Non-rhizobial nodule endophytes improve nodulation, change root exudation pattern and promote the growth of lentil, for prospective application in fallow soil.

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.

Treatment with atypical rhizobia, Pararhizobium giardinii and Ochrobactrum sp. modulate the rhizospheric bacterial community, and enhances Lens culinaris growth in fallow-soil.

Diazotrophic nodule isolates are acknowledged promoters of plant growth and rhizospheric community. Consequently, in the lentil agroecosystem, inoculation of atypical rhizobial isolates could be a viable alternative to chemical fertilizers for fallow land usage optimization. The aim of this study is to evaluate and select the rhizobial isolates of lentil nodules with plant-growth-promoting (PGP) attributes and to elucidate their application in rice-fallow soil for determining the growth of lentils and its impact on the rhizospheric bacterial community. Lentil's nodule isolates were identified and screened for their PGP attributes, biofilm, exopolysaccharide (EPS) formation, and early plant growth promotion. The pot experiment with the selected atypical rhizobial isolates Pararhizobium giardinii (P1) and Ochrobactrum sp. (42S) significantly enhanced germination, vigour index, nodule formation (P1 60%, 42S 42% increase), nodule fresh weight, shoot length (65% P1 & 35% 42S), and chlorophyll content as compared to the uninoculated control treatment. The genes for nitrogen fixation nifH and nifK were detected in both isolates. Scanning Electron Microscopy (SEM) revealed successful root and nodule colonization by both isolates, while Transmission Electron Microscopy (TEM) displayed nitrogen-fixing zones within root nodules. Proteobacteria predominated in the lentil rhizosphere of all the treatments. Whereas, application of either P1 or 42S increased Rhizobium, Mesorhizobium, and Bradyrhizobium genra, thus positively modulating rhizospheric community structure. The correlation network analysis revealed an abundance of some interdependent bacterial genera with a possible role in overall plant growth. Functional genes for siderophore biosynthesis and ABC transporter were positively modulated by application of either P1 or 42S. This study showed the significant effect of P. giardinii P1 and Ochrobactrum sp. 42S of L. culinaris on lentil growth, improving fallowsoil health for optimum usage, and modulated rhizospheric community structure which strongly manifest prospects of low-cost, eco-friendly and sustainable biofertilizers.

Effect of plant growth promoting Bacillus spp. on nutritional properties of Amaranthus hypochondriacus grains.

Amaranth (Amaranthus hypochondriacus Linn.) is an important pseudocereal crop having important nutrients along with the indispensable amino-acids. The present study was aimed to study the effect of plant growth promoting bacilli on proximate constituents of amaranth grains, including three of the essential amino acids (methionine, lysine and, tryptophan). The combination of Bacillus pumilus and Bacillus subtilis showed a significant increase in different proximate constituents, including crude protein (22.13%), dry matter (32.25%), fat (30.77%), and carbohydrate (49.08%) in amaranth grains. Similarly, a significant increase in essential amino-acids (methionine 47.68%, lysine 59.41% and, tryptophan 38.05%) was recorded. This study suggests that the combination of Bacillus pumilus BS-27 and Bacillus subtilis BS-58 provides the natural, persistent and durable potential to enhance the nutritive value of the crop. Therefore, present study was designed to explore the enhancement of most desirable amino acid synthesis in amaranth due to application of plant growth promoting Bacillus spp.

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.

Diverse role of fast growing rhizobia in growth promotion and enhancement of psoralen content in Psoralea corylifolia L.

BACKGROUND: Psoralea corylifolia (Bakuchi), a weed, which possesses a highly potent and medicinally important compound psoralen. P. corylifolia has been widely exploited since ages for its biological potential. MATERIALS AND METHODS: Fifteen root nodulating bacteria as pure culture collection (PCC) were isolated from P. corylifolia in India. Further, these strains were evaluated for their effect on the psoralen content in P. corylifolia. High performance liquid chromatography (HPLC) method was used for the estimation of psoralen in P. corylifolia seed extracts. The effectiveness of these rhizobial strains was assessed on the basis of screening of various plant growth promoting attributes. RESULTS: The 16S ribosomal RNA sequencing analysis revealed the identity of two most effective rhizobial isolates PCC2 and PCC7 as Rhizobium leguminosarum and Sinorhizobium meliloti, respectively. The R. leguminosarum PCC2 (JN546144) and Ensifer meliloti PCC7 (JN546145) strains showed solubilization of insoluble inorganic phosphate, secreted indole acetic acid (IAA), produced siderophore, showed ACC deaminase activity, and were positive for nodulation and nitrogen fixing genes. Seeds of P. corylifolia were bacterized with combination of R. leguminosarum PCC2 and Ensifer meliloti PCC7 along with their individual application that resulted in enhancement of various early vegetative and late reproduction parameters of plants in two consecutive field trials in the year 2009 and 2010. The psoralen content in the seeds of P. corylifolia was observed to be increased in the field trials where the combination of rhizobial strains PCC2 and PCC7 was used (2.79%) compared to control (1.91%). CONCLUSION: These findings indicate that rhizobial strains PCC2 and PCC7 showing good plant growth promoting attributes can be effective for increasing the psoralen content in the seeds of P. corylifolia to a certain level.

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.

Bacillus strains isolated from rhizosphere showed plant growth promoting and antagonistic activity against phytopathogens.

Seven bacterial isolates screened from rhizosphere of common bean growing at Uttarakhand Himalaya showed potential plant growth promoting (PGP) and antagonistic activities. Based on 16S rRNA gene sequence the isolate BPR7 was identified as Bacillus sp. BPR7. The strain BPR7 produced IAA, siderophore, phytase, organic acid, ACC deaminase, cyanogens, lytic enzymes, oxalate oxidase, and solubilized various sources of organic and inorganic phosphates as well as potassium and zinc. Strain BPR7 strongly inhibited the growth of several phytopathogens such as Macrophomina phaseolina, Fusarium oxysporum, F. solani, Sclerotinia sclerotiorum, Rhizoctonia solani and Colletotricum sp. in vitro. Cell-free culture filtrate of strain BPR7 also caused colony growth inhibition of all test pathogens. PGP and antifungal activities of Bacillus sp. BPR7 suggest that it may be exploited as a potential bioinoculant agent for P. vulgaris.

Transformation of pWWO in Rhizobium leguminosarum DPT to Engineer Toluene Degrading Ability for Rhizoremediation.

Rhizoremediation of organic xenobiotics is based on interactions between plants and their associated micro-organisms. The present work was designed to engineer a bacterial system having toluene degradation ability along with plant growth promoting characteristics for effective rhizoremediation. pWWO harboring the genes responsible for toluene breakdown was isolated from Pseudomonas putida MTCC 979 and successfully transformed in Rhizobium DPT. This resulted in a bacterial strain (DPT(T)) which had the ability to degrade toluene as well as enhance growth of host plant. The frequency of transformation was recorded 5.7 × 10(-6). DPT produced IAA, siderophore, chitinase, HCN, ACC deaminase, solubilized inorganic phosphate, fixed atmospheric nitrogen and inhibited the growth of Fusarium oxysporum and Macrophomina phaseolina in vitro. During pot assay, 50 ppm toluene in soil was found to inhibit the germination of Cajanus cajan seeds. However when the seeds bacterized with toluene degrading P. putida or R. leguminosarum DPT were sown in pots, again no germination was observed. Non-bacterized as well as bacterized seeds germinated successfully in toluene free soil as control. The results forced for an alternative mode of application of bacteria for rhizoremediation purpose. Hence bacterial suspension was mixed with soil having 50 ppm of toluene. Germination index in DPT treated soil was 100% while in P. putida it was 50%. Untreated soil with toluene restricted the seeds to germinate.

Suppression of charcoal rot of chickpea by fluorescent Pseudomonas under saline stress condition.

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.

Process of rock phosphate solubilization by Aspergillus sp PS 104 in soil amended medium.

Aspergillus sp PS 104, a soil isolate had excellent potential to solubilize rock phosphate in vitro. The process was influenced by the presence of various concentrations of local loess (red soil). The simultaneous occurrence, in our experiment, of high levels of solubilized phosphate and synthesized citric acid, together with the lowest reached pH values, confirmed the role of citric acid in the phosphate solubilization mechanism. When the soil was present, phosphate release was better correlated than citrate synthesis with H+ concentration. Changes in soluble phosphate concentration did not follow a sigmoid pattern. The ability of organism to release phosphatase was also studied. An interesting relationship was observed between the two processes of phosphate mobilization: citric acid synthesis and phosphatase production.

Bioformulation of Burkholderia sp. MSSP with a multispecies consortium for growth promotion of Cajanus cajan.

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.

Antagonistic effect of fluorescent pseudomonads against Macrophomina phaseolina that causes charcoal rot of groundnut.

Maximum colony growth inhibition was observed due to Pseudomonas PS2 (74%) as compared to PS1 (71%) on trypticase soy agar (TSM) plates after 5 days of incubation. Light and scanning electron microscopic examination showed hyphal coiling, vacuolation, coagulation and granulation of cytoplasm resulting in lysis of hyphae of M. phaseolina by pseudomonads. Cell free culture filtrates of strains PS1 and PS2 restricted the growth of mycelium of M. phaseolina. PS1 and PS2 caused maximum colony growth inhibition by 57 and 61% respectively at 20% concentration of culture filtrate after 4 days of incubation. Volatile substances produced by PS1 and PS2 also inhibited the colony growth of M. phaseolina by 25 and 32%, respectively. Inhibitory effect of volatile substances, however, decreased with advancing in incubation period. Colony growth of M. phaseolina was significantly decreased by PS1 and PS2 as compared to control both in iron- sufficient and iron-deficient conditions. PS2 showed higher antagonistic activity than PS1, as evidenced by pronounced colony growth inhibition.

Rhizobia as a biological control agent against soil borne plant pathogenic fungi.

Rhizobia promote the growth of plants either directly through N2 fixation, supply of nutrients, synthesis of phytohormones and solubilization of minerals, or indirectly as a biocontrol agent by inhibiting the growth of pathogens. The biocontrol effect of rhizobia is due to the secretion of secondary metabolites such as antibiotics and HCN. Siderophore production in iron stress conditions provides rhizobia an added advantage, resulting in exclusion of pathogens due to iron starvation.

Effect of metal ions on growth of Pseudomonas aeruginosa and siderophore and protein production.

Pseudomonas aeruginosa (GRC1) isolated from potato rhizosphere, grew better on succinate medium than tryptic soy medium and produced hydroxamate type of siderophore in iron-deficient succinate medium. When the strain GRC1 was grown in the presence of different metal ion compounds, viz. ZnSO4, MnSO4, MnCl2 and FeCl3 at 6 and 12 microM concentrations individually, ZnSO4 (12 microM) promoted siderophore production but suppressed the growth and protein content of test organism. MnCl2 and FeCl3 (12 microM) enhanced the growth, whereas MnCl2 and MnSO4 (12 microM) induced protein contents of strain GRC1.

Pseudomonas aeruginosa (GRC1) as a strong antagonist of Macrophomina phaseolina and Fusarium oxysporum.

A plant growth promotory bacterial strain, isolated from the potato rhizosphere, was characterized as Pseudomonas aeruginosa (GRC1). The isolate produced an hydroxamate type of siderophore after 48 h of incubation on tryptic soy medium under iron deficient conditions. The in vitro antifungal activity of P. aeruginosa was tested against two soil-borne plant pathogens, Macrophomina phaseolina and Fusarium oxysporum. The antagonistic behaviour of the isolate was tested by dual culture technique. The growth inhibition of M. phaseolina and F. oxysporum was 74.1% and 70.5%, respectively, after 5 days of incubation. The production of hydrocyanic acid and indole acetic acid was also recorded under normal growth conditions.

Microbial degradation of aquatic biomass by Trichoderma viride 992 and Aspergillus wentii 669 with reference to the physical structure.

There was a considerable change in the structural parameters of both the lignocellulosic substrates. This change was confirmed by infrared (IR) spectroscopy and X-ray diffraction data, which revealed the presence of phenolic groups in the peaks range 1310-1410 and 1200 cm-1 with large crystalline substrates. These were found to be reduced after treatment with alkali. Enzyme/s activity were found to be increased after pretreatment of hydrolysed substrates. The structural modifications in both the substrates were further evidenced by the results obtained after enzymatic hydrolysis of alkali-treated substrates.