Functional analysis and comparative genomics of Rahnella perminowiae S11P1 and Variovorax sp. S12S4, two plant growth-promoting rhizobacteria isolated from Crocus sativus L. (saffron) rhizosphere.

BACKGROUND: Rahnella perminowiae S11P1 and Variovorax sp. S12S4 are two plant growth-promoting rhizobacteria that were previously isolated from the rhizosphere of Crocus sativus L. (saffron), and have demonstrated interesting PGP activities and promising results when used as inoculants in field trials. To further elucidate the molecular mechanisms underlying their beneficial effects on plant growth, comprehensive genome mining of S11P1 and S12S4 and comparative genomic analysis with closely related strains were conducted. RESULTS: Functional annotation of the two strains predicted a large number of genes involved in auxin and siderophore production, nitrogen fixation, sulfur metabolism, organic acid biosynthesis, pyrroloquinoline quinone production, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, volatile organic compounds production, and polyamine biosynthesis. In addition, numerous genes implicated in plant-bacteria interactions, such as those involved in chemotaxis and quorum sensing, were predicted. Moreover, the two strains carried genes involved in bacterial fitness under abiotic stress conditions. Comparative genomic analysis revealed an open pan-genomic structure for the two strains. COG annotation showed that higher fractions of core and accessory genes were involved in the metabolism and transport of carbohydrates and amino acids, suggesting the metabolic versatility of the two strains as effective rhizosphere colonizers. Furthermore, this study reports the first comparison of Multilocus sequence analysis (MLSA) and core-based phylogenies of the Rahnella and Variovorax genera. CONCLUSIONS: The present study unveils the molecular mechanisms underlying plant growth promotion and biocontrol activity of S11P1 and S12S4, and provides a basis for their further biotechnological application in agriculture.

Plant growth promotion and biocontrol properties of a synthetic community in the control of apple disease.

BACKGROUND: Apple Replant Disease (ARD) is common in major apple-growing regions worldwide, but the role of rhizosphere microbiota in conferring ARD resistance and promoting plant growth remains unclear. RESULTS: In this study, a synthetic microbial community (SynCom) was developed to enhance apple plant growth and combat apple pathogens. Eight unique bacteria selected via microbial culture were used to construct the antagonistic synthetic community, which was then inoculated into apple seedlings in greenhouse experiments. Changes in the rhizomicroflora and the growth of aboveground plants were monitored. The eight strains, belonging to the genera Bacillus and Streptomyces, have the ability to antagonize pathogens such as Fusarium oxysporum, Rhizoctonia solani, Botryosphaeria ribis, and Physalospora piricola. Additionally, these eight strains can stably colonize in apple rhizosphere and some of them can produce siderophores, ACC deaminase, and IAA. Greenhouse experiments with Malus hupehensis Rehd indicated that SynCom promotes plant growth (5.23%) and increases the nutrient content of the soil, including soil organic matter (9.25%) and available K (1.99%), P (7.89%), and N (0.19%), and increases bacterial richness and the relative abundance of potentially beneficial bacteria. SynCom also increased the stability of the rhizosphere microbial community, the assembly of which was dominated by deterministic processes (|ß NTI| > 2). CONCLUSIONS: Our results provide insights into the contribution of the microbiome to pathogen inhibition and host growth. The formulation and manipulation of similar SynComs may be a beneficial strategy for promoting plant growth and controlling soil-borne disease.

Rhizosphere-xylem sap connections in the olive tree microbiome: implications for biostimulation approaches.

AIMS: Climate change is endangering olive groves. Farmers are adapting by exploring new varieties of olive trees and examining the role of microbiomes in plant health.The main objectives of this work were to determine the primary factors that influence the microbiome of olive trees and to analyze the connection between the rhizosphere and endosphere compartments. METHODS AND RESULTS: The rhizosphere and xylem sap microbiomes of two olive tree varieties were characterized by next-generation 16S rRNA amplicon sequencing, and soil descriptors were analyzed. Bacterial communities in the rhizosphere of olive trees were more diverse than those found in the xylem sap. Pseudomonadota, Actinobacteriota, Acidobacteriota, and Bacillota were the dominant phyla in both compartments. At the genus level, only very few taxa were shared between soil and sap bacterial communities. CONCLUSIONS: The composition of the bacteriome was more affected by the plant compartment than by the olive cultivar or soil properties, and a direct route from the rhizosphere to the endosphere could not be confirmed. The large number of plant growth-promoting bacteria found in both compartments provides promising prospects for improving agricultural outcomes through microbiome engineering.

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. A total of 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.

Diversity and Plant Growth Properties of Rhizospheric Bacteria Associated with Medicinal Plants.

Microbes in the rhizosphere play a significant role in the growth, development, and efficiency of plants and trees. The rhizospheric area's microbes are reliant on the soil's characteristics and the substances that the plants release. The majority of previous research on medicinal plants concentrated on their bioactive phytochemicals, but this is changing now that it is understood that a large proportion of phytotherapeutic substances are actually created by related microorganisms or through contact with their host. The roots of medicinal plants secrete a large number of secondary metabolites that determine the diversity of microbial communities in their rhizosphere. The dominant bacteria isolated from a variety of medicinal plants include various species of Bacillus, Rhizobium, Pseudomonas, Azotobacter, Burkholderia, Enterobacte, Microbacterium, Serratia, Burkholderia, and Beijerinckia. Actinobacteria also colonize the rhizosphere of medicinal plants that release low molecular weight organic solute that facilitate the solubilisation of inorganic phosphate. Root exudates of medicinal plants resist abiotic stress and accumulate in soil to produce autotoxic effects that exhibit strong obstacles to continuous cropping. Although having a vast bioresource that may be used in agriculture and modern medicine, medicinal plants' microbiomes are largely unknown. The purpose of this review is to (i) Present new insights into the plant microbiome with a focus on medicinal plants, (ii) Provide information about the components of medicinal plants derived from plants and microbes, and (iii) Discuss options for promoting plant growth and protecting plants for commercial cultivation of medicinal plants. The scientific community has paid a lot of attention to the use of rhizobacteria, particularly plant growth-promoting rhizobacteria (PGPR), as an alternative to chemical pesticides. By a variety of processes, these rhizobacteria support plant growth, manage plant pests, and foster resilience to a range of abiotic challenges. It also focuses on how PGPR inoculation affects plant growth and survival in stressful environments.

Assessing the role of ACC deaminase-producing bacteria in alleviating salinity stress and enhancing zinc uptake in plants by altering the root architecture of French bean (Phaseolus vulgaris) plants.

MAIN CONCLUSION: The consortium inoculation with strains R1 and R4 modified the root system to boost seedling growth, increase the zinc content of French bean pods, and reduce salinity stress. The present study demonstrated the effect of two 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase-producing plant growth-promoting rhizobacteria (Pantoea agglomerans R1 and Pseudomonas fragi R4) alone and consortia on the root system development, French bean growth, and zinc content as well as salinity stress tolerance. Both the strains were characterized for ACC utilization activity (426.23 and 380.54 nmol α-ketobutyrate mg protein-1 h-1), indole acetic acid (IAA) production, phosphate solubilization, ammonia, hydrogen cyanide (HCN), and siderophore production. The strains exhibited zinc solubilization in both plate and broth assays with zinc oxide and zinc carbonate as zinc sources as validated by atomic absorption spectroscopy (AAS). Single or combined inoculations with the selected strains significantly modulated the architectural and morphological traits of the root system of French bean plants. Furthermore, the application of R1and R4 consortia has enhanced zinc content in roots (60.83 mg kg-1), shoots (15.41 mg kg-1), and pods (30.04 mg kg-1) of French bean plants grown in ZnCO3 amended soil. In another set of pot experiments, the consortium bacterization has significantly enhanced length as well as fresh and dry biomass of roots and shoots of the French bean plant under saline stress conditions. Additionally, inoculation with ACC-degrading rhizobacterial strains has increased chlorophyll and carotenoid contents, osmoprotectant content, and antioxidative enzyme (catalase and peroxidase) activity in comparison to their counterparts exposed to salt treatments only. Current findings suggested ACC deaminase-producing rhizobacterial strains hold the potential to improve root architecture which in turn promotes plant growth under salt-stressed conditions as well as enhances micronutrient concentration in host plants.

Soil Protists Can Actively Redistribute Beneficial Bacteria along Medicago truncatula Roots.

The rhizosphere is the region of soil directly influenced by plant roots. The microbial community in the rhizosphere includes fungi, protists, and bacteria: all play significant roles in plant health. The beneficial bacterium Sinorhizobium meliloti infects growing root hairs on nitrogen-starved leguminous plants. Infection leads to the formation of a root nodule, where S. meliloti converts atmospheric nitrogen to ammonia, a bioavailable form. In soil, S. meliloti is often found in biofilms and travels slowly along the roots, leaving developing root hairs at the growing root tips uninfected. Soil protists are an important component of the rhizosphere system, able to travel quickly along roots and water films, who prey on soil bacteria and have been known to egest undigested phagosomes. We show that a soil protist, Colpoda sp., can transport S. meliloti down Medicago truncatula roots. Using model soil microcosms, we directly observed fluorescently labeled S. meliloti along M. truncatula roots and tracked the displacement of the fluorescence signal over time. Two weeks after co-inoculation, this signal extended 52 mm farther down plant roots when Colpoda sp. was also present versus treatments that contained bacteria but not protists. Direct counts also showed protists are required for viable bacteria to reach the deeper sections of our microcosms. Facilitating bacterial transport may be an important mechanism whereby soil protists promote plant health. IMPORTANCE Soil protists are an important part of the microbial community in the rhizosphere. Plants grown with protists fare better than plants grown without protists. Mechanisms through which protists support plant health include nutrient cycling, alteration of the bacterial community through selective feeding, and consumption of plant pathogens. Here, we provide data in support of an additional mechanism: protists act as transport vehicles for bacteria in soil. We show that protist-facilitated transport can deliver plant-beneficial bacteria to the growing tips of roots that may otherwise be sparsely inhabited with bacteria originating from a seed-associated inoculum. By co-inoculating Medicago truncatula roots with both S. meliloti, a nitrogen-fixing legume symbiont, and Colpoda sp., a ciliated protist, we show substantial and statistically significant transport with depth and breadth of bacteria-associated fluorescence as well as transport of viable bacteria. Co-inoculation with shelf-stable encysted soil protists may be employed as a sustainable agriculture biotechnology to better distribute beneficial bacteria and enhance the performance of inoculants.

Unravelling the potential plant growth activity of halotolerant Bacillus licheniformis NJ04 isolated from soil and its possible use as a green bioinoculant on Solanum lycopersicum L.

Immensely expanding world population and narrowing arable land for agriculture is a mighty concern faced by the planet at present. One of the major reasons for decline in arable lands is the increased soil salinity, making it unfavourable for crop cultivation. Utilisation of these saline land for agriculture is possible with suitable invention for improving the soil quality. Biofertizers manufactured out of Plant Growth Promoting Rhizobacteria is one such innovation. In the present study, Bacillus licheniformis NJ04 strain was isolated and studied for its halotolerance and other effective plant growth promoting traits. The NJ04 strain was able to tolerate salt up to 10% and highlighted remarkable antifungal activity against common fungal phytopathogens. The preliminary seed germination test in Solanum lycopersicum seeds revealed a significant increase in root length (16.29 ± 0.91 cm) and shoot length (9.66 ± 0.11 cm) of treated plants as compared with the control plants and thereby shows its possible use as a green bioinoculant in agriculture and an ideal candidate to compete with salt stress.

Effect of arbuscular mycorrhizal fungi (AMF) and plant growth-promoting rhizobacteria (PGPR) on microorganism of phenanthrene and pyrene contaminated soils.

A pot experiment was performed to investigate the effect of phytoremediation (CK, using tall fescue), fungi remediation (GV, using Glomus versiforme), bacterial remediation (PS, using Pseudomonas fluorescens Ps2-6), and microbial-phytoremediation (GVPS, using three species) on removing polycyclic aromatic hydrocarbons (PAHs) and the microbial diversity in soils. Inoculation with G. versiforme and P. fluorescens could increase the biomass of tall fescue and the accumulation of phenanthrene (PHE) and pyrene (PYR) in plants. Among them, the highest PHE and PYR removal efficiencies and highest biomass of tall fescue were observed in the GVPS treatment and the microbial diversity in contaminated soil was changed, the result revealed that Proteobacteria and Ascomycota were the dominant bacterial phylum and fungal phylum in all treatments, while more Proteobacteria were detected in GVPS treatment. At the genus level, the abundance of Sphingomonas (3.17%), Pseudomonas (2.05%), and Fusarium (8.65%) treated with GVPS increased compared with other treatments. These pieces of evidence contribute to a better understanding of the mechanisms involved in the combined microbial-phytoremediation strategies for PAHs-contaminated soils, especially the effects of microbial-phytoremediation on rhizosphere microbial diversity.

Inoculation with G. versiforme and P. fluorescens could increase PHE and PYR removal efficiencies, the biomass of tall fescue, microbial diversity, and the abundance of Proteobacteria, Sphingomonas, Pseudomonas, and Fusarium in (PHE + PYR)-contaminated soils.

Medicinal plant-associated rhizobacteria enhance the production of pharmaceutically important bioactive compounds under abiotic stress conditions.

Interest in cultivating valuable medicinal plants to collect bioactive components has risen extensively over the world to meet the demands of health care systems, pharmaceuticals, and food businesses. Farmers commonly use chemical fertilizers to attain maximal biomass and yield, which have negative effects on the growth, development, and bioactive constituents of such medicinally important plants. Because of its low cost, environmentally friendly behavior, and nondestructive impact on soil fertility, plant health, and human health, the use of beneficial rhizobial microbiota is an alternative strategy for increasing the production of useful medicinal plants under both standard and stressed conditions. Plant growth-promoting rhizobacteria (PGPR) associated with medicinal plants belong to the genera Azotobacter, Acinetobacter, Bacillus, Brevibacterium, Burkholderia, Exiguobacterium, Pseudomonas, Pantoea, Mycobacterium, Methylobacterium, and Serratia. These microbes enhance plant growth parameters by producing secondary metabolites, including enzymes and antibiotics, which help in nutrient uptake, enhance soil fertility, improve plant growth, and protect against plant pathogens. The role of PGPR in the production of biomass and their effect on the quality of bioactive compounds (phytochemicals) is described in this review. Additionally, the mitigation of environmental stresses including drought stress, saline stress, alkaline stress, and flooding stress to herbal plants is illustrated.

Synergistic Effect of Melatonin and Lysinibacillus fusiformis L. (PLT16) to Mitigate Drought Stress via Regulation of Hormonal, Antioxidants System, and Physio-Molecular Responses in Soybean Plants.

Drought is one of the most detrimental factors that causes significant effects on crop development and yield. However, the negative effects of drought stress may be alleviated with the aid of exogenous melatonin (MET) and the use of plant-growth-promoting bacteria (PGPB). The present investigation aimed to validate the effects of co-inoculation of MET and Lysinibacillus fusiformis on hormonal, antioxidant, and physio-molecular regulation in soybean plants to reduce the effects of drought stress. Therefore, ten randomly selected isolates were subjected to various plant-growth-promoting rhizobacteria (PGPR) traits and a polyethylene-glycol (PEG)-resistance test. Among these, PLT16 tested positive for the production of exopolysaccharide (EPS), siderophore, and indole-3-acetic acid (IAA), along with higher PEG tolerance, in vitro IAA, and organic-acid production. Therefore, PLT16 was further used in combination with MET to visualize the role in drought-stress mitigation in soybean plant. Furthermore, drought stress significantly damages photosynthesis, enhances ROS production, and reduces water stats, hormonal signaling and antioxidant enzymes, and plant growth and development. However, the co-application of MET and PLT16 enhanced plant growth and development and improved photosynthesis pigments (chlorophyll a and b and carotenoids) under both normal conditions and drought stress. This may be because hydrogen-peroxide (H2O2), superoxide-anion (O2-), and malondialdehyde (MDA) levels were reduced and antioxidant activities were enhanced to maintain redox homeostasis and reduce the abscisic-acid (ABA) level and its biosynthesis gene NCED3 while improving the synthesis of jasmonic acid (JA) and salicylic acid (SA) to mitigate drought stress and balance the stomata activity to maintain the relative water states. This may be possible due to a significant increase in endo-melatonin content, regulation of organic acids, and enhancement of nutrient uptake (calcium, potassium, and magnesium) by co-inoculated PLT16 and MET under normal conditions and drought stress. In addition, co-inoculated PLT16 and MET modulated the relative expression of DREB2 and TFs bZIP while enhancing the expression level of ERD1 under drought stress. In conclusion, the current study found that the combined application of melatonin and Lysinibacillus fusiformis inoculation increased plant growth and could be used to regulate plant function during drought stress as an eco-friendly and low-cost approach.

Mechanism of Salt Tolerance and Plant Growth Promotion in Priestia megaterium ZS-3 Revealed by Cellular Metabolism and Whole-Genome Studies.

Approximately one-third of agricultural land worldwide is affected by salinity, which limits the productivity and sustainability of crop ecosystems. Plant-growth-promoting rhizobacteria (PGPR) are a potential solution to this problem, as PGPR increases crop yield through improving soil fertility and stress resistance. Previous studies have shown that Priestia megaterium ZS-3(ZS-3) can effectively help plants tolerate salinity stress. However, how ZS-3 regulates its metabolic adaptations in saline environments remains unclear. In this study, we monitored the metabolic rearrangement of compatibilisers in ZS-3 and combined the findings with genomic data to reveal how ZS-3 survives in stressful environments, induces plant growth, and tolerates stress. The results showed that ZS-3 tolerated salinity levels up to 9%. In addition, glutamate and trehalose help ZS-3 adapt to osmotic stress under low NaCl stress, whereas proline, K+, and extracellular polysaccharides regulate the osmotic responses of ZS-3 exposed to high salt stress. Potting experiments showed that applying the ZS-3 strain in saline and neutral soils could effectively increase the activities of soil acid phosphatase, urease, and invertase in both soils, thus improving soil fertility and promoting plant growth. In addition, strain ZS-3-GFP colonised the rhizosphere and leaves of Cinnamomum camphora well, as confirmed by confocal microscopy and resistance plate count analysis. Genomic studies and in vitro experiments have shown that ZS-3 exhibits a variety of beneficial traits, including plant-promoting, antagonistic, and other related traits (such as resistance to saline and heavy metal stress/tolerance, amino acid synthesis and transport, volatile compound synthesis, micronutrient utilisation, and phytohormone biosynthesis/regulatory potential). The results support that ZS-3 can induce plant tolerance to abiotic stresses. These data provide important clues to further reveal the interactions between plants and microbiomes, as well as the mechanisms by which micro-organisms control plant health.

Genomic and Metabolic Characterization of Plant Growth-Promoting Rhizobacteria Isolated from Nodules of Clovers Grown in Non-Farmed Soil.

The rhizosphere microbiota, which includes plant growth-promoting rhizobacteria (PGPR), is essential for nutrient acquisition, protection against pathogens, and abiotic stress tolerance in plants. However, agricultural practices affect the composition and functions of microbiota, reducing their beneficial effects on plant growth and health. Among PGPR, rhizobia form mutually beneficial symbiosis with legumes. In this study, we characterized 16 clover nodule isolates from non-farmed soil to explore their plant growth-promoting (PGP) potential, hypothesizing that these bacteria may possess unique, unaltered PGP traits, compared to those affected by common agricultural practices. Biolog profiling revealed their versatile metabolic capabilities, enabling them to utilize a wide range of carbon and energy sources. All isolates were effective phosphate solubilizers, and individual strains exhibited 1-aminocyclopropane-1-carboxylate deaminase and metal ion chelation activities. Metabolically active strains showed improved performance in symbiotic interactions with plants. Comparative genomics revealed that the genomes of five nodule isolates contained a significantly enriched fraction of unique genes associated with quorum sensing and aromatic compound degradation. As the potential of PGPR in agriculture grows, we emphasize the importance of the molecular and metabolic characterization of PGP traits as a fundamental step towards their subsequent application in the field as an alternative to chemical fertilizers and supplements.

Fourier Transform Infrared (FTIR) Spectroscopic Study of Biofilms Formed by the Rhizobacterium Azospirillum baldaniorum Sp245: Aspects of Methodology and Matrix Composition.

Biofilms represent the main mode of existence of bacteria and play very significant roles in many industrial, medical and agricultural fields. Analysis of biofilms is a challenging task owing to their sophisticated composition, heterogeneity and variability. In this study, biofilms formed by the rhizobacterium Azospirillum baldaniorum (strain Sp245), isolated biofilm matrix and its macrocomponents have for the first time been studied in detail, using Fourier transform infrared (FTIR) spectroscopy, with a special emphasis on the methodology. The accompanying novel data of comparative chemical analyses of the biofilm matrix, its fractions and lipopolysaccharide isolated from the outer membrane of the cells of this strain, as well as their electrophoretic analyses (SDS-PAGE) have been found to be in good agreement with the FTIR spectroscopic results.

Transcriptome analysis of the growth-promoting effect of volatile organic compounds produced by Microbacterium aurantiacum GX14001 on tobacco (Nicotiana benthamiana).

BACKGROUND: Plant growth-promoting rhizobacteria (PGPR) release volatile organic compounds (VOCs), which promote plant growth. RESULTS: A potential PGPR strain GX14001 was isolated from marine samples, and the VOCs produced by GX14001 significantly promoted tobacco (Nicotiana benthamiana) growth in a plate experiment. Based on 16S rRNA sequence alignment and physiological and biochemical characterization, GX14001 was identified as Microbacterium aurantiacum. Comparative transcriptome analysis was conducted between GX14001 VOCs-treated tobacco and the control; it was found that 1286 genes were upregulated and 1088 genes were downregulated. Gene ontology (GO) analysis showed that upregulated genes were involved in three biological processes: polysaccharide metabolic, polysaccharide catabolic and carbohydrate metabolic. The downregulated genes were involved in six biological processes, namely cell redox homeostasis, cellular homeostasis, carbohydrate metabolic process, homeostatic process, obsolete electron transport, and regulation of biological quality. Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that 190 upregulated differentially expressed genes were mainly involved in plant hormone signal transduction, phenylpropyl biosynthesis, plant-pathogen interaction, and flavonoid biosynthesis. The 148 downregulated differentially expressed genes were mainly involved in plant hormone signal transduction and the metabolism of ascorbic, aldehyde, and pyruvate acids. Further analysis revealed that many genes were differentially expressed in the metabolic pathways of plant hormone signals, which were speculated to be the main reason why GX14001 VOCs promoted tobacco growth. To further study its regulatory mechanism, we found that GX14001 promoted plant growth through auxin, salicylic acid, and gibberellin in Arabidopsis mutant experiments. CONCLUSION: The VOCs produced by Microbacterium aurantiacum GX14001 may promote the growth of tobacco through the auxin, salicylic acid and gibberellin pathways.

Microbe- plant interaction as a sustainable tool for mopping up heavy metal contaminated sites.

BACKGROUND: Phytoremediation is a green technology that removes heavy metal (HM) contamination from the environment by using HM plant accumulators. Among soil microbiota, plant growth promoting bacteria (PGPR) have a role influencing the metal availability and uptake. METHODS: This current study evaluates the plant growth promoting qualities of microbial flora isolated from rhizosphere, plant roots, and marine aquatic HMs polluted environments in Alexandria through several biochemical and molecular traits. Metal contents in both collected soils and plant tissues were measured. Transcript levels of marker genes (HMA3 and HMA4) were analyzed. RESULTS: Three terrestrial and one aquatic site were included in this study based on the ICP-MS identification of four HMs (Zn, Cd, Cu, and Ni) or earlier reports of HMs contamination. Using the VITEK2 bacterial identification system, twenty-two bacteria isolated from these loci were biochemically described. Pseudomonas and Bacillus were the most dominant species. Furthermore, the soil microbiota collected from the most contaminated HMs site with these two were able to enhance the Helianthus annuus L. hyper-accumulation capacity significantly. Specifically, sunflower plants cultivated in soils with HMs adapted bacteria were able to accumulate about 1.7-2.5-folds more Zn and Cd in their shoots, respectively. CONCLUSION: The influence of PGPR to stimulate crop growth under stress is considered an effective strategy. Overall, our findings showed that plants cultivated in HMs contaminated sites in the presence of PGPR were able to accumulate significant amounts of HMs in several plant parts than those cultivated in soils lacking microbiota.

Pseudomonas putida and its close relatives: mixing and mastering the perfect tune for plants.

Plant growth-promoting rhizobacteria (PGPR) are a group of microorganisms of utmost interest in agricultural biotechnology for their stimulatory and protective effects on plants. Among the various PGPR species, some Pseudomonas putida strains combine outstanding traits such as phytohormone synthesis, nutrient solubilization, adaptation to different stress conditions, and excellent root colonization ability. In this review, we summarize the state of the art and the most relevant findings related to P. putida and its close relatives as PGPR, and we have compiled a detailed list of P. putida sensu stricto, sensu lato, and close relative strains that have been studied for their plant growth-promoting characteristics. However, the mere in vitro analysis of these characteristics does not guarantee correct plant performance under in vivo or field conditions. Therefore, the importance of studying adhesion and survival in the rhizosphere, as well as responses to environmental factors, is emphasized. Although numerous strains of this species have shown good performance in field trials, their use in commercial products is still very limited. Thus, we also analyze the opportunities and challenges related to the formulation and application of bioproducts based on these bacteria. KEY POINTS: •The mini-review updates the knowledge on Pseudomonas putida as a PGPR. • Some rhizosphere strains are able to improve plant growth under stress conditions. • The metabolic versatility of this species encourages the development of a bioproduct.

Drought-tolerant Bacillus megaterium isolated from semi-arid conditions induces systemic tolerance of wheat under drought conditions.

KEY MESSAGE: A detailed study of the response of wheat plants, inoculated with drought-tolerant PGPR is studied which would be beneficial to achieve genetic improvement of wheat for drought tolerance. Drought stress, a major challenge under current climatic conditions, adversely affects wheat productivity. In the current study, we observed the response of wheat plants, inoculated with drought-tolerant plant growth-promoting rhizobacteria (PGPR) Bacillus megaterium (MU2) and Bacillus licheniformis (MU8) under induced drought stress. In vitro study of 90 rhizobacteria exhibited 38 isolates showed one or more plant growth-promoting properties, such as solubilization of phosphorus, potassium, and exopolysaccharide production. Four strains revealing the best activities were tested for their drought-tolerance ability by growing them on varying water potentials (- 0.05 to - 0.73 MPa). Among them, two bacterial strains Bacillus megaterium and Bacillus licheniformis showed the best drought-tolerance potential, ACC deaminase activities, IAA production, and antagonistic activities against plant pathogens. Additionally, these strains when exposed to drought stress (- 0.73 MPa) revealed the induction of three new polypeptides (18 kDa, 35 kDa, 30 kDa) in Bacillus megaterium. We determined that 106 cells/mL of Bacillus megaterium and Bacillus licheniformis were enough to induce drought tolerance in wheat under drought stress. These drought-tolerant strains increased the germination index (11-46%), promptness index (16-50%), seedling vigor index (11-151%), fresh weight (35-192%), and dry weight (58-226%) of wheat under irrigated and drought stress. Moreover, these strains efficiently colonized the wheat roots and increased plant biomass, relative water content, photosynthetic pigments, and osmolytes. Upon exposure to drought stress, Bacillus megaterium inoculated wheat plants exhibited improved tolerance by enhancing 59% relative water content, 260, 174 and 70% chlorophyll a, b and carotenoid, 136% protein content, 117% proline content and 57% decline in MDA content. Further, activities of defense-related antioxidant enzymes were also upregulated. Our results revealed that drought tolerance was more evident in Bacillus megaterium as compared to Bacillus licheniformis. These strains could be effective bioenhancer and biofertilizer for wheat cultivation in arid and semi-arid regions. However, a detailed study at the molecular level to deduce the mechanism by which these strains alleviate drought stress in wheat plants needs to be explored.

Role of PGPR on the physiology of sunflower irrigated with produced water containing high total dissolved solids (TDS) and its residual effects on soil fertility.

The present study was conducted to evaluate the bioremediation potential of plant growth-promoting rhizobacteria (PGPR) PGPR isolates from high total dissolved solids (TDS) bearing produced water on the water quality, soil physicochemical properties and growth and physiology of sunflower irrigated with high TDS bearing produced water having salinity level 130 times higher above seawater and also containing traces of oil and grease. Seeds of sunflower hybrid Parsun 3 were soaked for 3-4 h prior to sowing in 72 h old culture of PGPR strains W1 and W2 isolated from high TDS bearing polluted water. The control plants were irrigated with 90% diluted TDS water supplemented with 5 ml LB media. Whereas, the inoculated plants were irrigated with 90% diluted TDS water supplemented with 5 ml PGPR inocula.in LB media. The plants were grown under natural conditions. The 16S rRNA sequence analyses identified the isolate W1 bearing 100% similarity with the plant growth-promoting rhizobacteria (PGPR) Ralstonia pickettii and W2 bearing 99.7% similarity with Brevibacillus invocatus. Both the isolate were catalase and oxidase positive. The Ralstonia pickettii and Brevibacillus invocatus treatments decreased the EC and TDS values significantly such that the EC and TDS values of 90% diluted TDS water were 29 times and 19 times higher than tap water. Sodium adsorption ratio (SAR), organic matter, nitrogen, potassium, magnesium and carbon content were 1.96, 1.10, 2.28 1.20, 6.63 and 1.00 times greater than control in the rhizosphere soil of Ralstonia pickettii inoculated plants irrigated with high TDS bearing water There were significant increases in plant growth, sugar, flavonoids and phenolics, chlorophyll b, total chlorophyll, carotenoids content and activities of superoxide dismutase, catalase and peroxidase in plants inoculated with Ralstonia pickettii and Brevibacillus invocatus. The flavonoids, phenolics and proline contents were 0.54, 0.72 and 0.30 times higher in Ralstonia pickettii inoculated plants. Shoot/root dry weight ratio was about (50%) lower than control in Ralstonia pickettii and Brevibacillus invocatus treatments. Ralstonia pickettii was more effective than Brevibacillus invocatus to combat oxidative and osmotic stresses. It is inferred that the high TDS bearing produced water from oil factory harbor Plant growth-promoting rhizobacteria (PGPR) having the potential to combat high salinity stress in plants when used as bioinoculant. The broth culture containing the bacteria may be supplemented with the saline water used for irrigation as it provides nutrients for the growth and proliferation of bacteria present in the saline water and hence the synergistic action of bacterial inocula with the indigenous bacteria present in saline water may better alleviate osmotic and oxidative stresses of plants encountered under salinity stress. The residual effect of Ralstonia pickettii on organic matter and Ca, Mg, K and P content of the rhizosphere soil was notably higher for succeeding crops. Novelty statement This is the first report demonstrating that rhizobacteria can proliferate in water containing salinity higher above seawater in addition to oil grease and TSS. Their efficiency to reduce TDS can be augmented by an exogenous supply of LB broth culture of PGPR isolated from the polluted water. These indigenous rhizobacteria when used as bioinoculant on the plant can act as plant growth promoters as well as bioremediation of salinity effects.

Antimicrobial activity screening of rhizosphere soil bacteria from tomato and genome-based analysis of their antimicrobial biosynthetic potential.

BACKGROUND: Tomato plant growth is frequently hampered by a high susceptibility to pests and diseases. Traditional chemical control causes a serious impact on both the environment and human health. Therefore, seeking environment-friendly and cost-effective green methods in agricultural production becomes crucial nowadays. Plant Growth Promoting Rhizobacteria (PGPR) can promote plant growth through biological activity. Their use is considered to be a promising sustainable approach for crop growth. Moreover, a vast number of biosynthetic gene clusters (BGCs) for secondary metabolite production are being revealed in PGPR, which helps to find potential anti-microbial activities for tomato disease control. RESULTS: We isolated 181 Bacillus-like strains from healthy tomato, rhizosphere soil, and tomato tissues. In vitro antagonistic assays revealed that 34 Bacillus strains have antimicrobial activity against Erwinia carotovora, Pseudomonas syringae; Rhizoctonia solani; Botrytis cinerea; Verticillium dahliae and Phytophthora infestans. The genomes of 10 Bacillus and Paenibacillus strains with good antagonistic activity were sequenced. Via genome mining approaches, we identified 120 BGCs encoding NRPs, PKs-NRPs, PKs, terpenes and bacteriocins, including known compounds such as fengycin, surfactin, bacillibactin, subtilin, etc. In addition, several novel BGCs were identified. We discovered that the NRPs and PKs-NRPs BGCs in Bacillus species are encoding highly conserved known compounds as well as various novel variants. CONCLUSIONS: This study highlights the great number of varieties of BGCs in Bacillus strains. These findings pave the road for future usage of Bacillus strains as biocontrol agents for tomato disease control and are a resource arsenal for novel antimicrobial discovery.