Plant Growth-promoting Effects Research Articles

Evaluation of hemp stem retting water for carbon dots production and their phyto-activity

Water-retting hemp stem yields high-quality fiber but generates wastewater causing environmental pollution. In the present study, the retting water of hemp stem was used to synthesize carbon dots (HP-CDs). The obtained HP-CDs not only exhibited similar fluorescence emission characteristics to that of widely studied citric acid-based CDs (CA-CDs) but also contained nitrogen elements, primarily presented in the form of graphitic and pyridinic structures. Furthermore, the effect of HP-CDs on hemp growth and physiological characteristics were examined at six concentration levels (0, 10, 100, 500, 1000, and 2000 mg⋅L-1) in comparison with those of CA-CDs. Results indicated that HP-CDs were non-phytotoxic across the tested concentration levels. In contrast, CA-CDs exhibited concentration-dependent phytotoxicity, with concentration higher than 500mgL−1 resulted in adverse effects, including increased activity of superoxide dismutase (SOD) and increased concentration of malondialdehyde (MDA) in leaves, necrotic symptom at the leaf edges, reduced root volume, and decreased dry weight accumulation. In a subsequent experiment, plants grown aeroponically with 100mgL−1 of HP-CDs promoted nitrogen uptake, and resulted in a higher leaf and stem dry weight than the plants at 0mgL−1 by 36.4% and 37.8%, respectively, which was comparable with the effects of CA-CDs. In conclusion, CDs derived from hemp retting water exhibited low biological toxicity and demonstrated similar plant growth-promoting effect as chemically derived CDs. Therefore, hemp stem retting water holds potentioal as a source for carbon dot production, and thereby supporting the sustainable hemp industry.

Manipulation in root-associated microbiome via carbon nanosol for plant growth improvements

BackgroundModulating the microbiome with nanomaterials has been proposed to improve plant growth, and reduce reliance on external inputs. Carbon Nanosol (CNS) was attracted for its potential to improve plant productivity. However, the mechanism between CNS and rhizosphere microorganisms remained largely elusive.ResultsHere, we tried to systematically explore the effects of CNS (600 and 1200 mg/L by concentration) on tobacco growth, soil physical properties, and root-associated microbiome. The influence of CNS on soil physicochemical properties and plant growth was significant and dose-dependent, leading to a 28.82% increase in biomass accumulation by 600 mg/L CNS. Comparison between the CNS-treated and control plants revealed significant differences in microbiome composition, including 1148 distinct ASVs (923 bacteria and 225 fungi), microbiome interactions, and metabolic function of root-associated microbiomes. Fungal and bacterial communities had different response patterns for CNS treatment, with phased and dose-dependent effects, with the most significant changes in microbial community structure observed at 1200 mg/L after 10 days of treatment. Microbial networks of CNS-treated plants had more nodes and edges, higher connectivity, and more hub microorganisms than those of control plants. Compared with control, CNS significantly elevated abundances of various bacterial biomarkers (such as Sphingomonas and Burkholderia) and fungi biomarkers (including Penicillium, Myceliophthora, and Talaromyces), which were potential plant-beneficial organisms. Functional prediction based on metagenomic data demonstrated pathways related to nutrient cycling being greatly enriched under CNS treatment. Furthermore, 391 culturable bacteria and 44 culturable fungi were isolated from soil and root samples. Among them, six bacteria and two fungi strains enriched upon CNS treatment were validated to have plant growth promotion effect, and two fungi (Cladosporium spp. and Talaromyces spp.) played their roles by mediating volatile organic compounds (VOCs). To some extent, the driving and shaping of the microbiome by CNS contributed to its impact on plant growth and development.ConclusionOur results revealed the key role of root-associated microbiota in mediating the interaction between CNS and plants, thus providing valuable insights and strategies for harnessing CNS to enhance plant growth.Graphical

Optimization of fermentation conditions for Bacillus velezensis TCS001 and evaluation of its growth promotion and disease prevention effects on strawberries

Bacillus velezensis TCS001 is a novel biocontrol bacterium with broad-spectrum antifungal activity and plant growth-promoting effects, holding great potential for development in agricultural production. This study optimized the fermentation conditions for B. velezensis TCS001 through single-factor experiments combined with response surface methodology, and developed a formulation for TCS001 suspension concentrate (TCS001-SC). The efficacy of TCS001-SC to promote the growth of strawberries and to prevent and control strawberry anthracnose was evaluated. The optimal liquid fermentation condition for TCS001 was determined to be 2.89 % soluble peanut cake powder, 3.0 % glucose, 3.0 % soluble starch, 0.002 % FePO4, 0.006 % KCl, 0.6 % NaCl, 0.05 % MgCl2·6H2O, 0.3 % K2HPO4, 0.15 % KH2PO4, 0.05 % CaCO3, 0.005 % MnSO4, a working volume of 31 % (77.5 mL/250 mL), a rotation speed of 173 r/min, a cultivation temperature of 28°C, an inoculum volume of 1.0 %, and a pH of 7.0. The 15 L fermenter upscale culture achieved a spore count of 9.46 × 109 CFU/mL, which was 2.01 times the spore count of 4.7 × 109 CFU/mL before optimization, and a preliminary TCS001-SC was developed with the fermented broth as the main component. Agar plate confrontation tests showed that TCS001 had antifungal activity against five types of anthracnose fungi, with inhibition rates ranging from 70.3 % to 87.2 %. TCS001-SC could promote the growth of strawberries and induce a rapid defense enzyme response in their leaves, enhancing the plant's resistance to pathogens. After treatment, individual strawberry plants showed significant increases in the number of leaves, fresh weight of stems and leaves, root fresh weight, leaf area, plant height, and the content of POD, SOD, CAT enzymes, GA, IAA, and ABA compared to the control.Additionally, it shows good prevention and control effects against strawberry anthracnose, with a control efficacy of 60.45% after five spray treatments at a concentration of 2 × 107 CFU/mL, which is not significantly different from the efficacy of commercial microbial agents such as Bacillus subtilis wettable powder, subsequent field trials will be conducted to determine its potential as a microbial pesticide. This study provides important support for the future industrial production and application of the strain TCS001.

Microbiological mechanism of hydrogen fertilizer effect in soil.

For a long time, intercropping and rotation of leguminous with non-leguminous crops is widely used to reduce the application of nitrogen fertilizer and increase yield in agroecosystems. At present, most researchers considered that this management measure is helpful for reducing fertilizer consumption and increasing its efficiency, as it can improve nutrient supply of legumestonon-legumes, the spatial nutrient utilization efficiency by enhancing soil spatial heterogeneity, and improve soil structure and disease resistance. However, current theories cannot fully explain the positive effect of crop rotation and inter-cropping systems involving legumes. A large amount of hydrogen (H2) can be produced as an obligatory by-product of nitrogenase responsible for nitrogen (N2) fixation in the root nodules of leguminous plants. Despite of substantial amounts of H2 enriched in the rhizosphere of legumes, only a minor proportion of H2 is found to leak to soil surface. Increasing evidence showed that most H2 released in soil is immediately depleted in the surrounding of N2-fixing nodules by H2-oxidizing bacteria (HOB) thriving in soil. HOB can use H2 as an electron donor to assimilate and fix CO2 through redox reactions to synthesize cellular substances and consequently promote plant growth. To date, however, little is known about the biological mechanism and ecological process behind the "hydrogen fertilizer effect". Therefore, we review the H2-induced plant growth-promoting effects and its microbiological mechanisms. Our aims were to explore a new way for enhancing agroecosystem production, and to provide scientific basis for future utilization of H2 in agricultural production practices.

Exploring native arsenic (As)-resistant bacteria: unveiling multifaceted mechanisms for plant growth promotion under As stress.

This study explores the plant growth-promoting effect (PGPE) and potential mechanisms of the arsenic (As)-resistant bacterium Flavobacterium sp. A9 (A9 hereafter). The influences of A9 on the growth of Arabidopsis thaliana, lettuce, and Brassica napus under As(V) stress were investigated. Additionally, a metabolome analysis was conducted to unravel the underlying mechanisms that facilitate PGPE. Results revealed that A9 significantly enhanced the fresh weight of Arabidopsis seedlings by 62.6%-135.4% under As(V) stress. A9 significantly increased root length (19.4%), phosphorus (25.28%), chlorophyll content (59%), pod number (24.42%), and weight (18.88%), while decreasing As content (48.33%, P≤.05) and oxidative stress of Arabidopsis. It also significantly promoted the growth of lettuce and B. napus under As(V) stress. A9 demonstrated the capability to produce≥31 beneficial substances contributing to plant growth promotion (e.g. gibberellic acid), stress tolerance (e.g. thiamine), and reduced As accumulation (e.g. siderophores). A9 significantly promoted the plant growth under As stress and decreased As accumulation by decreasing oxidative stress and releasing beneficial compounds.

Plant extract-mediated biosynthesis of sulphur nanoparticles and their antibacterial and plant growth-promoting activity

This study reports green synthesis of sulphur nanoparticles using sodium thiosulfate pentahydrate (Na2S2O35H2O) and Cannabis sativa leaf extracts. X-ray diffraction (XRD) pattern and scanning electron microscopy (SEM) was employed to examine the crystallinity of the particles and morphological characteristics, proved both spherical and rod-shaped morphology of the S NPs having porous nature. The FTIR spectra revealed the interaction of the synthesized SNPs with the biomolecules present in the leaf extract. UV–VIS spectral investigations confirmed the production of SNPs from C. sativa leaf extract and that these SNPs can be used for visible region photocatalysis for the removal of pollutants from wastewater. Energy dispersive X-ray (EDX) spectrum of the SNP shows a single peak around 2.4 keV, confirmed S NPs purity. TEM image revealed the formation of mainly nanorods having a width of ∼20–25 nm and a length of 50–100 nm. Furthermore, some spherical particles (∼20–30 nm) were also formed. HRTEM image of the rod-shaped particles clearly shows the crystal fringe spacing of 0.38 nm. Further, disc diffusion method (DDM) was used to check the antibacterial activity of S NPs against gram-positive S. aureus (MTCC737) 18 ± 0.12 mm and gram-negative bacteria against E. coli (MTCC443) 21.5 ± 0.12 mm, A. salmonicida (MTCC1522) 19.1 ± 0.12 mm, K. pneumoniae (MTCC3384) 17.8 ± 0.10 mm. Among all the strains of bacteria, E. coli (MTCC443) showed a maximum zone of inhibition of 21.5 ± 0.12 mm and its antibacterial activity is somewhat like streptomycin sulfate. These SNPs also promote growth of C. sativa in pot experiment, resulting in a 30 % increase in biomass, 90 cm in shoot length and 28 cm in root length and higher fresh and dry weight (50g and 20g, respectively) with 1.0 mg mL−1 NPs treatment. In addition, SEM-EDX confirmed the accumulation of nanomaterial in plant leaves. This environmentally friendly approach to SNP synthesis using C. sativa extracts demonstrates both potent antibacterial properties and plant growth-promoting effects, making it a promising solution for agriculture and biomedicine.

Biosynthesized Zinc Nanoparticles and their Plant Growth-Promoting Effect on Wheat (Triticum aestivum L.) Cultivation

The zinc nanoparticles were biosynthesized using Pseudomonas and actinobacteria and characterized through UV-Visible spectroscopy, Particle Size Analyzer (PSA), Scanning Electron Microscope (SEM), (EDX), X-Ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR). After biosynthesis and characterization of the nanoparticles (NPs), a field experiment was conducted to know the effect of biosynthesized zinc nanoparticles on wheat crop. In wheat seed priming at 500 ppm and foliar spraying at 500 ppm at panicle initiation stage with zinc nanoparticles biosynthesized through actinobacteria (T6) increased the plant height (25.75%), number of tillers per meter row length (35.98%), leaf area (25.84%), leaf area index (25.75%), total dry matter production (36.46%), productive spikes per square meter (16.98%), number of grains per spike (27.29%), grain weight per spike (30.71), 1,000 grain weight (10.43%), grain yield (17.58%) and straw yield (14.59%) than seed priming at 500 ppm and foliar spraying at 500 ppm at panicle initiation stage with zinc nanoparticles biosynthesized through Pseudomonas (T3) and commercial zinc nanoparticles (T9). Farmers can replace the conventional zinc source with nano forms to obtain the higher yield and yield attributing characteristics, where biosynthesized nanoparticles could be alternative to chemical nanoparticles in terms of high cost and pollution hazards.

Plant growth-promoting abilities of Methylobacterium sp. 2A involve auxin-mediated regulation of the root architecture.

Methylobacterium sp. 2A, a plant growth-promoting rhizobacteria (PGPR) able to produce indole-3-acetic acid (IAA), significantly promoted the growth of Arabidopsis thaliana plants in vitro. We aimed to understand the determinants of Methylobacterium sp. 2A-A. thaliana interaction, the factors underlying plant growth-promotion and the host range. Methylobacterium sp. 2A displayed chemotaxis to methanol and formaldehyde and was able to utilise 1-aminocyclopropane carboxylate as a nitrogen source. Confocal microscopy confirmed that fluorescent protein-labelled Methylobacterium sp. 2A colonises the apoplast of A. thaliana primary root cells and its inoculation increased jasmonic and salicylic acid in A. thaliana, while IAA levels remained constant. However, inoculation increased DR5 promoter activity in root tips of A. thaliana and tomato plants. Inoculation of this PGPR partially restored the agravitropic response in yucQ mutants and lateral root density was enhanced in iaa19, arf7, and arf19 mutant seedlings. Furthermore, Methylobacterium sp. 2A volatile organic compounds (VOCs) had a dose-dependent effect on the growth of A. thaliana. This PGPR is also able to interact with monocots eliciting positive responses upon inoculation. Methylobacterium sp. 2A plant growth-promoting effects can be achieved through the regulation of plant hormone levels and the emission of VOCs that act either locally or at a distance.

A novel Microbacterium strain SRS2 promotes the growth of Arabidopsis and MicroTom (S. lycopersicum) under normal and salt stress conditions.

Microbacterium strain SRS2 promotes growth and induces salt stress resistance in Arabidopsis and MicroTom in various growth substrates via the induction of the ABA pathway. Soil salinity reduces plant growth and development and thereby decreases the value and productivity of soils. Plant growth-promoting rhizobacteria (PGPR) have been shown to support plant growth such as in salt stress conditions. Here, Microbacterium strain SRS2, isolated from the root endosphere of tomato, was tested for its capability to help plants cope with salt stress. In a salt tolerance assay, SRS2 grew well up to medium levels of NaCl, but the growth was inhibited at high salt concentrations. SRS2 inoculation led to increased biomass of Arabidopsis and MicroTom tomato in various growth substrates, in the presence and in the absence of high NaCl concentrations. Whole-genome analysis revealed that the strain contains several genes involved in osmoregulation and reactive oxygen species (ROS) scavenging, which could potentially explain the observed growth promotion. Additionally, we also investigated via qRT-PCR, promoter::GUS and mutant analyses whether the abscisic acid (ABA)-dependent or -independent pathways for tolerance against salt stress were involved in the model plant, Arabidopsis. Especially in salt stress conditions, the plant growth-promotion effect of SRS2 was lost in aba1, abi4-102, abi3, and abi5-1 mutant lines. Furthermore, ABA genes related to salt stress in SRS2-inoculated plants were transiently upregulated compared to mock under salt stress conditions. Additionally, SRS2-inoculated ABI4::GUS and ABI5::GUS plants were slightly more activated compared to the uninoculated control under salt stress conditions. Together, these assays show that SRS2 promotes growth in normal and in salt stress conditions, the latter possibly via the induction of ABA-dependent and -independent pathways.

A Synergistic Indole-3-Acetic Acid-Producing Synthetic Bacterial Consortium Benefits Walnut Seedling Growth

Synthetic microbial communities (SynComs) have been shown to be an ecofriendly alternative for promoting plant growth. However, the mechanisms by which SynCom inoculants drive plant growth promotion in rhizosphere soil are still not fully explored. Herein, we designed a three-strain consortium based on the biocompatibility among strains and indole-3-acetic acid (IAA) production. The consortium containing Bacillus safensis 5-49, Bacillus stratosphericus 5-54, and Bacillus halotolerans 6-30 possessed a synergistic effect on IAA production and biofilm formation. Genetic analysis suggested that IAA was synthesized through tryptophan-dependent pathways in the strains. The consortium outperformed the plant growth-promoting effect observed with single strains, showing an increase in walnut (Juglans regia) seedling dry weight by 92.3% over the non-inoculated plants after 60 days of cultivation. This effect was underpinned by the synergistic interactions of the consortium, which was evidenced by the significantly increased relative abundance of Bacillus and tryptophan metabolism-associated genes in the rhizosphere of consortium-inoculated plants. Meanwhile, the consortium increased the relative abundance of indigenous Pseudomonas in rhizosphere soil, providing a synergistic effect on improving soil enzyme activities and thus available nutrients. The available N, P, and K contents in the consortium-inoculated plant rhizosphere were 3.77–28.4% higher than those in non-inoculated samples. This work provided an efficient bacterial consortium and proposed the mode of action by which this consortium improved plant growth and soil fertility.

Urease-producing bacteria with plant growth-promoting ability that may tolerate and remove cadmium from aqueous solution

Urease-producing bacteria (UPB) are widely present in soil and play an important role in soil ecosystems. In this study, 65 UPB strains were isolated from cadmium (Cd)-polluted soil around a lead-zinc mine in Yunnan Province, China. The Cd tolerance, removal of Cd from aqueous solution, production of indoleacetic acid (IAA) and plant growth-promoting effects of these materials were investigated. The results indicate that among the 65 UPB strains, four strains with IAA-producing ability were screened and identified as Bacillus thuringiensis W6-11, B. cereus C7-4, Serratia marcescens W11-10, and S. marcescens C5-6. Among the four strains, B. cereus C7-4 had the highest Cd tolerance, median effect concentration (EC50) of 59.94 mg/L. Under Cd 5 mg/L, S. marcescens C5-6 had the highest Cd removal from aqueous solution, up to 69.83%. Under Cd 25 mg/kg, inoculation with B. cereus C7-4 significantly promoted maize growth in a sand pot by increasing the root volume, root surface area, and number of root branches by 22%, 29%, and 20%, respectively, and plant height and biomass by 16% and 36%, respectively, and significantly increasing Cd uptake in the maize roots. Therefore, UPB is a potential resource for enhancing plant adaptability to Cd stress in plants with Cd-polluted habitats.

A novel bio-based film-forming helper derived from Leuconostoc mesenteroides: A promising alternative to chemicals for the preparation of biomass film

Some microorganisms produce biopolymers with adhesive, film-forming, and plant-promoting properties in their fermentation broths. These biopolymers hold significant potential as substitutes for chemical film-forming helpers in liquid mulch. The present study involved the preparation of a novel bio-based film-forming helper (BFH-L) through the process of bio-fermentation utilizing Leuconostoc mesenteroides. The successful preparation of BFH-L was confirmed through the assessment of adhesive properties and plant growth-promoting efficacy exhibited by L. mesenteroides and BFH-L. The comprehensive genomic analysis of L. mesenteroides provided compelling evidence supporting the effectiveness of BFH-L, which can be attributed to the functional contribution of genes encoding extracellular glycoside polymers from L. mesenteroides. Through the evaluation and comparison of a triangle model, BFH-L can effectively replace chemical film-forming helpers in the preparation of biomass film (BF). The optimal performance of BF was achieved when 3.0 g/600 mL of BFH-L and 30 g/600 mL of cow dung were added. BF exhibited exceptional water retention ability (water loss: 38.00 ± 3.46 %), superior plant growth promotion effect (germination rate: 100.00 ± 0 %, plant growth: 0.34 ± 0.03 g), and minimal environmental impact (pH value = 8.7). This research provides a new strategy for reducing plastic film pollution while achieving green recycling of organic solid waste from agriculture and animal husbandry.

Characterization of Bacillus velezensis DCJ 2 isolated from the rhizosphere soil of aglaonema ‘Maria’ and investigation of its plant growth-promoting effects

Aglaonema ‘Maria’ is one of the key foliage plants that has gained high demand in foreign destinations in the last few years. This study was conducted to characterize Bacillus velezensis DCJ 2, assess its plant growth promotion effect and investigate the effectiveness of different delivery methods of the bacterial inoculum for the growth promotion of Aglaonema ‘Maria’. In this context, DCJ 2, isolated from the rhizosphere soil of A. Maria was screened to investigate its potential as a plant growth-promoting bacterium. The pure culture of DCJ 2 maintained on nutrient agar was utilized for morphological, biochemical and physiological examinations. The molecular characterization of the strain was performed with different primer pairs targeting 16SrRNA, gyrA, pgsB and sugar kinase genes of DCJ 2. A pot experiment was carried out to investigate the plant growth promotion effect. It was conducted in the greenhouse as per the complete randomized block design with eight treatments and five replicates. . Healthy plants of A. ‘Maria’ established in field soil and solarized soil were treated with 100 ml of the suspension of the DCJ 2 at the rate of 10 8 CFU/ml either as a soil drench or foliar spray. The partial 16SrRNA, gyrA, pgsB and sugar kinase gene sequences of DCJ 2, BLAST searched in the NCBI database revealed that it has a high similarity to Bacillus velezensis. The pot experiment showed that the application of strain DCJ 2 as a soil drench to solarized soil (Treatment 6) yielded a significant increase in growth parameters at p=0.05. Based on molecular analysis, morphological, biochemical, and physiological reactions, DCJ 2 was identified as Bacillus velezensis. This study revealed that B. velezensis DCJ 2 has the potential to boost the growth of A. Maria and hence serve as a base for developing a biofertilizer.

Comparative Analysis of Plant Growth-Promoting Rhizobacteria's Effects on Alfalfa Growth at the Seedling and Flowering Stages under Salt Stress.

Alfalfa (Medicago sativa L.), a forage legume known for its moderate salt-alkali tolerance, offers notable economic and ecological benefits and aids in soil amelioration when cultivated in saline-alkaline soils. Nonetheless, the limited stress resistance of alfalfa could curtail its productivity. This study investigated the salt tolerance and growth-promoting characteristics (in vitro) of four strains of plant growth-promoting rhizobacteria (PGPR) that were pre-selected, as well as their effects on alfalfa at different growth stages (a pot experiment). The results showed that the selected strains belonged to the genera Priestia (HL3), Bacillus (HL6 and HG12), and Paenibacillus (HG24). All four strains exhibited the ability to solubilize phosphate and produce indole-3-acetic acid (IAA) and 1-aminocyclopropane-1-carboxylate (ACC) deaminase. Among them, except for strain HG24, the other strains could tolerate 9% NaCl stress. Treatment with 100 mM NaCl consistently decreased the IAA production levels of the selected strains, but inconsistent changes (either enhanced or reduced) in terms of phosphate solubilization, ACC deaminase, and exopolysaccharides (EPS) production were observed among the strains. During the various growth stages of alfalfa, PGPR exhibited different growth-promoting effects: at the seedling stage, they enhanced salt tolerance through the induction of physiological changes; at the flowering stage, they promoted growth through nutrient acquisition. The current findings suggest that strains HL3, HL6, and HG12 are effective microbial inoculants for alleviating salt stress in alfalfa plants in arid and semi-arid regions. This study not only reveals the potential of indigenous salt-tolerant PGPR in enhancing the salt tolerance of alfalfa but also provides new insights into the mechanisms of action of PGPR.

Bacillus Species: Excellent Biocontrol Agents against Tomato Diseases.

Tomatoes encounter many pathogens, such as fungi and bacteria, which reduce the yield and quality of plants and lead to large losses in production. The application of plant protection products (PPPs) is still an important and most effective measure to control plant diseases. However, the use of chemicals in agriculture contributes to environmental pollution and biodiversity loss, and it can also threaten non-target living organisms. Biological control is a widely accessible, environmentally safe, and cost-efficient alternative for the prevention and suppression of plant diseases. Bacillus species with antimicrobial and plant growth-promoting effects are most frequently used as biocontrol agents to increase the resilience of agricultural production against biotic stresses. The present review discusses the antagonistic mechanisms and the biocontrol potential of Bacillus spp. against tomato diseases caused by different pathogens. The main mechanisms of Bacillus spp. include the production of antimicrobial compounds (antibiotics, extracellular enzymes, siderophores, and volatile compounds), competition for nutrients and space, and induced systemic resistance (ISR). Although Bacillus-based PPPs have been developed and commercialised worldwide for various crops and pathogens, the efficiency issues are still subject to debate. Additionally, a combined strategy for controlling tomato diseases based on Bacillus spp. and other available methods (conventional or natural-based) is a promising research field.

Deciphering the interaction of bacteria inoculants with the recipient endophytic community in grapevine micropropagated plants.

Engineering the plant microbiome with beneficial endophytic bacteria can improve the growth, health, and productivity of the holobiont. Here, we administered two beneficial bacterial strains, Kosakonia VR04 sp. and Rhizobium GR12 sp., to micropropagated grapevine cuttings obtained via somatic embryogenesis. While both strains colonized the plant endosphere, only Rhizobium GR12 sp. increased root biomass under nutritional-deficit conditions, as supported by the plant growth promotion traits detected in its genome. Phylogenetic and co-occurrence analyses revealed that the plant native bacterial community, originally dominated by Streptococcaceae and Micrococcaceae, dramatically changed depending on the inoculation treatments, as invading strains differently affected the relative abundance and the interactions of pre-existing taxa. After 30 days of plantlets' growth, Pantoea became a predominant taxon, and considering untreated plantlets as references, Rhizobium sp. GR12 showed a minor impact on the endophytic bacterial community. On the other hand, Kosakonia sp. VR04 caused a major change in community composition, suggesting an opportunistic colonization pattern. Overall, the results corroborate the importance of preserving the native endophytic community structure and functions during plant microbiome engineering.IMPORTANCEA better comprehension of bacterial colonization processes and outcomes could benefit the use of plant probiotics in the field. In this study, we applied two different beneficial bacteria to grapevine micropropagated plantlets and described how the inoculation of these strains impacts endophytic microbiota assembly. We showed that under nutritional deficit conditions, the response of the receiving endophytic bacterial communities to the invasion of the beneficial strains related to the manifestation of plant growth promotion effects by the inoculated invading strains. Rhizobium sp. GR12 was able to preserve the native microbiome structure despite its effective colonization, highlighting the importance of the plant-endophyte associations for the holobiont performance. Moreover, our approach showed that the use of micropropagated plantlets could be a valuable strategy to study the interplay among the plant, its native microbiota, and the invader on a wider portfolio of species besides model plants, facilitating the application of new knowledge in agriculture.

The PGPR Bacillus aryabhattai promotes soybean growth via nutrient and chlorophyll maintenance and the production of butanoic acid.

Plant growth-promoting rhizobacteria (PGPR) colonize plant roots, establish a mutualistic relationship with the plants and help them grow better. This study reports novel findings on the plant growth-promoting effects of the PGPR Bacillus aryabhattai. Soil was collected from a soybean field, PGPR were isolated, identified, and characterized for their ability to promote plant growth and development. The bacterium was isolated from the soybean rhizosphere and identified as B. aryabhattai strain SRB02 via 16s rRNA sequencing. As shown by SEM, the bacterium successfully colonized rice and soybean roots within 2 days and significantly promoted the growth of the GA-deficient rice cultivar Waito-C within 10 days, as well as the growth of soybean plants with at least six times longer shoots, roots, higher chlorophyll content, fresh, and dry weight after 10 days of inoculation. ICP analysis showed up to a 100% increase in the quantity of 18 different amino acids in the SRB02-treated soybean plants. Furthermore, the 2-DE gel assay indicated the presence of several differentially expressed proteins in soybean leaves after 24 hrs of SRB02 application. MALDI-TOF-MS identified β-conglycinin and glycinin along with several other proteins that were traced back to their respective genes. Analysis of bacterial culture filtrates via GCMS recorded significantly higher quantities of butanoic acid which was approximately 42% of all the metabolites found in the filtrates. The application of 100 ppm butanoic acid had significantly positive effects on plant growth via chlorophyll maintenance. These results establish the suitability of B. aryabhattai as a promising PGPR for field application in various crops.

Facilitating Growth of Maize (Zea mays L.) by Biostimulants: A Perspective from the Interaction between Root Transcriptome and Rhizosphere Microbiome.

The plant growth-promoting effects of biostimulants have been widely documented, while little is known about the intrinsic mechanism. In our study, a pot experiment was conducted to investigate the effects of biostimulants on maize, and the maize root transcriptome and rhizosphere microbiome were assessed. The physicochemical properties of the soil were significantly altered with various trends, and the growth and yield of maize were promoted by biostimulants. Sampling time and maize strain were the strongest factors that altered the rhizosphere microorganisms. Rhizosphere microbiota with biostimulant application exhibited high community robustness. Root transcriptome analysis suggested an altered expression profile induced by biostimulants and maize strains. An integrated correlation analysis demonstrated that phosphate and nitrate metabolism genes are tightly associated with some rhizosphere microbiota. These results implied the plant growth-promoting effects of biostimulants might act in a rhizosphere microorganism-dependent manner and help to expand the use of biostimulants in sustainable agriculture.

Humic acid improves wheat growth by modulating auxin and cytokinin biosynthesis pathways.

Humic acids have been widely used for centuries to enhance plant growth and productivity. The beneficial effects of humic acids have been attributed to different functional groups and phytohormone-like compounds enclosed in macrostructure. However, the mechanisms underlying the plant growth-promoting effects of humic acids are only partially understood. We hypothesize that the bio-stimulatory effect of humic acids is mainly due to the modulation of innate pathways of auxin and cytokinin biosynthesis in treated plants. A physiological investigation along with molecular characterization was carried out to understand the mechanism of bio-stimulatory effects of humic acid. A gene expression analysis was performed for the genes involved in auxin and cytokinin biosynthesis pathways in wheat seedlings. Furthermore, Arabidopsis thaliana transgenic lines generated by fusing the auxin-responsive DR5 and cytokinin-responsive ARR5 promoter to ß-glucuronidase (GUS) reporter were used to study the GUS expression analysis in humic acid treated seedlings. This study demonstrates that humic acid treatment improved the shoot and root growth of wheat seedlings. The expression of several genes involved in auxin (Tryptophan Aminotransferase of Arabidopsis and Gretchen Hagen 3.2) and cytokinin (Lonely Guy3) biosynthesis pathways were up-regulated in humic acid-treated seedlings compared to the control. Furthermore, GUS expression analysis showed that bioactive compounds of humic acid stimulate endogenous auxin and cytokinin-like activities. This study is the first report in which using ARR5:GUS lines we demonstrate the biostimulants activity of humic acid.

Interspecific interactions facilitate keystone species in a multispecies biofilm that promotes plant growth.

Microorganisms colonizing plant roots co-exist in complex, spatially structured multispecies biofilm communities. However, little is known about microbial interactions and the underlying spatial organization within biofilm communities established on plant roots. Here, a well-established four-species biofilm model (Stenotrophomonas rhizophila, Paenibacillus amylolyticus, Microbacterium oxydans, and Xanthomonas retroflexus, termed as SPMX) was applied to Arabidopsis roots to study the impact of multispecies biofilm on plant growth and the community spatial dynamics on the roots. SPMX co-culture notably promoted root development and plant biomass. Co-cultured SPMX increased root colonization and formed multispecies biofilms, structurally different from those formed by monocultures. By combining 16S rRNA gene amplicon sequencing and fluorescence in situ hybridization with confocal laser scanning microscopy, we found that the composition and spatial organization of the four-species biofilm significantly changed over time. Monoculture P. amylolyticus colonized plant roots poorly, but its population and root colonization were highly enhanced when residing in the four-species biofilm. Exclusion of P. amylolyticus from the community reduced overall biofilm production and root colonization of the three species, resulting in the loss of the plant growth-promoting effects. Combined with spatial analysis, this led to identification of P. amylolyticus as a keystone species. Our findings highlight that weak root colonizers may benefit from mutualistic interactions in complex communities and hereby become important keystone species impacting community spatial organization and function. This work expands the knowledge on spatial organization uncovering interspecific interactions in multispecies biofilm communities on plant roots, beneficial for harnessing microbial mutualism promoting plant growth.