Occurrence of Sphingomonas olei with elemental S oxidation capability in sodic soil: Potential role in sodicity reclamation and plant growth promotion.

The success of the sodic soil reclamation using elemental S (S°) depends on the population of the native S° oxidizers. Augmenting the native flora of the sodic soils with effective S° oxidizers can enhance the success of the sodic soil reclamation. Present study reports for the first time the S° oxidation potential of the Sphingomonas olei strain 20UP7 isolated from sodic soils with pHs 9.8 and ECe 3.6 dS m-1. Inoculation with S. olei strain 20UP7 caused 13.0-24.2 % increase in S° oxidation in different sodic soils (pHs 9.1-10.5). It improved the concentration of the Ca2+, Mg2+, PO43- and declined the HCO3- and total alkalinity of the soil solution. This isolate also showed appreciable P and Zn solubilization, indole acetic acid, ammonia, and titratable acidity production in the growth media. It tended to the formation of biofilm around sulphur particles. The PCR amplification with gene-specific primers showed the occurrence of soxA, soxB, and soxY genes with a single band corresponding to length of 850, 460, and 360 base pairs, respectively. The integration of the S. olei strain 20UP7 with S° caused 21.7-25.4 % increase in the rice and wheat yield compared to the soil treated with S° alone. This study concludes that the S. olei, native to high saline-sodic soils can be utilized for improving the sodicity reclamation and plant growth promotion using elemental S based formulations.

Unusual concurrence of P-solubilizing and biocontrol traits under P-limitation in plant-beneficial Pseudomonas aeruginosa P4: insights from in vitro metabolic and gene expression analysis.

Plant-beneficial fluorescent Pseudomonas species with concurrent P-solubilizing and biocontrol traits could have improved rhizospheric survival and efficacy; this rare ability being subject to diverse environmental and endogenous regulations. This study correlates growth patterns, time-course analysis of selected metabolites, non-targeted metabolomics of exometabolites and selected gene expression analysis to elucidate P-limitation-induced physiological shifts enabling co-production of metabolites implied in P-solubilization and biocontrol by P. aeruginosa P4 (P4). P-limited culture supernatants showed enhanced production of selected biocontrol metabolites such as pyocyanin, pyoverdine and pyochelin and IAA while maintaining biomass yield despite reduced growth rate and glucose consumption. Non-targeted exometabolomics further indicated that P-limitation positively impacted pentose phosphate pathway as well as pyruvate, C5-branched dibasic acid and amino acid metabolism. Its correlation with unusually reduced aroC expression and growth phase-dependent changes in the expression of key biosynthetic genes pchA, pchE, pchG, pvdQ and phzM implied a probable regulation of biosynthesis of chorismate-derived secondary metabolites, not neglecting the possibility of multiple factors influencing the gene expression profiles. Similar increase in biocontrol metabolite production was also observed in Artificial Root Exudates (ARE)-grown P4 cultures. While such metabolic flexibility could impart physiological advantage in sustaining P-starvation stress, it manifests as unique coexistence of P-solubilizing and biocontrol abilities.

Altered microbial P cycling genes drive P availability in soil after afforestation.

Soil Phosphorous (P) availability is a limiting factor for plant growth and regulates biological metabolism in plantation ecosystems. The effect of variations in soil microbial P cycling potential on the availability of soil P during succession in plantation ecosystems is unclear. In this study, a metagenomics approach was used to explore variations in the composition and diversity of microbial P genes along a 45-year recovery sequence of Robinia pseudoacacia on the Loess Plateau, as well soil properties were measured. Our results showed that the diversity of P cycling genes (inorganic P solubilization and organic P mineralization genes) increased significantly after afforestation, and the community composition showed clear differences. The gcd and ppx genes were dominant in inorganic P transformation, whereas phnM gene dominated the transformation of organic P. The abundance of genes involved in inorganic P solubilization and organic P mineralization was significantly positively correlated with P availability, particularly for phnM, gcd, ppx, and phnI genes, corresponding to the phyla Gemmatimonadetes, Acidobacteria, Bacteroidetes, and Planctomycetes. The critical drivers of the microbial main genes of soil P cycling were available P (AP) and total N (TN) in soil. Overall, these findings highlight afforestation-induced increases in microbial P cycling genes enhanced soil P availability. and help to better understand how microbial growth metabolism caused by vegetation restoration in ecologically fragile areas affects the soil P cycling.

Exploring plant growth-promoting rhizobacteria as stress alleviators: a methodological insight.

Rhizospheric and root-endophyte bacteria can stimulate plant growth through diverse biochemical mechanisms and pathways, particularly under biotic and abiotic stresses. For this reason, biotechnological trends on plant growth-promoting rhizobacteria (PGPR) application as biofertilizers, bioremediators, and stress alleviators are gaining increasing interest as ecofriendly strategies for sustainable agriculture management and soil restoration. The first steps needed to implement these technologies are isolation, screening, and characterization of PGPR that can be potentially applied as bioinoculants to alleviate biotic and/or abiotic stresses. Therefore, a complete and accurate methodological study and laboratory techniques are required to warrant the correct achievement of these steps. This review compiles and details the fundamentals, methods, and procedures of key protocols used in isolation and characterization of PGPR for plant stress alleviation.

Sequential uptake of aldoses over fructose and enhanced phosphate solubilization in Rhizobium sp. RM.

Rhizobium sp. RM solubilized tri-calcium phosphate (TCP: 324-463 µg ml-1) and rock phosphate (RP: 36-46.58 µg ml-1) in the presence of common rhizospheric sugars-glucose, arabinose, xylose and their combinations. Fructose, though did not support RP solubilization individually, surprisingly solubilized significantly higher phosphate when combined with aldoses. The highest TCP (644 µg ml-1) and RP (75 µg ml-1) solubilization was achieved in fructose + glucose combination. Presence of gluconate, malate and oxalate in culture supernatant indicated functioning of periplasmic glucose oxidation, the non-phosphorylative arabinose dehydrogenase pathway and the tricarboxylate (TCA) cycle, respectively. Aldoses, when present together, were co-utilized (monoauxic growth) however, when added with fructose, prevented the uptake of fructose yielding a typical diauxic growth. This presented an unusual sequential utilization of aldoses over a ketose (fructose) in strain RM. The prevention of fructose uptake by aldoses was investigated through real-time expression of key genes coding fructose transport proteins and initial enzymes of sugar metabolism. Fructose was actively transported via fructose-specific ABC transporters as suggested by upregulation of frcB and frcC only in fructose and fructose growth phases of fructose + aldose combinations. The probable route of initial fructose metabolism involved either fructokinase and/or xylose isomerase, as confirmed by enzyme activities. The upregulation of hfq and hprK genes only in aldose phase of fructose + aldose combinations suggested their possible involvement in governing the preferential utilization. The novel aspects of this study are enhanced organic acid mediated P solubilization in fructose + aldose combinations and a rare hierarchy of aldoses over fructose which is possibly regulated at the level of fructose transport and fructokinase. KEY POINTS: • Sugars when provided in different dual combinations, supported enhanced P solubilization from complex phosphate sources like TCP and RP in Rhizobium sp. RM. • Transcriptional status of genes in cells of RM when grown in different individual sugars and their combinations suggested that fructose might be a less preferred carbon source and hence was utilized after aldoses with the possible regulation by Hfq and HPrK. • First study to present a unique phenomenon of sequential utilization of aldoses (glucose, arabinose and xylose) over fructose in a concentration-independent manner in Rhizobium sp. RM. and to present the effect of dual combinations of sugars on organic acid mediated P solubilization trait of rhizobia.

Purple acid phosphatases: roles in phosphate utilization and new emerging functions.

Plants strive for phosphorus (P), which is an essential mineral for their life. Since P availability is limiting in most of the world's soils, plants have evolved with a complex network of genes and their regulatory mechanisms to cope with soil P deficiency. Among them, purple acid phosphatases (PAPs) are predominantly associated with P remobilization within the plant and acquisition from the soil by hydrolyzing organic P compounds. P in such compounds remains otherwise unavailable to plants for assimilation. PAPs are ubiquitous in plants, and similar enzymes exist in bacteria, fungi, mammals, and unicellular eukaryotes, but having some differences in their catalytic center. In the recent past, PAPs' roles have been extended to multiple plant processes like flowering, seed development, senescence, carbon metabolism, response to biotic and abiotic stresses, signaling, and root development. While new functions have been assigned to PAPs, the underlying mechanisms remained understood poorly. Here, we review the known functions of PAPs, the regulatory mechanisms, and their relevance in crop improvement for P-use-efficiency. We then discuss the mechanisms behind their functions and propose areas worthy of future research. Finally, we argue that PAPs could be a potential target for improving P utilization in crops. In turn, this is essential for sustainable agriculture.

Recycling of silicon-rich agro-wastes by their combined application with phosphate solubilizing microbe to solubilize the native soil phosphorus in a sub-tropical Alfisol.

It is imperative to find suitable strategies to utilize the native soil phosphorus (P), as natural rock phosphate deposits are at a verge of depletion. We explored two such cost-effective and eco-friendly strategies for native soil P solubilization: silicon (Si)-rich agro-wastes (as Si source) and phosphate solubilizing microorganism (PSM). An incubation study was conducted in a sub-tropical Alfisol for 90 days at 25 °C under field capacity moisture. A factorial completely randomized design with 3 factors, namely: Si sources (three levels: sugarcane bagasse ash, rice husk ash, and corn cob ash), PSM (two levels: without PSM, and with PSM); and Si doses [three levels: no Si (Si0), 125 (Si125) and 250 (Si250) mg Si kg-1 soil] was followed. The PSM increased solution P and soluble Si level by ∼22.2 and 1.88%, respectively, over no PSM; whereas, Si125 and Si250 increased solution P by ∼60.4 and 77.1%, as well as soluble Si by ∼41.5 and 55.5%, respectively, over Si0. Also, interaction of PSM × Si doses was found significant (P<0.05). Activities of soil enzymes (dehydrogenase, acid phosphatase) and microbial biomass P also increased significantly both with PSM and Si application. Overall, PSM solubilized ∼4.18 mg kg-1 of inorganic P and mineralized ∼5.92 mg kg-1 of organic P; whereas, Si125 and Si250 solubilized ∼3.85 and 5.72 mg kg-1 of inorganic P, and mineralized ∼4.15 and 5.37 mg kg-1 of organic P, respectively. Path analysis revealed that inorganic P majorly contributed to total P solubilization; whereas, soluble and loosely bound, iron bound and aluminium bound P significantly influenced the inorganic P solubilization. Thus, utilization of such wastes as Si sources will not only complement the costly P fertilizers, but also address the waste disposal issue in a sustainable manner.

Can dark septate endophytic fungi (DSE) mobilize selectively inorganic soil phosphorus thereby promoting sorghum growth? A preliminary study.

Phosphate fertilizers tend to precipitate with soil components, affecting fertilization efficiency and causing negative environmental effects. Soil microorganisms have been used to solve this problem. However, the ability of dark septate endophytic fungi (DSE) to dissolve phosphates and increase crop yield are not well known. The activity of DSE fungi capable of solubilizing reagent grade phosphates was studied in a Typic Hapludoll (Hapludol típico). The effect of the fungi on the inorganic phosphorus fractions was evaluated and an experiment was conducted in pots with sorghum as a crop. No fungal structures were found in the roots. Curvularia sp. aerial biomass and root length increased; however, P concentration was not affected. Although the results are not conclusive, they represent an advance in the potential use of DSE fungi as P solubilizers to treat crop nutrition.

The fungicide "fluopyram" promotes pepper growth by increasing the abundance of P-solubilizing and N-fixing bacteria.

Fluopyram, as a reasonably good fungicide and nematicide, is widely used to control agricultural pests worldwide. However, its effects on soil microbial communities and plant growth remain controversial. Therefore, in this study, we investigated the effects of three concentrations (0.5, 1.5, and 5.0 mg/kg) of the fluopyram (Lufuda 41.7% a.i., suspension concentrate, SC) on the pepper rhizosphere microorganisms and pepper seedlings growth in a plant growth room. Moreover, we also investigated the dissipation of fluopyram in the soil, pepper roots, and leaves across a time interval of 45 days. The results showed that fluopyram application increased the number of pepper rhizosphere phosphate (P)-solubilizing bacteria, the abundance of nitrogen (N)-fixing nifH genes, and the pepper seedling growth. The results of terminal restriction fragment length polymorphism (T-RFLP) analysis demonstrated that fluopyram did not alter rhizosphere bacterial community structure and diversity. However, fluopyram did increase the relative abundances of 138 bp and 400 bp T-RFs closely representing Bacillus and Rhizobium genera that were known as efficient plant growth promoting bacteria with P-solubilization and N-fixation properties. Corresponding to the increase of plant growth and beneficial microbes, the half-lives of fluopyram in soil and plant tissues also decreased that nevertheless suggested the role of plant-microbe interactions in the faster removal of fluopyram after application. Our results suggest that short-lived and easily degradable pesticides may have less toxicological effects on soil health while their judicious use may reshape plant-microbe interactions in favor of the plant growth.

High-Throughput Sequencing and Expression Analysis Suggest the Involvement of Pseudomonas putida RA-Responsive microRNAs in Growth and Development of Arabidopsis.

Beneficial soil microorganisms largely comprise of plant growth-promoting rhizobacteria (PGPR), which adhere to plant roots and facilitate their growth and development. Pseudomonas putida (RA) strain MTCC5279 is one such PGPR that exhibits several characteristics of plant growth promotion, such as P-solubilization, and siderophores and IAA production. Plant-PGPR interactions are very complex phenomena, and essentially modulate the expression of numerous genes, consequently leading to changes in the physiological, biochemical, cellular and molecular responses of plants. Therefore, in order to understand the molecular bases of plant-PGPR interactions, we carried out the identification of microRNAs from the roots of Arabidopsis upon P. putida RA-inoculation, and analyses of their expression. MicroRNAs (miRNAs) are 20- to 24-nt non-coding small RNAs known to regulate the expression of their target genes. Small RNA sequencing led to the identification of 293 known and 67 putative novel miRNAs, from the control and RA-inoculated libraries. Among these, 15 known miRNAs showed differential expression upon RA-inoculation in comparison to the control, and their expressions were corroborated by stem-loop quantitative real-time PCR. Overall, 28,746 and 6931 mRNAs were expected to be the targets of the known and putative novel miRNAs, respectively, which take part in numerous biological, cellular and molecular processes. An inverse correlation between the expression of RA-responsive miRNAs and their target genes also strengthened the crucial role of RA in developmental regulation. Our results offer insights into the understanding of the RA-mediated modulation of miRNAs and their targets in Arabidopsis, and pave the way for the further exploitation and characterization of candidate RA-responsive miRNA(s) for various crop improvement strategies directed towards plant sustainable growth and development.

Characterization of phosphate solubilizing bacteria isolated from heavy metal contaminated soils and their potential for lead immobilization.

Phosphate solubilizing bacteria (PSB) were isolated from heavy metal contaminated soils and their potentials for lead (Pb) immobilization in soil were studied in this paper. 53 PSB strains were isolated and their minimum inhibitory concentrations (MICs) for lead were determined. The results showed that strains B3, B4, B10, F2-1 and L1-5 had relatively high lead resistant capabilities with MICs ranged from 0.5 to 8 mM, and were identified as species of Leclercia adecarboxylata and Pseudomonas putida by 16S rRNA. The most efficient PSB strains could solubilize 200 mg L-1 of P, and phosphate solubilizing capacity of those PSBs was related to the concentrations of organic acids, acid phosphatase activity and pH. Furthermore, the growth pattern of L1-5 strain with the presence of Pb(NO3)2 was observed and a mechanism of biomineralization of lead ions in bacteria biomass was determined by FT-IR and XRD analyses. Results showed that high concentration of lead can inhibit the growth of PSBs, and L1-5 isolate can transform lead ions into lead hydroxyapatite and pyromorphite. The PSBs, which possessing the properties of immobilizing lead through phosphate solubilization and biomineralization could be exploited for bioremediation of Pb polluted soils in future.

Co-existence of Leclercia adecarboxylata (LSE-1) and Bradyrhizobium sp. (LSBR-3) in nodule niche for multifaceted effects and profitability in soybean production.

The present study was designed with the objective of improving growth and nodulation of soybean [Glycine max (L.) Merill] with co-inoculation of native Bradyrhizobium sp. (LSBR-3) (KF906140) and non-rhizobial nodule endophytic diazotroph Leclercia adecarboxylata (LSE-1) (KX925974) with multifunctional plant growth promoting (PGP) traits in cereal based cropping system (Rice-Wheat). A total of 40 endophytic bacteria from cultivated and wild sp. of soybean were screened for multifarious PGP traits and pathogenicity test. Based on PGP traits, antagonistic activities and bio-safety test; L. adecarboxylata (LSE-1) was identified with 16 S rRNA gene sequencing along with the presence of nifH (nitrogen fixation) and ipdc (IAA production) genes. Dual inoculant LSE-1 and LSBR-3 increased indole acetic acid (IAA), P & Zn-solubilization, 1-aminocyclopropane-1-carboxylate deaminase (ACCD) activity, siderophore, biofilm formation and exo-polysaccharides in contrast to single inoculation treatment. Further, assessment of dual inoculant LSBR-3 + LSE-1 improved growth parameters, nodulation, soil enzymes activities, nutrient accumulation and yield as compared to single as well as un-inoculated control treatment under field conditions. Single inoculant LSBR-3 improved yield by 8.84% over control. Further, enhancement of 4.15% grain yield was noticed with LSBR-3 + LSE-1 over LSBR-3 alone treatment. Application of LSBR-3 + LSE-1 gave superior B:C ratio (1.29) and additional income approximately 116 USD ha-1 in contrast to control treatment. The present results thus, is the first report of novel endophytic diazotroph L. adecarboxylata (LSE-1) as PGPR from Indian conditions particularly in Punjab region for exploiting as potential PGPR along with Bradyrhizobium sp. (LSBR-3) in soybean.

Synergism of Pseudomonas aeruginosa (LSE-2) nodule endophyte with Bradyrhizobium sp. (LSBR-3) for improving plant growth, nutrient acquisition and soil health in soybean.

The present study was aimed to assess the scope of native potential endophyte Pseudomonas aeruginosa (LSE-2) strain (KX925973) with recommended Bradyrhizobium sp. (LSBR-3) (KF906140) for synergistic effect to develop as consortium biofertilizer of soybean. A total of 28 non-rhizobial endophytic bacteria were isolated from cultivated and wild sp. of soybean. All isolates were screened for multifarious PGP traits viz. Indole-3-acetic acid (IAA), phosphate (P) and zinc (Zn) solubilization, siderophore, cell wall degrading enzymes and pathogenicity. Compatible of LSBR-3 and LSE-2 enhanced IAA, P-solubilization, 1-aminocyclopropane-carboxylate deaminase and biofilm formation over the single inoculant treatment. Further, consortium was evaluated in vivo for growth, symbiotic traits, nutrient acquisition, soil quality parameters and yield attributes of soybean. Improvement in growth parameters were recorded with dual inoculant LSBR-3 + LSE-2 as compared to LSBR-3 alone and un-inoculated control treatments. Significantly (p ≥ 0.05) high symbiotic and soil quality parameters (phosphatase and soil dehydrogenase activity) was recorded with LSBR-3 + LSE-2 at vegetative and flowering stage as compared to LSBR-3 alone and un-inoculated control treatments. Single inoculation of LSBR-3 improved grain yield by 4.25% over the un-inoculated control treatment, further, enhancement in yield was recorded with consortium inoculant (LSBR-3 and LSE-2) by 3.47% over the LSBR-3 alone. Application of consortium inoculant (LSBR-3 + LSE-2) gave an additional income of Rs. 5089/ha over the un-inoculated control treatment. The results, thus strongly suggest that endophytic diazotroph LSE-2 can be used as potent bio-inoculant along with LSBR-3 as bio-enhancer for improving soybean productivity in a sustainable system.

Fungal extracellular phosphatases: their role in P cycling under different pH and P sources availability.

AIMS: The aim of this work is to analyse the effect of pH, fungal identity and P chemical nature on microbial development and phosphatase release, discussing solubilization and mineralization processes in P cycling. METHODS AND RESULTS: P solubilizing fungi (Talaromyces flavus, T. helicus L, T. helicus N, T. diversus and Penicillium purpurogenum) were grown under three pH conditions (6, 6·5 and 8·5) and with different inorganic (calcium, iron, aluminium and rock) and organic (lecithin and phytate) P sources. P solubilization, mineralization, growth and phosphatase production were recorded. Acid and neutral environments maximized fungal development and P recycling. P chemical nature changed the phosphatases release pattern depending on the fungal identity. Acid phosphatase activity was higher than alkaline phosphatases, regardless of pH or sample times. Alkaline phosphatases were affected by a combination of those factors. CONCLUSIONS: P chemical nature and pH modify fungal growth, P mineralization and solubilization processes. The underlying fungal identity-dependent metabolism governs the capacity and efficiency of P solubilization and mineralization. P solubilization and mineralization processes are interrelated and simultaneously present in soil fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: This study constitutes a reference work to improve the selection of fungal bioinoculants in different environmental conditions, highlighting their role in P cycling.

Pseudomonas sesami sp. nov., a plant growth-promoting Gammaproteobacteria isolated from the rhizosphere of Sesamum indicum L.

A novel Gram-stain negative, aerobic, motile, rod-shaped bacterium was isolated from Sesame (Sesamum indicum L.) rhizosphere soil. Based on the 16S rRNA gene similarity value (99.4-98.6%) obtained with phylogenetically closely related strains and through analyses of their house keeping genes (atpD, infB and rpoB), the strain SI-P133T was delineated among the species of the genus Pseudomonas and was subjected to polyphasic taxonomic analysis. It was a chemoorganotroph which grew at wide range of temperature (4-45 °C), pH (5.5-9.5) and NaCl concentrations (0-7% (w/v). DNA-DNA hybridization values with closely related type strains DSM 9751T, DSM 19095T, DSM 21509T, ICMP 9151T and DSM 6929T ranged from 23.1 to 44.2%. The most abundant fatty acids were C16:0, C10:0 3-OH, summed feature 3 (comprising C16:1 ω7c and/or C16:1 ω6c), C17:0 cyclo and C12:0 3-OH. The major isoprenoid quinone system was ubiquinone 9 (Q-9) and the G+C content was 61.3 mol%. The major polar lipids of the strain SI-P133T were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. With regard to prospective use in agriculture, plant growth-promoting properties of the strain were tested and plant growth-promotion was demonstrated under in vitro conditions. Based on the various polyphasic taxonomic traits analysed, the strain SI-P-133T was novel and placed within the genus Pseudomonas. Hence we propose a novel species named Pseudomonas sesami sp. nov., for which the type strain is SI-P133T (=NCIMB 14519T = KCTC 22518T).

Efficiency of plant growth-promoting P-solubilizing Bacillus circulans CB7 for enhancement of tomato growth under net house conditions.

P-solubilizing bacterial isolate CB7 isolated from apple rhizosphere soil of Himachal Pradesh, India was identified as Bacillus circulans on the basis of phenotypic characteristics, biochemical tests, fatty acid methyl esters analysis, and 16S rRNA gene sequence. The isolate exhibited plant growth-promoting traits of P-solubilization, auxin, 1-aminocyclopropane-1-carboxylate deaminase activity, siderophore, nitrogenase activity, and antagonistic activity against Dematophora necatrix. In vitro studies revealed that P-solubilization and other plant growth-promoting traits were dependent on the presence of glucose in PVK medium and removal of yeast extract had no significant effect on plant growth-promoting traits. Plant growth-promoting traits of isolate CB7 were repressed in the presence of KH2 PO4 . P-solubilization activity was associated with the release of organic acids and a drop in the pH of the Pikovskaya's medium. HPLC analysis detected gluconic and citric acid as major organic acids in the course of P-solubilization. Remarkable increase was observed in seed germination (22.32%), shoot length (15.91%), root length (25.10%), shoot dry weight (52.92%) and root dry weight (31.4%), nitrogen (18.75%), potassium (57.69%), and phosphorus (22.22%) content of shoot biomass over control. These results demonstrate that isolate CB7 has the promising PGPR attributes to be developed as a biofertilizer to enhance soil fertility and promote plant growth.

Co-existence of halo-tolerant Pseudomonas fluorescens and Enterococcus hirae with multifunctional growth promoting traits to ameliorate salinity stress in Vigna radiata.

Soil salinization has become a prominent obstacle in diverse arid and semi-arid region damaging agricultural productivity globally. From this perspective, present investigation was aimed to compare the potential compatible consortium of bio-inoculants for improving Plant Growth Promoting (PGP) attributes, anti-oxidative enzymes, grain yield and profitability of Vigna radiata in saline soil conditions. A total of 101 rhizobacterium isolated from salt affected regions of Punjab, India were screened for their ability to induce salt tolerance, multifunctional PGP traits and antagonistic activities. The 16S rRNA sequencing identified the strains LSMR-29 and LSMRS-7 as Pseudomonas flourescens and Enterococcus hirae, respectively. In-vitro compatible halo-tolerant dual inoculant (LSMR-29 + LSMRS-7) as bio-inoculants mitigated salt stress in Vigna radiata (spring mungbean) seedling with improved seed germination, biomass and salt tolerance index together with the presence of nifH, acds, pqq and ipdc gene under salinity stress as compared to single inoculants. Further, the potential of single and dual bio-inoculants were also exploited for PGP attributes in pot and field experiments. Results indicated that a significant improvement in chlorophyll content (2.03 fold), nodulation (1.24 fold), nodule biomass (1.23 fold) and leghemoglobin content (1.13 fold) with dual inoculant of LSMR-29 + LSMRS-7 over the LSMR-29 alone. The concentrations of macro & micronutrients, proline, soil enzyme activities i.e. soil dehydrogenase, acid & alkaline phosphatases and antioxidant enzymes such as superoxide dismutase, catalase and peroxidase also found to be high for LSMR-29 + LSMRS-7 as compared to un-inoculated control. The high grain yield thereby leading to Benefit: Cost (B: C) ratio at field scale was indicative of the commercial use bio-inoculants under salt affected Vigna radiata (spring mungbean) to improvement of productivity and soil health. The current finding reveals a co-inoculation of halo-tolerating Pseudomonas fluorescens and Enterococcus hirae containing ACC deaminase could prove to be novel approach for inducing salt tolerance and improving productivity of Vigna radiata (spring mungbean).

Efficacy of DAP coated with bacterial strains and their metabolites for soil phosphorus availability and maize growth.

Phosphorus (P) use efficiency in alkaline/calcareous soils is only 20% due to precipitation of P2O5 with calcium and magnesium. However, coating Diammonium Phosphate (DAP) with phosphorus solubilizing bacteria (PSB) is more appropriate to increase fertilizer use efficiency. Therefore, with the aim to use inorganic fertilizers more effectively present study was conducted to investigate comparative effect of coated DAP with PSB strains Bacillus subtilis ZE15 (MN003400), Bacillus subtilis ZR3 (MN007185), Bacillus megaterium ZE32 (MN003401) and Bacillus megaterium ZR19 (MN007186) and their extracted metabolites with uncoated DAP under axenic conditions. Gene sequencing was done against various sources of phosphorus to analyze genes responsible for phosphatase activity. Alkaline phosphatase (ALP) gene amplicon of 380bp from all tested strains was showed in 1% w/v gel. Release pattern of P was also improved with coated fertilizer. The results showed that coated phosphatic fertilizer enhanced shoot dry weight by 43 and 46% under bacterial and metabolites coating respectively. Shoot and root length up to 44 and 42% with metabolites coated DAP and 41% with bacterial coated DAP. Physiological attributes also showed significant improvement with coated DAP over conventional. The results supported the application of coated DAP as a useful medium to raise crop yield even at lower application rates i.e., 50 and 75% DAP than non-coated 100% DAP application which advocated this coating technique a promising approach for advancing circular economy and sustainable development in modern agriculture.

A Comprehensive Approach Combining Short-Chain Polyphosphate and Bacterial Biostimulants for Effective Nutrient Solubilization and Enhanced Wheat Growth.

Phosphorus constitutes a crucial macronutrient for crop growth, yet its availability often limits food production. Efficient phosphorus management is crucial for enhancing crop yields and ensuring food security. This study aimed to enhance the efficiency of a short-chain polyphosphate (PolyP) fertilizer by integrating it with plant growth-promoting bacteria (PGPB) to improve nutrient solubilization and wheat growth. Specifically, the study investigated the effects of various bacterial strains on wheat germination and growth when used in conjunction with PolyP. To achieve this, a greenhouse experiment was conducted in which the wheat rhizosphere was amended with a short-chain PolyP fertilizer. Based on the morphological aspect, eight bacteria, designated P1 to P8, were isolated and further characterized. Plant growth-promoting traits were observed in all bacterial strains, as they presented the ability to produce Indole Acetic Acid (IAA) in significant amounts ranging from 7.5 ± 0.3 µg/mL to 44.1 ± 2 µg/mL, expressed by B. tropicus P4 and P. soyae P1, respectively. They also produced ammonia, hydrogen cyanide (HCN), and siderophores. Their effect against the plant pathogen Fusarium culmorum was also assessed, with P. reinekei P2 demonstrating the highest biocontrol activity as it presented a total inhibitory effect. Additionally, some strains exhibited the ability to solubilize/hydrolyze phosphorus, potassium, and zinc. In vivo, the initial growth potential of wheat seeds indicated that those inoculated with the isolated strains exhibited elevated germination rates and enhanced root growth. Based on their plant growth-promoting traits and performance in the germination assay, three strains were selected for producing the best results, specifically phosphorus hydrolyzation/solubilization, zinc solubilization, IAA production, HCN, and siderophores production. Wheat seeds were inoculated by drenching in a bacterial suspension containing 1010 CFU/mL of log phase culture, and an in planta bioassay was conducted in a growth chamber using three selected strains (Pseudomonas soyae P1, Pseudomonas reinekei P2, and Bacillus tropicus P4), applied either individually or with PolyP on a P-deficient soil (28 mg/kg of P Olsen). Our findings demonstrated that the combination of Pseudomonas soyae P1 and PolyP achieved the highest shoot biomass, averaging 41.99 ± 0.87 g. Notably, applying P. soyae P1 or Bacillus tropicus P4 alone yielded similar results to the use of PolyP alone. At the heading growth stage, the combination of Bacillus tropicus P4 and PolyP significantly increased the Chlorophyll Content Index (CCI) to 37.02 µmol/m2, outperforming both PolyP alone (24.07 µmol/m2) and the control (23.06 µmol/m2). This study presents an innovative approach combining short-chain PolyP with bacterial biostimulants to enhance nutrient availability and plant growth. By identifying and characterizing effective bacterial strains, it offers a sustainable alternative to conventional fertilizers.

Alleviation of cold stress in wheat with psychrotrophic phosphorus solubilizing Acinetobacter rhizosphaerae EU-KL44.

Low-temperature stress can seriously impair plant physiology. Chilling injury leads to a complex array of cellular dysfunctions, and symptoms include chlorosis, sterility, loss of vigor, wilting, and even death of the plants. Furthermore, phosphorus limitations additionally halt the growth of plants. Low-temperature adaptive plant growth-promoting microbes through various direct and indirect mechanisms help in the survival of plants under stress conditions. The present investigation deals with isolation of P-solubilizing psychrotrophic bacteria from diverse cultivars of wheat grown in the Keylong region of Himachal Pradesh. A total of 33 P-solubilizing bacterial isolates were obtained. P-solubilizers were screened for different plant growth-promoting (PGP) attributes of K and Zn solubilization, production of IAA, siderophores, and different hydrolytic enzymes. Among 33 P-solubilizers, 8 efficient strains exhibiting multiple PGP attributes were used as bioinoculants for wheat under low-temperature stress in different in vitro and in vivo experiments. The psychrotrophic bacterial isolates positively influenced the growth and physiological parameters as well as nutrient uptake and yield of wheat and efficiently alleviated low-temperature stress. The potential of low-temperature stress adaptive and PGP microbes can be utilized in agricultural sector for amelioration of low-temperature stress and plant growth promotion. The present study deals with the isolation of psychrotrophic P-solubilizers with multiple PGP attributes and their role in alleviation of cold stress in wheat.