Next-generation biofertilizers and novel biostimulants: documentation and validation of mechanism of endophytic plant growth-promoting rhizobacteria in tomato.

A study was conducted to determine the suitability of the endophytes as probable next-generation biofertilizers and novel biostimulants. Enterobacter turicensis RCT5 and Stenotrophomonas maltophilia RCT31 showed a zone of solubilization, of phosphate, potassium, silicate, and zinc, produced phytase. Among the three media used for phosphate solubilization, the rhizospheric medium turned out to be the best-producing results in less than 24 h, while others took a longer time to give the same results. The strains exhibited differential ability to produce organic acids in the plate assay and eight of these were profuse producers of exopolysaccharides. We were able to partially elucidate the mechanism of solubilization of insoluble salts that included organic acids and protein activity in the cell-free culture filtrates of endophytes. All the root nodule endophytes showed potential as novel biostimulants and next-generation biofertilizers as found in the germination assay of tomato, a non-host crop using different methodologies. It proved that the endophytes have different mechanism of expressions of their plant growth-promoting traits as well as can promote the growth of tomato plant irrespective of the method used.

Seed bio-priming with tri-species consortia of phosphate solubilizing rhizobacteria (PSR) and its effect on plant growth promotion.

Three potential rhizobacteria namely Burkholderia gladioli (MTCC 10216), Pseudomonas sp. (MTCC 9002) and Bacillus subtilis (MTCC 8528) procured from IMTECH, Chandigarh (India) were evaluated individually and as consortia for its phosphate (P) solubilizing ability and effect of growth of fenugreek (Trigonella foenum-graecum L.) and tomato (Lycopersicon esculentum L.). Phosphate solubilizing ability of these strains individually and as consortia was tested on Pikovskayas agar medium, Phosphate solubilizing agar medium and National Botanical Research Institute phosphate agar medium containing six different sources of insoluble inorganic phosphate such as tri-calcium phosphate (TCP), di-calcium phosphate (DCP), zinc phosphate (ZP), ferric phosphate (FP), sodium di-hydrogen phosphate (SP), and aluminum phosphate (AP), and two organic P such as calcium and sodium phytate. The maximum P solubilizing ability was recorded in consortium-4 having all three potential bacterial strains. Phosphate solubilization after 7th day of incubation was 37.9 mg/100 ml of TCP, 40.01 mg/100 ml of DCP, 15.79 mg/100 ml of FP, 43.02 mg/100 ml of SP, no solubilization of ZP and AP, 39.75 mg/100 ml of calcium phytate and 24.01mg/100 ml of sodium phytate. Seed germination and the other plant parameters such as plant height and weight significantly increased in fenugreek and tomato seeds, bio-primed with consortium-4 followed by consortium-3. After bio-priming of seeds in pot assay, the level of phosphorus in soil got increased by 54% in consortium-4 treated soil followed by consortium-3 (47%) over untreated control soil. Based on these findings, consoritium-4 could be recommended as a good bio-inoculant for fenugreek, tomato and other crops in comparison to individual strains and other consortia.

Characterization of a plant-growth-promoting non-nodulating endophytic bacterium (Stenotrophomonas maltophilia) from the root nodules of Mucuna utilis var. capitata L. (Safed Kaunch).

Nonrhizobial root nodule endophytic bacteria are known to have beneficial effects on host plants and are also considered contaminants or opportunists. They grow either individually or as a co-occupant of the root nodules of legumes. In this study, a nonrhizobial endophytic bacterial strain was isolated from the root nodules of the medicinal legume Mucuna utilis var. capitata L.; phenotypic, genotypic, and agricultural characterization was performed using a HiMedia kit and 16S rRNA sequencing. This strain showed tremendous seedling growth potential (30%), compared with the control, as well as a strong antagonistic nature against the plant pathogenic fungus Fusarium udum when plant growth parameters were analyzed. The strain, identified by 16S rRNA as Stenotrophomonas maltophilia, showed a multitude of plant-growth-promoting attributes both direct (IAA, phosphate solubilization) and indirect (ACC deaminase, siderophore) and enhanced the growth of host plant in field trials. This is the first report of the plant-growth-promoting potential of this endophytic bacterium from the nodules of M. utilis var. capitata L.; hence, it has potential for use in various biotechnological applications in various industries.

Endophytic bacteria promote growth of the medicinal legume Clitoria ternatea L. by chemotactic activity.

Part of the native root nodule endophytic microflora referring to members of the genera Proteobacteria and Sphingobacteria were used to test their bioefficacy as seed biopriming. These were quantified for their plant growth promoting (PGP) attributes such as IAA production, P and K-solubilization and ACC deaminase production. Results showed that significantly highest IAA was produced by E. hormaechi RCT10. The highest P-solubilization was observed with S. maltophila RCT31 it was solubilizing all the substrate tri-calcium phosphate, di-calcium phosphate, and zinc phosphate. Significantly highest K-solubilization was observed with S. maltophila RCT31 followed by E. turicensis RCT5. However, the maximum zinc solubilization was reported with S. maltophila RCT31 followed by E. turicensis RCT5. The maximum ACC deaminase was quantified in the bacterium. Results revealed that the E. hormaechi RCT10 utilized seed leechates most effectively while root exudates were maximally utilized by S. maltophila RCT31. The pots experiment proves that S. maltophila RCT31 was the most effective bacterium and it was replication vis-à-vis field experiment. In particular, S. maltophila RCT31 holds strong potential to be possibly used as a bioformulation for the medicinal legume, as an economical and eco-friendly alternative to agrochemicals.

Revisiting the plant growth-promoting rhizobacteria: lessons from the past and objectives for the future.

Plant beneficial rhizobacteria (PBR) is a group of naturally occurring rhizospheric microbes that enhance nutrient availability and induce biotic and abiotic stress tolerance through a wide array of mechanisms to enhance agricultural sustainability. Application of PBR has the potential to reduce worldwide requirement of agricultural chemicals and improve agro-ecological sustainability. The PBR exert their beneficial effects in three major ways; (1) fix atmospheric nitrogen and synthesize specific compounds to promote plant growth, (2) solubilize essential mineral nutrients in soils for plant uptake, and (3) produce antimicrobial substances and induce systemic resistance in host plants to protect them from biotic and abiotic stresses. Application of PBR as suitable inoculants appears to be a viable alternative technology to synthetic fertilizers and pesticides. Furthermore, PBR enhance nutrient and water use efficiency, influence dynamics of mineral recycling, and tolerance of plants to other environmental stresses by improving health of soils. This report provides comprehensive reviews and discusses beneficial effects of PBR on plant and soil health. Considering their multitude of functions to improve plant and soil health, we propose to call the plant growth-promoting bacteria (PGPR) as PBR.

Ghost probiotics with a combined regimen: a novel therapeutic approach against the Zika virus, an emerging world threat.

The Zika virus (ZIKV) used to be an obscure flavivirus closely related to dengue virus (DENV). Transmission of this epidemic pathogen occurs mainly via mosquitoes, but it is also capable of placental and sexual transmission. Although the characteristics of these viruses are well defined, infections are unpredictable in terms of disease severity, unusual clinical manifestations, unexpected methods of transmission, long-term persistence, and the development of new strains. Recently, ZIKV has gained huge medical attention following the large-scale epidemics around the world, and reported cases of congenital abnormalities associated with Zika virus infections which have created a public health emergency of international concern. Despite continuous research on ZIKV, no specific treatment or vaccine has been developed, excepting a preventive strategy for congenital ZIKV infection. Probiotics, known as GRAS, are bacteria that confer various health beneficial effects, and have been shown to be effective at curing a number of viral diseases by modulating the immune system. Furthermore, probiotic preparations consisting of dead cells and cellular metabolites, so-called "Ghost probiotics", can also act as biological response modifiers. Here, we review available information on the epidemiology, transmission, and clinical features of ZIKV, and on treatment and prevention strategies. In addition, we emphasize the use of probiotics and plant-based natural remedies and describe their action mechanisms, and the green technologies for microbial conversion, which could contribute to the development of novel therapies that may reduce the pathogenicity of ZIKV. Accordingly, we draw attention to new findings, unanswered questions, unresolved issues, and controversies regarding ZIKV.

Root nodule bacteria from Clitoria ternatea L. are putative invasive nonrhizobial endophytes.

In this study, bacteria (8 species and 5 genera) belonging to the classes Betaproteobacteria, Gammaproteobacteria, and Sphingobacteria were isolated from root nodules of the multipurpose legume Clitoria ternatea L. and identified on the basis of partial 16S rRNA sequencing. The root nodule bacteria were subjected to phenotypic clustering and diversity studies using biochemical kits, including Hi-Media Carbokit™, Enterobacteriaceae™ identification kit, ERIC-PCR, and 16S ARDRA. All the strains showed growth on Ashby's N-free media over 7 generations, indicative of presumptive nitrogen fixation and further confirmed by amplification of the nifH gene. None of the strains showed the capability to renodulate the host plant, neither alone nor in combination with standard rhizobial strains, which was further confirmed by the absence of nodC bands in PCR assay. The results clearly indicate the common existence of nonrhizobial microflora inside the root nodules of legumes, which were thought to be colonized only by rhizobia and were responsible for N2 fixation in leguminous crops. However, with the recent discovery of nodule endophytes from a variety of legumes, as also observed here, it can be assumed that symbiotic rhizobia are not all alone and that these invasive endophytes belonging to various bacterial genera are more than just opportunistic colonizers of specialized nodule niche.

Integrated approach for disease management and growth enhancement of Sesamum indicum L. utilizing Azotobacter chroococcum TRA2 and chemical fertilizer.

Azotobacter chroococcum TRA2, an isolate of wheat rhizosphere displayed plant growth promoting attributes including indole acetic acid, HCN, siderophore production, solubilization of inorganic phosphate and fixation of atmospheric nitrogen. In addition, it showed strong antagonistic effect against Macrophomina phaseolina and Fusarium oxysporum. It also caused degradation and digestion of cell wall components, resulting in hyphal perforations, empty cell (halo) formation, shrinking and lysis of fungal mycelia along with significant degeneration of conidia. Fertilizer adaptive variant strain of A. chroococcum TRA2 was studied with Tn5 induced streptomycin resistant transconjugants of wild type tetracycline-resistant TRA2 (designated TRA2(tetra+strep+)) after different durations. The strain was significantly competent in rhizosphere, as its population increased by 15.29 % in rhizosphere of Sesamum indicum. Seed bacterization with the strain TRA2 resulted in significant increase in vegetative growth parameters and yield of sesame over the non-bacterized seeds. However, application of TRA2 with half dose of fertilizers showed sesame yield almost similar to that obtained by full dose treatment. Moreover, the oil yield increased by 24.20 %, while protein yield increased by 35.92 % in treatment receiving half dose of fertilizer along with TRA2 bacterized seeds, as compared to untreated control.

Practical use of CMC-amended rhizobial inoculant for Mucuna pruriens cultivation to enhance the growth and protection against Macrophomina phaseolina.

In many parts of the world Mucuna pruriens is used as an important medicinal, forage and green manure crop. In the present investigation the effect of the addition of CMC in carrier during development of bioformulation on shelflife, plant growth promotive and biocontrol activity against Macrophomina phaseolina was screened taking M. pruriens as a test crop. Ensifer meliloti RMP6(Ery+Kan+) and Bradyrhizobium sp. BMP7(Tet+Kan+) (kanamycin resistance engineered by Tn5 transposon mutagenesis) used in the study showed production of siderophore, IAA, solubilizing phosphate and biocontrol of M. phaseolina. RMP6(Ery+Kan+) also showed ACC deaminase activity. The survival of both the strains in sawdust-based bioformulation was enhanced with an increase in the concentration of CMC from 0 to 1%. At 0% CMC Bradyrhizobium sp. BMP7(Tet+Kan+) showed more increase in nodule number/plant (500.00%) than E. meliloti RMP6(Ery+Kan+) (52.38%), over the control in M. phaseolina-infested soil. There was 185.94% and 59.52% enhancement in nodule number/plant by RMP6(Ery+Kan+) and BMP7(Tet+Kan+) with an increase in the concentration of CMC from 0% to 1% in the bioformulations. However further increase in concentration of CMC did not result in enhancement in survival of either the strains or nodule number/plant.