Evaluation of inorganic phosphate solubilizing efficiency and multiple plant growth promoting properties of endophytic bacteria isolated from root nodules Erythrina brucei.

BACKGROUND: In soils, phosphorous (P) mostly exists in fixed/insoluble form and unavailable for plants use in soil solution, hence it is in scarcity. P is fixed in the form of aluminium, iron and manganese phosphates in acidic soils and calcium phosphate in alkaline soils. Phosphate solubilizing bacteria, the ecological engineers play a pivotal role in the mobilization of fixed forms of P by using different mechanisms. The objectives of this study were to evaluate inorganic phosphate solubilizing efficiency and other multiple plant growth promoting traits of Erythrina brucei root nodule endophytic bacteria and to investigate effects of the selected endophytic bacteria on the growth of wheat plant under phosphorous deficient sand culture at greenhouse conditions. RESULTS: Among a total of 304 passenger endophytic bacteria, 119 (39%) exhibited tricalcium phosphate (TCP) solubilization; however, none of them were formed clear halos on solid medium supplemented with aluminum phosphate (Al-P) or iron phosphate (Fe-P). Among 119 isolates, 40% exhibited IAA production. The selected nine potential isolates also exhibited potentials of IAA, HCN, NH3 and/or hydrolytic enzymes production. All the selected isolates were potential solubilizers of the three inorganic phosphates (Al-P, Fe-P and TCP) included in liquid medium. The highest values of solubilized TCP were recorded by isolates AU4 and RG6 (A. soli), 108.96 mg L-1 and 107.48 mg L-1, respectively at sampling day3 and 120.36 mg L-1 and 112.82 mg L-1, respectively at day 6. The highest values of solubilized Al-P and Fe-P were recorded by isolate RG6, 102.14 mg L-1 and 96.07 mg L-1, respectively at sampling days 3 and 6, respectively. The highest IAA, 313.61 µg mL-1 was recorded by isolate DM17 (Bacillus thuringiensis). Inoculation of wheat with AU4, RG6 and RG5 (Acinetobacter soli) increased shoot length by 11, 17.4 and 14.6%, respectively compared to the negative control. Similarly, 76.9, 69.2 and 53.8% increment in shoot dry weight is recorded by inoculation with RG6, AU4 and RG5, respectively. These nine potential endophytic isolates are identified to Gluconobacter cerinus (4), Acinetobacter soli (3), Achromobacter xylosoxidans (1) and Bacillus thuringiensis (1). CONCLUSION: AU4, RG6 and RG5 can be potential bio-inoculants candidates as low cost agricultural inputs in acidic and/or alkaline soils for sustainable crop production.

Recent insights into oceanic dimethylsulfoniopropionate biosynthesis and catabolism.

Dimethylsulfoniopropionate (DMSP), a globally important organosulfur compound is produced in prodigious amounts (2.0 Pg sulfur) annually in the marine environment by phytoplankton, macroalgae, heterotrophic bacteria, some corals and certain higher plants. It is an important marine osmolyte and a major precursor molecule for the production of climate-active volatile gas dimethyl sulfide (DMS). DMSP synthesis take place via three pathways: a transamination 'pathway-' in some marine bacteria and algae, a Met-methylation 'pathway-' in angiosperms and bacteria and a decarboxylation 'pathway-' in the dinoflagellate, Crypthecodinium. The enzymes DSYB and TpMMT are involved in the DMSP biosynthesis in eukaryotes while marine heterotrophic bacteria engage key enzymes such as DsyB and MmtN. Several marine bacterial communities import DMSP and degrade it via cleavage or demethylation pathways or oxidation pathway, thereby generating DMS, methanethiol, and dimethylsulfoxonium propionate, respectively. DMSP is cleaved through diverse DMSP lyase enzymes in bacteria and via Alma1 enzyme in phytoplankton. The demethylation pathway involves four different enzymes, namely DmdA, DmdB, DmdC and DmdD/AcuH. However, enzymes involved in the oxidation pathway have not been yet identified. We reviewed the recent advances on the synthesis and catabolism of DMSP and enzymes that are involved in these processes.

Detection of Diverse N-Acyl Homoserine Lactone Signalling Molecules Among Bacteria Associated with Rice Rhizosphere.

Bacterial communities communicate, regulate and coordinate their cooperative activities and physiological process by releasing, sensing and responding to small diffusible signal molecules such as acyl homoserine lactones (AHLs), auto-inducing oligo-peptides (AIPs) and autoinducer-2, a process referred to as Quorum sensing (QS). The QS mediated communication in rhizosphere associated bacterial communities significantly influence traits governing plant-microbe interactions. This study aimed to identify AHL-mediated QS signals in bacterial communities associated with rice rhizosphere using two AHL biosensors reporter strains Chromobacterium violaceum CV026 and Agrobacterium tumefaciens NTL4 (pZLR4). Approximately 375 bacterial isolates isolated from rice rhizosphere and screened using both the biosensors, detected 49 (13%) AHL positive isolates. The BOX-Polymerase Chain reaction (BOX-PCR) fingerprinting profiles of the 49 AHL positive isolates represented 11 distinct cluster groups. Subsequent 16S rRNA gene sequence analysis identified 11 different species affiliated to two different phyla; predominantly γ-proteobacteria, representing 5 genera and 1 genus in α-proteobacteria. Thin-layer chromatography (TLC) analysis detected diverse AHL profiles among the 11 AHL positive isolates with both substituted and unsubstituted acyl side chains of C4, C6 and C8 carbon. Further, AHL production in Acinetobacter lactucae, Aeromonas popoffii, Serratia oryzae, and Rhizobium wuzhouense is being reported for the first time. Detection of diverse AHLs from different groups of rhizobacteria associated with rice indicates that these signalling molecules may be involved in the regulation of rhizobacterial behaviour and symbiotic plant-microbe interactions. Future research on the role of AHLs in trans-kingdom communication particularly plant-microbe interaction using synthetic microbial community will enable in evaluating and developing potential plant specific bioproducts.

Kinetics of Phenol Biodegradation by Heavy Metal Tolerant Rhizobacteria Glutamicibacter nicotianae MSSRFPD35 From Distillery Effluent Contaminated Soils.

Biodegradation of phenol using bacteria is recognized as an efficient, environmentally friendly and cost-effective approach for reducing phenol pollutants compared to the current conventional physicochemical processes adopted. A potential phenol degrading bacterial strain Glutamicibacter nicotianae MSSRFPD35 was isolated and identified from Canna indica rhizosphere grown in distillery effluent contaminated sites. It showed high phenol degrading efficiency up to 1117 mg L-1 within 60 h by the secretion of catechol 1,2-dioxygenase via ortho intradial pathway. The strain MSSRFPD35 possess both the catechol 1,2 dioxygenase and catechol 2,3 dioxygenase coding genes that drive the ortho and meta pathways, but the enzymatic assay revealed that the strain cleaves catechol via ortho pathway. Haldane's kinetic method was well fit to exponential growth data and the following kinetic parameter was obtained: µ∗ = 0.574 h-1, K i = 268.1, K s = 20.29 mg L-1. The true µmax and S m were calculated as 0.37 h-1 and 73.76 mg L-1, respectively. The Haldane's constant values were similar to earlier studies and healthy fitness depicted in correlation coefficient value R 2 of 0.98. Phenol degrading kinetic's was predicted using Haldane's model as q max 0.983, K i' 517.5 and K s' 9.152. Further, MSSRFPD35 was capable of utilizing different monocyclic and polycyclic aromatic hydrocarbons and to degrade phenol in the presence of different heavy metals. This study for the first time reports high phenol degrading efficiency of G. nicotianae MSSRFPD35 in the presence of toxic heavy metals. Thus, the strain G. nicotianae MSSRFPD35 can be exploited for the bioremediation of phenol and its derivatives polluted environments, co-contaminated with heavy metals.

Potential of Finger Millet Indigenous Rhizobacterium Pseudomonas sp. MSSRFD41 in Blast Disease Management-Growth Promotion and Compatibility With the Resident Rhizomicrobiome.

Finger millet [Eleusine coracona (L). Gaertner] "Ragi" is a nutri-cereal with potential health benefits, and is utilized solely for human consumption in semi-arid regions of Asia and Africa. It is highly vulnerable to blast disease caused by Pyricularia grisea, resulting in 50-100% yield loss. Chemical fungicides are used for the management of blast disease, but with great safety concern. Alternatively, bioinoculants are widely used in promoting seedling efficiency, plant biomass, and disease control. Little is known about the impact of introduced indigenous beneficial rhizobacteria on the rhizosphere microbiota and growth promotion in finger millet. Strain MSSRFD41 exhibited a 22.35 mm zone of inhibition against P. grisea, produces antifungal metabolites, siderophores, hydrolytic enzymes, and IAA, and solubilizes phosphate. Environmental SEM analysis indicated the potential of MSSRFD41 to inhibit the growth of P. grisea by affecting cellular functions, which caused deformation in fungal hyphae. Bioprimed finger millet seeds exhibited significantly higher levels of germination, seedling vigor index, and enhanced shoot and root length compared to control seeds. Cross streaking and RAPD analysis showed that MSSRFD41 is compatible with different groups of rhizobacteria and survived in the rhizosphere. In addition, PLFA analysis revealed no significant difference in microbial biomass between the treated and control rhizosphere samples. Field trials showed that MSSRFD41 treatment significantly reduced blast infestation and enhanced plant growth compared to other treatments. A liquid formulated MSSRFD41 product maintained shelf life at an average of 108 CFU ml-1 over 150 days of storage at 25°C. Overall, results from this study demonstrated that Pseudomonas sp. MSSRFD41, an indigenous rhizobacterial strain, is an alternative, effective, and sustainable resource for the management of P. grisea infestation and growth promotion of finger millet.

Salinicola rhizosphaerae sp. nov., isolated from the rhizosphere of the mangrove Avicennia marina L.

A Gram-stain-negative, motile, rod-shaped bacterium, designated MSSRFH1T, was isolated from the rhizosphere of the mangrove, Avicennia marina, in Pichavaram, Tamil Nadu, India. Phylogenetic analysis, based on the 16S rRNA gene sequence of MSSRFH1T, indicated that it clustered in the genus Salinicola and was most closely related to Salinicola peritrichatus JCM18795T (96.7 % similarity). The 16S rRNA gene sequence similarity was < 96.5 % with other strains of species of the genus Salinicola. The distinctiveness of strain MSSRFH1T was also shown by low similarities of its rpoD ( < 87 % similarity) and gyrB ( < 85 %) gene sequences with those of other members of the genus Salinicola. Strain MSSRFH1T could tolerate NaCl concentrations of up to 30 % (w/v). The main fatty acids of MSSRFH1T included C18 : 1ω7c, C16 : 0 and C19 : 0ω8c. The polar lipids present included diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, an unidentified amino lipid and unidentified phospholipids. Comparison of BOX-PCR fingerprints indicated that MSSRFH1T showed a unique DNA profile and its genomic G+C content was 64 mol%. On the basis of the data presented, strain MSSRFH1T represents a novel species of the genus Salinicola, for which the name Salinicola rhizosphaerae sp. nov. is proposed. The type strain is MSSRFH1T ( = KCTC 32998T = NBRC 110222T).

Novel Phl-producing genotypes of finger millet rhizosphere associated pseudomonads and assessment of their functional and genetic diversity.

Genetic diversity of phlD gene, an essential gene in the biosynthesis of 2,4-diacetylphloroglucinol, was studied by restriction fragment length polymorphism (RFLP) in 20 Phl-producing pseudomonads isolated from finger millet rhizosphere. RFLP analysis of phlD gene displayed three patterns with HaeIII and TaqI enzymes. phlD gene sequence closely correlated with RFLP results and revealed the existence of three new genotypes G, H and I. Further, the phylogenetic and concatenated sequence analysis of the 16S rRNA, rpoB, gyrB, rpoD genes supported the hypothesis that these genotypes G, H and I were different from reported genotypes A-F. In all phylogenetic studies, the genotype G formed a distant clade from the groups of Pseudomonas putida and P. aeruginosa (sensu strictu), but the groups H and I were closely related to P. aeruginosa/P. stutzeri group. The Phl-producing pseudomonads exhibited antagonistic activity against Pyricularia grisea (TN508), Gaeumannomyces graminis (DSM1463), Fusarium oxysporum (DSM62297), Xanthomonas campestris (DSM3586) and Erwinia persicina (HMGU155). In addition, these strains exhibited various plant growth-promoting traits. In conclusion, this study displays the existence of novel Phl-producing pseudomonads genotypes G, H and I from finger millet rhizosphere, which formed taxonomically outward phylogenetic lineage from the groups of P. putida and P. aeruginosa (sensu strictu).