Bacteria currently included in Rhizobium leguminosarum are too diverse to be considered a single species, so we can refer to this as a species complex (the Rlc). We have found 429 publicly available genome sequences that fall within the Rlc and these show that the Rlc is a distinct entity, well separated from other species in the genus. Its sister taxon is R. anhuiense. We constructed a phylogeny based on concatenated sequences of 120 universal (core) genes, and calculated pairwise average nucleotide identity (ANI) between all genomes. From these analyses, we concluded that the Rlc includes 18 distinct genospecies, plus 7 unique strains that are not placed in these genospecies. Each genospecies is separated by a distinct gap in ANI values, usually at approximately 96% ANI, implying that it is a ‘natural’ unit. Five of the genospecies include the type strains of named species: R. laguerreae, R. sophorae, R. ruizarguesonis, “R. indicum” and R. leguminosarum itself. The 16S ribosomal RNA sequence is remarkably diverse within the Rlc, but does not distinguish the genospecies. Partial sequences of housekeeping genes, which have frequently been used to characterize isolate collections, can mostly be assigned unambiguously to a genospecies, but alleles within a genospecies do not always form a clade, so single genes are not a reliable guide to the true phylogeny of the strains. We conclude that access to a large number of genome sequences is a powerful tool for characterizing the diversity of bacteria, and that taxonomic conclusions should be based on all available genome sequences, not just those of type strains.

Soil fertility is one of the major limiting factors for crop’s productivity in Egypt and the world in general. Biological nitrogen fixation (BNF) has a great importance as a non-polluting and a cost-effective way to improve soil fertility through supplying N to different agricultural systems. Faba bean (Vicia faba L.) is one of the most efficient nitrogen-fixing legumes that can meet all of their N needs through BNF. Therefore, understanding the impact of rhizobial inoculation and contrasting soil rhizobia on nodulation and N₂ fixation in faba bean is crucial to optimize the crop yield, particularly under low fertility soil conditions. This study investigated the symbiotic effectiveness of 17 Rhizobium/Agrobacterium strains previously isolated from different Egyptian governorates in improving the nodulation and N₂ fixation in faba bean cv. Giza 843 under controlled greenhouse conditions. Five strains that had a high nitrogen-fixing capacity under greenhouse conditions were subsequently tested in field trials as faba bean inoculants at Ismaillia Governorate in northeast Egypt in comparison with the chemical N-fertilization treatment (96 kg N·ha⁻¹). A starter N-dose (48 kg N·ha⁻¹) was applied in combination with different Rhizobium inoculants. The field experiments were established at sites without a background of inoculation under low fertility sandy soil conditions over two successive winter growing seasons, 2012/2013 and 2013/2014. Under greenhouse conditions, inoculated plants produced significantly higher nodules dry weight, plant biomass, and shoot N-uptake than non-inoculated ones. In the first season (2012/2013), inoculation of field-grown faba bean showed significant improvements in seed yield (3.73–4.36 ton·ha⁻¹) and seed N-yield (138–153 Kg N·ha⁻¹), which were higher than the uninoculated control (48 kg N·ha⁻¹) that produced 2.97 Kg·ha⁻¹ and 95 kg N·ha⁻¹, respectively. Similarly, in the second season (2013/2014), inoculation significantly improved seed yield (3.16–4.68 ton·ha⁻¹) and seed N-yield (98–155 Kg N·ha⁻¹) relative to the uninoculated control (48 kg N·ha⁻¹), which recorded 2.58 Kg·ha⁻¹ and 80 kg N·ha⁻¹, respectively. Interestingly, faba bean inoculated with strain Rlv NGB-FR 126 showed significant increments in seed yield (35%–48%) and seed N-yield (34%–49%) compared to the inorganic N fertilizers treatment (96 kg N·ha⁻¹) over the two cropping seasons, respectively. These results indicate that inoculation of faba bean with effective rhizobial strains can reduce the need for inorganic N fertilization to achieve higher crop yield under low fertility soil conditions.

Bacteria colonizing the rhizosphere, a narrow zone of soil surrounding the root system, are known to have beneficial effects in improving the growth and stress tolerance of plants. However, most bacteria in natural environments, especially those in rhizosphere soils, are recalcitrant to cultivation using traditional techniques, and thus their roles in soil health and plant growth remain unexplored.

Wheat crops require effective nitrogen fertilization to produce high yields. Only half of chemical N₂ fertilizers are absorbed into plants while the rest remains in the soil, causing environmental problems. Fungi could maximize nitrogen absorption, and from an environmental and biodiversity point of view, there is an urgent necessity for bioprospecting native fungi associated with wild plants growing in harsh environments, e.g., St. Katherine Protectorate (SKP) in the arid Sinai. Recovered taxa, either endophytic and/or rhizospheric, were screened for their plant growth-promoting (PGP) traits. Eighteen fungal isolates (15 rhizospheric and 3 endophytic) belonging to anamorphic ascomycetes were recovered from 9 different wild plants, and their PGP traits (indole-3-acetic acid [IAA] production, phosphate solubilization, siderophore production, and hydrolytic enzyme production) were measured. Rhizospheric isolate NGB-WS14 (Chaetosphaeronema achilleae) produced high levels of IAA (119.1 μg mL⁻¹) in the presence of tryptophan, while NGB-WS 8 (Acrophialophora levis) produced high IAA levels (42.4 μg mL⁻¹) in the absence of tryptophan. The highest phosphate-solubilizing activity (181.9 μg mL⁻¹) was recorded by NGB-WFS2 (Penicillium chrysogenum). Endophytic isolate NGB-WFE16 (Fusarium petersiae) exhibited a high percentage level of Siderophore Unit (96.5% SU). All isolates showed variability in the secretion of extracellular hydrolytic enzymes. Remarkably, all isolates had antagonistic activity (55.6% to 87.3% suppression of pathogen growth) against the pathogenic taxon Alternaria alternata (SCUF00001378) in the dual-assay results. Out of the 18 isolates, 4 rhizospheric and 1 endophytic isolate showed significant increases in shoot dry weight and shoot nitrogen and chlorophyll content of wheat plants subjected to low inputs of chemical nitrogen (N) fertilizers (50% reduction) compared with the non-inoculated control in a pot experiment. Potent taxa were subjected to sequencing for molecular confirmation of phenotypic identification. The retrieved ITS sequences in this study have been deposited in GenBank under accession numbers from LC642736 to LC642740. This study considered the first report of endophytic fungi of Cheilanthes vellea, a wild plant with PGPF which improves wheat growth. These results recommend using PGPF as inoculants to alleviate low nitrogen fertilization.