Bioformulation of Bacillus proteolyticus MITWPUB1 and its biosurfactant to control the growth of phytopathogen Sclerotium rolfsii for the crop Brassica juncea var local, as a sustainable approach.

Bacillus proteolyticus MITWPUB1 is a potential producer of biosurfactants (BSs), and the organism is also found to be a producer of plant growth promoting traits, such as hydrogen cyanide and indole acetic acid (IAA), and a solubilizer of phosphate. The BSs were reportedly a blend of two classes, namely glycolipids and lipopeptides, as found by thin layer chromatography and Fourier-transform infrared spectroscopy analysis. Furthermore, semi-targeted metabolite profiling via liquid chromatography mass spectroscopy revealed the presence of phospholipids, lipopeptides, polyamines, IAA derivatives, and carotenoids. The BS showed dose-dependent antagonistic activity against Sclerotium rolfsii; scanning electron microscopy showed the effects of the BS on S. rolfsii in terms of mycelial deformations and reduced branching patterns. In vitro studies showed that the application of B. proteolyticus MITWPUB1 and its biosurfactant to seeds of Brassica juncea var local enhanced the seed germination rate. However, sawdust-carrier-based bioformulation with B. proteolyticus MITWPUB1 and its BS showed increased growth parameters for B. juncea var L. This study highlights a unique bioformulation combination that controls the growth of the phytopathogen S. rolfsii and enhances the plant growth of B. juncea var L. Bacillus proteolyticus MITWPUB1 was also shown for the first time to be a prominent BS producer with the ability to control the growth of the phytopathogen S. rolfsii.

Community-forming traits play role in effective colonization of plant-growth-promoting bacteria and improved plant growth.

Community-forming traits (CFts) play an important role in the effective colonization of plant-growth-promoting bacterial communities that influence host plants positively by modulating their adaptive functions. In this study, by considering plant-growth-promoting traits (PGPts) and community-forming traits (CFts), three communities were constructed, viz., SM1 (PGPts), SM2 (CFts), and SM3 (PGPts+CFts). Each category isolates were picked up on the basis of their catabolic diversity of different carbon sources. Results revealed a distinctive pattern in the colonization of the communities possessed with CF traits. It was observed that the community with CFts colonized inside the plant in groups or in large aggregations, whereas the community with only PGPts colonized as separate individual and small colonies inside the plant root and leaf. The effect of SM3 in the microcosm experiment was more significant than the uninoculated control by 22.12%, 27.19%, and 9.11% improvement in germination percentage, chlorophyll content, and plant biomass, respectively. The significant difference shown by the microbial community SM3 clearly demonstrates the integrated effect of CFts and PGPts on effective colonization vis-à-vis positive influence on the host plant. Further detailed characterization of the interaction will take this technology ahead in sustainable agriculture.

Unleashing Bacillus species as versatile antagonists: Harnessing the biocontrol potentials of the plant growth-promoting rhizobacteria to combat Macrophomina phaseolina infection in Gloriosa superba.

Charcoal rot caused by Macrophomina phaseolina is one of the most devastating diseases that cause severe yield loss in Gloriosa superba cultivation. Plant growth-promoting rhizobacteria (PGPR) are extensively harnessed as biocontrol agents due to their effectiveness in combating a wide array of plant pathogens through a multifaceted approach. The present study delved into the mechanisms underlying its ability to inhibit root rot pathogen and its capacity to promote plant growth in G. superba, commonly known as glory lily. PGPR isolated from the rhizosphere of glory lily were subjected to in vitro assessments using the dual plate technique. The isolated Bacillus subtilis BGS-10 and B. velezensis BGS-21 showed higher mycelial inhibition (61%) against M. phaseolina. These strains also promote plant growth by producing indole-3-acetic acid, siderophore, ammonia, amylase, cellulase, pectinase, xylanase, and lipase chemicals. Genome screening of BGS-10 and BGS-21 revealed the presence of antimicrobial peptide genes such as Iturin (ituD gene), surfactin (srfA and sfp genes) along with the mycolytic enzyme ß-1,3-glucanase. Further, the presence of secondary metabolites in the bacterial secretome was identified through gas chromatography-mass spectrometry (GC/MS) analysis. Notably, pyrrolo[1,2-a] pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl), 9 H-pyrido[3,4-b] indole and L-leucyl-D-leucine exhibited the highest docking score against enzymes responsible for pathogen growth and plant cell wall degradation. Under glasshouse conditions, tuber treatment and soil application of talc-based formulation of B. subtilis BGS-10 and B. velezensis BGS-21 suppress the root rot incidence with a minimal disease incidence of 27.78% over untreated control. Concurrently, there was a notable induction of defense-related enzymes, including peroxidase (PO), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL), in glory lily. Therefore, it can be concluded that plant growth-promoting Bacillus strains play a significant role in fortifying the plant's defense mechanisms against the root rot pathogen.

Pseudomonas aeruginosa Strain 91: A Multifaceted Biocontrol Agent against Banana Fusarium Wilt.

Banana Fusarium wilt (BFW), caused by the soil-borne fungus Fusarium oxysporum f. sp. cubense (Foc), poses significant threats to banana cultivation. Currently, effective control methods are lacking, and biological control has emerged as a possible strategy to manage BFW outbreaks. In this investigation, 109 bacterial strains were isolated from the rhizospheric soil surrounding banana plants in search of potent biological agents against Foc. Strain 91 exhibited the highest antifungal activity against the causal agent of Foc and was identified as Pseudomonas aeruginosa through 16S rRNA gene sequencing and scanning electron microscopy (SEM). Elucidation of strain 91's inhibitory mechanism against Foc revealed a multifaceted antagonistic approach, encompassing the production of bioactive compounds and the secretion of cell wall hydrolytic enzymes. Furthermore, strain 91 displayed various traits associated with promoting plant growth and showed adaptability to different carbon sources. By genetically tagging with constitutively expressing GFP signals, effective colonization of strain 91 was mainly demonstrated in root followed by leaf and stem tissues. Altogether, our study reveals the potential of P. aeruginosa 91 for biocontrol based on inhibition mechanism, adaptation, and colonization features, thus providing a promising candidate for the control of BFW.

Genome analysis of a halophilic Virgibacillus halodenitrificans ASH15 revealed salt adaptation, plant growth promotion, and isoprenoid biosynthetic machinery.

Globally, due to widespread dispersion, intraspecific diversity, and crucial ecological components of halophilic ecosystems, halophilic bacteria is considered one of the key models for ecological, adaptative, and biotechnological applications research in saline environments. With this aim, the present study was to enlighten the plant growth-promoting features and investigate the systematic genome of a halophilic bacteria, Virgibacillus halodenitrificans ASH15, through single-molecule real-time (SMRT) sequencing technology. Results showed that strain ASH15 could survive in high salinity up to 25% (w/v) NaCl concentration and express plant growth-promoting traits such as nitrogen fixation, plant growth hormones, and hydrolytic enzymes, which sustain salt stress. The results of pot experiment revealed that strain ASH15 significantly enhanced sugarcane plant growth (root shoot length and weight) under salt stress conditions. Moreover, the sequencing analysis of the strain ASH15 genome exhibited that this strain contained a circular chromosome of 3,832,903 bp with an average G+C content of 37.54%: 3721 predicted protein-coding sequences (CDSs), 24 rRNA genes, and 62 tRNA genes. Genome analysis revealed that the genes related to the synthesis and transport of compatible solutes (glycine, betaine, ectoine, hydroxyectoine, and glutamate) confirm salt stress as well as heavy metal resistance. Furthermore, functional annotation showed that the strain ASH15 encodes genes for root colonization, biofilm formation, phytohormone IAA production, nitrogen fixation, phosphate metabolism, and siderophore production, which are beneficial for plant growth promotion. Strain ASH15 also has a gene resistance to antibiotics and pathogens. In addition, analysis also revealed that the genome strain ASH15 has insertion sequences and CRISPRs, which suggest its ability to acquire new genes through horizontal gene transfer and acquire immunity to the attack of viruses. This work provides knowledge of the mechanism through which V. halodenitrificans ASH15 tolerates salt stress. Deep genome analysis, identified MVA pathway involved in biosynthesis of isoprenoids, more precisely "Squalene." Squalene has various applications, such as an antioxidant, anti-cancer agent, anti-aging agent, hemopreventive agent, anti-bacterial agent, adjuvant for vaccines and drug carriers, and detoxifier. Our findings indicated that strain ASH15 has enormous potential in industries such as in agriculture, pharmaceuticals, cosmetics, and food.

Changes of microbiome in response to supplements with silver nanoparticles in cotton rhizosphere.

The current study focuses on analyzing the effects of supplements containing silver nanoparticles (AgNPs) on plant growth and rhizospheric bacterial communities. Specifically, the impact of AgNP supplements was assessed on both plant growth promoting traits and bacterial communities in the soil. To do this, a screening process was conducted to select bacteria capable of synthesizing AgNPs through extracellular biosynthesis. UV-Visible spectrophotometer, Fourier transform infrared, X-ray diffraction, scanning electron microscope, and field emission scanning electron microscopy all confirmed, produced AgNPs is in agglomerates form. The resulting AgNPs were introduced into soil along with various supplements and their effects were evaluated after 10 days using next generation sequencing (Illumina-16S rDNA V3-V4 region dependent) to analyze changes in bacterial communities. Seed germination, root-shoot biomass and chlorophyll content were used to assess the growth of the cotton plant, whereas the bacterial ability to promote growth was evaluated by measuring its culturable diversity including traits like phosphate solubilization and indole acetic acid production. The variance in Bray-Curtis ß diversity among six selected combinations including control depends largely on the type of added supplements contributing to 95%-97% of it. Moreover, seed germination improves greatly between 63% and 100% at a concentration range of 1.4 to 2.8 mg/L with different types of supplements. Based on the results obtained through this study, it is evident that using AgNPs along with fructose could be an effective tool for promoting Gossypium hirsutum growth and enhancing plant growth traits like profiling rhizospheric bacteria. The results that have been obtained endorse the idea of boosting the growth of rhizospheric bacteria in a natural way when AgNPs are present. Using these supplements in fields that have been contaminated will lead to a better understanding of how ecological succession occurs among rhizospheric bacteria, and what effect it has on the growth of plants.

In vitro assessment of Bacillus subtilis FJ3 affirms its biocontrol and plant growth promoting potential.

Bacillus species and their metabolites have potential alternative uses as chemical pesticides that can limit the growth of potential plant pathogens and enhance crop productivity. The aim of this study was to investigate the potential of Bacillus subtilis FJ3 for promoting plant growth and controlling fungal plant pathogens. The study evaluated the ability of the strain to promote plant growth in vitro by characterizing its growth-promoting traits, which included the production of hydrolytic enzymes, indole acetic acid, siderophores, biofilm formation, and phosphate solubilization. Polymerase Chain Reaction (PCR) testing revealed that strain FJ3 has the potential to produce lipopeptides such as fengycin, surfactin, mycosubtilin, and pilpastatin. Through in vitro antagonism testing it was demonstrated that strain FJ3 is able to inhibit Fusarium oxysporum by 52% compared to the untreated control and was antagonistic against Aspergillus flavus, Aspergillus niger, and Rhizopus oryzae using a dual method. The minimum inhibitory concentration of Bacillus crude extract resulted in a 92%, 90%, 81.5%, and 56% growth inhibition of Fusarium oxysporum, A. niger, A. flavus, and Rhizopus oryzae, respectively. In FT-IR and GC-MS analysis of crude LPs extract, the transmission and mass spectrum confirmed the existence of aforesaid lipopeptides containing ß-fatty acids with chain lengths ranging from C14 to C21 in which the majority were saturated fatty acids. Greenhouse experimentation revealed that Bacillus strain FJ3 and its metabolites significantly diminished the disease incidence with an average reduction of 31.56%. In sterilized soil, FJ3 and its metabolites caused 24.01% and 10.46% growth promotion, respectively, in chickpea. The results demonstrated that Bacillus strain FJ3 has broad-spectrum antifungal and plant growth-promoting applications and could be a promising candidate for development into a commercialized biobased product for use in sustainable agriculture practice.

The Plant Growth-Promoting Potential of Halotolerant Bacteria Is Not Phylogenetically Determined: Evidence from Two Bacillus megaterium Strains Isolated from Saline Soils Used to Grow Wheat.

(1) Background: Increasing salinity, further potentiated by climate change and soil degradation, will jeopardize food security even more. Therefore, there is an urgent need for sustainable agricultural practices capable of maintaining high crop yields despite adverse conditions. Here, we tested if wheat, a salt-sensitive crop, could be a good reservoir for halotolerant bacteria with plant growth-promoting (PGP) capabilities. (2) Methods: We used two agricultural soils from Algeria, which differ in salinity but are both used to grow wheat. Soil halotolerant bacterial strains were isolated and screened for 12 PGP traits related to phytohormone production, improved nitrogen and phosphorus availability, nutrient cycling, and plant defence. The four 'most promising' halotolerant PGPB strains were tested hydroponically on wheat by measuring their effect on germination, survival, and biomass along a salinity gradient. (3) Results: Two halotolerant bacterial strains with PGP traits were isolated from the non-saline soil and were identified as Bacillus subtilis and Pseudomonas fluorescens, and another two halotolerant bacterial strains with PGP traits were isolated from the saline soil and identified as B. megaterium. When grown under 250 mM of NaCl, only the inoculated wheat seedlings survived. The halotolerant bacterial strain that displayed all 12 PGP traits and promoted seed germination and plant growth the most was one of the B. megaterium strains isolated from the saline soil. Although they both belonged to the B. megaterium clade and displayed a remarkable halotolerance, the two bacterial strains isolated from the saline soil differed in two PGP traits and had different effects on plant performance, which clearly shows that PGP potential is not phylogenetically determined. (4) Conclusions: Our data highlight that salt-sensitive plants and non-saline soils can be reservoirs for halotolerant microbes with the potential to become effective and sustainable strategies to improve plant tolerance to salinity. However, these strains need to be tested under field conditions and with more crops before being considered biofertilizer candidates.

Multifaceted plant growth-promoting traits of indigenous rhizospheric microbes against Phomopsis theae, a causal agent of stem canker in tea plants.

Phomopsis canker is one of the major devastating stem diseases that occur in tea plants caused by the fungal pathogen Phomopsis theae. Rapid development of this disease leads to a capital loss in the tea industry which demands an ecofriendly disease management strategy to control this aggressive pathogen. A total of 245 isolates were recovered from the tea rhizosphere and screened for in vitro plant growth promoting (PGP) traits and antagonism against P. theae. Among them, twelve isolates exhibited multifarious PGP traits including phytohormones, siderophore, hydrogen cyanide, salicylic acid production, phosphate solubilization, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, and antifungal activity. In vitro studies on morphological, biochemical, and phylogenetic analyses classified the selected isolates as Pseudomonas fluorescens (VPF5), Bacillus subtilis (VBS3), Streptomyces griseus (VSG4) and Trichoderma viride (VTV7). Specifically, P. fluorescens VPF5 and B. subtilis VBS3 strains showed the highest level of PGP activities. On the other hand, VBS3 and VTV7 strains showed higher biocontrol efficacy in inhibiting mycelia growth and spore germination of P. theae. A detailed investigation on hydrolytic enzymes produced by antagonistic strains, which degrade the fungus cell wall, revealed that highest amount of chitinase and ß-1,3- glucanase in VTV7 and VBS3 strains. Further, the key antifungal secondary metabolites from these biocontrol agents associated with suppression of P. theae were identified using gas chromatography mass spectrometry. The above study clearly recognized the specific traits in the isolated microbes, which make them good candidates as plant growth-promoting rhizobacteria (PGPR) and biocontrol agents to improve plant growth and health. However, greenhouse trials and field application of these beneficial microbes is required to further confirm their efficacy for the management of stem canker in tea cultivation.

Genetic diversity, symbiotic efficiency, stress tolerance, and plant growth promotion traits of rhizobia nodulating Vachellia tortilis subsp. raddiana growing in dryland soils in southern Morocco.

In the present study, we analyzed the genetic diversity, phylogenetic relationships, stress tolerance, phytobeneficial traits, and symbiotic characteristics of rhizobial strains isolated from root nodules of Vachellia tortilis subsp. raddiana grown in soils collected in the extreme Southwest of the Anti-Atlas Mountains in Morocco. Subsequent to Rep-PCR fingerprinting, 16S rDNA gene sequencing of 15 representative strains showed that all of them belong to the genus Ensifer. Phylogenetic analysis and concatenation of the housekeeping genes gyrB, rpoB, recA, and dnaK revealed that the entire collection (except strain LMR678) shared 99.08 % to 99.92% similarity with Ensifer sp. USDA 257 and 96.92% to 98.79% with Sinorhizobium BJ1. Phylogenetic analysis of nodC and nodA sequences showed that all strains but one (LMR678) formed a phylogenetic group with the type strain "E. aridi" LMR001T (similarity over 98%). Moreover, it was relevant that most strains belong to the symbiovar vachelliae. In vitro tests revealed that five strains produced IAA, four solubilized inorganic phosphate, and one produced siderophores. All strains showed tolerance to NaCl concentrations ranging from 2 to 12% and grew at up to 10% of PEG6000. A greenhouse plant inoculation test conducted during five months demonstrated that most rhizobial strains were infective and efficient. Strains LMR688, LMR692, and LMR687 exhibited high relative symbiotic efficiency values (respectively 231.6 %, 171.96 %, and 140.84 %). These strains could be considered as the most suitable candidates for inoculation of V. t. subsp. raddiana, to be used as a pioneer plant for restoring arid soils threatened with desertification.

Genome mining, antimicrobial and plant growth-promoting potentials of halotolerant Bacillus paralicheniformis ES-1 isolated from salt mine.

Salinity severely affects crop yield by hindering nitrogen uptake and reducing plant growth. Plant growth-promoting bacteria (PGPB) are capable of providing cross-protection against biotic/abiotic stresses and facilitating plant growth. Genome-level knowledge of PGPB is necessary to translate the knowledge into a product as efficient biofertilizers and biocontrol agents. The current study aimed to isolate and characterize indigenous plant growth-promoting strains with the potential to promote plant growth under various stress conditions. In this regard, 72 bacterial strains were isolated from various saline-sodic soil/lakes; 19 exhibited multiple in vitro plant growth-promoting traits, including indole 3 acetic acid production, phosphate solubilization, siderophore synthesis, lytic enzymes production, biofilm formation, and antibacterial activities. To get an in-depth insight into genome composition and diversity, whole-genome sequence and genome mining of one promising Bacillus paralicheniformis strain ES-1 were performed. The strain ES-1 genome carries 12 biosynthetic gene clusters, at least six genomic islands, and four prophage regions. Genome mining identified plant growth-promoting conferring genes such as phosphate solubilization, nitrogen fixation, tryptophan production, siderophore, acetoin, butanediol, chitinase, hydrogen sulfate synthesis, chemotaxis, and motility. Comparative genome analysis indicates the region of genome plasticity which shapes the structure and function of B. paralicheniformis and plays a crucial role in habitat adaptation. The strain ES-1 has a relatively large accessory genome of 649 genes (~ 19%) and 180 unique genes. Overall, these results provide valuable insight into the bioactivity and genomic insight into B. paralicheniformis strain ES-1 with its potential use in sustainable agriculture.

Monilinia fructigena Suppressing and Plant Growth Promoting Endophytic Pseudomonas spp. Bacteria Isolated from Plum.

Brown rot caused by Monilinia spp. fungi causes substantial losses in stone and pome fruit production. Reports suggest that up to 90% of the harvest could be lost. This constitutes an important worldwide issue in the food chain that cannot be solved by the use of chemical fungicides alone. Biocontrol agents (BCAs) based on microorganisms are considered a potential alternative to chemical fungicides. We hypothesized that endophytic bacteria from Prunus domestica could exhibit antagonistic properties towards Monilinia fructigena, one of the main causative agents of brown rot. Among the bacteria isolated from vegetative buds, eight isolates showed antagonistic activity against M. fructigena, including three Pseudomonas spp. isolates that demonstrated 34% to 90% inhibition of the pathogen's growth when cultivated on two different media in vitro. As the stimulation of plant growth could contribute to the disease-suppressing activity of the potential BCAs, plant growth promoting traits (PGPTs) were assessed for bacterial isolates with M. fructigena-suppressing activity. While all isolates were capable of producing siderophores and indole-3-acetic acid (IAA), fixating nitrogen, mineralizing organic phosphate, and solubilizing inorganic phosphate and potassium, only the Pseudomonas spp. isolates showed 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. Overall, our study paves the way for the development of an eco-friendly strategy for managing M. fructigena pathogens by using BCAs including Pseudomonas spp. bacteria, which could also serve as growth stimulators.

Plant growth promoting activities and effect of fermented panchagavya isolate Klebsiella sp. PG-64 on Vigna radiata.

A natural bacterial isolate from fermented panchagavya named as PG-64, exhibits multiple plant growth-promoting traits. This Gram-negative bacteria was identified as Klebsiella sp. PG-64 by 16S rRNA gene sequencing. The Klebsiella sp. PG-64 has shown production of indole acetic acid (106.0 µg/ml), gibberellic acid (20.0 µg/ml), ammonia (7.12 µmol/ml), exopolysaccharide (2.04% w/v) and phosphate solubilization (106.0 µg/ml). It produced 437 µg/ml IAA with 0.75% (w/v) L-tryptophan supplementation and was increased to 575 µg/ml in a laboratory-scale fermenter. The PG-64 has shown tolerance to abiotic stress conditions like pH (5.0-12.0), temperature (28-46 °C), salt (0.5-10.0% w/v NaCl) and osmotic resistance (1-10% w/v PEG-6000). The PG-64 also produced 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase (0.3 ng α-ketobutyrate/mg protein/h) indicating its potential for drought tolerance. Owing to its diverse properties, the effect of Klebsiella sp. PG-64 on Vigna radiata (Mung bean) was examined. The seeds treated with PG-64 culture showed 92% germination with a good seedling vigour index (202). In the pot study, Vigna radiata growth showed 2.23, 1.55, 2.00, 1.65, 1.73, 1.88, 5.00, 5.00, 1.57 times increase in primary root length, dry root weight, root hair numbers, leaf width, leaf numbers, leaf area, fruits number, flower number and chlorophyll content, respectively after 75 days. The application of Klebsiella sp. PG-64 culture resulted in substantial growth enhancement of Vigna radiata. The Klebsiella sp. PG-64 has multiple plant growth-promoting properties along with capabilities to tolerate abiotic stresses, making it a promising liquid biofertilizer contender for various crops.

Description of a novel species of Leclercia, Leclercia tamurae sp. nov. and proposal of a novel genus Silvania gen. nov. containing two novel species Silvania hatchlandensis sp. nov. and Silvania confinis sp. nov. isolated from the rhizosphere of oak.

BACKGROUND: Acute Oak Decline (AOD) is a decline disease first reported on native oaks in the UK, but in recent years reports from further afield such as Europe and the Middle East, indicate that the distribution and host range is increasing at an alarming rate. The stem weeping symptoms of the disease partially develop due to polymicrobial-host interaction, caused by several members of the order Enterobacterales. While investigating the rhizosphere soil of AOD-unaffected trees, termed 'healthy' trees, and diseased oaks suffering from Acute Oak Decline (AOD), an enrichment method designed for enhanced recovery of Enterobacterales led to the recovery of several isolates that could not be classified as any existing species. These isolates showed a close relationship to the genus Leclercia, of which both species are of clinical importance, but the type species Leclercia adecarboxylata also displays plant growth-promoting properties in the rhizosphere. RESULTS: Partial sequencing of four housekeeping genes revealed similarity to the genus Leclercia with varying degrees of relatedness. As such a complete polyphasic approach was used to determine the true taxonomic position of these isolates. This involved whole genome sequencing, phylogenomic analysis, phylogenetic analysis of both the 16S rRNA and four housekeeping gene sequences, combined with phenotypic testing and fatty acid analysis. Both the phylogenomic and phylogenetic analyses separated the isolates into four clusters, two of which were contained in the Leclercia clade. The remaining two clusters formed a separate lineage far removed from any currently defined species. Further investigation into the role of the isolates as plant growth-promoting bacteria as well as plant pathogens was investigated computationally, revealing a number of plant growth-promoting traits as well as virulence genes related to motility, adhesion and immune modulation. CONCLUSION: Based on the genotypic and phenotypic data presented here, these isolates could be differentiated from each other and their closest neighbours. As such we propose the description of Leclercia tamurae sp. nov. (type strain H6S3T = LMG 32609T = CCUG 76176T), Silvania gen. nov., Silvania hatchlandensis sp. nov. (type strain H19S6T = LMG 32608T = CCUG 76185T) and Silvania confinis sp. nov. (type strain H4N4T = LMG 32607T = CCUG 76175T). Due to their interesting protein annotations and alignments, these species warrant further investigation for their role in relation to plant health.

Biocontrol of wilt disease of rice seedlings incited by Fusarium oxysporum through soil application of Streptomyces chilikensis RC1830.

The genus Streptomyces includes many antifungal metabolite-producing novel strains. Fusarium oxysporum soil-inhabiting pathogenic fungi, that affects rice to cause wilt disease. This work demonstrates the efficacy of novel Streptomyces chilikensis strain RC1830, previously isolated from estuarine habitat Chilika Lake in preventing the F. oxysporum wilting/root rot disease and promoting the growth of rice (Var. Swarna) seedlings. A total of 25 different compounds were identified from crude extracts of S. chilikensis RC1830 by GC-MS. In pot trial experiments, Streptomyces-treated rice seedlings showed significantly reduced disease severity index by 80·51%. The seedlings growth parameters (root length, root fresh weight and root dry weight) were also increased by 53·91, 62·5 and 73·46%, respectively, in Streptomyces-treated groups of seedlings compared to Fusarium-infected seedlings. Similarly, the shoot length, shoot dry weight and shoot fresh weight were also increased by 26, 58 and 34·4%, respectively, in Streptomyces-treated groups of seedlings compared to Fusarium-infected seedlings. Formulations of the strain were prepared using seven organic and inorganic wastes as the carrier material and the shelf lives of the propagules were also monitored. Vermiculite and activated charcoal formulations stored at 4°C exhibited a higher viable cell count after 3 months of storage.

A manganese-oxidizing bacterium-Enterobacter hormaechei strain DS02Eh01: Capabilities of Mn(II) immobilization, plant growth promotion and biofilm formation.

While biogenic Mn oxides (BioMnOx) generated by Mn(II)-oxidizing bacteria (MOB) have attracted increasing attention, a MOB strain isolated from Mn-polluted sediments was identified and assigned as Enterobacter hormaechei DS02Eh01. Its Mn(II) immobilization activity, plant growth-promoting traits, and biofilm formation capability were investigated. The results showed that strain DS02Eh01 was found to be able to tolerate Mn(II) up to 122 mM. The strain immobilized Mn(II) in aquatic media mainly through extracellular adsorption, bio-oxidation and pH-induced precipitation as well as manganese oxidation. DS02Eh01-derived BioMnOx are negatively charged and have a larger specific surface area (86.70 m2/g) compared to the previously reported BioMnOx. The strain can immobilize Mn(II) at extreme levels, for instance, when it was exposed to 20 mM Mn(II), about 59% of Mn(II) were found immobilized and 17% of Mn(II) were converted to MnOx. The SEM and TEM observation revealed that the DS02Eh01-derived BioMnOx were aggregates doped with granules and microbial pellets. The precipitated Mn(II) and the Mn(III)/Mn(IV) oxides co-existed in BioMnOx, in which Mn(II) and Mn(IV) were found dominant with Mn(II) accounting for 49.6% and Mn(IV) accounting for 41.3%. DS02Eh01 possesses plant growth-promoting traits and biofilm formation capacity even under Mn(II) exposure. Mn(II) exposure at 5 mM was found to stimulate strain DS02Eh01 to form biofilms, from which, the extracted EPS was mainly composed of aromatic proteins. This study reveals that E. hormaechei strain DS02Eh01 possesses the potential in environmental ecoremediation via coupling processes of macrophytes extraction, biochemical immobilization and biosorption.

Effects of urease-producing bacteria and eggshell on physiological characteristics and Cd accumulation of pakchoi (Brassica chinensis L.) plants.

Soil cadmium (Cd) contamination resulting from anthropogenic activity poses severe threats to food safety and human health. In this study, a pot experiment was performed to evaluate the possibility of using urease-producing bacterium UR21 and eggshell (ES) waste for improving the physiological characteristics and reducing Cd accumulation of pakchoi (Brassica chinensis L.) plants. UR21 has siderophore and IAA production ability. The application of UR21 and ES individually or in combination could improve the root and shoot length, and fresh and dry weight of pakchoi plants under Cd stress. In Cd + ES + UR21-treated plants, the dry weight of shoot and root were increased by 61.54% and 72.73%, respectively. The chlorophyll a, chlorophyll b, and carotenoid content were increased by 52.19%, 42.95%, and 95.56% in Cd + ES + UR21-treated plants. Meanwhile, the H2O2 and MDA content were decreased while the SOD and POD activity were increased, and an increase of soluble protein level in pakchoi plants was observed under Cd + ES + UR21 treatment. Importantly, eggshell and UR21 alone or in combination induced a decline of Cd content in pakchoi plants, especially that Cd + ES + UR21 treatment decreased Cd content in shoot and root by 26.96% and 42.91%, respectively. Meanwhile, the soil urease and sucrase activities were enhanced. Generally, the combined application of ureolytic bacteria UR21 and eggshell exhibited better effects than applied them individually in terms of alleviating Cd toxicity in pakchoi plants. Our findings may give a unique perspective for an eco-friendly and sustainable strategy to remediate heavy metal-polluted soils.

Variation of the seed endophytic bacteria among plant populations and their plant growth-promoting activities in a wild mustard plant species, Capsella bursa-pastoris.

Recent studies have revealed that some bacteria can inhabit plant seeds, and they are likely founders of the bacterial community in the rhizosphere of or inside plants at the early developmental stage. Given that the seedling establishment is a critical fitness component of weedy plant species, the effects of seed endophytic bacteria (SEB) on the seedling performance are of particular interest in weed ecology. Here, we characterized the SEB in natural populations of Capsella bursa-pastoris, a model species of weed ecology. The composition of endophytic bacterial community was evaluated using deep sequencing of a 16S rDNA gene fragment. Additionally, we isolated bacterial strains from seeds and examined their plant growth-promoting traits. Actinobacteria, Firmicutes, Alpha-, and Gammaproteobacteria were major bacterial phyla inside seeds. C. bursa-pastoris natural populations exhibited variable seed microbiome such that the proportion of Actinobacteria and Alphaproteobacteria differed among populations, and 60 out of 82 OTUs occurred only in a single population. Thirteen cultivable bacterial species in six genera (Bacillus, Rhodococcus, Streptomyces, Staphylococcus, Paenibacillus, Pseudomonas) were isolated, and none of them except Staphylococcus haemolyticus were previously reported as seed endophytes. Eight isolates exhibited plant growth-promoting traits like phosphate solubilization activity, indole-3-acetic acid, or siderophore production. Despite the differences in the bacterial communities among plant populations, at least one isolated strain from each population stimulated shoot growth of either C. bursa-pastoris or its close relative A. thaliana when grown with plants in the same media. These results suggest that a weedy plant species, C. bursa-pastoris, contains bacterial endophytes inside their seeds, stimulating seedling growth and thereby potentially affecting seedling establishment.

In vitro screening of siderophore-producing rice root endophytic bacteria from up-land paddies in north-western Vietnam for plant growth-promoting activities

Aims@#Endophytic bacteria (EB) living inside plant tissues possess different beneficial traits including siderophore production and other plant growth-promoting (PGP) activities. Siderophore-producing EB promote host plant growth by secreting ferrum in iron-deficient conditions. This study screened 19 siderophore producers in vitro, isolated from upland rice roots grown in mountain farms of Tung Village, Nậm Có Commune, Mù Cang Chải District, Yên Bái Province, Vietnam, for PGP traits, including phosphate solubilisation, indole-3-acetic acid (IAA), ammonia, gelatinase, amylase and catalase production.@*Methodology and results@#The bacteria were identified by Matrix assisted Laser Desorption Ionization Time of Flight mass spectrometry (MALDI-TOF MS). All 19 isolates were identified as genera Pseudomonas, Enterobacter, Pantoe, Bacillus, Burkholderia, Staphylococcus, Ralstonia and Cronotacter. The isolates produced catalase and ammonia. The amount of ammonia ranged from 60.74 ± 0.14 to 466.72 ± 0.18 mg/L. Out of the 19 siderophore producers, 17 (89.47%) were able to solubilise phosphate with solubilisation index (PSI) ranging from 1.12 ± 0.07 to 2.14 ± 0.15. The qualitative assays identified 12 isolates (63.15%) positive for IAA production with a tryptophan concentration of 5 mM, whereas 15 (78.94%) and 17 (89.47%) isolates were positive for gelatin and starch hydrolysis, respectively. Especially, 7 isolates were found to be positive for all tested assays in vitro including Pseudomonas rhodesiae (NC2), Enterobacter asburiae (NC50), Pantoea ananatis (NC63), Bacillus cereus (NC64), Burkholderia cenocepacia (NC110), Staphylococcus sciuri (NC112) and Ralstonia pickettii (NC122).@*Conclusion, significance and impact of study@#This study serves as crucial findings of multi-trait plant growth-promoting endophytic bacteria isolated from upland rice root in north-western Vietnam. The seven potential isolates positive for all tested assays could be effective PGP bacteria for bio-inoculants.

Psychrotolerant Mesorhizobium sp. Isolated from Temperate and Cold Desert Regions Solubilizes Potassium and Produces Multiple Plant Growth Promoting Metabolites.

Soil potassium (K) supplement depends intensively on the application of chemical fertilizers, which have substantial harmful environmental effects. However, some bacteria can act as inoculants by converting unavailable and insoluble K forms into plant-accessible forms. Such bacteria are an eco-friendly approach for enhancing plant K absorption and consequently reducing utilization of chemical fertilization. Therefore, the present research was undertaken to isolate, screen, and characterize the K solubilizing bacteria (KSB) from the rhizosphere soils of northern India. Overall, 110 strains were isolated, but only 13 isolates showed significant K solubilizing ability by forming a halo zone on solid media. They were further screened for K solubilizing activity at 0 °C, 1 °C, 3 °C, 5 °C, 7 °C, 15 °C, and 20 °C for 5, 10, and 20 days. All the bacterial isolates showed mineral K solubilization activity at these different temperatures. However, the content of K solubilization increased with the upsurge in temperature and period of incubation. The isolate KSB (Grz) showed the highest K solubilization index of 462.28% after 48 h of incubation at 20 °C. The maximum of 23.38 µg K/mL broth was solubilized by the isolate KSB (Grz) at 20 °C after 20 days of incubation. Based on morphological, biochemical, and molecular characterization (through the 16S rDNA approach), the isolate KSB (Grz) was identified as Mesorhizobium sp. The majority of the strains produced HCN and ammonia. The maximum indole acetic acid (IAA) (31.54 µM/mL) and cellulase (390 µM/mL) were produced by the isolate KSB (Grz). In contrast, the highest protease (525.12 µM/mL) and chitinase (5.20 µM/mL) activities were shown by standard strain Bacillus mucilaginosus and KSB (Gmr) isolate, respectively.