Genome mining of Bacillus thuringiensis strain SY49.1 reveals novel candidate pesticidal and bioactive compounds.

BACKGROUND: Bacillus thuringiensis SY49.1 (Bt SY49.1) strain has promising insecticidal and fungicidal activity against phytopathogens and pests. Therefore, we selected this strain for whole-genome sequencing (WGS), annotation and analysis, with the aim of identifying genes responsible for producing putative pesticidal toxins, antimicrobial metabolites and plant growth-promoting features. RESULTS: Our results showed that the SY49.1 genome is 6. 32 Mbp long with a GC content of 34.68%. Genome mining revealed the presence of multiple gene inventories for the biosynthesis of bioactive compounds such as insecticidal delta endotoxins, secondary metabolites, and several plant growth-promoting proteins. Multiple sequence alignment revealed residue variations in the toxic core of Cry1Ab when compared with known Cry1Ab sequences from Bt nomenclature databases. This suggests that the cry1Ab of SY49.1 is a new kind of its group. Among the predicted secondary metabolites, we found a kurstakin with a predicted peptide that differs from the known kurstakin peptide available in the NORINE database. In addition, lipopeptides extracted from SY49.1 suppressed the growth of Verticillium dahliae and Fusarium oxysporum. CONCLUSION: We anticipate that the complete genome of Bt SY49.1 may provide a model for properly understanding and studying antimicrobial compound mining, genetic diversity among the B. cereus group, and pathogenicity against insects. This is the first report on the WGS and mining of the Bt strain isolated from Turkey. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Plant growth-promoting microorganisms drive K strategists through deterministic processes to alleviate biological stress caused by Fusarium oxysporum.

Soybean root rot, caused by soil-borne pathogens such as Fusarium oxysporum, frequently occurs in Northeast China and leads to a decline in soil health and becoming a bottleneck for soybean yield in the region. To address this issue, applying beneficial microorganisms and altering soil microbial community structure have become effective strategies. In this study, the 90-day soybean pot experiment was conducted to explore the assembly process and life strategy selection of bacterial communities in the rhizosphere of healthy (inoculated with Funneliformis mosseae, F group and treated with Pseudomonas putida, P group) and diseased (inoculated with F. oxysporum, O group) soybean plants, as well as the recovery effect of beneficial microorganisms on soil-borne diseases (combined treatments OP and OF). Results indicated that in healthy soils (P and F), microbial community assembly process in the soybean rhizosphere was entirely governed by heterogeneous selection (HeS, 100 %). However, inoculated with P. putida (OP) was primarily driven by stochastic processes (HeS 40 %, dispersal limitation (DL) 60 %), and the F. mosseae treatment (OF) predominantly followed a deterministic process (HeS 89 %, DL 11 %) in diseased soils. Inoculation of plant growth-promoting microorganisms (PGPMs) in diseased soil drove the life strategy of the rhizosphere bacterial community from r- to K-strategy, evident from the lower rRNA operon (rrn) copy numbers (O 3.7, OP 2.1, OF 2.3), higher G+ to G- ratios (O 0.47, OP 0.58, OF 0.57), and a higher abundance of oligotrophs (O 50 %, OP 53 %, OF 54 %). In healthy (P and F) and diseased (O, OP, OF) rhizosphere soils, OTU820, OTU6142, and OTU8841 under the K-strategy, and OTU6032 and OTU6917 under the r-strategy, which served as keystone species, had a significant promoting relationship with plant biomass and defense capabilities ( p ＜0.05). Additionally, inoculation of PGPMs improved autotoxin degradation and positively correlated with bacterial life strategies in both healthy and diseased soils (P, F, OP and OF) ( p ＜0.05). These findings enhance our understanding of soil-microbe interactions and offer new insights and precise control measures for soybean disease management and soil environment remediation.

Antagonism of rhizosphere Trichoderma brevicompactum DTN19 against the pathogenic fungi causing corm rot in saffron (Crocus sativus L.) in vitro.

Introduction: Corm rot in saffron (Crocus sativus L.) significantly impacts yield and quality. Non-toxic fungi, particularly Trichoderma species, are valuable for biological control due to their production of diverse and biologically active secondary metabolites. Methods: This study aimed to isolate an effective antagonistic fungus against the pathogenic fungi causing corm rot in saffron. Four pathogenic fungi (Fusarium oxysporum, Fusarium solani, Penicillium citreosulfuratum, and Penicillium citrinum) were isolated from diseased saffron bulbs in Chongming. Initial screening through dual culture with these pathogens re-screening from rhizosphere soil samples of C. sativus based on its inhibitory effects through volatile, nonvolatile, and fermentation broth metabolites. The inhibitory effect of biocontrol fungi on pathogenic fungi in vitro was evaluated by morphological observation and molecular biology methods. Results: Antagonistic fungi were identified as Trichoderma brevicompactum DTN19. F. oxysporum was identified as the most severe pathogen. SEM (scanning electron microscope) and TEM (transmission electron microscope) observations revealed that T. brevicompactum DTN19 significantly inhibited the growth and development of F. oxysporum mycelium, disrupting its physiological structure and spore formation. Additionally, T. brevicompactum DTN19 demonstrated nitrogen fixation and production of cellulase, IAA (Indole acetic acid), and siderophores. Whole-genome sequencing of strain DTN19 revealed genes encoding protease, cellulase, chitinase, ß-glucosidase, siderophore, nitrogen cycle, and sulfate transporter-related proteins. Discussion: T. brevicompactum DTN19 may inhibit the propagation of pathogenic fungi by destroying their cell walls or producing antibiotics. It can also produce IAA and iron carriers, which have the potential to promote plant growth. Overall, T. brevicompactum DTN19 showed the development prospect of biological agents.

Soil and root microbiome analysis and isolation of plant growth-promoting bacteria from hybrid buffaloberry (Shepherdia utahensis 'Torrey') across three locations.

The effects of climate change are becoming increasingly hazardous for our ecosystem. Climate resilient landscaping, which promotes the use of native plants, has the potential to simultaneously decrease the rate of climate change, enhance climate resilience, and combat biodiversity losses. Native plants and their associated microbiome form a holo-organism; interaction between plants and microbes is responsible for plants' growth and proper functioning. In this study, we were interested in exploring the soil and root microbiome composition associated with Shepherdia utahensis, a drought hardy plant proposed for low water use landscaping, which is the hybrid between two native hardy shrubs of Utah, S. rotudifolia and S. argentea. The bulk soil, rhizosphere, root, and nodule samples of the hybrid Shepherdia plants were collected from three locations in Utah: the Logan Campus, the Greenville farm, and the Kaysville farm. The microbial diversity analysis was conducted, and plant growth-promoting bacteria were isolated and characterized from the rhizosphere. The results suggest no difference in alpha diversity between the locations; however, the beta diversity analysis suggests the bacterial community composition of bulk soil and nodule samples are different between the locations. The taxonomic classification suggests Proteobacteria and Actinobacteriota are the dominant species in bulk soil and rhizosphere, and Actinobacteriota is solely found in root and nodule samples. However, the composition of the bacterial community was different among the locations. There was a great diversity in the genus composition in bulk soil and rhizosphere samples among the locations; however, Frankia was the dominant genus in root and nodule samples. Fifty-nine different bacteria were isolated from the rhizosphere and tested for seven plant growth-promoting (PGP) traits, such as the ability to fix nitrogen, phosphates solubilization, protease activity, siderophore, Indole Acetic Acid (IAA) and catalase production, and ability to use ACC as nitrogen source. All the isolates produced some amount of IAA. Thirty-one showed at least four PGP traits and belonged to Stenotrophomonas, Chryseobacterium, Massilia, Variovorax, and Pseudomonas. We shortlisted 10 isolates that showed all seven PGP traits and will be tested for plant growth promotion.

Identification of Stutzerimonas stutzeri volatile organic compounds that enhance the colonization and promote tomato seedling growth.

AIMS: Volatile organic compounds (VOCs) have an important function in plant growth-promoting rhizobacteria (PGPR) development and plant growth. This study aimed to identify VOCs of the PGPR strain, Stutzerimonas stutzeri NRCB010, and investigate their effects on NRCB010 biofilm formation, swarming motility, colonization, and tomato seedling growth. METHODS AND RESULTS: Solid-phase microextraction and gas chromatography-mass spectrometry were performed to identify the VOCs produced during NRCB010 fermentation. 28 VOCs were identified. Among them, seven (e.g. γ-valerolactone, 3-octanone, mandelic acid, 2-heptanone, methyl palmitate, S-methyl thioacetate, and 2,3-heptanedione), which smell well, are beneficial for plant, or as food additives, and without serious toxicities were selected to evaluate their effects on NRCB010 and tomato seedling growth. It was found that most of these VOCs positively influenced NRCB010 swarming motility, biofilm formation, and colonization, and the tomato seedling growth. Notably, γ-valerolactone and S-methyl thioacetate exhibited the most positive performances. CONCLUSION: The seven NRCB010 VOCs, essential for PGPR and crop growth, are potential bioactive ingredients within microbial fertilizer formulations. Nevertheless, the long-term sustainability and replicability of the positive effects of these compounds across different soil and crop types, particularly under field conditions, require further investigation.

Biodegradation of monocrotophos, cypermethrin & fipronil by Proteus myxofaciens VITVJ1: A plant - microbe based remediation.

Current study was focused on the degradation of pesticides such as Monocrotophos, Cypermethrin & Fipronil (M, C & F) using phyto and rhizoremediation strategies. The isolate Proteus myxofaciens (VITVJ1) obtained from agricultural soil was capable of degrading M, C & F. The bacteria exhibited resistance to all the pesticides (M, C & F) up to 1500 ppm and was also capable of forming biofilms. The degraded products identified using Gas Chromatography-Mass Spectroscopy (GC-MS) and FTIR was further used for deriving the degradation pathway. The end product of M, C & F was acetic acid and 3-phenoxy benzoic acid which was confirmed by the presence of functional groups such as C=O and OH. Seed germination assay revealed the non-toxic nature of the degraded products with increased germination index in the treatments augmented with degraded products. The candidate genes such as opdA gene, Est gene and MnP1gene was amplified with the amplicon size of 700bp, 1200bp and 500bp respectively. P. myxofaciens not only degraded M, C & F, but was also found to be a plant growth promoting rhizobacteria. Since, it was capable of producing Indole Acetic acid (IAA), siderophore and was able to solubilize insoluble phosphate. Therefore, VITVJ1 upon augmentation to the rhizoremediation setup aided the degradation of pesticides with increase in plant growth as compared to that of the phytoremediation setup. To our knowledge this is the first study where P. myxofaciens has been effectively used for the degradation of three different classes of pesticides, which could also enhance the growth of plants and simultaneously degrade M, C & F.

Plant growth-promoting rhizobacterial secondary metabolites in augmenting heavy metal(loid) phytoremediation: An integrated green in situ ecorestorative technology.

In recent times, increased geogenic and human-centric activities have caused significant heavy metal(loid) (HM) contamination of soil, adversely impacting environmental, plant, and human health. Phytoremediation is an evolving, cost-effective, environment-friendly, in situ technology that employs indigenous/exotic plant species as natural purifiers to remove toxic HM(s) from deteriorated ambient soil. Interestingly, the plant's rhizomicrobiome is pivotal in promoting overall plant nutrition, health, and phytoremediation. Certain secondary metabolites produced by plant growth-promoting rhizobacteria (PGPR) directly participate in HM bioremediation through chelation/mobilization/sequestration/bioadsorption/bioaccumulation, thus altering metal(loid) bioavailability for their uptake, accumulation, and translocation by plants. Moreover, the metallotolerance of the PGPR and the host plant is another critical factor for the successful phytoremediation of metal(loid)-polluted soil. Among the phytotechniques available for HM remediation, phytoextraction/phytoaccumulation (HM mobilization, uptake, and accumulation within the different plant tissues) and phytosequestration/phytostabilization (HM immobilization within the soil) have gained momentum in recent years. Natural metal(loid)-hyperaccumulating plants have the potential to assimilate increased levels of metal(loid)s, and several such species have already been identified as potential candidates for HM phytoremediation. Furthermore, the development of transgenic rhizobacterial and/or plant strains with enhanced environmental adaptability and metal(loid) uptake ability using genetic engineering might open new avenues in PGPR-assisted phytoremediation technologies. With the use of the Geographic Information System (GIS) for identifying metal(loid)-impacted lands and an appropriate combination of normal/transgenic (hyper)accumulator plant(s) and rhizobacterial inoculant(s), it is possible to develop efficient integrated phytobial remediation strategies in boosting the clean-up process over vast regions of HM-contaminated sites and eventually restore ecosystem health.

Phosphate solubilization potential of PSB: an advance approach to enhance phosphorous availability for phytostimulation.

Rhizosphere engineering approach is considered a quantum leap in plant sciences. The current study focused on investigating rhizobacterial efficiency to mobilize bioavailable phosphate from insoluble-phosphate source. Four efficient phosphate-solubilizing bacterial isolates, i.e., Pseudomonas songnenensis (GR3), Stutzerimonas stutzeri (HH2), Bacillus bingmayongensis (KH3), and Achromobacter aegrifaciens (MH1) were selected for the current study. Interactions between various physiological parameters and phosphate solubilization efficiency of isolates revealed that glucose significantly facilitated phosphorus solubilization at 37 ℃, with media having pH 7 and 0.5% phosphorous. Additionally, positive correlation among P-solubilization potential, acids produced, and pH was observed. Plant microbe-interaction analysis was performed to evaluate the efficiency of these bacterial isolates on various morpho-physiological responses of Zea mays L. For this purpose, various concentrations of tricalcium phosphate (TCP) (0, 10, 20, 30, 40, and 50 mM) were applied to plants in the presence and absence of bacterial isolates. The results showed that lower phosphate levels (10 and 20 mM) trigger shoot development and improve plant weight and leaf formation whereas higher phosphate concentrations (30 mM and above) stimulated the development of longer root system. The bacterial isolates, KH3 and HH2, were observed as efficient phosphate-solubilizing bacteria (PSB) that positively stimulated various plant growth and biochemical attributes over untreated plants. At lower phosphate levels, substantial increase of 92, 65, and 200% in shoot length, fresh weight, and number of leaves was recorded with bacterial isolate HH2, whereas, at 30 mM TCP, increase of 165% was observed in root length of plants treated with bacterial isolate KH3 compared to control. Similarly, at lower phosphate levels, increment of 57.3, 76.7, and 217% in phosphate, protein, and auxin content was recorded in plants treated with bacterial isolate HH2, and increase of 188.8% in total soluble carbohydrates was observed in plants treated with bacterial isolate KH3 as compared to control. Contrarily, increment in total chlorophyll content was most substantial (207%) by the bacterial isolate KH3 when provided with 30 mM TCP. Hence, the current study reviled that the use of these phosphates (KH3 and HH2)-solubilizing PGPR, as an efficient phytostimulator used for crop production in the replacement of chemical fertilizers, is carcinogenic and deteriorating our eco-system.

Plant growth-promoting rhizobacteria enhance active ingredient accumulation in medicinal plants at elevated CO2 and are associated with indigenous microbiome.

Introduction: Plant growth-promoting rhizobacteria (PGPR) and elevated CO2 (eCO2) have demonstrated their individual potential to enhance plant yield and quality through close interaction with rhizosphere microorganisms and plant growth. However, the efficacy of PGPR under eCO2 on rhizosphere microbiome and, ultimately, plant yield and active ingredient accumulation are not yet fully understood. Methods: This study investigated how the medicinal plant Pseudostellaria heterophylla (P. heterophylla) and its rhizosphere microbes respond to PGPR (Bacillus subtilis and Pseudomonas fluorescens) at eCO2 (1,000 ppm). Results and Discussion: It was found that the yield and active ingredient polysaccharides accumulation in the tuber of P. heterophylla were significantly increased by 38 and 253%, respectively. This promotion has been associated with increased root development and changes in the indigenous microbial community. Metagenomics analysis revealed a significant reduction in pathogenic Fusarium abundance in the rhizosphere. Potential biocontrol bacteria Actinobacteria and Proteobacteria were enriched, especially the genera Bradyrhizobium and Rhodanobacter. The reshaping of the rhizosphere microbiome was accompanied by the upregulation of biological pathways related to metabolite biosynthesis in the rhizosphere. These modifications were related to the promotion of the growth and productivity of P. heterophylla. Our findings highlighted the significant role played by PGPR in medicinal plant yield and active ingredient accumulation when exposed to eCO2.

Genomic insights and biocontrol potential of ten bacterial strains from the tomato core microbiome.

Introduction: Despite their adverse environmental effects, modern agriculture relies heavily on agrochemicals to manage diseases and pests and enhance plant growth and productivity. Some of these functions could instead be fulfilled by endophytes from the plant microbiota, which have diverse activities beneficial for plant growth and health. Methods: We therefore used a microbiome-guided top-down approach to select ten bacterial strains from different taxa in the core microbiome of tomato plants in the production chain for evaluation as potential bioinoculants. High-quality genomes for each strain were obtained using Oxford Nanopore long-read and Illumina short-read sequencing, enabling the dissection of their genetic makeup to identify phyto-beneficial traits. Results: Bacterial strains included both taxa commonly used as biofertilizers and biocontrol agents (i.e. Pseudomonas and Bacillus) as well as the less studied genera Leclercia, Chryseobacterium, Glutamicibacter, and Paenarthorbacter. When inoculated in the tomato rhizosphere, these strains promoted plant growth and reduced the severity of Fusarium Crown and Root Rot and Bacterial Spot infections. Genome analysis yielded a comprehensive inventory of genes from each strain related to processes including colonization, biofertilization, phytohormones, and plant signaling. Traits directly relevant to fertilization including phosphate solubilization and acquisition of nitrogen and iron were also identified. Moreover, the strains carried several functional genes putatively involved in abiotic stress alleviation and biotic stress management, traits that indirectly foster plant health and growth. Discussion: This study employs a top-down approach to identify new plant growth-promoting rhizobacteria (PGPRs), offering an alternative to the conventional bottom-up strategy. This method goes beyond the traditional screening of the strains and thus can expand the range of potential bioinoculants available for market application, paving the way to the use of new still underexplored genera.

Improved salinity tolerance in cucumber seedlings inoculated with halotolerant bacterial isolates with plant growth-promoting properties.

To address salinity stress in plants in an eco-friendly manner, this study investigated the potential effects of salinity-resistant bacteria isolated from saline agricultural soils on the growth of cucumber (Cucumis sativus, cv. Royal) seedlings. A greenhouse factorial experiment was conducted based on a completely randomized design (CRD) with two factors, salinity at four levels and five bacterial treatments, with three replications (n = 3). Initially, fifty bacterial isolates were screened for their salinity and drought tolerance, phosphate solubilization activity, along with production of auxin, siderophore and hydrogen cyanide. Isolates K4, K14, K15, and C8 exhibited the highest resistance to salinity and drought stresses in vitro. Isolates C8 and K15 demonstrated the highest auxin production capacity, generating 2.95 and 2.87 µg mL- 1, respectively, and also exhibited significant siderophore production capacities (by 14% and 11%). Additionally, isolates C8 and K14 displayed greater phosphate solubilization activities, by 184.64 and 122.11 µg mL- 1, respectively. The statistical analysis revealed that the selected four potent isolates significantly enhanced all growth parameters of cucumber plants grown under salinity stress conditions for six weeks. Plant height increased by 41%, fresh and dry weights by 35% and 7%, respectively, and the leaf area index by 85%. The most effective isolate, C8, was identified as Bacillus subtilis based on the 16 S rDNA amplicon sequencing. This study demonstrated that inoculating cucumber seedlings with halotolerant bacterial isolates, such as C8 (Bacillus subtilis), possessing substantial plant growth-promoting properties significantly alleviated salinity stress by enhancing plant growth parameters. These findings suggest a promising eco-friendly strategy for improving crop productivity in saline agricultural environments.

Mechanisms of ROS-mediated interactions between Bacillus aryabhattai LAD and maize roots to promote plant growth.

BACKGROUND: Plant growth-promoting rhizobacteria (PGPR), as a group of environmentally friendly bacteria growing in the rhizosphere of plants, play an important role in plant growth and development and resistance to environmental stresses. However, their limited understanding has led to the fact that their large-scale use in agriculture is still scarce, and the mechanisms by which beneficial bacteria are selected by plants and how they interact with them are still unclear. METHOD: In this study, we investigated the interaction between the auxin-producing strain Bacillus aryabhattai LAD and maize roots, and performed transcriptomic and metabolomic analyses of Bacillus aryabhattai LAD after treatment with maize root secretions(RS). RESULTS: Our results show that there is a feedback effect between the plant immune system and bacterial auxin. Bacteria activate the immune response of plant roots to produce reactive oxygen species(ROS), which in turn stimulates bacteria to synthesize IAA, and the synthesized IAA further promotes plant growth. Under the condition of co-culture with LAD, the main root length, seedling length, root surface area and root volume of maize increased by 197%, 107%, 89% and 75%, respectively. In addition, the results of transcriptome metabolome analysis showed that LAD was significantly enriched in amino acid metabolism, carbohydrate metabolism and lipid metabolism pathways after RS treatment, including 93 differentially expressed genes and 45 differentially accumulated metabolites. CONCLUSION: Our findings not only provide a relevant model for exploring the effects of plant-soil microbial interactions on plant defense functions and thereby promoting plant growth, but also lay a solid foundation for the future large-scale use of PGPR in agriculture for sustainable agricultural development.

Genome of Dietzia cinnamea 55, a desert-isolated microbe with plant growth-promoting properties for grain crops.

Here, we report the genome sequence of Dietzia cinnamea 55, isolated from the Negev Desert, Israel. D. cinnamea 55 was found to promote the growth of several cereal crops (corn, wheat, and pearl millet) in greenhouse and field studies.

Method for evaluation of Streptomyces growth and metabolism in the presence of glyphosate-based herbicide.

The use of pesticides, such as glyphosate, has increased due to population growth and the rising demand for food. Plant growth-promoting rhizobacteria (PGPR), such as Streptomyces, offer a more ecologically friendly alternative to the excessive use of pesticides. However, these bacteria undergo a complex life cycle involving the formation of hyphae, mycelia, and spores, which makes standardizing laboratory cultures challenging. In this context, we tested three methods for cultivating a Streptomyces isolate (CLV322) in the presence of the stressor agent glyphosate, denoted as M1, M2, and M3. These methods involved the simultaneous addition of the herbicide 24-48 h after the start of cultivation. We evaluated the growth and cell viability of CLV322 using the 2,3,5-triphenyl tetrazolium chloride (TTC) assay under glyphosate-based herbicide stress (Roundup® Original DI) at concentrations ranging from 0.002 to 7.2 mg mL- 1. We also assessed the ability of CLV322 to maintain PGPR characteristics in the presence of the herbicide by quantifying indolic compounds, siderophores, and phenazines. The cultivation method significantly influenced the production of metabolites by CLV322, with M3 yielding more consistent results across the evaluated parameters. Our findings suggest that germinating Streptomyces spores for 48 h before introducing glyphosate (M3) enables the analysis of bacterial tolerance to herbicide stress. This methodology may also apply to evaluate other abiotic stresses on Streptomyces strains.

PGPR isolated from hot spring imparts resilience to drought stress in wheat (Triticum aestivum L.).

Drought is a major abiotic stress that occurs frequently due to climate change, severely hampers agricultural production, and threatens food security. In this study, the effect of drought-tolerant PGPRs, i.e., PGPR-FS2 and PGPR-VHH4, was assessed on wheat by withholding water. The results indicate that drought-stressed wheat seedlings treated with PGPRs-FS2 and PGPR-VHH4 had a significantly higher shoot and root length, number of roots, higher chlorophyll, and antioxidant enzymatic activities of guaiacol peroxidase (GPX) compared to without PGPR treatment. The expression study of wheat genes related to tryptophan auxin-responsive (TaTAR), drought-responsive (TaWRKY10, TaWRKY51, TaDREB3, and TaDREB4) and auxin-regulated gene organ size (TaARGOS-A, TaARGOS-B, and TaARGOS-D) exhibited significantly higher expression in the PGPR-FS2 and PGPR-VHH4 treated wheat under drought as compared to without PGPR treatment. The results of this study illustrate that PGPR-FS2 and PGPR-VHH4 mitigate the drought stress in wheat and pave the way for imparting drought in wheat under water deficit conditions. Among the two PGPRs, PGPR-VHH4 more efficiently altered the root architecture to withstand drought stress.

Engineering the cell wall reactive groups of Plant Growth Promoting Rhizobacteria by culture strategy for heavy metal removal.

This research delved into the effects of nutrient limitation on the level of sporulation and the cadmium adsorption capacity of the bacterium Bacillus sp. isolated from the rhizosphere of endemic soils in the Region of Valparaiso, Chile. The bacteria were subjected to nitrogen limitation in fed-batch mode and were compared to bacteria grown in batch culture without nutrient limitation. The cultures were carried out in a 3 L bioreactor with an external nitrogen supply of ammonium at a flow of 0.123 L h-1. The specific maximum growth rate was 0.42 h-1 in batch and 0.45 h-1 in the exponential phase of the fed-batch. The analysis of sporulation did not show any significant difference between the biomass coming from the fed-batch and batch cultures. It was found that maximum cadmium adsorption capacity varied with culture strategy. The dry biomass grown without nutrient limitation exhibited a maximum adsorption capacity for cadmium of 65.0 mgCd g-1biomass. Conversely, the limited biomass achieved a lower cadmium adsorption capacity of approximately 36.0 mgCd g-1biomass. FTIR analysis showed that nitrogen limitation induced changes in the composition of the outer cell wall, specifically an increase of deacetlylated polysaccharides, reducing the relative amount of secondary amines and proteins from the peptidoglycan matrix. Amino groups from acetylated polysaccharides and proteins have been associated elsewhere with greater cadmium affinity, which could explain the poor results obtained with the nitrogen-restricted biomass. This study shows that new physiological states displaying different adsorption capabilities were effectively obtained by engineering the cell coverage of the bacteria using varying culture strategies. The fed-batch culture proved to be a valuable tool for studying PGPR strains for biosorption and other applications. Exploring diverse nutrient limitations and other pollutants in this bacterium and other members of the PGPR family offer great opportunities to tailor biosorption strategies based on specific conditions, ultimately contributing to sustainable environmental solutions.

Alleviated cadmium toxicity in wheat (Triticum aestivum L.) by the coactive role of zinc oxide nanoparticles and plant growth promoting rhizobacteria on TaEIL1 gene expression, biochemical and physiological changes.

Cadmium (Cd) contamination in agricultural soil is a major global concern among the multitude of human health and food security. Zinc oxide nanoparticles (ZnO-NPs) and plant growth promoting rhizobacteria (PGPR) have been known to combat heavy metal toxicity in crops. Herein, the study intended to explore the interactive effect of treatments mediated by inoculation of PGPR and foliar applied ZnO-NPs to alleviate Cd induced phytotoxicity in wheat plants which is rarely investigated. For this purpose, TaEIL1 expression, morpho-physiological, and biochemical traits of wheat were examined. Our results revealed that Cd reduced growth and biomass, disrupted plant physiological and biochemical traits, and further expression patterns of TaEIL1. The foliar application of ZnO-NPs improved growth attributes, photosynthetic pigments, and gas exchange parameters in a dose-additive manner, and this effect was further amplified with a combination of PGPR. The combined application of ZnO-NPs (100 mg L-1) with PGPR considerably increased the catalase (CAT; 52.4%), peroxidase (POD; 57.4%), superoxide dismutase (SOD; 60.1%), ascorbate peroxidase (APX; 47.4%), leading to decreased malondialdehyde (MDA; 47.4%), hydrogen peroxide (H2O2; 38.2%) and electrolyte leakage (EL; 47.3%) under high Cd (20 mg kg-1) stress. Furthermore, results revealed a significant reduction in roots (56.3%), shoots (49.4%), and grains (59.4%) Cd concentration after the Combined treatment of ZnO-NPs and PGPR as compared to the control. Relative expression of TaEIL1 (two homologues) was evaluated under control (Cd 0), Cd, ZnO-NPs, PGPR, and combined treatments. Expression profiling revealed a differential expression pattern of TaEIL1 under different treatments. The expression pattern of TaEIL1 genes was upregulated under Cd stress but downregulated under combined ZnO-NPs and PGPR, revealing its crucial role in Cd stress tolerance. Inferentially, ZnO-NPs and PGPR showed significant potential to alleviate Cd toxicity in wheat by modulating the antioxidant defense system and TaEIL1 expression. By inhibiting Cd uptake, and facilitating their detoxification, this innovative approach ensures food safety and security.

Plant-Growth-Promoting Rhizobacteria Improve Seeds Germination and Growth of Argania spinosa.

Argania spinosa is among the most important species of the Moroccan forest in terms of ecological, environmental, and socio-economic aspects. However, it faces a delicate balance between regeneration and degradation in its natural habitat. Hence, the efforts to preserve and regenerate argan forests are crucial for biodiversity, soil quality, and local livelihoods, yet they face challenges like overgrazing and climate change. Sustainable management practices, including reforestation and community engagement, are vital for mitigating degradation. Similarly, exploiting the argan tree's rhizosphere can enhance soil quality by leveraging its rich microbial diversity. This approach not only improves crop growth but also maintains ecosystem balance, ultimately benefiting both agriculture and the environment. This enrichment can be achieved by different factors: mycorrhizae, plant extracts, algae extracts, and plant growth-promoting rhizobacteria (PGPR). The benefits provided by PGPR may include increased nutrient availability, phytohormone production, shoot, root development, protection against several plant pathogens, and disease reduction. In this study, the effect of rhizobacteria isolated from the Agran rhizosphere was evaluated on germination percentage and radicle length for Argania spinosa in vitro tests, growth, collar diameter, and branching number under greenhouse conditions. One hundred and twenty (120) bacteria were isolated from the argan rhizosphere and evaluated for their capacity for phosphate solubilization and indole acetic acid production. The results showed that 52 isolates could solubilize phosphorus, with the diameters of the solubilization halos varying from 0.56 ± 0.14 to 2.9 ± 0.08 cm. Among 52 isolates, 25 were found to be positive for indole acetic acid production. These 25 isolates were first tested on maize growth to select the most performant ones. The results showed that 14 isolates from 25 tested stimulated maize growth significantly, and 3 of them by 28% (CN005, CN006, and CN009) compared to the control. Eight isolates (CN005, CN006, CN004, CN007, CN008, CN009, CN010, and CN011) that showed plant growth of more than 19% were selected to evaluate their effect on argan germination rate and radicle length and were subjected to DNA extraction and conventional Sanger sequencing. The 8 selected isolates were identified as: Brevundimonas naejangsanensis sp2, Alcaligenes faecalis, Brevundimonas naejangsanensis sp3, Brevundimonas naejangsanensis sp4, Leucobacter aridicollis sp1, Leucobacter aridicollis sp2, Brevundimonas naejangsanensis sp1, and Staphylococcus saprophyticus. The results showed that Leucobacter aridicollis sp2 significantly increased the germination rate by 95.83%, and the radicle length with a value of 2.71 cm compared to the control (1.60 cm), followed by Brevundimonas naejangsanensis sp3 and Leucobacter aridicollis sp1 (2.42 cm and 2.11 cm, respectively). Under greenhouse conditions, the results showed that the height growth increased significantly for Leucobacter aridicollis sp1 (42.07%) and Leucobacter aridicollis sp2 (39.99%). The isolates Brevundimonas naejangsanensis sp3 and Leucobacter aridicollis sp1 increased the gain of collar diameter by 41.56 and 41.21%, respectively, followed by Leucobacter aridicollis sp2 and Staphyloccocus saprophyticus (38.68 and 22.79%). Leucobacter aridicollis sp1 increased the ramification number per plant to 12 compared to the control, which had 6 ramifications per plant. The use of these isolates represents a viable alternative in sustainable agriculture by improving the germination rate and root development of the argan tree, as well as its development, while increasing the availability of nutrients in the soil and consequently improving fertilization.

Isolation and Characterization of Plant Growth-Promoting Bacteria from the Rhizosphere of Medicinal and Aromatic Plant Minthostachys verticillata.

This study aimed to isolate and characterize Pseudomonas native strains from the rhizospheric soil of Minthostachys verticillata plants to evaluate their potential as plant growth-promoting rhizobacteria (PGPR). A total of 22 bacterial isolates were obtained and subjected to various biochemical tests, as well as assessments of plant growth-promoting traits such as phosphate solubilization, hydrogen cyanide production, biocontrol properties through antibiosis, and indole acetic production. Genotypic analysis via 16S rRNA gene sequencing and phylogenetic tree construction identified the strains, with one particular strain named SM 33 showing significant growth-promoting effects on M. verticillata seedlings. This strain, SM 33, showed high similarity to Stutzerimonas stutzeri based on 16S rRNA gene sequencing and notably increased both shoot fresh weight and root dry weight of the plants. These findings underscore the potential application of native Pseudomonas strains in enhancing plant growth and health, offering promising avenues for sustainable agricultural practices.