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Non-rhizobial endophytes (NREs) are active colonizers inhabiting the root nodules. Though their active role in the lentil agroecosystem is not well defined, here we observed that these NREs might promote the growth of lentils, modulate rhizospheric community structure and could be used as promising organisms for optimal use of rice fallow soil. NREs from root nodules of lentils were isolated and examined for plant growth-promoting traits, exopolysaccharide (EPS) and biofilm production, root metabolites, and the presence of nifH and nifK elements. The greenhouse experiment with the chosen NREs, i.e., Serratia plymuthica 33GS and Serratia sp. R6 significantly increased the germination rate, vigour index, development of nodules (in non-sterile soil) and fresh weight of nodules (33GS 94%, R6 61% growth) and length of the shoot (33GS 86%, R6 51.16%) as well as chlorophyll levels when compared to the uninoculated control. Scanning Electron Microscopy (SEM) revealed that both isolates could successfully colonize the roots and elicit root hair growth. The inoculation of the NREs resulted in specific changes in root exudation patterns. The plants with 33GS and R6 treatment significantly stimulated the exudation of triterpenes, fatty acids, and their methyl esters in comparison to the uninoculated plants, altering the rhizospheric microbial community structure. Proteobacteria dominated the rhizospheric microbiota in all the treatments. Treatment with 33GS or R6 also enhanced the relative abundance of other favourable microbes, including Rhizobium, Mesorhizobium, and Bradyrhizobium. The correlation network analysis of relative abundances resulted in numerous bacterial taxa, which were in cooperation with each other, having a possible role in plant growth promotion. The results indicate the significant role of NREs as plant growth promoters, which also includes their role in root exudation patterns, enhancement of soil nutrient status and modulation of rhizospheric microbiota, suggesting their prospects in sustainable, and bio-based agriculture.

Background and aims Amaranth is an important underutilized crop with high nutritional value but due to acidic soils and low nutrient availability, its production is low in Uttarakhand hills. Use of inorganic chemicals is restricted as they may disturb the fragile Himalayan ecosystem. Therefore, native cold tolerant Bacillus spp. isolates were used to assess their effect on growth and yield enhancement in amaranth. Methods A repository of 120 isolates of Bacillus spp. was developed and evaluated for their cold tolerance ability and PGP (Plant Growth Promoting) traits. Among these isolates, three potential isolates (BS-27, BS-41, BS-58) were selected for the field experiments to assess their effects on plant growth, soil nutrients (N, P and K) availability, nutrient uptake and yield of amaranth. Effect of different treatments on the nutrient use efficiency of amaranth was also determined. Results Biopriming of amaranth seeds by the selected three isolates exhibited a significant enhancement in plant growth and yield under field conditions. Among the different treatments, T-8 (BS-27 + BS-58) exhibited a maximum increase in grain yield (36.94%) closely followed by T-4 (BS-27; 33.80%). A significant increase (36.18%, 32.45 and 17.11%) in soil nutrients (NPK) and their uptake (68.01%, 104.16 and 116.46%) was recorded in different treatments. Principal component analysis revealed a positive correlation between soil nutrients and their uptake by host plants. The rpoB gene sequencing and phylogenetic analysis suggested the identity of BS-27 and BS-41 as B. pumilus and BS-58 as B. subtilis. Conclusion Our study shows that plant growth promoting bacteria (PGPB) can enhance the nutrient use efficiency of the host plant that further contributes to increase the crop yield. A significant increase in crop yield suggests that regular use of such effective PGPBs by farmers may increase the soil fertility and crop yield. Ultimately, this will lead towards food security under the organic farming framework in general, and sustainable crop production in particular.

Allium hookeri is an indigenous perennial herb known for its therapeutic properties. It’s grown in the eastern Himalayas and East Asia, where it is used as a flavoring agent in local cuisines. This research aims to enhance soil phosphorus mobilization and promote A. hookeri growth using a consortium of phosphate-solubilizing bacteria (PSB). The synergistic effect of a bacterial consortium containing multiple PSBs (Arthrobacter luteolus and several Klebsiella spp.) combined with tricalcium phosphate (TCP), was investigated to enhance the growth of A. hookeri plants, and its influence on modulating the rhizosphere microbiome was also assessed. The greenhouse experiment revealed that the bacterial consortium with tricalcium phosphate (BTCP) treatment enhanced the dry shoot weight by 70%. Proteobacteria dominated the rhizosphere’s microbiome in all treatments. BTCP treatment enhanced the relative abundance of several beneficial genera such Bacillus, Mesorhizobium, Pseudomonas, Ensifer, Hyphomicrobium, Planctomyces, and Bradyrhizobium. The augmentation of bacterial consortium increased P in shoots (4.36 ± 0.63 mg/g) and in roots (2.34 ± 0.27 mg/g), which was more than 500% higher as compared to the uninoculated control. Canonical correspondence analysis (CCA) indicated significant correlations (p ≤ 0.05) between phosphorus content in the shoot, fresh weight, and dry weight, with higher relative abundances of Bacteroidetes, Cyanobacteria, and Fibrobacteres. Functional genes related to siderophore biosynthesis, ABC transporters, phosphatenate, and phosphinate metabolism exhibited positive modulation, indicating higher relative abundances associated with the BTCP treatment. The findings demonstrate the crucial contribution of the bacterial consortium in promoting plant development, improving soil nutrient levels, and influencing the rhizospheric microbiota, implying its significance in sustainable agriculture.

Bacterial isolates having antifungal and good plant growth-promoting attributes were isolated from chir-pine (Pinus roxburghii) rhizosphere. An isolate, Bacillus subtilis BN1 exhibited strong antagonistic activity against Macrophomina phaseolina, and other phytopathogens including Fusarium oxysporum and Rhizoctonia solani. It was characterized and selected for the present studies. BN1 resulted in vacuolation, hyphal squeezing, swelling, abnormal branching and lysis of mycelia. The cell-free culture filtrate of BN1 inhibited the growth of M. phaseolina. Pot trial study resulted in statistically significant increase in seedling biomass besides reduction in root rot symptoms in chir-pine seedlings. BN1 treatment resulted in 43.6% and 93.54% increases in root and shoot dry weights respectively, as compared to control. Also, 80-85% seed viability was recorded in treatments receiving BN1 either alone or in the presence of M. phaseolina, compared to 54.5% with M. phaseolina. Bioinoculant formulation study suggested that maximum viability of bacteria was in a sawdust-based carrier. B. subtilis BN1 produced lytic enzymes, chitinase and β-1,3-glucanase, which are known to cause hyphal degradation and digestion of the cell wall component of M. phaseolina. In the presence of M. phaseolina, population of B1 was 1.5 x 10⁴ c.f.u. g⁻¹ root after one month, which increased to 4.5 x 10⁴ c.f.u. g⁻¹ root in three months. Positive root colonization capability of B. subtilis BN1 proved it as a potent biocontrol agent.

The ability of fluorescent Pseudomonas strain EKi, in production of biocontrol and plant growth promotory (PGP) metabolites under saline stress was evaluated. Strain EKi could tolerate NaCl up to 1,550 mM and showed biocontrol of Macrophomina phaseolina (76.19%) in the presence of up to 400 mM NaCl. Strain EKi was able to produce IAA, siderophore and pyocyanin with gradual reduction of up to 76.31, 45.46, and 48.99%, respectively, as NaCl concentration increased from 0 to 500 mM. Reduced growth rate resulted in delayed induction of IAA, siderophore and pyocyanin by the PGPR. Thin layer chromatography of chloroform extract from non-stressed and salt stressed EKi, and inhibition of M. phaseolina by purified pyocyanin clearly indicated its role in biocontrol. In vitro and in vivo results showed the growth promotion and charcoal rot disease suppression of chickpea by strain EKi under both non-stressed and saline stress. There was 76.75 and 65.25% reduction of disease incidence in non-saline and saline conditions, respectively, in vitro conditions. In presence of M. phaseolina strain EKi brought about 67.65 and 58.45% reduction of disease incidence in non-saline and saline soil, respectively.

Most of the legume crops are affected by metal stress present in the soil mainly due to contaminated agrochemicals and sewage sludge. The effect of aluminium, and heavy metals copper, iron and molybdenum on growth and activity of enzymes of fast and slow growing rhizobial sps. was studied. Sinorhizobium meliloti RMP₅ was found to be more tolerant to metal stress than Bradyrhizobium BMP₁. Both the strains were extremely sensitive to Al than other metals. Al was much more deleterious for the enzymatic activities (nitrate reduction, nitrite reduction, nitrogenase and uptake hydrogenase) of strain RMP₅ and BMP₁. Cu showed inhibitory effect on growth and enzyme activities of Bradyrhizobium strain at all concentrations. However, in S. meliloti RMP₅ all the tested enzymatic activities increased up to the concentration of 0.1 mM Cu. Fe enhanced the growth and enzyme activities of S. meliloti RMP₅ and Bradyrhizobium BMP₁ up to 100 mM concentration. Mo enhanced all the tested enzymatic activities of S. meliloti RMP₅ up to 1 mM. Nitrate and nitrite reduction activities of Bradyrhizobium BMP₁ increased up to 1 mM concentration. However, nitrogenase and hydrogenase activities of Bradyrhizobium BMP₁ got enhanced only up to 0.5 mM Mo. Both Fe and Mo are the key components of the enzyme nitrogenase and nitrate reductase and enhanced the growth and enzyme activities of both the sps. The study of physiology of nitrogen fixing ability of both fast and slow growing rhizobial strains reported that the supplementation of Mo and Fe in soils along with the biological formulations will enhance the process of symbiotic nitrogen fixation.

The present work was undertaken to formulate an effective bioformulation using Burkholderia sp. strain MSSP, a known plant-growth-promoting rhizobacterium. MSSP was tagged with the reporter gene of green fluorescent protein (gfp) to monitor its population in cost-effective solid carriers, including sugarcane-bagasse, sawdust, cocoa peat, rice husk, wheat bran, charcoal, and rock phosphate, and paneer-whey as liquid carrier. Physical and chemical properties of different low-cost carrier materials were studied. The viability of the green fluorescent tagged variant of MSSP was estimated in different sterile carrier materials. Whey and wheat bran proved to be efficient carrier materials for the bioformulation. Sawdust, rock phosphate, rice husk, and cocoa peat were average, while charcoal and sugarcane-bagasse proved to be inferior carriers. The viability of strain MSSP was also assessed in wheat bran and whey-based consortium, having three other bacterial strains, namely Sinorhizobium meliloti PP3, Rhizobium leguminosarum Pcc, and Bacillus sp. strain B1. Presence of other plant-growth-promoting bacteria did not have any detrimental effect on the viability of MSSP. Efficiency of the wheat-bran-based multispecies consortium was studied on the growth of pigeonpea in field conditions. A considerable increase in plant biomass, nodule number and weight, and number of pods was recorded as compared with individual trials and with the control.

PGPR strain Pseudomonas fluorescens PS1 was evaluated to formulate carrier based bioformulations. The viability of P. fluorescens PS1 was monitored at different time intervals during the period of storage at room temperature in different carriers such as soil, charcoal, sawdust and sawdust-soil. Sawdust-soil was found to be the most efficient carrier material for P. fluorescens PS1 followed by other carriers. After 1 year of storage, P. fluorescens PS1 was re-isolated and assayed for its antifungal activity against Sclerotinia sclerotiorum a phytopathogenic fungus causing stem blight in Indian mustard, Brassica campestris . Results of scanning electron microscopy exhibited that P. fluorescens PS1 caused morphological alteration in mycelia of S. sclerotiorum as evident by hyphal perforation, and fragmented lysis. Seed bacterization of B. campestris with P. fluorescens PS1 induced enhanced seed germination, increased overall plant growth as well as reduced stem blight in mustard with improved yield. These findings demonstrate that P. fluorescens PS1 has significant potential to raise disease-free crops due to the presence of a wide array of PGP characteristics.

Endophytic bacteria reside within plant tissues and have often been reported to promote plant growth. Rhizobia are particularly known for their symbiotic relationship with legumes. A bacterial strain MSSP was isolated from surface-sterilized root nodules of Mimosa pudica. MSSP was Gram-negative, capsulated, motile, non-endospore forming rod with free nitrogen (N) fixation ability. Unlike N-fixing bacteria forming symbiotic relationship with legumes that largely exist in α-subclass of proteobacteria, MSSP belongs to β-class of proteobacteria. Phylogenetic analysis of 16 S rDNA demonstrated that MSSP belongs to the genus Burkholderia. This isolate secretes phytohormone, ACC deaminase, solubilizes phosphate and is antagonistic against phytopathogens.

Pseudomonas aeruginosa GRC1 exhibited strong antagonistic activity against Sclerotinia sclerotiorum, in vitro and in vivo. Scanning electron microscopic (SEM) studies showed morphological abnormalities such as perforation, lysis and fragmentation of hyphae of S. sclerotiorum caused by P. aeruginosa GRC1. This strain produced extracellular chitinase enzyme, the role of which was clearly demonstrated through Tn5 mutagenesis. Bacterization of peanut seeds with GRC1 resulted in increased seed germination and reduced stem-rot of peanut in S. sclerotiorum-infested soil by 97%. Other vegetative and yield plant parameters such as nodules per plant, pods and grain yield per plant were enhanced with a statistical significance in comparison to control. Neomycin resistant (GRC1neo+) bacterium was a good root colonizer and frequently isolated from rhizosphere of peanut plants. These findings showed P. aeruginosa GRC1 as a potential biocontrol agent against S. sclerotiorum.