobutyric acid (GABA), which serves as bacterial nutrient. In this work, we found that RipI can also suppress plant immune responses to bacterial elicitors, which seems to be unrelated to the ability of RipI to induce GABA accumulation and plant cell death. A detailed characterization of the RipI features that contribute to its virulence activities identified two residues at the C-terminal domain that mediate RipI interaction with plant GADs and the subsequent promotion of GABA accumulation. These residues are also required for the appropriate homeostasis of RipI in plant cells and the induction of cell death, although they are partially dispensable for the suppression of plant immune responses. Altogether, we decipher and uncouple the virulence activities of an important bacterial effector at the biochemical level.

Nonphytotoxic and pH-responsive ZnO-ZIF‑8 loaded with honokiol as a “nanoweapon” effectively controls the soil-borne bacterial pathogen Ralstonia solanacearum

The development of intelligently released and environmentally safe nanocarriers not only aligns with the sustainable agricultural strategy but also offers a potential solution for controlling severe soil-borne bacterial diseases. Herein, the core-shell structured nanocarrier loaded with honokiol bactericide (honokiol@ZnO-ZIF-8) was synthesized via a one-pot method for the targeted control of Ralstonia solanacearum, the causative agent of tobacco bacterial wilt disease. Results indicated that honokiol@ZnO-ZIF-8 nanoparticles induced bacterial cell membrane and DNA damage through the production of excessive reactive oxygen species (ROS), thereby reducing bacterial cell viability and ultimately leading to bacterial death. Additionally, the dissociation mechanism of the nanocarriers was elucidated for the first time through thermodynamic computational simulation. The nanocarriers dissociate primarily due to H+ attacking the N atom on imidazole, causing the rupture of the Zn-N bond under acidic conditions and at room temperature. Furthermore, honokiol@ZnO-ZIF-8 exhibited potent inhibitory effects against other prominent Solanaceae pathogenic bacteria (Pseudomonas syringae pv. tabaci), demonstrating its broad-spectrum antibacterial activity. Biosafety assessment results indicated that honokiol@ZnO-ZIF-8 exhibited non-phytotoxicity towards tobacco and tomato plants, with its predominant accumulation in the roots and no translocation to aboveground tissues within a short period. This study provides potential application value for the intelligent release of green pesticides. Environment implicationThe indiscriminate use of agrochemicals poses a significant threat to environmental, ecological security, and sustainable development. Slow-release pesticides offer a green and durable strategy for crop disease control. In this study, we developed a non-phytotoxic and pH-responsive honokiol@ZnO-ZIF-8 nano-bactericide based on the pathogenesis of Ralstonia solanacearum. Thermodynamic simulation revealed the dissociation mechanism of ZIF-8, with different acidity controlling the dissociation rate. This provides a theoretical basis for on-demand pesticide release while reducing residue in the. Our findings provide strong evidence for effective soil-borne bacterial disease control and on-demand pesticide release.

A robust high-throughput functional screening assay for plant pathogen effectors using the TMV-GFP vector.

Uncovering the function of phytopathogen effectors is crucial for understanding mechanisms of pathogen pathogenicity and for improving our ability to protect plants from diseases. An increasing number of effectors have been predicted in various plant pathogens. Functional characterization of these effectors has become a major focus in the study of plant-pathogen interactions. In this study, we designed a novel screening system that combines the TMV (tobacco mosaic virus)-GFP vector and Agrobacterium-mediated transient expression in the model plant Nicotiana benthamiana. This system enables the rapid identification of effectors that interfere with plant immunity. The biological function of these effectors can be easily evaluated by observing the GFP fluorescence signal using a UV lamp within just a few days. To evaluate the TMV-GFP system, we initially tested it with well-described virulence and avirulence type III effectors from the bacterial pathogen Ralstonia solanacearum. After proving the accuracy and efficiency of the TMV-GFP system, we successfully screened a novel virulence effector, RipS1, using this approach. Furthermore, using the TMV-GFP system, we reproduced consistent results with previously known cytoplasmic effectors from a diverse array of pathogens. Additionally, we demonstrated the effectiveness of the TMV-GFP system in identifying apoplastic effectors. The easy operation, time-saving nature, broad effectiveness, and low technical requirements of the TMV-GFP system make it a promising approach for high-throughput screening of effectors with immune interference activity from various pathogens.

Protective role of biomass endophytic fungi to enhance potato plant responses against wilt disease caused by Ralstoniasolanacearum

Globally, Ralstonia solanacearum wilt disease significantly reduces the value of tomatoes, eggplants, bananas, and potatoes. In this study, endophytic fungi were isolated and used as biopesticides against the bacterial pathogen Ralstonia solanacearum in potato plants, offering a promising alternative to chemical pesticides that harm the environment. Aspergillus niger, Alternaria burnsii, and Aspergillus niveus were used in this study to support potato growth, combat Ralstonia solanacearum wilt (RSW) disease, and induce defense mechanisms and growth-promoting tactics in potato plants. The mix of A. burnsii, A. niveus, and A. niger was the most effective treatment in reducing the percent disease index (PDI) with an 18 % decrease compared to the infected control. This was followed by treatments with A. niger and A. niveus, which had PDIs of 32 % and 35 %, respectively. The next best treatment was with A. burnsii alone, resulting in a PDI of 61 %. Treatment with mixed filtrates from A. burnsii, A. niveus, and A. niger showed the greatest significant increase in shoot length by 251.9 %, root length by 38.1 %, and the number of leaves by 40.5 %. A clear and noticeable improvement in the levels of chlorophyll and carotene pigments was observed in plants treated with the tested endophytic fungi compared to those in infected plants. The infected plants treated with a mix of endophytic fungi showed a 94.1 % increase in free proline compared to untreated infested plants. Endophytic fungi can be applied alone or in combination to increase the number and density of antioxidant enzymes isozymes (PPO and POD)in both infected and uninfected plants. The study results suggested using mixed filtrates from A. burnsii, A. niveus, and A. niger as growth inducers for potato plants and as bioagents to control R. solanacearum wilt disease.

Illumina 16S rRNA Gene Sequencing Dataset of Bacterial Communities of Soil Associated with Ironwood Trees (Casuarina equisetifolia) in Guam

Ironwood trees, which are of great importance for the economy and environment of tropical areas, were first discovered to suffer from a slow progressive dieback in Guam in 2002, later referred to as ironwood tree decline (IWTD). A variety of biotic factors have been shown to be associated with IWTD, including putative bacterial pathogens Ralstonia solanacearum and Klebsiella species (K. variicola and K. oxytoca), the fungus Ganoderma australe, and termites. Due to the soilborne nature of these pathogens, soil microbiomes have been suggested to be a significant factor influencing tree health. In this project, we sequenced the microbiome in the soil collected from the root region of healthy ironwood trees and those showing signs of IWTD to evaluate the association between the bacterial community in soil and IWTD. This dataset contains 4,782,728 raw sequencing reads present in soil samples collected from thirty-nine ironwood trees with varying scales of decline severity in Guam obtained via sequencing the V1–V3 region of the 16S rRNA gene on the Illumina NovaSeq (2 × 250 bp) platform. Sequences were taxonomically assigned in QIIME2 using the SILVA 132 database. Firmicutes and Actinobacteria were the most dominant phyla in soil. Differences in soil microbiomes were detected between limestone and sand soil parent materials. No putative plant pathogens of the genera Ralstonia or Klebsiella were found in the samples. Bacterial diversity was not linked to parameters of IWTD. The dataset has been made publicly available through NCBI GenBank under BioProject ID PRJNA883256. This dataset can be used to compare the bacterial taxa present in soil associated with ironwood trees in Guam to bacteria communities of other geographical locations to identify microbial signatures of IWTD. In addition, this dataset can also be used to investigate the relationship between soil microbiomes and the microbiomes of ironwood trees as well as those of the termites which attack ironwood trees.

New Antibacterial Peptaibiotics against Plant and Fish Pathogens from the Deep-Sea-Derived Fungus Simplicillium obclavatum EIODSF 020.

Bacterial diseases could severely harm agricultural production. To develop new antibacterial agents, the secondary metabolites of a deep-sea-derived fungus Simplicillium obclavatum EIODSF 020 with antibacterial activities against plant and fish pathogens were investigated by a bioassay-guided approach, which led to the isolation of 11 new peptaibiotics, simplicpeptaibs A-K (1-11). They contain 16-19 residues, including β-alanine, tyrosine, or tyrosine O-sulfate, that were rarely present in peptaibiotics. Their structures were elucidated by spectroscopic analyses (NMR, HRMS, HRMS2, and ECD) and Marfey's method. The primary and secondary structures of novel sulfated peptaibiotic 9 were reconfirmed by single-crystal X-ray diffraction analysis. Genome sequencing of S. obclavatum EIODSF 020 allowed the detection of a gene cluster encoding two individual NRPSs (totally containing 19 modules) that was closely related to simplicpeptaib biosynthesis. Antibacterial investigations of 1-11 together with the previously isolated linear and cyclic peptides from this strain suggested the antibacterial property of this fungus was attributed to the peptaibiotics and cyclic lipopeptides. Among them, compounds 4, 6, 7, and 9 showed significant activity against the tobacco pathogen Ralstonia solanacearum or tilapia pathogens Streptococcus iniae and Streptococcus agalactiae. The antibacterial activity of 6 against R. solanacearum could be enhanced by the addition of 1% NaCl. The structure-bioactivity relationship of simplicpeptaibs was discussed.

Lipid transfer protein StLTPa enhances potato disease resistance against different pathogens by binding and disturbing the integrity of pathogens plasma membrane.

Potato is the third most important food crop worldwide. Potato production suffers from severe diseases caused by multiple detrimental plant pathogens, and broad-spectrum disease resistance genes are rarely identified in potato. Here we identified the potato non-specific lipid transfer protein StLTPa, which enhances species none-specific disease resistance against various pathogens, such as the oomycete pathogen Phytophthora infestans, the fungal pathogens Botrytis cinerea and Verticillium dahliae, and the bacterial pathogens Pectobacterium carotovorum and Ralstonia solanacearum. The StLTPa overexpression potato lines do not show growth penalty. Furthermore, we provide evidence that StLTPa binds to lipids present in the plasma membrane (PM) of the hyphal cells of P. infestans, leading to an increased permeability of the PM. Adding of PI(3,5)P2 and PI(3)P could compete the binding of StLTPa to pathogen PM and reduce the inhibition effect of StLTPa. The lipid-binding activity of StLTPa is essential for its role in pathogen inhibition and promotion of potato disease resistance. We propose that StLTPa enhances potato broad-spectrum disease resistance by binding to, and thereby promoting the permeability of the PM of the cells of various pathogens. Overall, our discovery illustrates that increasing the expression of a single gene in potato enhances potato disease resistance against different pathogens without growth penalty.

Metagenomic insights into the response of soil microbial communities to pathogenic Ralstonia solanacearum.

Understanding the response of soil microbial communities to pathogenic Ralstonia solanacearum is crucial for preventing bacterial wilt outbreaks. In this study, we investigated the soil physicochemical and microbial community to assess their impact on the pathogenic R.solanacearum through metagenomics. Our results revealed that certain archaeal taxa were the main contributors influencing the health of plants. Additionally, the presence of the pathogen showed a strong negative correlation with soil phosphorus levels, while soil phosphorus was significantly correlated with bacterial and archaeal communities. We found that the network of microbial interactions in healthy plant rhizosphere soils was more complex compared to diseased soils. The diseased soil network had more linkages, particularly related to the pathogen occurrence. Within the network, the family Comamonadaceae, specifically Ramlibacter_tataouinensis, was enriched in healthy samples and showed a significantly negative correlation with the pathogen. In terms of archaea, Halorubrum, Halorussus_halophilus (family: Halobacteriaceae), and Natronomonas_pharaonis (family: Haloarculaceae) were enriched in healthy plant rhizosphere soils and showed negative correlations with R.solanacearum. These findings suggested that the presence of these archaea may potentially reduce the occurrence of bacterial wilt disease. On the other hand, Halostagnicola_larseniia and Haloterrigena_sp._BND6 (family: Natrialbaceae) had higher relative abundance in diseased plants and exhibited significantly positive correlations with R.solanacearum, indicating their potential contribution to the pathogen's occurrence. Moreover, we explored the possibility of functional gene sharing among the correlating bacterial pairs within the Molecular Ecological Network. Our analysis revealed 468 entries of horizontal gene transfer (HGT) events, emphasizing the significance of HGT in shaping the adaptive traits of plant-associated bacteria, particularly in relation to host colonization and pathogenicity. Overall, this work revealed key factors, patterns and response mechanisms underlying the rhizosphere soil microbial populations. The findings offer valuable guidance for effectively controlling soil-borne bacterial diseases and developing sustainable agriculture practices.

EVALUATION OF MICROBIAL POTENTIAL OF RHIZOBACTERIAL ISOLATES ASSOCIATED WITH SPENT MUSHROOM COMPOST AGAINST BACTERIAL WILT OF TOMATO

Bacterial wilt, caused by the soil-borne pathogen Ralstonia solanacearum, poses a significant threat to tomato crops worldwide. This study aims to assess the microbial potential of rhizobacterial isolates obtained from spent mushroom compost in suppressing bacterial wilt of tomatoes. Spent mushroom compost is a byproduct of mushroom cultivation and is known to harbor diverse microbial communities with potential plant-beneficial properties. Tomato leaves that were contaminated were collected from a number of places in the Rawalpindi Area, Punjab, Pakistan. Rhizobacterial isolates were collected from the rhizosphere of tomato plants grown in the presence of spent mushroom compost. These isolates were then subjected to laboratory evaluations for their antagonistic activity against R. solanacearum. Selected rhizobacterial isolates were further characterized for their plant growth-promoting traits. The potential of these isolates to enhance tomato plant growth and confer resistance against bacterial wilt was evaluated through greenhouse experiments. Results indicated that certain rhizobacterial isolates exhibited substantial antagonistic activity against R. solanacearum. Additionally, these isolates demonstrated multiple plant growth-promoting traits, suggesting a potential dual role in both pathogen suppression and plant enhancement. Greenhouse experiments revealed a significant reduction in the incidence of bacterial wilt in tomato plants and increase in the growth promotion traits were observed while treated with the selected rhizobacterial isolates alone and in combination compared to control groups. The findings from this study highlight the promising role of rhizobacterial isolates associated with spent mushroom compost in managing bacterial wilt in tomatoes.

Silica nanoparticles conferring resistance to bacterial wilt in peanut (Arachis hypogaea L.)

Peanut bacterial wilt (PBW) caused by the pathogen Ralstonia solanacearum severely affects the growth and yield potential of peanut crop. In this study, we synthesized silica nanoparticles (SiO2 NPs), a prospective efficient management approach to control PBW, and conducted a hydroponic experiment to investigate the effects of different SiO2 NPs treatments (i.e., 0, 100, and 500 mg L−1 as NP0, NP100, and NP500, respectively) on promoting plant growth and resistance to R. solanacearum. Results indicated that the disease indices of NP100 and NP500 decreased by 51.5 % and 55.4 % as compared with NP0 under R. solanacearum inoculation, respectively, while the fresh and dry weights and shoot length of NP100 and NP500 increased by 7.62–42.05 %, 9.45–32.06 %, and 2.37–17.83 %, respectively. Furthermore, SiO2 NPs induced an improvement in physio-biochemical enzymes (superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, and lipoxygenase) which eliminated the excess production of hydrogen peroxide, superoxide anions, and malondialdehyde to alleviate PBW stress. Notably, the targeted metabolomic analysis indicated that SiO2 NPs enhanced salicylic acid (SA) contents, which involved the induction of systemic acquired resistance (SAR). Moreover, the transcriptomic analysis revealed that SiO2 NPs modulated the expression of multiple transcription factors (TFs) involved in the hormone pathway, such as AHLs, and the identification of hormone pathways related to plant defense responses, such as the SA pathway, which activated SA-dependent defense mechanisms. Meanwhile, the up-regulated expression of the SA-metabolism gene, salicylate carboxymethyltransferase (SAMT), initiated SAR to promote PBW resistance. Overall, our findings revealed that SiO2 NPs, functioning as a plant elicitor, could effectively modulate physiological enzyme activities and enhance SA contents through the regulation of SA-metabolism genes to confer the PBW resistance in peanuts, which highlighted the potential of SiO2 NPs for sustainable agricultural practices.

HrpY protein of Ralstonia solanacearum exhibits spontaneous formation of pilus like assembly: analysis of its stability

Type 3 secretory system (T3SS), a complex protein machinery has a unique virulence mechanism that involves injecting effector proteins directly into host cells. The T3SS effector proteins are transported through an extracellular long hollow needle made up of multiple copies of a small protein. In T3SS of the plant pathogen Ralstonia solanacearum, the 8.6 kDa HrpY protein assembles into a large needle like apparatus (pilus) for transporting effector proteins. To study structural details of HrpY, we recombinantly expressed and purified HrpY in E. coli. The dynamic light scattering (DLS) analysis showed that rHrpY has spontaneously formed oligomers of large order (>100 nm). Transmission electron microscopy of rHrpY samples revealed that the large structures are tube like assembly having dimensions 86.3–166.6 nm and 5.8–6.8 nm in length and width respectively. Different molecular sizes of the purified rHrpY hindered the crystallization of the protein. The stability of oligomer assembly was studied with denaturants and surfactants. Denaturants like urea and guanidine HCl could not break them apart; however, detergents like SDS, sarkosyl, Octyl-β-Glucoside, CHAPS, Tween 20, Tween 80 and Triton X-100 showed disassembly of the oligomer. rHrpY assembly was found to withstand up to 50 °C and the circular dichroism analysis revealed that there is no significant change in the secondary structural composition with increase in temperature. However, change in the secondary structure was observed with the addition of SDS. Communicated by Ramaswamy H. Sarma