Bacterial endophytes inhabiting desert plants provide protection against seed rot caused by Fusarium verticillioides and promote growth in maize.

BACKGROUND: Fusarium maize ear and root rot disease caused by Fusarium verticillioides has become one of the most serious fungal diseases associated with maize production. Due to their abilities to promote plant development and manage diseases, bacterial endophytes provide a more promising approach for treating this vascular disease. RESULTS: This work was undertaken for the selection and identification of promising isolates as plant growth promoters and biocontrol agents against F. verticillioides in maize agroecosystems. A screening procedure consisting of in vitro and in situ tests was applied to 27 endophytic strains originating from desert plants: Euphorbia antiquorum, Calotropis procera, and Alcasia albida. In vitro studies indicated that the bacteria exhibited variable results in biocontrol, endophytism, and plant growth-promoting traits. In addition, in situ plant growth promotion and biocontrol experiments allowed the identification of the most promising bacterial endophytes. In vitro and in situ comparative study results indicated a low correlation. Our data revealed that in situ screening must be used as the method of selection of biocontrol agents against Fusarium ear and root rot disease. Based on in situ results, seven potent strains were selected and identified as Bacillus subtilis, Bacillus velezensis, Bacillus tequilensis, Stenotrophomonas maltophilia, and Klebsiella pneumoniae. CONCLUSION: The results of this study showed that the selected strains seem to be promising candidates to be exploited as biofertilizers and biocontrol agents against Fusarium maize ear and root rot disease. © 2023 Society of Chemical Industry.

Synthesis of silver nanoparticles using Bacillus velezensis M3-7 lipopeptides: Enhanced antifungal activity and potential use as a biocontrol agent against Fusarium crown rot disease of wheat seedlings.

Bacillus velezensis M3-7 is a hyperactive mutant, 12-fold improved in its antifungal activity, obtained during a previous study from the wild strain BLB371 after a combination of random mutagenesis and medium component optimization. This study explores the use of this mutant in synthesizing silver nanoparticles (Ag-NPs) for the control of Fusarium crown rot disease (FCR) in wheat seedlings. LC-MS/MS analysis proved that both strains co-produced different families of lipopeptides and that mutagenesis caused the hyper-production of iturin A C14 and C15, the liberation of iturin A C10 and C12, and the inhibition of fengycin release. Our aim was a further improvement in the antifungal activity of the wild strain and the mutant M3-7 in order to control Fusarium crown rot disease (FCR) in wheat seedlings. Therefore, a nanotechnology approach was adopted, and different lipopeptide concentrations produced by the wild strain and the mutant M3-7 were used as capping agents to synthesize silver nanoparticles (Ag-NPs) with enhanced antifungal activity. Ag-NPs formed using 3 mg·mL-1 of the mutant lipopeptides were found to exhibit a good distribution, improved antifungal activity, a promising potential to be used as a biofortified agent for seed germination, and an effective compound to control FCR in wheat seedlings.

Halobacteria-Based Biofertilizers: A Promising Alternative for Enhancing Soil Fertility and Crop Productivity under Biotic and Abiotic Stresses-A Review.

Abiotic and biotic stresses such as salt stress and fungal infections significantly affect plant growth and productivity, leading to reduced crop yield. Traditional methods of managing stress factors, such as developing resistant varieties, chemical fertilizers, and pesticides, have shown limited success in the presence of combined biotic and abiotic stress factors. Halotolerant bacteria found in saline environments have potential as plant promoters under stressful conditions. These microorganisms produce bioactive molecules and plant growth regulators, making them a promising agent for enhancing soil fertility, improving plant resistance to adversities, and increasing crop production. This review highlights the capability of plant-growth-promoting halobacteria (PGPH) to stimulate plant growth in non-saline conditions, strengthen plant tolerance and resistance to biotic and abiotic stressors, and sustain soil fertility. The major attempted points are: (i) the various abiotic and biotic challenges that limit agriculture sustainability and food safety, (ii) the mechanisms employed by PGPH to promote plant tolerance and resistance to both biotic and abiotic stressors, (iii) the important role played by PGPH in the recovery and remediation of agricultural affected soils, and (iv) the concerns and limitations of using PGHB as an innovative approach to boost crop production and food security.

Desirable traits for a good biocontrol agent against fire blight disease.

AIMS: This study aimed to improve the screening strategy for the selection of biocontrol agents with high biocontrol efficacy against fire blight disease. METHODS AND RESULTS: A two-step screening procedure consisting of in-vitro laboratory tests and an ex-vivo test system using detached pear leaves was applied to 43 Bacillus strains originated from the rhizosphere and the aerial parts of apple and pear plants. The grouping of the studied strains and the tested traits based on the principal component analysis and the two-way hierarchical cluster analysis showed that siderophore production and biofilm formation are the most desirable traits in a Bacillus biocontrol agent to control fire blight disease and that rhizospheric originating strains are the most effective. CONCLUSIONS: In contrast to the previous screening strategies that are often insufficient to select the most suitable microorganisms, this study reported an improved strategy based on the microbial competition traits to select potential Bacillus biocontrol agents with high biocontrol efficacy against fire blight disease.

Detection of Clones B2-ST131-C2 and A-ST617 in Escherichia coli Producing Both CTX-M-15 and CTX-M-27 from Tunisian Community Patients.

During a two-month period (2017-2018), 336 urine samples positive for Escherichia coli were collected from Tunisian patients. Of the 336 samples, 266 were collected from community patients and 70 from hospital settings. In all, 15 ESBL producers were identified (8 and 7, respectively) and assigned to 13 pulsotypes, including four ESBL-producing E. coli (ESBL-E) with E1 and E2 profiles (2 isolates each) from community patients. The two strains E1 were identified as B2-ST131 subclade C2 and the two isolates E2, A-ST617. The four strains carrying both CTX-M-15 and CTX-M-27, exhibited the multireplicon IncFII/F1A/F1B with the same formula F31:A4:B1. Two isolates with patterns E3 and E4 (Dice coefficient, 78.7%) isolated from community and hospital settings of two geographic areas were assigned to the emerging ST131 C1-M27 subclade and contained the replicon F1:A-:B20. The remaining ESBL-E divided into different sequence types/associated CTX-M: 2 ST131-C2/CTX-M-15 and ST744/CTX-M-55, ST617/CTM-15, ST2973/CTX-M-55, ST6448/CTX-M-15, ST224/CTX-M-15, ST1431/CTX-M-15, and ST38/CTX-M-27, one isolate each. Our study reports for the first time the presence in the Tunisian community of two clones of E. coli, including the virulent clone ST131-C2 harboring both CTX-M-15 and CTX-M-27, and confirms the spread of the emergent clone ST131-C1-M-27, notably in community urinary tract infections.

Thymol-enriched extract from Thymus vulgaris L leaves: Green extraction processes and antiaggregant effects on human platelets.

This work focuses on the selection and the optimization of an efficient green-extraction method, used to recover a thymol-enriched extract from thyme (Thymus vulgaris L), as well as the evaluation of the inhibitory effect of this latter on the human platelet aggregation. Different innovative extraction techniques, namely bead milling extraction, ultrasound and microwave assisted extraction, were tested for their ability to recover a high added value extract from thyme. Among all tested eco-extraction techniques, microwave extraction (MAE) was the best method in term of its extraction yield (20.84% ± 0.51), thymol concentration (731.71 mg/g) and total phenolic (23.53 ± 1.83 mg (GAE)/g of extract) and flavonoid (6.22 ± 0.35 mg of QE/g of extract) contents. Moreover, thyme extract obtained by microwave assisted extraction (TMAE) showed the most active antioxidant effect comparing to the other tested extracts. Based on these results, TMAE was chosen to be evaluated for its antiplatelet effect. Thereby, arachidonic acid, collagen and ADP were used to induce the platelet aggregation on human platelet rich plasma taken from healthy controls and results revealed that TMAE strongly inhibited the induced platelet aggregation. Indeed, TMAE exhibited potent antiaggregant activity by inhibiting platelet activation, secretion and aggregation. Additionally, cytotoxicity assay on normal HEK-293 cells showed that TMAE has no cytotoxic effect even at high concentration (8 mg/ml) and can further be taken up to various biomedical applications mainly in the prevention of cardiovascular diseases.

Optimization of Bacillus amyloliquefaciens BLB369 Culture Medium by Response Surface Methodology for Low Cost Production of Antifungal Activity.

Bacillus amyloliquefaciens BLB369 is an important plant growth-promoting bacterium, which produces antifungal compounds. A statistics-based experimental design was used to optimize a liquid culture medium using inexpensive substrates for increasing its antifungal activity. A Plackett-Burman design was first applied to elucidate medium components having significant effects on antifungal production. Then the steepest ascent method was employed to approach the experimental design space, followed by an application of central composite design. Three factors were retained (candy waste, peptone, and sodium chloride), and polynomial and original trigonometric models fitted the antifungal activity. The trigonometric model ensured a better fit. The contour and surface plots showed concentric increasing levels pointing out an optimized activity. Hence, the polynomial and trigonometric models showed a maximal antifungal activity of 251.9 (AU/mL) and 255.5 (AU/mL) for (19.17, 19.88, 3.75) (g/L) and (19.61, 20, 3.7) (g/L) of candy waste, peptone, and NaCl, respectively. This study provides a potential strategy for improving the fermentation of B. amyloliquefaciens BLB369 in low-cost media for large-scale industrial production.

Two Novel Bacillus Strains (subtilis and simplex Species) with Promising Potential for the Biocontrol of Zymoseptoria tritici, the Causal Agent of Septoria Tritici Blotch of Wheat.

Two novel Algerian field-collected isolates were selected for their antifungal activity against Zymoseptoria tritici (teleomorph Mycosphaerella graminicola). The novel strains, termed Alg.24B1 and Alg.24B2, were identified as Bacillus subtilis and Bacillus simplex since their respective nucleotide sequences of the 16S rRNA gene were 100% and 99.93% identical to those of B. subtilis and B. simplex, respectively. The antifungal activities of Alg.24B1 and Alg.24B2 were evaluated by the well diffusion method and compared to those of other Bacillus species. The maximum activity was obtained after two days of confrontation of the bacterial strain supernatants with the fungus for Alg.24B1 and three days for Alg.24B2. Furthermore, the metabolites responsible for the antifungal activity of both strains were detected by the investigation of either gene presence (PCR) or molecule production (activity detection of lytic enzymes and HPLC detection of lipopeptides). Overall, this study showed that in addition to their ability to produce lytic enzymes (protease and ß-glucanase), both strains coproduce three types of lipopeptides viz. surfactin, iturin, and fengycin. Thus, the biofungicide activity of both strains may be a result of a combination of different mechanisms. Therefore, they had a great potential to be used as biocontrol agents to effectively manage septoria tritici blotch of wheat (STB).

Endophytic halotolerant Bacillus velezensis FMH2 alleviates salt stress on tomato plants by improving plant growth and altering physiological and antioxidant responses.

Salinity stress has significant deleterious effects on agricultural lands and plant yields. Plants undergo a series of physiological and molecular changes to reduce salt-induced damage. However, these mechanisms remain insufficient. The inoculation of plant growth promoting bacteria to improve plant health under stress conditions offers promise. Bacillus velezensis FMH2 has been shown to protect tomato fruits against black mold disease and to improve seed tolerance to abiotic stresses. During this study, the major physiological and metabolic changes connected with FMH2 mitigation of abiotic stress tolerance in tomato plants were explored. In presence of different salt levels, FMH2 showed a high potentiality to colonize internal plant tissues and to produce several plant growth promoting metabolites such as siderophores, indole acetic acid, and hydrolytic enzymes. FMH2-treatment promoted plant growth (root structure, plant elongation, leaf emission, fresh and dry weights, water content, etc.) in absence as well as in presence of salt stress. FMH2 treatment decreased endogenous Na+ accumulation and increased K+ and Ca2+ uptake. Furthermore, B. velezensis FMH2-treatment improved chlorophyll contents, membrane integrity and phenol peroxidase concentrations, and reduced malondialdehyde and hydrogen peroxide levels under saline conditions with a significant salinity × strain interaction. The present study suggests the endophytic strain FMH2 involved different mechanisms and regulatory functions to enhance plant oxidative systems and regulate ion uptake mechanisms supporting both growth and stress management.

Halotolerant Bacillus spizizenii FMH45 promoting growth, physiological, and antioxidant parameters of tomato plants exposed to salt stress.

KEY MESSAGE: Bacillus spizizenii is for the first time described as a plant growth salt-tolerant bacterium able to alleviate salt stress in crop plants by improving physiological parameters and antioxidant defense mechanisms. Agricultural soil salinization is a serious issue worldwide affecting agricultural yield. Plant growth promoting bacteria can enhance salt tolerance and plant yield. Bacillus spizizenii FMH45 has been shown to inhibit fungal attacks in tomato fruits and to augment tomato seed germination in presence of abiotic stresses. During this study, we reported for the first time B. spizizenii as a salt-tolerant bacterium able to alleviate salt stress in tomato plants. B. spizizenii FMH45 was examined in vitro for its potential to produce several plant growth promoting characters (siderophores, IAA, and phosphate solubilization) and hydrolytic enzymes (cellulase, glucanase and protease) in the presence of saline conditions. FMH45 was also investigated in vivo in pot experiments to evaluate its ability to promote tomato plant growth under salt stress condition. FMH45 inoculation, enhanced tomato seedling length, vigor index, and plant fresh and dry weights when compared to the non-inoculated controls exposed and not exposed to a regular irrigation with salt solutions containing: 0; 3.5; 7; and 10 g L-1 of NaCl. FMH45-treated plants also presented improved chlorophyll content, membrane integrity (MI), and phenol peroxidase (POX) concentrations, as well as reduced malondialdehyde (MDA) and hydrogen peroxide (H2O2) levels under saline conditions with a significant salinity × strain interaction. Furthermore, FMH45 inoculation significantly decreased endogenous Na+ accumulation, increased K+ and Ca2+ uptake, and thereby improved K+/Na+ and Ca2+/Na+ ratios. This study proves that bio-inoculation of FMH45 efficiently increases salt tolerance in tomato plants. This sustainable approach can be applied to other stressed plant species in affected soils.

Development of a cost-effective medium for Photorhabdus temperata bioinsecticide production from wastewater and exploration of performance kinetic.

This study investigates the optimization of the culture conditions for enhancing Photorhabdus temperata biopesticide production using wastewater (WS4) as a raw material. Box-Behnken design (BBD) was used to evaluate the effects of carbon to nitrogen ratio (C/N), sodium chloride concentration and inoculum size on P. temperata biomass production and insecticidal activity. For an enhanced biopesticide production, the optimum operating conditions were as follows: inoculum size = 4%; C/N ratio = 12.5 and [NaCl] = 4 g/L for two responses. 1.95 and 2.75 fold improvements in oral toxicity and biomass production were respectively obtained in the cost-effective medium developed in this study (WS4 I) using the three variables at their optimal values. Under the optimized conditions, WS4 I-grown cells exhibited higher membrane integrity according to flow cytometry analysis since dead cells presented only 9.2% compared to 29.2% in WS4. From batch fermentations carried out in WS4 I and WS4, P. temperata kinetic parameters in terms of biomass production and substrate consumption rates were modeled. The obtained results showed that the maximum specific growth rate in WS4 I was of 0.43 h-1 while that obtained in WS4 was of 0.14 h-1. In addition, the efficiency of P. temperata to metabolize organic carbon was enhanced by optimizing the culture conditions. It reached 72.66% instead of 46.18% in the control fermentation after 10 h of incubation. Under the optimized conditions, P. temperata cells showed the highest specific consumption rate resulting in a toxin synthesis improvement.

Acidic pretreatment as a chemical approach for enhanced Photorhabdus temperata bioinsecticide production from industrial wastewater.

The chemical treatment of the wastewater used for the bioinsecticide production by the bacterium Photorhabdus temperata was investigated in this study. An improvement of the volatile suspended solids (VSS) solubilization along with an increase in protein, carbohydrate, reducing sugar and nitrogen concentrations were demonstrated after alkali and thermo-alkali hydrolysis. In contrast, the application of acidic and thermo-acidic pretreatments reduced the organic matter hydrolysis. Compared to untreated wastewater, the chemical oxygen demand (COD) solubilization and the heavy metal concentration, except manganese, were enhanced in all the chemically pretreated wastewaters. Although its low contribution in the solubilization of the wastewater organic matter, the acidic-pretreated wastewater showed the highest performance in supporting P. temperata biopesticide production. Indeed, using the acidic-pretreated wastewater as a fermentation medium decreased the lag phase, enhanced the growth of the strain K122 to reach a final biomass production of 20 × 108 cells/mL, increased culturable cell count to 262 × 106 cells/mL and improved oral toxicity against Ephestia kuehniella larvae by 68.4%. Among chemical pretreatments performed, the acidic hydrolysis was demonstrated to be the unique promising one for P. temperata bioinsecticide production due to its ability to reduce aromatic compounds as shown by Gas Chromatography-Mass Spectrometry (GC-MS) analysis.

Biocontrol and plant growth-promoting potentiality of bacteria isolated from compost extract.

The use of compost extracts is steadily increasing, offering an attractive way for plant growth enhancement and disease management replacing chemical pesticides. In this study, potential mechanisms involved in plant growth promotion and suppressive activity against fungal diseases, of a compost extract produced from poultry manure/olive husk compost, were investigated. Results of physico-chemical and microbiological investigations showed high ability to reduce Fusarium oxysporum, Alternaria alternata, Aspergillus niger and Botrytis cinerea growth. The suppressive ability detected using confrontation test and the phytostimulatory effect tested on tomato seeds were related mainly to its microbial population content. Among 150 bacterial strains, isolated from the compost extract, 13 isolates showed antifungal activity against the four tested plant pathogenic fungi. Their identification based on 16S rRNA gene sequence revealed they belonged to different species of the genus Bacillus, Alcaligenes, Providencia and Ochrobactrum. When tested for their ability to produce cell wall degradation enzymes using specific media, the majority of the 13 isolates were shown to synthesize proteases, lipases and glucanases. Similarly, the best part of them showed positive reaction for plant growth promoting substances liberation, biosurfactant production and biofilm formation. In vivo tests were carried out using tomato seeds and fruits and proved that 92% of strains improved tomato plants vigor indexes when compared to the control and 6 among them were able to reduce decay severity caused by B. cinerea over 50%. Principal component analysis showed an important correlation between in vitro and in vivo potentialities and that Bacillus siamensis CEBZ11 strain was statistically the most effective strain in protecting tomato plants from gray mould disease. This study revealed the selected strains would be useful for plant pathogenic fungi control and plant growth promotion.

Overproduction of Glucose Oxidase by Aspergillus tubingensis CTM 507 Randomly Obtained Mutants and Study of Its Insecticidal Activity against Ephestia kuehniella.

In order to enhance the production of glucose oxidase (GOD), random mutagenesis of Aspergillus tubingensis CTM 507 was performed using the chemical and physical mutagens: nitric acid and UV irradiation, respectively. The majority of the isolated mutants showed good GOD production, but only some mutants presented a significant overproduction, as compared with the parent strain. The selected mutants (19 strains), showing an overproduction larger than 200%, are quite stable after three successive subcultures. Among these, six strains revealed an important improvement in submerged fermentation. The insecticidal activity of GOD produced by the wild and the selected mutant strains was evaluated against the third larval instars of E. kuehniella. Mutant strains U11, U12, U20, and U21, presenting the most important effect, displayed an LC50 value of 89.00, 88.51, 80.00, and 86.00 U/cm2, respectively, which was 1.5-fold more important than the wild strain (61 U/cm2). According to histopathology observations, the GOD enzyme showed approximately similar damage on the E. kuehniella midgut including rupture and disintegration of the epithelial layer and cellular vacuolization. The data supports, for the first time, the use of GOD as a pest control agent against E. kuehniella.

Agrobacterium tumefaciens C58 presence affects Bacillus velezensis 32a ecological fitness in the tomato rhizosphere.

The persistence of pathogenic Agrobacterium strains as soil-associated saprophytes may cause an inconsistency in the efficacy of the biocontrol inoculants under field condition. The study of the interaction occurring in the rhizosphere between the beneficial and the pathogenic microbes is thus interesting for the development of effective biopesticides for the management of crown gall disease. However, very little is still known about the influence of these complex interactions on the biocontrol determinants of beneficial bacteria, especially Bacillus strains. This study aimed to evaluate the effect of the soil borne pathogen Agrobacterium tumefaciens C58 on root colonization and lipopeptide production by Bacillus velezensis strain 32a during interaction with tomato plants. Results show that the presence of A. tumefaciens C58 positively impacted the root colonization level of the Bacillus strain. However, negative impact on surfactin production was observed in Agrobacterium-treated seedling, compared with control. Further investigation suggests that these modulations are due to a modified tomato root exudate composition during the tripartite interaction. Thus, this work contributes to enhance the knowledge on the impact of interspecies interaction on the ecological fitness of Bacillus cells living in the rhizosphere.

MgrB Inactivation Is Responsible for Acquired Resistance to Colistin in Enterobacter hormaechei subsp. steigerwaltii.

Multidrug-resistant strains belonging to the Enterobacter cloacae complex (ECC) group, and especially those belonging to clusters C-III, C-IV, and C-VIII, have increasingly emerged as a leading cause of health care-associated infections, with colistin used as one of the last lines of treatment. However, colistin-resistant ECC strains have emerged. The aim of this study was to prove that MgrB, the negative regulator of the PhoP/PhoQ two-component regulatory system, is involved in colistin resistance in ECC of cluster C-VIII, formerly referred to as Enterobacter hormaechei subsp. steigerwaltii An in vitro mutant (Eh22-Mut) was selected from a clinical isolate of Eh22. The sequencing analysis of its mgrB gene showed the presence of one nucleotide deletion leading to the formation of a truncated protein of six instead of 47 amino acids. The wild-type mgrB gene from Eh22 and that of a clinical strain of Klebsiella pneumoniae used as controls were cloned, and the corresponding recombinant plasmids were used for complementation assays. The results showed a fully restored susceptibility to colistin and confirmed for the first time that mgrB gene expression plays a key role in acquired resistance to colistin in ECC strains.

Abiotic stress resistance, plant growth promotion and antifungal potential of halotolerant bacteria from a Tunisian solar saltern.

The uses of halotolerant bacteria isolated from naturally saline habitats have the potential to be useful crop protection agents for plants in stressful conditions. These beneficial microbes generate several plant growth regulators and bioactive molecules, which enhance plant protection from adversities, such as plant pathogens, salts and metals stresses. In this study, 15 halotolerant bacterial strains endowed with important antimicrobial activities were isolated from Sfax solar saltern (Tunisia). All of these strains were characterized by biochemical and molecular tools aiming to investigate their in-vitro and in-vivo antifungal potentialities, plant growth promotion capabilities and metal tolerance abilities under saline stress condition. The 16S rRNA gene sequencing showed that the isolated strains were affiliated to different phylum and three species were described for the first time as plant growth promoting strains (Idiomarina zobelli FMH6v, Nesterenkonia halotolerans FMH10 and Halomonas janggokensis FMH54). The tested strains exhibited several potentialities: to tolerate high salt and heavy metal concentrations, to produce biosurfactants, exopolysaccharides and extracellular hydrolytic enzymes, to form biofilms and to liberate plant promoting substances. Eight strains were able to protect tomatoes fruits from the proliferation of the fungal disease caused by Botrytis cinerea and six strains improved plant vigor indexes. Principal component analysis showed an important correlation between in-vitro and in-vivo potentialities and two strains Bacillus velezensis FMH2 and Bacillus subtilis subsp. spizizenii FMH45 were statistically considered as the most effective strains in protecting plants from fungal pathogens attack and promoting the growth of tomatoes seedlings under saline and multi heavy-metals stress conditions.

Olea europaea L. Root Endophyte Bacillus velezensis OEE1 Counteracts Oomycete and Fungal Harmful Pathogens and Harbours a Large Repertoire of Secreted and Volatile Metabolites and Beneficial Functional Genes.

Oomycete and fungal pathogens, mainly Phytophthora and Fusarium species, are notorious causal agents of huge economic losses and environmental damages. For instance, Phytophthora ramorum, Phytophthora cryptogea, Phytophthora plurivora and Fusarium solani cause significant losses in nurseries and in forest ecosystems. Chemical treatments, while harmful to the environment and human health, have been proved to have little or no impact on these species. Recently, biocontrol bacterial species were used to cope with these pathogens and have shown promising prospects towards sustainable and eco-friendly agricultural practices. Olive trees prone to Phytophthora and Fusarium disease outbreaks are suitable for habitat-adapted symbiotic strategies, to recover oomycetes and fungal pathogen biocontrol agents. Using this strategy, we showed that olive trees-associated microbiome represents a valuable source for microorganisms, promoting plant growth and healthy benefits in addition to being biocontrol agents against oomycete and fungal diseases. Isolation, characterization and screening of root microbiome of olive trees against numerous Phytophthora and other fungal pathogens have led to the identification of the Bacillus velezensis OEE1, with plant growth promotion (PGP) abilities and strong activity against major oomycete and fungal pathogens. Phylogenomic analysis of the strain OEE1 showed that B. velezensis suffers taxonomic imprecision that blurs species delimitation, impacting their biofertilizers' practical use. Genome mining of several B. velezensis strains available in the GenBank have highlighted a wide array of plant growth promoting rhizobacteria (PGPR) features, metals and antibiotics resistance and the degradation ability of phytotoxic aromatic compounds. Strain OEE1 harbours a large repertoire of secreted and volatile secondary metabolites. Rarefaction analysis of secondary metabolites richness in the B. velezenis genomes, unambiguously documented new secondary metabolites from ongoing genome sequencing efforts that warrants more efforts in order to assess the huge diversity in the species. Comparative genomics indicated that B. velezensis harbours a core genome endowed with PGP features and accessory genome encoding diverse secondary metabolites. Gas Chromatography-Mass Spectrometry (GC-MS) analysis of OEE1 Volatile Organic Compounds (VOCs) and Liquid Chromatography High Resolution Mas Spectrometry (LC-HRMS) analysis of secondary metabolites identified numerous molecules with PGP abilities that are known to interfere with pathogen development. Moreover, B. velezensis OEE1 proved effective in protecting olive trees against F. solani in greenhouse experiments and are able to inhabit olive tree roots. Our strategy provides an effective means for isolation of biocontrol agents against recalcitrant pathogens. Their genomic analysis provides necessary clues towards their efficient implementation as biofertilizers.

Chemical Composition and Acaricidal Activity of the Essential Oils of Some Plant Species of Lamiaceae and Myrtaceae against the Vector of Tropical Bovine Theileriosis: Hyalomma scupense (syn. Hyalomma detritum).

The present study aimed to investigate the acaricidal properties of six essential oils. They were extracted from some plant species (Lamiaceae and Myrtaceae) using the technique of hydrodistillation with the Clevenger apparatus. The chemical compositions of the essential oils under study were determined by gas chromatography-mass spectrometer (GC-MS). An Adult Immersion Test (AIT) and a Larval Immersion Test (LIT) were used to evaluate the acaricidal activity of these essential oils against the adults and larvae of Hyalomma scupense. GC-MS analysis showed the major constituents of each essential oil: 25.49% of α-thujone (lavender); 46.82% of carvacrol (oregano); 78.78% of carvacrol (thyme); 40.27% of 1,8-cineole (blue gum); 17.45% of p-cymene (river red gum); and 26.96% of 1,8-cineole (rosemary). The biotests on the essential oils revealed that they inhibit the reproduction of H. scupense engorged females at a rate of 100 % with doses of 0.781 µl/ml of rosemary, 1.562 µl/ml of thyme, 3.125 µl/ml of lavender and oregano, and 6.250 µl/ml of blue gum and river red gum. After a treatment that lasted for 24 hours, essential oils showed a larvicidal activity with respective values of lethal concentrations (LC): LC50, LC90, and LC95 (0.058, 0.358, and 0.600 µl/ml for thyme; 0.108, 0.495, and 0.761 µl/ml for rosemary; 0.131, 0.982, and 1.740 µl/ml for oregano; 0.155, 2.387, and 5.183 µl/ml for blue gum; 0.207, 1.653, and 2.978 µl/ml for river red gum; and 0.253, 2.212, and 4.092 µl/ml for lavender). This is the first report on the acaricidal activity of these essential oils against H. scupense. The results obtained showed that the essential oils with chemotype carvacrol, 1,8-cineole, α-thujone, and p-cymene are highly acaricidal, and they can be used for ticks control. However, further studies on their toxicity in nontarget organisms are required.