Plant Growth-promoting Bacteria Species Research Articles

Biotechnological potential of growth-promoting bacteria in cotton (Gossypium hirsutum L.) crop

Studies involving plant growth-promoting bacteria are attracting increasing attention in the agricultural sector due to their potential to improve growth and production, and to protect plants from biotic and abiotic stresses. The present study aimed to evaluate the effects of three species of plant growth-promoting bacteria (Bacillus subtilis, Priestia megaterium, and Priestia aryabhattai) on the growth and morphological and biochemical aspects of Gossypium hirsutum L. (cotton) seedlings. The experiment was conducted in a greenhouse with four treatments (one control and three inoculations) and five replications per treatment. The seeds were inoculated by immersion in bacterial suspensions (109CFU/mL) and then sown in pots. The plants were monitored for 60 days. During collection, the plants were measured for the fresh mass of roots and shoots, the height of the shoots, stem diameter, and number of leaves. Leaf samples were collected and used for biochemical analyses. The results obtained showed that seeds treated with P. aryabhattaihad significant improvements in the parameters of fresh mass, plant height, stem diameter, and number of leaves, and in the contents of chlorophyll (a, b, and total), nitrogen, and proteins concerning plants in the control treatment. Plants treated with P. megaterium also achieved improvements in fresh mass, stem diameter, nitrogen, and protein contents. These results indicate the potential of these plant growth-promoting bacteria for use in cotton crops and can be employed in the preparation of biostimulants and biofertilizers.

Technological Quality of Sugarcane Inoculated with Plant-Growth-Promoting Bacteria and Residual Effect of Phosphorus Rates.

Phosphate fertilization in highly weathered soils has been a major challenge for sugarcane production. The objective of this work was to evaluate the foliar levels of phosphorus (P) and nitrogen (N) and the technological quality and productivity of second ratoon cane as a function of inoculation with plant-growth-promoting bacteria (PGPBs) together with the residual effect of phosphate fertilization. The experiment was carried out at the research and extension farm of Ilha Solteira, state of São Paulo, Brazil. The experiment was designed in a randomized block with three replications in a 5 × 8 factorial scheme. The treatments consisted of five residual doses of phosphorus (0, 45, 90, 135 and 180 kg ha-1 of P2O5, 46% P) applied at planting from the source of triple superphosphate and eight inoculations from three species of PGPB (Azospirillum brasilense, Bacillus subtilis and Pseudomonas fluorescens), applied in single or co-inoculation at the base of stems of sugarcane variety RB92579. Inoculation with PGPBs influenced leaf N concentration, while inoculations with Pseudomonas fluorescens and combinations of bacteria together with the highest doses exerted a positive effect on leaf P concentration. Co-inoculation with A. brasilense + Pseudomonas fluorescens associated with a residual dose of 135 kg ha-1 of P2O5 increased stem productivity by 42%. Thus, it was concluded that inoculations with Pseudomonas fluorescens and their combinations are beneficial for the sugarcane crop, reducing phosphate fertilization and increasing productivity.

Comparative analysis of adenylate isopentenyl transferase genes in plant growth-promoting bacteria and plant pathogenic bacteria

Cytokinin is a major phytohormone that has been used in agriculture as a plant-growth stimulating compound since its initial discovery in the 1960s. Isopentenyl transferase (IPT) is a rate-limiting enzyme for cytokinin biosynthesis, which is produced by plants as well as bacteria including both plant pathogenic species and plant growth-promoting bacteria (PGPB). It has been hypothesized that there may be differences in IPT function between plant pathogens and PGPB. However, a comprehensive comparison of IPT genes between plant pathogenic and PGPB species has not been performed. Here, we performed a global comparison of IPT genes across bacteria, analyzing their DNA sequences, codon usage, phyletic distribution, promoter structure and genomic context. We found that adenylate type IPT genes are highly specific to plant-associated bacteria and subdivide into two major clades: clade A, largely composed of proteobacterial plant pathogens; and clade B, largely composed of actinomycete PGPB species. Besides these phylogenetic differences, we identified several genomic features that suggest differences in IPT regulation between pathogens and PGPB. Pathogen-associated IPTs tended to occur in predicted virulence loci, whereas PGPB-associated IPTs tended to co-occur with other genes involved in cytokinin metabolism and degradation. Pathogen-associated IPTs also showed elevated gene copy numbers, significant deviation in codon usage patterns, and extended promoters, suggesting differences in regulation and activity levels. Our results are consistent with the hypothesis that differences in IPT regulation and activity exist between plant pathogens and PGPB, which determine their effect on plant host phenotypes through the control of cytokinin levels.

Virulence Factors of Plant Growth-Promoting Bacteria: Analysis of Pseudomonas protegens Genomes

Pseudomonas protegens is the species of plant growthpromoting bacteria, which is widely used in agriculture. In article, previously unknown virulence factors of this microorganism are revealed. When researching the three bacterial genomes P. protegens, genes encoding adherence, antiphagocytosis, iron uptake, biofilm formation, immune evasion, serum resistance, and other virulence traits have been found. More research is needed to learn the role of predicted virulence factors in phytopathology and medicine.

Assessment of plant growth promoting bacteria strains on growth, yield and quality of sweet corn

The use of plant growth promoting bacteria (PGPB) is increasingly gaining acceptance from all the stakeholders of the agricultural production. Different strains of PGPB species had been found to have a vast variety of mechanisms of action, while at the same time, affect differently a variety of crops. This study investigated the effectiveness of ten PGPB strains, on sweet corn cultivation under Mediterranean soil and climatic conditions. A field experiment that followed a completely randomized design was conducted at the region of Attica at Oropos. The results indicated that B. mojavensis increased yield by 16%, B. subtilis by 13.8%, B. pumilus by 11.8% and B. pseudomycoides by 9.8% compared to control. In addition, the harvested grains of the plants treated with B. mojavensis, B. subtilis and B. pumilus presented the highest values of protein and fiber content. Moreover, in most of the cases, high values of photosynthetic rate, transpiration rate and stomatal conductance during the cultivation period, resulted in high productivity. Regarding the texture, the size, the sphericity and the ash content of corn grains, it was found that they were not influenced by the application of different treatments of PGPB. The use of certain strains of PGPB, under specific soil and climatic conditions could contribute to better understand which strains are better suited to certain crops.

Diversity, function and assembly of the Trifolium repens L. root-associated microbiome under lead stress

Root-associated microbial layers provide unique niches that drive specific microbe assemblies. While the rhizosphere microbiome has long received much attention, endophytic microbes remain largely elusive. Characterizing metal-tolerant plants’ strategies for assembling different root-associated microbial layers is important for optimizing phytoremediation. Here, a pre-stratified rhizo-box assay was conducted with Trifolium repens L. under greenhouse conditions with artificial Pb-contaminated soil. Cultivation compensated for the pollution-driven loss of soil microbial biomass carbon, enzyme activities and abundance. The acid-soluble Pb proportion increased in the rhizosphere (from 6.5–13.7% to 7.1–18.0%) compared with bulk soil. Under stress, root-layer variants were a considerable source of variation in the microbiome, with the endosphere representing a unique and independent niche. A core set of root microbes were selected by T. repens, with Proteobacteria and Actinobacteria composed of diverse plant-growth-promoting bacteria (PGPBs) and metal-tolerant members. Cluster analysis revealed endosphere-enriched genera, with Rhizobium, Nocardioides, Novosphingobium, Phyllobacterium, and Sphingomonas being the most dominant. Finally, inferred microbial metabolic pathways suggested that these potential metal-tolerant PGPB species provide critical services to hosts, enabling them to tolerate and even flourish in contaminated soil. Our results provide novel insights for understanding how root-associated microbes help metal-tolerant plants cope with abiotic stress.

Inoculation with Plant Growth-Promoting Bacteria to Reduce Phosphate Fertilization Requirement and Enhance Technological Quality and Yield of Sugarcane.

Phosphorus (P) is a critical nutrient for high sugarcane yields throughout its cultivation cycles, however, a higher amount of P becomes rapidly unavailable to plants due to its adsorption to soil colloids. Some plant growth-promoting bacteria (PGPBs) may be able to enhance P availability to plants and produce phytohormones that contribute to crop development, quality, and yield. Thus, this study aimed to evaluate leaf concentrations of nitrogen (N) and P, yield, and technological quality of sugarcane as a function of different levels of phosphate fertilization associated with inoculation of PGPBs. The experiment was carried out at Ilha Solteira, São Paulo—Brazil. The experimental design was randomized blocks with three replications, consisting of five phosphorus rates (0, 25, 50, 75, and 100% of the recommended P2O5 rate) and eight inoculations, involving three species of PGPBs (Azospirillum brasilense, Bacillus subtilis, and Pseudomonas fluorescens) which were applied combined or in a single application into the planting furrow of RB92579 sugarcane variety. The inoculation of B. subtilis and P. fluorescens provided a higher concentration of leaf P in sugarcane. The P2O5 rates combined with inoculation of bacteria alter technological variables and stalk yield of sugarcane. The excess and lack of phosphate fertilizer is harmful to sugarcane cultivation, regardless of the use of growth-promoting bacteria. We recommend the inoculation with A. brasilense + B. subtilis associated with 45 kg ha−1 of P2O5 aiming at greater stalk yield. This treatment also increases sugar yield, resulting in a savings of 75% of the recommended P2O5 rate, thus being a more efficient and sustainable alternative for reducing sugarcane crop production costs.

PGPB Improve Photosynthetic Activity and Tolerance to Oxidative Stress in Brassica napus Grown on Salinized Soils

Soil salinization, one of the most common causes of soil degradation, negatively affects plant growth, reproduction, and yield in plants. Saline conditions elicit some physiological changes to cope with the imposed osmotic and oxidative stresses. Inoculation of plants with some bacterial species that stimulate their growth, i.e., plant growth-promoting bacteria (PGPB), may help plants to counteract saline stress, thus improving the plant’s fitness. This manuscript reports the effects of the inoculation of a salt-sensitive cultivar of Brassica napus (canola) with five different PGPB species (separately), i.e., Azospirillum brasilense, Arthrobacter globiformis, Burkholderia ambifaria, Herbaspirillum seropedicae, and Pseudomonas sp. on plant salt stress physiological responses. The seeds were sown in saline soil (8 dS/m) and inoculated with bacterial suspensions. Seedlings were grown to the phenological stage of rosetta, when morphological and physiological features were determined. In the presence of the above-mentioned PGPB, salt exposed canola plants grew better than non-inoculated controls. The water loss was reduced in inoculated plants under saline conditions, due to a low level of membrane damage and the enhanced synthesis of the osmolyte proline, the latter depending on the bacterial strain inoculated. The reduction in membrane damage was also due to the increased antioxidant activity (i.e., higher amount of phenolic compounds, enhanced superoxide dismutase, and ascorbate peroxidase activities) in salt-stressed and inoculated Brassica napus. Furthermore, the salt-stressed and inoculated plants did not show detrimental effects to their photosynthetic apparatus, i.e., higher efficiency of PSII and low energy dissipation by heat for photosynthesis were detected. The improvement of the response to salt stress provided by PGPB paves the way to further use of PGPB as inoculants of plants grown in saline soils.

In silico search for the hallmarks of plant‐associated phenotypes in Bacillus strains

Gram-positive bacteria of Bacillus genus exploit various ecological strategies, ranging from soil saprotrophy to pathogenecity in animal species. Some of the Bacillus strains dwelling in the soil or associated with the plant tissues have been demonstrated to belong to the so-called plant growth-promoting bacteria (PGPB), ameliorating plant growth by either phytohormone production, nutrient uptake enhancement, or response to pathogens. The molecular bases for these features have been thoroughly characterized in different PGPB species; however, the actual distribution of growth promoting potential among the bacterial species has yet to be estimated. Recently, our research group has elaborated a tool named CryProcessor designed to rapidly mine genes encoding for Cry entomocidal toxins. Here we demonstrate the extension of its utility to find the genes determining PGPB-related phenotypes in Bacillus strains. Gene sequences associated with plant growth promotion and/or pathogen suppression were selected based on the literature evidence and presence in the confirmed PGPB strains’ genomes. These sequences were further used to construct hidden Markov models (HMMs) using HMMer v3.3.1, which were subsequently wrapped in a master script similar to that of CryProcessor. The resulting tool is capable of assessing the presence of PGPB genetic features in both raw sequencing data and complete genome assemblies. The preliminary tests of its functionality revealed the diversity of genes involved in allelopathic interactions with bacterial and fungal species in Bacillus strains. What is more, a hidden presence of genes encoding for the primary steps of phytohormones in the non-PGPB strains was observed, prompting further evaluation of their putative beneficial effects on plant growth. Although the resulting tool was not tested on genomic data for species outside the Bacillus genus, we suggest that it can be used for the assessment of the PGPB-associated traits in various bacterial species, facilitating the discovery of novel beneficial strains.

Plant Growth Stimulation by Microbial Consortia

Plant-associated microorganisms play an important role in agricultural production. Although various studies have shown that single microorganisms can exert beneficial effects on plants, it is increasingly evident that when a microbial consortium—two or more interacting microorganisms—is involved, additive or synergistic results can be expected. This occurs, in part, due to the fact that multiple species can perform a variety of tasks in an ecosystem like the rhizosphere. Therefore, the beneficial mechanisms of plant growth stimulation (i.e., enhanced nutrient availability, phytohormone modulation, biocontrol, biotic and abiotic stress tolerance) exerted by different microbial players within the rhizosphere, such as plant-growth-promoting bacteria (PGPB) and fungi (such as Trichoderma and Mycorrhizae), are reviewed. In addition, their interaction and beneficial activity are highlighted when they act as part of a consortium, mainly as mixtures of different species of PGPB, PGPB–Mycorrhizae, and PGPB–Trichoderma, under normal and diverse stress conditions. Finally, we propose the expansion of the use of different microbial consortia, as well as an increase in research on different mixtures of microorganisms that facilitate the best and most consistent results in the field.

Plant-Growth-Promoting Bacteria (PGPB) against Insects and Other Agricultural Pests

The interest in using plant-growth-promoting bacteria (PGPB) as biopesticides is significantly growing as a result of the discovery of new properties of certain beneficial microbes in protecting agricultural crops. While several rhizobial species have been widely exploited for their ability to optimize plant use of environmental resources, now the focus is shifted to species that are additionally capable of improving plant health and conferring resistance to abiotic stress and deleterious biotic agents. In some cases, PGPB species may directly act against plant pathogens and parasites through a variety of mechanisms, including competition, protective biofilm formation, and the release of bioactive compounds. The use of this type of bacteria is in line with the principles of ecosustainability and integrated pest management, including the reduction of employing chemical pesticides. Several strains of Bacillus, Paenibacillus, Brevibacillus, Pseudomonas, Serratia, Burkholderia, and Streptomyces species have been the subject of specific studies in this direction and are under evaluation for further development for their use in biological control. Accordingly, specific case studies are presented and discussed.

Growth-promoting bacteria and arbuscular mycorrhizal fungi differentially benefit tomato and corn depending upon the supplied form of phosphorus.

The ability of plants to take up phosphorus (P) from soil depends on root morphology and root exudates release and can be modulated by beneficial soil microbes. These microbes can solubilize P, affect root elongation and branching, and lead to a higher uptake of P and other nutrients. However, coordination of these mechanisms is unclear, especially the mechanism for changing the available form of P. We aimed to dissect the effects of two different beneficial microbial taxa (plant growth-promoting bacteria (PGPB) and arbuscular mycorrhizal fungi (AMF)) on root morphological traits, plant nutrient content, and growth in tomato and corn fertilized with either Gafsa rock phosphate (RP) or triple superphosphate (TSP), which have contrasting solubility levels. Tomato and corn were grown in pots and inoculated with one of three PGPB species or a mix of two AMF species or were not inoculated. Root traits, botanical fractions, and the contents of various mineral nutrients were measured. TSP stimulated tomato biomass accumulation compared to RP but did not stimulate corn biomass accumulation. PGPB improved the growth of both plant species under RP, with limited differences among the strains, whereas AMF only improved tomato growth under TSP. These differences between microbial systems were explained by a bacterial effect on the total root length but not on the mean root diameter and by the ability of AMF to improve the mineral nutrient content. The effects of PGPB were less dependent on the plant species and on P form than the effects of AMF.These results have implications for the improvement of the early plant growth through the management of beneficial microbes.

Medicago truncatula root developmental changes by growth-promoting microbes isolated from Fabaceae, growing on organic farms, involve cell cycle changes and WOX5 gene expression.

Both root nodules and the rhizosphere of Fabaceae plants grown on organic farms are a rich source of bacteria, mainly from the families Enterobacteriaceae and Pseudomonadaceae. The enhanced root system growth in M. truncatula after inoculation with selected bacteria includes an increase of nuclei in the cell cycle S phase and a reduction in phase G2 as well as an enhanced expression of the WOX5 gene. Synthetic fertilizers and pesticides are commonly used to improve plant quality and health. However, it is necessary to look for other efficient and also environmentally safe methods. One such method involves the use of bacteria known as plant growth-promoting bacteria (PGPB). Seventy-two bacterial isolates from the rhizospheric soil and root nodule samples of legumes, including bean, alfalfa, lupine and barrel medic, grown on an organic farm in Western Pomerania (Poland) were screened for their growth-promoting capacities and 38 selected isolates were identified based on 16S rRNA gene sequencing. The analysis showed the isolates to represent 17 strains assigned to 6 families: Enterobacteriaceae, Pseudomonadaceae, Xanthomonadaceae, Rhizobiaceae, Bacillaceae and Alcaligenaceae. Pot experiments showed that 13 strains, capable of producing indole compounds from tryptophan in vitro, could significantly enhance the root and shoot weight of 10-week-old Medicago truncatula seedlings. Compared to non-inoculated seedlings, the root system of inoculated ones was more branched; in addition, the root length, surface area and, especially, the root volume were higher. The 24-h root inoculation with the three selected strains increased the nuclei population in the G1 and S phases, decreased it in the G2 phase and enhanced the WUSCHEL-related Homeobox5 (WOX5) gene expression in root tips and lateral zones. The "arrest" of nuclei in the S phase and the enhancement of the WOX5 gene expression were observed to gradually disappear once the bacterial suspension was rinsed off the seedling roots and the roots were transferred to water for further growth. This study shows that the nodules and rhizosphere of legumes grown on organic farms are a rich source of different PGPB species and provides new data on the ability of these bacteria to interfere with cell cycle and gene expression during the root development.

Seed Coating: A Tool for Delivering Beneficial Microbes to Agricultural Crops.

Plant beneficial microbes (PBMs), such as plant growth-promoting bacteria, rhizobia, arbuscular mycorrhizal fungi, and Trichoderma, can reduce the use of agrochemicals and increase plant yield, nutrition, and tolerance to biotic–abiotic stresses. Yet, large-scale applications of PBM have been hampered by the high amounts of inoculum per plant or per cultivation area needed for successful colonization and consequently the economic feasibility. Seed coating, a process that consists in covering seeds with low amounts of exogenous materials, is gaining attention as an efficient delivery system for PBM. Microbial seed coating comprises the use of a binder, in some cases a filler, mixed with inocula, and can be done using simple mixing equipment (e.g., cement mixer) or more specialized/sophisticated apparatus (e.g., fluidized bed). Binders/fillers can be used to extend microbial survival. The most reported types of seed coating are seed dressing, film coating, and pelleting. Tested in more than 50 plant species with seeds of different dimensions, forms, textures, and germination types (e.g., cereals, vegetables, fruits, pulses, and other legumes), seed coating has been studied using various species of plant growth-promoting bacteria, rhizobia, Trichoderma, and to a lesser extent mycorrhizal fungi. Most of the studies regarding PBM applied via seed coating are aimed at promoting crop growth, yield, and crop protection against pathogens. Studies have shown that coating seeds with PBM can assist crops in improving seedling establishment and germination or achieving high yields and food quality, under reduced chemical fertilization. The right combination of biological control agents applied via seed coating can be a powerful tool against a wide number of diseases and pathogens. Less frequently, studies report seed coating being used for adaptation and protection of crops under abiotic stresses. Notwithstanding the promising results, there are still challenges mainly related with the scaling up from the laboratory to the field and proper formulation, including efficient microbial combinations and coating materials that can result in extended shelf-life of both seeds and coated PBM. These limitations need to be addressed and overcome in order to allow a wider use of seed coating as a cost-effective delivery method for PBM in sustainable agricultural systems.

Differential impacts of plant growth-promoting bacteria (PGPB) on seeds of neotropical tree species with contrasting tolerance to shade

The inoculation of seeds from neotropical tree species with PGPB revealed an active role of inoculant strains in modulating the germination and physiology of seedlings, although the specific responses were subject to specific plant–bacteria and genotype–genotype combinations. Beneficial microbial species encompass different phylogenetic groups, and growth-promoting mechanisms can be direct (nutritional effects) or indirect (resistance against biotic and abiotic stresses). Seed inoculation with plant growth-promoting bacteria (PGPB) can increase the growth and productivity of agricultural crops. However, few studies have been conducted to determine these effects on the seed germination and seedling growth of neotropical tree species. Thus, this study aimed to determine the effects of inoculation with PGPB on the germination, initial development and activities of the enzymes phenylalanine ammonia-lyase and polyphenol oxidase of seedlings of neotropical tree species. Shade-intolerant (Cecropia pachystachya, Heliocarpus popayanensis and Trema micrantha) and shade-tolerant tree species (Cabralea canjerana, Cariniana estrellensis and Trichilia elegans) were inoculated with different species of PGPB (Azospirillum brasilense, Bacillus sp. and Azomonas sp.) and allowed to germinate and grow for 45 days under greenhouse conditions. Inoculation of C. pachystachya with all types of PGPB increased the percentage of germinated seeds (GS) and the vigor index (VI), while the mean time to germination (MTG) decreased. The VI of T. micrantha increased in response to Bacillus sp. and Azomonas sp. inoculation. Cabralea canjerana showed a decreased GS and VI and an increased MTG when inoculated with PGPB and increased activity of phenylalanine ammonia-lyase and polyphenol oxidase when inoculated with Azomonas sp. Inoculation of PGPB interfered with seed germination and seedling physiology, and shade-intolerant species were more likely to benefit from inoculation. Inoculation with PGPB can modulate the seed germination patterns of neotropical tree species, with indications that the ecological group may be involved in the magnitude of the response.

Induction of essential oil production in Mentha x piperita by plant growth promoting bacteria was correlated with an increase in jasmonate and salicylate levels and a higher density of glandular trichomes

Plant growth promoting bacteria (PGPB) are agriculturally important soil bacteria that increase plant growth. We subjected peppermint to inoculation with three species of PGPB. After inoculation, the plants were sprayed with methyl jasmonate solution (MeJA) or SA (salicylic acid). Then, the plants were harvested and the plant growth parameters, trichome density, EO content and endogenous phytohormones were measured. Shoot fresh weight was reduced in plants inoculated and treated with MeJA whereas EO content varied depending on the MeJA concentration applied. Plants inoculated and treated with MeJA 2 mM showed the maximum increase in EO production, revealing a synergism between PGPB and MeJA. SA treatments also enhanced EO yield. The increased growth and EO production observed upon PGPB application were at least partly due to an increase in the JA and SA concentrations in the plant, as well as to an associated rise in the glandular trichome density.

Co-inoculation Effects of Rhizobium sullae and Pseudomonas sp. on Growth, Antioxidant Status, and Expression Pattern of Genes Associated with Heavy Metal Tolerance and Accumulation of Cadmium in Sulla coronaria

Recently, phytoremediation assisted by soil bacteria has emerged as a potential tool to clean up the metal-contaminated/polluted environment. Three plant-growth-promoting bacteria (PGPBs): Rhizobium sullae, Pseudomonas fluorescens, and Pseudomonas sp. were found to tolerate cadmium (Cd) stress. Sulla coronaria inoculated with these PGPBs, and grown under different Cd concentrations (0, 100, and 200 µM), showed increases in dry biomass and proline content. Notable increases in different gas-exchange characteristics such as photosynthesis rate (A), transpiration rate (E), and water-use efficiency (WUE), as well as increases in nitrogen (N) and Cd accumulations were also recorded in inoculated plants compared to non-inoculated Cd stressed plants. The activities of antioxidant enzymes superoxide dismutase (SOD), guaiacol peroxidase (GPOX), catalase (CAT), and ascorbate peroxidase (APX) in S. coronaria roots increased under Cd stress after PGPB co-inoculation, suggesting that these PGPB species could be used for amelioration of stress tolerance in S. coronaria. The expression patterns of ScPCS, ScMT, ScF-box, ScGR, and ScGST in roots of S. coronaria indicated that these genes are differentially expressed under Cd treatments, suggesting their possible roles in Cd and heavy metal stress responses. The results indicate that co-inoculation with R. sullae and Pseudomonas sp. could alleviate Cd toxicity in S. coronaria. In the present study, the obtained data suggest that the application of PGPBs could be a promising strategy for enhancing the phytostabilization efficiency of Cd-contaminated soils.

Ammonium excretion, auxin production and effects of maize inoculation with ethylenediamine-resistant mutants of Pseudomonas sp.

ABSTRACT Plant growth-promoting bacteria (PGPB) comprise part of plant microbiome of biotechnological interest due to their potential to decrease the use of agrochemicals in agriculture. Among the commonly found PGPB species, the Pseudomonas genus is known for high competitiveness and efficiency in expressing growth-promotion traits. To increase the contribution of diazotrophic Pseudomonas sp. to the plant nitrogen nutrition, the strain AZM-01 was chemically mutagenized with methyl methanesulfonate (MMS), following the selection for resistance to ethylenediamine (EDA). From the 13 EDA-resistant mutant strains selected, four showed increased the ammonium excretion, with the highest value reaching up to 284% increase as compared to the wild strain, and six strains were found to produce significantly more auxins than the wild strain. Two independent inoculation trials with the wild and EDA-resistant Pseudomonas were performed on maize, with the objective to study the influence of bacteria on seed germination and its potential to promote maize growth under N-limiting condition. In general, Pseudomonas inoculation modified the root architecture of germinating seeds, and increased biomass of maize plants grown under N-limiting conditions. Shoot dry weight of maize was increased by inoculation with several EDA-resistant mutants as compared to the strain AZM-01, with emphasis on the EDA-5 strain which supports biomass accumulation at equivalent amount of plants grown under full N supply. Significant correlations between in vitro and in vivo parameters were found although low coefficient values predominate. The strategy of random mutagenesis was found suitable to develop PGPB strains with higher potential to supply maize plants with nitrogen.

Structural development and assembly patterns of the root-associated microbiomes during phytoremediation

Successful in situ phytoremediation depends on beneficial interactions between roots and microbes. However, the assembly strategies of root-associated microbiome during phytoremediation are not well known. Here we investigated the assembly patterns of root-associated microbiomes during phytoremediation as well as its regulation by both plants and heavy metals. Plant cultivation and soil amendment increased microbial diversity and restructured microbial communities. Rhizo-compartmentalization was the largest source of variation in root-associated microbiomes, with endosphere being the most independent and exclusive compartment. Soil type explained a larger amount of microbiomes variation in bulk soil and rhizosphere than that in endosphere. A specific core root microbiome was likely to be selected by the metal-tolerant plant H. cannabinus, with Enterobacteriaceae, Pseudomonadaceae and Comamonadaceae which contain a large number of metal-tolerant and plant growth-promoting bacteria (PGPB) being the most abundant families. The root-associated microbial community tended to proceed a niche-assembled patterns and formed a smaller bacterial pool dominant by Proteobacteria, Actinobacteria and Chloroflexi under metal-contaminated conditions. Among these genera, potential metal-tolerant PGPB species have taken up the keystone positions in the microbial co-occurrence networks, revealing their key roles in metal-contaminated environment due to niche selection. We also detected a keystone functional group reducing metal bioavailability which might work as vanguards and devote to maintaining the structure and function of the whole microbial community. In conclusion, this study suggested a specific assembly pattern of root-associated microbiomes of the metal-tolerant plant H. cannabinus during phytoremediation, showing the directional selections of the associated microbiomes by both the plant and metal-contaminated conditions in such a system.

Enhanced drought tolerance in seedlings of Neotropical tree species inoculated with plant growth-promoting bacteria

The inoculation of tree species with plant growth-promoting bacteria (PGPB) has emerged as an important strategy for the acclimation of seedlings by improving plant tolerance to biotic and abiotic stresses. This study aimed to evaluate the effects of inoculation with bacterial species (Azospirillum brasilense - Ab-V5, Bacillus sp., Azomonas sp. and Azorhizophillus sp.) on the growth and physiology of the Neotropical tree species Trema micrantha and Cariniana estrellensis under drought conditions. When associated with Ab-V5 and Azomonas sp., T. micrantha showed increased protein in the leaves, starch in the leaves and roots, photosynthesis, instantaneous carboxylation efficiency and root and shoot dry mass. Moreover, there were reductions in hydrogen peroxide, lipid peroxidation, water potential and proline. In C. estrellensis associated with Ab-V5, higher values of photosynthesis and instantaneous carboxylation efficiency were observed, in addition to higher starch content in the leaves and roots and higher protein content in the leaves; lower hydrogen peroxide and lipid peroxidation contents were also observed. The associations of T. micrantha with Ab-V5 and Azomonas sp. and C. estrellensis with Ab-V5 favored the activation of metabolic processes under drought, leading to greater drought tolerance. This work demonstrates the effects of compatible associations of Neotropical tree and PGPB species and suggests that the identification of compatible PGPB strains can result in tree seedlings with increased tolerance to abiotic stresses, such as drought.