Plant Pathogenic Bacteria Research Articles

Bioinspired total synthesis and biological activity of Pegaharine A.

Phytopathogens cause various diseases by parasitizing crops, reducing crop yield and resulting in substantial economic losses in agricultural production. A novel type isolated from the perennial herbaceous Peganum harmala L. seeds, β-carboline alkaloids pegaharine A (PA), has become a hot topic in developing plant-originated green pesticides owing to their significant physiological activities. A scalable bioinspired total synthesis of PA is accomplished in the present work. The systematical biological assay study showed that PA exhibited moderate inhibitory activity against nine tested plant pathogenic fungi and showed significant inhibitory activity in vitro against the three tested plant pathogenic bacteria. Most noteworthy is the inhibitory rates of PA on Xanthomonas oryzae pv. oryzae (Xoo), X. oryzae pv. oryzicola (Xoc) and X. axonopodis pv. citri (Xac) of 93.6%, 92.1% and 86.1%, respectively, which are better than the control drug, bismerthiazol (63.4%, 61.2% and 53.7% at 100 μg mL-1 concentration). Furthermore, the EC50 value of PA against Xoo, Xoc and Xac was 52.2, 60.0 and 65.1 μg mL-1 , respectively, superior to 72.9, 64.2 and 70.1 μg mL-1 of the control drug. Moreover, the anti-Xoo mechanistic studies revealed that PA exerted its antibacterial effects by increasing the permeability of the bacterial membrane, reducing the extracellular polysaccharide content and inducing morphological changes in bacterial cells. A novel β-carboline alkaloid, PA, was prepared by biomimetic total synthesis. Its significant antibacterial activity was closely related to the permeation of bacterial cell membranes, which was confirmed by anti-Xoo mechanistic studies. More importantly, the structure could be regarded as a model for developing novel bactericides. © 2023 Society of Chemical Industry.

Transcriptome profiling of type VI secretion system core gene tssM mutant of Xanthomonas perforans highlights regulators controlling diverse functions ranging from virulence to metabolism.

T6SS has received attention due to its significance in mediating interorganismal competition through contact-dependent release of effector molecules into prokaryotic and eukaryotic cells. Reverse-genetic studies have indicated the role of T6SS in virulence in a variety of plant pathogenic bacteria, including the one studied here, Xanthomonas. However, it is not clear whether such effect on virulence is merely due to a shift in the microbiome-mediated protection or if T6SS is involved in a complex virulence regulatory network. In this study, we conducted in vitro transcriptome profiling in minimal medium to decipher the signaling pathways regulated by tssM-i3* in X. perforans AL65. We show that TssM-i3* regulates the expression of a suite of genes associated with virulence and metabolism either directly or indirectly by altering the transcription of several regulators. These findings further expand our knowledge on the intricate molecular circuits regulated by T6SS in phytopathogenic bacteria.

The application of entomopathogenic nematode modified microbial communities within nesting mounds of the red imported fire ants, Solenopsis invicta

Entomopathogenic microorganisms (e.g., fungi, bacteria, nematodes) have been widely used in biological control of soil-dwelling pests, including the red imported fire ant (RIFA), Solenopsis invicta, a notorious invasive pest worldwide. The application of large amounts of entomopathogenic microorganisms to soil may affect the indigenous soil microbial communities. However, reports about the effect of entomopathogenic nematodes (EPN) on soil microbial communities are very few. In this study, the effects of EPN on RIFA populations and microbial communities in mounds were investigated. Our results showed that the application of the EPN Steinernema carpocapsae. All strain on mounds efficaciously suppressed RIFA worker populations, without forming significantly more satellite mounds compared with the control treatment. The application of EPN did not impact the bacterial and fungal diversity in soils derived from the RIFA mounds. However, it slightly altered the taxonomic make-up of the bacterial communities, but significantly altered the taxonomic composition of fungal communities at the phylum, family, and genus levels. The abundances of some beneficial bacteria and fungi, such as Streptomyces, decreased, while those of plant and animal pathogenic bacteria and fungi, dramatically increased, after EPN treatment. On the other hand, the abundances of some entomopathogenic fungi, such as Fusicolla, Clonostachys, and Mortierella, increased. Redundancy analysis or canonical correspondence analysis revealed a positive correlation between the efficacious EPN control and the presence of the insect-resistant bacteria, Sinomonas, as well as entomopathogenic fungi Fusicolla and Mortierella. This suggests that the interactions between EPN and entomopathogenic fungi may play a role in the biological control of RIFA. Our discoveries shed light on the interactions among EPN, RIFA, and soil microbial communities, and emphasize a possible mutualistic relationship between EPN and entomopathogenic fungi in the biological control of RIFA.

The Pseudomonas syringae pv. tomato DC3000 effector HopD1 interferes with cellular dynamics associated with the function of the plant immune protein AtNHR2B

The plant pathogenic bacterium Pseudomonas syringae pv tomato DC3000 (Pst DC3000) causes disease in tomato, in the model plant Arabidopsis thaliana, and conditionally in Nicotiana benthamiana. The pathogenicity of Pst DC3000 is mostly due to bacterial virulence proteins, known as effectors, that are translocated into the plant cytoplasm through the type III secretion system (T3SS). Bacterial type III secreted effectors (T3SEs) target plants physiological processes and suppress defense responses to enable and support bacterial proliferation. The Pst DC3000 T3SE HopD1 interferes with plant defense responses by targeting the transcription factor NTL9. This work shows that HopD1 also targets the immune protein AtNHR2B (Arabidopsis thaliana nonhost resistance 2B), a protein that localizes to dynamic vesicles of the plant endomembrane system. Live-cell imaging of Nicotiana benthamiana plants transiently co-expressing HopD1 fused to the epitope haemagglutinin (HopD1-HA) with AtNHR2B fused to the red fluorescent protein (AtNHR2B-RFP), revealed that HopD1-HA interferes with the abundance and cellular dynamics of AtNHR2B-RFP-containing vesicles. The results from this study shed light into an additional function of HopD1 while contributing to understanding how T3SEs also target vesicle trafficking-mediated processes in plants.

Phage selection drives resistance-virulence trade-offs in Ralstonia solanacearum plant-pathogenic bacterium irrespective of the growth temperature.

While temperature has been shown to affect the survival and growth of bacteria and their phage parasites, it is unclear if trade-offs between phage resistance and other bacterial traits depend on the temperature. Here, we experimentally compared the evolution of phage resistance-virulence trade-offs and underlying molecular mechanisms in phytopathogenic Ralstonia solanacearum bacterium at 25 °C and 35 °C temperature environments. We found that while phages reduced R. solanacearum densities relatively more at 25 °C, no difference in the final level of phage resistance was observed between temperature treatments. Instead, small colony variants (SCVs) with increased growth rate and mutations in the quorum-sensing (QS) signaling receptor gene, phcS, evolved in both temperature treatments. Interestingly, SCVs were also phage-resistant and reached higher frequencies in the presence of phages. Evolving phage resistance was costly, resulting in reduced carrying capacity, biofilm formation, and virulence in planta, possibly due to loss of QS-mediated expression of key virulence genes. We also observed mucoid phage-resistant colonies that showed loss of virulence and reduced twitching motility likely due to parallel mutations in prepilin peptidase gene, pilD. Moreover, phage-resistant SCVs from 35 °C-phage treatment had parallel mutations in type II secretion system (T2SS) genes (gspE and gspF). Adsorption assays confirmed the role of pilD as a phage receptor, while no loss of adsorption was found with phcS or T2SS mutants, indicative of other downstream phage resistance mechanisms. Additional transcriptomic analysis revealed upregulation of CBASS and type I restriction-modification phage defense systems in response to phage exposure, which coincided with reduced expression of motility and virulence-associated genes, including pilD and type II and III secretion systems. Together, these results suggest that while phage resistance-virulence trade-offs are not affected by the growth temperature, they could be mediated through both pre- and postinfection phage resistance mechanisms.

The Antibacterial Effect of Selected Essential Oils and Their Bioactive Constituents on Pseudomonas savastanoi pv. savastanoi: Phytotoxic Properties and Potential for Future Olive Disease Control.

Plant pathogenic bacteria pose a significant threat to olive cultivation, leading to substantial economic losses and reduced yield. The efficacy of antimicrobial agents against these pathogens is of great interest for sustainable disease management strategies. As such, the management of olive knot disease is one of the major challenges in olive protection. In the presented study, through a series of in vitro assays, we investigated the antimicrobial effect of six essential oils (EOs) and their most concentrated constituents against causative agent of olive knot disease-Pseudomonas savastanoi pv. savastanoi, highlighting the high potential of Origanum compactum EO and its constituent carvacrol. Carvacrol exhibited the highest potential for practical application, demonstrating membrane disruption as its mechanism of action even at the lowest concentration. The bactericidal effect of antimicrobials was confirmed in a time-kill assay, where concentrations of MIC, 2× MIC, and 4× MIC were evaluated. Some of the applied treatments resulted in inhibition equal or higher than copper-based treatment. Additionally, we assessed the phytotoxicity of carvacrol by foliar application on olive cv. Leccino. The appearance of phytotoxic injuries majorly occurred on the young leaves of olive plants, with the highest proportion of damaged canopy observed when the 2× MIC concentration was applied. Due to its great efficiency against P. savastanoi pv. savastanoi in vitro, these findings highlight the potential of carvacrol as a molecule of interest for the development of environmentally friendly biopesticides. This study also contributes to the advancement of disease management practices in olive cultivation, leading to enhanced crop protection.

Novel approach toward the understanding of genetic diversity based on the two types of amino acid repeats in Erwinia amylovora

Erwinia amylovora is a notorious plant pathogenic bacterium of global concern that has devastated the apple and pear production industry worldwide. Nevertheless, the approaches available currently to understand the genetic diversity of E. amylovora remain unsatisfactory because of the lack of a trustworthy index and data covering the globally occurring E. amylovora strains; thus, their origin and distribution pattern remains ambiguous. Therefore, there is a growing need for robust approaches for obtaining this information via the comparison of the genomic structure of Amygdaloideae-infecting strains to understand their genetic diversity and distribution. Here, the whole-genome sequences of 245 E. amylovora strains available from the NCBI database were compared to identify intraspecific genes for use as an improved index for the simple classification of E. amylovora strains regarding their distribution. Finally, we discovered two kinds of strain-typing protein-encoding genes, i.e., the SAM-dependent methyltransferase and electron transport complex subunit RsxC. Interestingly, both of these proteins carried an amino acid repeat in these strains: SAM-dependent methyltransferase comprised a single-amino-acid repeat (asparagine), whereas RsxC carried a 40-amino-acid repeat, which was differentially distributed among the strains. These noteworthy findings and approaches may enable the exploration of the genetic diversity of E. amylovora from a global perspective.

Complete genome sequence of "Candidatus Phytoplasma cynodontis" GY2015, a plant pathogen associated with Bermuda grass white leaf disease in Taiwan.

The complete genome sequence of "Candidatus Phytoplasma cynodontis" strain GY2015, which consists of one 498,922-bp circular chromosome, is presented in this work. This uncultivated plant-pathogenic bacterium is associated with Bermuda grass white leaf disease in Taoyuan, Taiwan.

Phytochemical analysis and antimicrobial activity of Tamarindus indica extracts against Fusarium oxysporum and Xanthomonas campestris

Abstract. Gitari FM, Githae EW, Kuria EK. 2023. Phytochemical analysis and antimicrobial activity of Tamarindus indica extracts against Fusarium oxysporum and Xanthomonas campestris. Intl J Trop Drylands 7: 73-82. Plant-pathogenic bacteria and fungi are a major threat to biodiversity and food security worldwide. The pathogens are difficult to control using cultural methods and have sometimes acquired resistance to conventional pesticides. This has necessitated the search for more efficient active compounds against them. One promising source of such compounds is tropical-medicinal plants such as Tamarindus indica L. This study first determined the phytochemical composition of T. indica extracts from different parts (leaves, bark, roots, and pods). Then it evaluated in-vitro the antimicrobial activity against plant pathogenic bacteria (Xanthomonas campestris) and fungi (Fusarium oxysporum). Crude extracts were obtained using different solvents (dichloromethane, methanol, and acetone). The analysis revealed the presence of nine pharmaco-active phytochemicals; methanol extracts had the highest concentrations of these phytochemicals. All extracts demonstrated inhibitory effects against F. oxysporum. However, the extracts did not show any antimicrobial effect against X. campestris. There was a significant difference (p<0.05) in the percentage of inhibition of F. oxysporum growth by different extracts. Generally, high growth inhibition was observed in media containing different plant extracts at 250 and 500 ppm concentrations. For acetone extracts, the highest inhibition (71.042%) was induced by root extract at a 250 ppm concentration, whereas for dichloromethane extracts, the highest inhibition (68.811%) was induced by 500 ppm of leaf extract. Methanol extracts from stem recorded the highest inhibition of 86.953% at concentrations of 125 ppm. This was followed by root extracts (75.169% inhibition) at 500 ppm. The T. indica, therefore, has great potential as a source of bio-pesticide for use in integrated pest management of F. oxysporum.

Bioinformatic Analysis Reveals the Role of Translation Elongation Efficiency Optimisation in the Evolution of Ralstonia Genus.

Translation efficiency modulates gene expression in prokaryotes. The comparative analysis of translation elongation efficiency characteristics of Ralstonia genus bacteria genomes revealed that these characteristics diverge in accordance with the phylogeny of Ralstonia. The first branch of this genus is a group of bacteria commonly found in moist environments such as soil and water that includes the species R. mannitolilytica, R. insidiosa, and R. pickettii, which are also described as nosocomial infection pathogens. In contrast, the second branch is plant pathogenic bacteria consisting of R. solanacearum, R. pseudosolanacearum, and R. syzygii. We found that the soil Ralstonia have a significantly lower number and energy of potential secondary structures in mRNA and an increased role of codon usage bias in the optimization of highly expressed genes' translation elongation efficiency, not only compared to phytopathogenic Ralstonia but also to Cupriavidus necator, which is closely related to the Ralstonia genus. The observed alterations in translation elongation efficiency of orthologous genes are also reflected in the difference of potentially highly expressed gene' sets' content among Ralstonia branches with different lifestyles. Analysis of translation elongation efficiency characteristics can be considered a promising approach for studying complex mechanisms that determine the evolution and adaptation of bacteria in various environments.

Identification of the chemical profile and evaluation of the antimicrobial effect of Eryngium billardieri Delar essential oil component against bacterial species of agricultural and food interest.

Studies on the antibacterial activity of the essential oil of E. billardieri are limited. In this study, we identified this herb as a natural complex effective against several bacteria by employing disk diffusion and broth microdilution susceptibility methods. Primary estimation of the antimicrobial effect of this herbal compound by disk diffusion method showed that the oil could inhibit the growth of the tested bacteria by the appearance of haloes between 8.25 and 21.25 mm. In the next step, the oil was found to be active against all 24 tested Gram-negative and Gram-positive bacteria in the broth media, at minimum inhibitory concentrations ranging from 0.67 to 34.17 g L-1. Furthermore, Enterococcus faecalis and Curtobacterium flaccumfaciens pv. flaccumfaciens were the most sensitive food and plant pathogenic bacteria, respectively. Gas chromatography-mass spectrometry analysis was conducted to assign the ingredients present in the oil; 34 different components representing 95.71% of the total oil were identified, with n-hexadecanoic acid being the dominant component, followed by 2-Pentadecanone, 6,10,14-trimethyl, 1H-Indene, 1-ethylideneoctahydro-, and Cinnamyl tiglate. These findings demonstrate, for the first time, a broad spectrum of the antibacterial capacity of E. billardieri. Based on these observations, the oil could be applied as a natural preservative with the potential for designing novel products. Its bioactive agents can also be isolated for further use in the food and agricultural industries.

Short Pathways to Highly Active Antimicrobial: Structurally Diverse Polyamines Derivatives from Amino-Aldehyde Condensation Strategy.

Chemical fungicides are the mainstay of plant disease control in agricultural production, but there are a very limited number of drugs that can effectively control plant diseases. Two series of secondary amine derivatives were synthesized using the diamine skeleton combined with saturated aromatic and aliphatic aldehydes, and their antibacterial and antifungal activities against plant pathogens were determined. In addition, the antimicrobial mechanism of the highly active compound A26 was preliminarily examined against Xanthomonas oryzae (Xoo). Compound A26 exhibited the highest antibacterial potency among all the target compounds, with MIC values of 3.12, 3.12, and 12.5 μg/mL against Xoo, Xanthomonas axonopodis pv. Citri (Xac) and Pseudomonas sollamacearum (Ps), respectively. In addition, compound A26 had powerful curative and protective effects against Xoo at 200 μg/mL, and was better than the control agent Xinjunan. Preliminary mechanistic studies showed that compound A26 reduced the bacterial pathogenicity by targeting cell membranes and inhibiting the secretion of extracellular polysaccharides (EPS). Meanwhile, the toxicity of compound A26 to Human Embryonic Kidney 293 cells and Human Liver-7702 was similar to that of Xinjunan, and it had the moderate toxicity according to the World Health Organization classification standard of oral exogenous toxicity with the LD50 of 245.47 mg/kg. Secondary amines have efficient and broad-spectrum antibacterial activity against plant pathogenic bacteria and are expected to be a new class of candidate compounds for antibacterial drugs. This article is protected by copyright. All rights reserved.

Antibacterial and plant growth promoting attributes of Limosilactobacillus sp. MYSN3 isolated from noni fruit

The aim of the study was to determine the antibacterial, plant growth promotion and bio-control activity of Limosilactobacillus sp. MYSN3 isolated from noni fruit (Morinda citrifolia). The noni fruit yielded eight distinct isolates and one effective isolate MYSN3 was chosen considering their antibacterial activity for further studies. The isolate MYSN3 recorded good inhibitory effect toward the tested plant pathogenic bacteria. The CFS of MYSN3 showed 69.12, 69.98 and 63.60 % inhibition against, Pectobacterium carotovorum sub sp. carotovorum, Ralstonia solanacearum and Xanthomonas oryzae pv. oryzae respectively in microplate assay. The isolate MYSN3 was identified as Limosilactobacillus sp. with accession number OL347895 and was verified using 16sRNA gene sequencing and had 95.23 % similarity with the earlier deposited sequence of Limosilactobacillus sp. - KJ865402. The effective isolate MYSN3 was characterized for growth promotion in plants and biological control attributes. Isolate MYSN3 showed IAA production (35.18 µg/ml), gibberellic acid production (23.45 µg/ml), solubilization of phosphate, potash, zinc and silicon and gave positive outcomes for protease and siderophore production and negative for cellulose degradation activity, HCN production activity and amylase activity. Seed treatment with Limosilactobacillus sp. MYSN3 greatly enhanced the standard of seed germination and seedling vigour of tomato by paper towel method and significantly increased plant height of tomato, chilli and okra under glasshouse condition when compared to control. The findings of the current research revealed that isolated Limosilactobacillus sp. MYSN3 possess promising antibacterial activity, plant growth promoting characteristics and bio-control traits. The potentiality of the isolate as bioinoculants and bio-control agents deserves further investigation.

A Systematic Review and Quantitative Synthesis of the Efficacy of Quaternary Ammonium Compounds in Disinfesting Non-fungal Plant Pathogens.

This quantitative review and systematic analysis of the effectiveness of quaternary ammonium compounds (QACs) in disinfesting non-fungal plant pathogens in agricultural and horticultural cropping systems is a complementary follow-up to a previous study that evaluated the efficacy of QACs against fungal plant pathogens. In the present study, a meta-analysis involving 67 studies was conducted to assess the overall efficacy of QACs against plant pathogenic bacteria, oomycetes and viruses, and to identify factors associated with observed differences in product efficacy. Across all studies, QACs resulted in a significant (P < 0.0001) reduction in either disease intensity or propagule viability with a mean Hedges' g (g+) of 1.75, indicating that overall QAC treatments were moderately effective against non-fungal pathogens. Significant differences in product efficacy were observed between organism types (P = 0.0001), with QAC interventions resulting in higher efficacy (P = 0.0002) against oomycetes ( g+ = 4.20) than against viruses ( g+ = 1.42) and bacteria ( g+ = 1.07), which were not different (P = 0.2689) from each other. As a result, bacterium and virus types were combined into a composite set (BacVir). QAC intervention against BacVir resulted in significant differences in efficacy within categorical moderator subgroups for genus (P = 0.0133), target material (P = 0.0001) and QAC product generation (P = 0.0281). QAC intervention against oomycetes resulted in significant differences in efficacy only for genus (P < 0.0001). For the BacVir composite, five random effects (RE) meta-regression models were significant (P = 0.05), where models with dose and time, dose and genus, time and genus, dose and target; and time and target accounted for 62%, 61%, 52%, 83% and 88%, respectively, of the variance in true effect sizes (R2) associated with . For oomycetes, three RE meta-regression models were significant (P = 0.05), where models with dose and time; dose and genus, and time and genus accounted for 64%, 86% and 90 %, respectively, of R2 associated with g+. These results show that while QACs are moderately effective against non-fungal plant pathogens, the observed variability in their efficacy due to dose of active ingredient and contact time of these products can be influenced by organism type, genus within organism type, the target being treated, and the generation of QAC products.

Not All Acidovorax Are Created Equal: Gibberellin Biosynthesis in the Turfgrass Pathogen Acidovorax avenae subsp. avenae.

In recent years Acidovorax avenae subsp. avenae was identified as a major cause of bacterial etiolation and decline (BED) in turfgrasses and has become a growing economical concern for the turfgrass industry. The symptoms of BED resemble those of "bakanae," or foolish seedling disease, of rice (Oryzae sativa), in which the gibberellins produced by the infecting fungus, Fusarium fujikuroi, contribute to the symptom development. Additionally, an operon coding for the enzymes necessary for bacterial gibberellin production was recently characterized in plant-pathogenic bacteria belonging to the γ-proteobacteria. We therefore investigated whether this gibberellin operon might be present in A. avenae subsp. avenae. A homolog of the operon has been identified in two turfgrass-infecting A. avenae subsp. avenae phylogenetic groups but not in closely related phylogenetic groups or strains infecting other plants. Moreover, even within these two phylogenetic groups, the operon presence is not uniform. For that reason, the functionality of the operon was examined in one strain of each turfgrass-infecting phylogenetic group (A. avenae subsp. avenae strains KL3 and MD5). All nine operon genes were functionally characterized through heterologous expression in Escherichia coli and enzymatic activities were analyzed by liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometry. All enzymes were functional in both investigated strains, thus demonstrating the ability of phytopathogenic β-proteobacteria to produce biologically active GA4. This additional gibberellin produced by A. avenae subsp. avenae could disrupt phytohormonal balance and be a leading factor contributing to the pathogenicity on turf grasses. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Broad-Spectrum Antimicrobial Activity of Synthetic Peptides GV185 and GV187.

Optimizing synthetic antimicrobial peptides for safe and enhanced activity against fungal and bacterial pathogens is useful for genetic engineering of plants for resistance to plant pathogens and their associated mycotoxins. Nine synthetic peptides modeled after lytic peptides tachyplesin 1, D4E1 from cecropin A, and protegrin 1 were added to germinated spores of fungal species Aspergillus flavus, Rhizopus stolonifer, Fusarium oxysporum f. sp. vasinfectum, F. verticillioides, F. graminearum, Claviceps purpurea, Verticillium dahliae, and Thielaviopsis basicola and bacterial cultures of Pseudomonas syringae pv. tabaci and Xanthomonas campestris pv. campestris at different doses and inhibitory dose response curves, and were modeled to assess antimicrobial activity. Peptides GV185 and GV187, modified from tachyplesin 1, had superior abilities to inhibit fungal and bacterial growth (50% inhibitory concentrations [IC50] ranging from 0.1 to 8.7 µM). R. stolonifer (IC50 = 8.1 µM), A. flavus (IC50 = 3.1 µM), and F. graminearum (IC50 = 2.2 µM) were less inhibited by GV185 and GV187 than all the remaining fungi (IC50 = 1.4 µM) and bacteria (IC50 = 0.1 µM). Of the remaining peptides, GV193, GV195, and GV196 (IC50 range of 0.9 to 6.6 µM) inhibited fungal growth of A. flavus, F. verticillioides, and F. graminearum less than GV185 and GV187 (IC50 range of 0.8 to 3.9 µM), followed by GV197 (IC50 range of 0.8 to 9.1 µM), whereas GV190 and GV192 inhibited poorly (IC50 range of 28.2 to 36.6 µM and 15.5 to 19.4 µM, respectively) and GV198 stimulated growth. GV185 and GV187 had slightly weaker hydrophobic and cationic residues than other tachyplesin 1 modified peptides but still had unexpectedly high lytic activity. Germinated fungal spores of R. stolonifer and F. graminearum exposed to these two peptides and D4E1 and AGM182 appeared wrinkled, with perforations near potential cytoplasmic leakage, which provided evidence of plasma membrane and cell wall lysis. We conclude that peptides GV185 and GV187 are promising candidates for genetic engineering of crops for resistance to plant-pathogenic bacteria and fungi, including A. flavus and aflatoxin contamination.

Monoterpenoids and Their Synthesized Brominate Derivatives as Eco-Friendly Measures to Control Some Plant Pathogenic Fungi and Bacteria

Monoterpenoids and Their Synthesized Brominate Derivatives as Eco-Friendly Measures to Control Some Plant Pathogenic Fungi and Bacteria

The plant pathogenic bacterium Candidatus Liberibacter solanacearum induces calcium-regulated autophagy in midgut cells of its insect vector Bactericera trigonica.

Autophagy plays an important role against pathogen infection in many organisms; however, little has been done with regard to vector-borne plant and animal pathogens, that sometimes replicate and cause deleterious effects in their vectors. Candidatus Liberibacter solanacearum (CLso) is a fastidious gram-negative phloem-restricted plant pathogen and vectored by the carrot psyllid, Bactericera trigonica. The plant disease caused by this bacterium is called carrot yellows and has recently gained much importance due to worldwide excessive economical losses. Here, we demonstrate that calcium ATPase, cytosolic calcium, and most importantly Beclin-1 have a role in regulating autophagy and its association with Liberibacter inside the psyllid. The presence of CLso generates reactive oxygen species and induces the expression of detoxification enzymes in the psyllid midguts, a main site for bacteria transmission. CLso also induces the expression of both sarco/endoplasmic reticulum Ca2+pump (SERCA) and 1,4,5-trisphosphate receptors (ITPR) in midguts, resulting in high levels of calcium in the cellular cytosol. Silencing these genes individually disrupted the calcium levels in the cytosol and resulted in direct effects on autophagy and subsequently on Liberibacter persistence and transmission. Inhibiting Beclin1-phosphorylation through different calcium-induced kinases altered the expression of autophagy and CLso titers and persistence. Based on our results obtained from the midgut, we suggest the existence of a direct correlation between cytosolic calcium levels, autophagy, and CLso persistence and transmission by the carrot psyllid. IMPORTANCE Plant diseases caused by vector-borne Liberibacter species are responsible for the most important economic losses in many agricultural sectors. Preventing these diseases relies mostly on chemical sprays against the insect vectors. Knowledge-based interference with the bacteria-vector interaction remains a promising approach as a sustainable solution. For unravelling how Liberibacter exploits molecular pathways in its insect vector for transmission, here, we show that the bacterium manipulates calcium levels on both sides of the endoplasmic reticulum membrane, resulting in manipulating autophagy. Silencing genes associated with these pathways disrupted the calcium levels in the cytosol and resulted in direct effects on autophagy and Liberibacter transmission. These results demonstrate major pathways that could be exploited for manipulating and controlling the disease transmission.

Uracil hydrazones: design, synthesis, antimicrobial activities, and putative mode of action.

Crop diseases caused by plant pathogenic fungi and bacteria have led to substantial losses in global food production. Chemical pesticides have been widely used as a primary means to mitigate these issues. Nevertheless, the persistent and excessive use of pesticides has resulted in the emergence of microbial resistance. Moreover, the improper application and excessive utilization of pesticides can contribute to environmental pollution and the persistence of pesticide residues. Consequently, the development of novel and highly effective bactericides and fungicides to combat plant pathogens holds immense practical importance. A series of uracil hydrazones IV-B was deliberately designed and evaluated for their antimicrobial efficacy. The results of bioassays indicated that most IV-B exhibited >80% inhibition against the fungal species Monilia fructigena and Sclerotium rolfsii, as well as the bacterial species Clavibacter michiganensis subsp. michiganensis, Xanthomonas oryzae pv. oryzae, and Ralstonia solanacearum, at 50 μg/mL in vitro. In vivo, IV-B20 showed 89.9% of curative and 71.8% of protective activities against C. michiganensis subsp. michiganensis at 100 μg/mL superior to thiodiazole copper and copper hydroxide. IV-B20 also showed excellent protective activity against M. fructigena (96.3% at 200 μg/mL) and S. rolfsii (80.4% at 1000 μg/mL), which were greater than chlorothalonil and equivalent to thifluzamide. Mechanistic studies revealed that IV-B20 induced oxidative damage in pathogenic bacteria and promoted the leakage of cellular contents. This study suggests that IV-B20 with uracil hydrazone skeleton has great potential as an antimicrobial candidate. These findings lay a foundation for practical application in agriculture. © 2023 Society of Chemical Industry.

Antibacterial activities of coumarin-3-carboxylic acid against Acidovorax citrulli.

Coumarin-3-carboxylic acid (3-CCA), previously screened from natural coumarins, was found to possess strong antibacterial activity against Acidovorax citrulli (Ac). In order to further evaluate the activity of this compound against plant bacterial pathogens and explore its potential value as a bactericidal lead compound, the activity of 3-CCA against 14 plant pathogenic bacteria in vitro and in vivo was tested. Results showed that 3-CCA exhibited strong in vitro activities against Ac, Ralstonia solanacearum, Xanthomonas axonopodis pv. manihotis, X. oryzae pv. oryzae, and Dickeya zeae with EC50 values ranging from 26.64 μg/mL to 40.73 μg/mL. Pot experiment results showed that 3-CCA had powerful protective and curative effects against Ac. In addition, the protective efficiency of 3-CCA was almost equivalent to that of thiodiazole copper at the same concentration. The results of SEM and TEM observation and conductivity tests showed that 3-CCA disrupted the integrity of the cell membrane and inhibited polar flagella growth. Furthermore, 3-CCA resulted in reductions in motility and extracellular exopolysaccharide (EPS) production of Ac while inhibiting the biofilm formation of Ac. These findings indicate that 3-CCA could be a promising natural lead compound against plant bacterial pathogens to explore novel antibacterial agents.