Multifarious Indigenous Diazotrophic Rhizobacteria of Rice (Oryza sativa L.) Rhizosphere and Their Effect on Plant Growth Promotion

A diverse group of rhizobacteria persists in the rhizospheric soil, on the surface of roots, or in association with rice plants. These bacteria colonize plant root systems, enhance plant growth and crop yield. Indigenous rhizobacteria are known to promote soil health, grain production quality and serve as sustainable bioinoculant. The present study was aimed to isolate, identify and characterize indigenous plant growth promoting (PGP) diazotrophic bacteria associated with the rhizosphere of rice fields from different areas of Jammu and Kashmir, India. A total of 15 bacteria were isolated and evaluated for various PGP traits, antagonistic activity against phytopathogens, production of hydrolytic enzymes and biofilm formation under in-vitro conditions. The majority of the isolated bacteria were Gram-negative. Out of 15 bacterial isolates, nine isolates produced IAA (12.24 ± 2.86 to 250.3 ± 1.15 μg/ml), 6 isolates exhibited phosphate solubilization activity (36.69 ± 1.63 to 312.4 ± 1.15 μg/ml), 7 isolates exhibited rock phosphate solubilization while 5 isolates solubilized zinc (10–18 mm), 7 isolates showed siderophore production, 8 isolates exhibited HCN production, 6 isolates exhibited aminocyclopropane-1-carboxylate (ACC) deaminase activity, 13 isolates exhibited cellulase activity, nine isolates exhibited amylase and lipase activity and six isolates exhibited chitinase activity. In addition, 5 isolates showed amplification with the nifH gene and showed a significant amount of nitrogenase activity in a range of 0.127–4.39 μmol C2H4/mg protein/h. Five isolates viz., IHK-1, IHK-3, IHK-13, IHK-15 and IHK-25 exhibited most PGP attributes and successfully limited the mycelial growth of Rhizoctonia solani and Fusarium oxysporum in-vitro. All the five bacterial isolates were identified based on morphological, biochemical and 16S rDNA gene sequencing study, as Stenotrophomonas maltophilia, Enterobacter sp., Bacillus sp., Ochrobactrum haematophilum and Pseudomonas aeruginosa. Rice plants developed from seeds inoculated with these PGP strains individually had considerably higher germination percentage, seed vigor index and total dry biomass when compared to control. These findings strongly imply that the PGP diazotrophic bacteria identified in this work could be employed as plant growth stimulators in rice.

Soil organic matter is essential for colony growth in subterranean termites

Intrinsic dinitrogen (N2) fixation by diazotrophic bacteria in termite hindguts has been considered an important pathway for nitrogen acquisition in termites. However, studies that supported this claim focused on measuring instant N2 fixation rates and failed to address their relationship with termite colony growth and reproduction over time. We here argue that not all wood-feeding termites rely on symbiotic diazotrophic bacteria for colony growth. The present study looks at dietary nitrogen acquisition in a subterranean termite (Rhinotermitidae, Coptotermes). Young termite colonies reared with wood and nitrogen-rich organic soil developed faster, compared to those reared on wood and inorganic sand. More critically, further colony development was arrested if access to organic soil was removed. In addition, no difference of relative nitrogenase expression rates was found when comparing the hindguts of termites reared between the two conditions. We therefore propose that subterranean termite (Rhinotermitidae) colony growth is no longer restricted to metabolically expensive intrinsic N2 fixation, as the relationship between diazotrophic bacteria and subterranean termites may primarily be trophic rather than symbiotic. Such reliance of Rhinotermitidae on soil microbial decomposition activity for optimal colony growth may also have had a critical mechanistic role in the initial emergence of Termitidae.

Productivity and growth in cowpea inoculated with rhizobia under different light environments

The objective of this work was to verify the influence of light environments combined with rhizobia inoculation on cowpea growth and productivity. A completely random design was used in a 4x4 factorial scheme, with four light environments, four nitrogen sources and eight replicates in split plot parcels. Light environments were set by means of photo-conversion and thermo-conversion nettings (Aluminet®, red net and black net) and control treatment without shading (full sun). Nitrogen sources were constituted by the strains INPA 03-11B - SEMIA 6462 (Bradyrhizobium elkanni) and UFLA 03-84 - SEMIA 6461 (Bradyrhizobium viridifuturi), and two control treatments: with 70 kg ha-1 of mineral nitrogen and without N. Plant height, indices of chlorophyll a, b and total chlorophill, the number of leaves, number of nodules, dry matter of nodules, dry matter of the aerial portion, dry matter of roots and total dry matter, relative efficiency, gathering of nitrogen in the aerial portion, number, length and matter of pods per plant and dry matter of 100 grains, were evaluated. . There was interaction between light conditions and nitrogen sources for the number of nodules. Individual effect was observed in all other variables. Strain INPA 03-11B was able to promote higher nodulation in cowpea plants in light environments under full sun and Aluminet and the strain UFLA 03-84 only under full sun conditions. However, the efficiency of diazotrophic bacteria to promote vegetative growth, nitrogen gathering and production was not influenced by different light environments. Thus, full sun cultivation is recommended, independently of the nitrogen source used.

A Simple in situ Assay to Assess Plant-Associative Bacterial Nitrogenase Activity

Assessment of plant-associative bacterial nitrogen (N) fixation is crucial for selection and development of elite diazotrophic inoculants that could be used to supply cereal crops with nitrogen in a sustainable manner. Although diazotrophic bacteria possess diverse oxygen tolerance mechanisms, most require a sub 21% oxygen environment to achieve optimal stability and function of the N-fixing catalyst nitrogenase. Consequently, assessment of N fixation is routinely carried out on “free-living” bacteria grown in the absence of a host plant and such experiments may not accurately divulge activity in the rhizosphere where the availability and forms of nutrients such as carbon and N, which are key regulators of N fixation, may vary widely. Here, we present a modified in situ acetylene reduction assay (ARA), utilizing the model cereal barley as a host to comparatively assess nitrogenase activity in diazotrophic bacteria. The assay is rapid, highly reproducible, applicable to a broad range of diazotrophs, and can be performed with simple equipment commonly found in most laboratories that investigate plant-microbe interactions. Thus, the assay could serve as a first point of order for high-throughput identification of elite plant-associative diazotrophs.

Redox Regulation in Diazotrophic Bacteria in Interaction with Plants

Plants interact with a large number of microorganisms that greatly influence their growth and health. Among the beneficial microorganisms, rhizosphere bacteria known as Plant Growth Promoting Bacteria increase plant fitness by producing compounds such as phytohormones or by carrying out symbioses that enhance nutrient acquisition. Nitrogen-fixing bacteria, either as endophytes or as endosymbionts, specifically improve the growth and development of plants by supplying them with nitrogen, a key macro-element. Survival and proliferation of these bacteria require their adaptation to the rhizosphere and host plant, which are particular ecological environments. This adaptation highly depends on bacteria response to the Reactive Oxygen Species (ROS), associated to abiotic stresses or produced by host plants, which determine the outcome of the plant-bacteria interaction. This paper reviews the different antioxidant defense mechanisms identified in diazotrophic bacteria, focusing on their involvement in coping with the changing conditions encountered during interaction with plant partners.

The Evolution of Molybdenum Dependent Nitrogenase in Cyanobacteria

Nitrogen fixation plays a crucial role in the nitrogen cycle by helping to convert nitrogen into a form usable by other organisms. Bacteria capable of fixing nitrogen are found in six phyla including Cyanobacteria. Molybdenum dependent nitrogenase (nif) genes are thought to share a single origin as they have homologs in various phyla. However, diazotrophic bacteria have a mosaic distribution within the cyanobacterial lineage. Therefore, the aim of this study was to determine the cause of this mosaic distribution. We identified nif gene operon structures in the genomes of 85 of the 179 cyanobacterial strains for which whole genome sequences were available. Four nif operons were conserved in each diazotroph Cyanobacterium, although there were some gene translocations and insertions. Phylogenetic inference of these genes did not reveal horizontal gene transfer from outside the phylum Cyanobacteria. These results support the hypothesis that the mosaic distribution of diazotrophic bacteria in the cyanobacterial lineage is the result of the independent loss of nif genes inherited from common cyanobacterial ancestors in each lineage.

Can inoculation with diazotrophic bacteria decrease the productivity loss of defoliated Vigna unguiculata (L.) Walp?

ABSTRACT The objective of this study was to evaluate the effect of artificial defoliation on vegetative and productive stages of cowpea inoculated with diazotrophic bacteria, in two experiments. The first experiment was performed in a greenhouse with 5 × 5 factorial (five defoliation percentages and five N sources), with four repetitions. N sources consisted of bacterial inoculation with strains INPA 03-11B, UFLA 03-84, UFRB FA34C2-2, and two control treatments: i - with N fertilization and ii - blank control, without N fertilization and without inoculation. The second experiment was performed in the field, in a 3 × 5 factorial scheme, with three repetitions. Treatments consisted of three N sources: with N fertilization, with bacterial strain INPA 03-11B that was selected in the first experiment, and five percentages of artificial defoliation. Defoliation percentages for both experiments were 0, 25, 50, 75, and 100%. Artificial defoliation during the vegetative stage caused reduction in the dry mass of bacterial nodules. Inoculation increased plant tolerance to defoliation and enhanced grain nutrient concentration (N and P). Inoculation with the strain INPA 03-11B allowed cowpea plants to tolerate 50% defoliation in the vegetative stage. The mean productivity of cowpea was reduced under > 50% defoliation during the productive stage; therefore, control of defoliating pests until the productive stage is not necessary under field conditions.