Peganum harmala (L.) Schrad., a perennial medicinal plant thriving in arid Moroccan soils, represents a natural reservoir of unexplored bacterial diversity. To uncover its hidden foliar endosphere microbiota, we isolated and characterized two Acinetobacter strains: a novel endophytic bacterium, AGC35, and another strain, AGC59, newly reported from this host. Both are non-halophilic, aerobic, Gram-negative bacteria exhibiting optimal growth at 30–35 °C, pH5, and with 1% NaCl. An integrative genomic, phylogenetic, functional, and phenotypic characterization classified both strains within the genus Acinetobacter (class Gamma-pseudomonadota). However, Average Nucleotide Identity (<96%) and digital DNA-DNA Hybridization (<70%) values distinguished the AGC35 strain as a novel species, for which the name Acinetobacter endophylla sp. nov. is proposed. A comparative genomic and phenotypic analysis with the co-isolated Acinetobacter pittii strain AGC59 revealed extensive genome rearrangements, reflecting distinct evolutionary lineage and ecological strategies. While both genomes share core metabolic pathways, A. endophylla harbors specialized genes for the degradation of plant-derived aromatic compounds (e.g., catechol) but shows reduced capacities in carbohydrate metabolism and osmotic stress tolerance, traits indicative of a metabolic specialist with plant-growth-promotion potential, including phosphorus, potassium, and silicon solubilization and indole-3-acetic acid production. In contrast, A. pittii exhibits a more generalist genome enriched in stress functions. Analysis using the Antibiotics and Secondary Metabolite Analysis Shell revealed multiple biosynthetic gene clusters in both strains, showing ≤26% similarity to known references, suggesting the potential for novel antimicrobial secondary metabolite biosynthesis, including antifungal lipopeptides and thiopeptide antibiotics. Altogether, functional specialization and ecological coherence of these findings support the recognition of A. endophylla sp. nov. as a genomically and functionally distinct species, highlighting niche partitioning and adaptive metabolism within the P. harmala holobiont. These results underscore the plant’s value as a reservoir of untapped microbial diversity with significant ecological and biotechnological relevance. Finally, future work will focus on elucidating the novel metabolites encoded by the biosynthetic gene clusters in both isolates and exploring their applications in crop-improvement strategies and natural-product discovery.

Plants inhabiting arid regions often harbor microbial communities that contribute to their resilience under extreme conditions. Yet, the genomic diversity and functional potential of bacterial endophytes associated with desert-adapted plants, particularly in Africa, remain largely unexplored. In this study, we investigated Microbacterium endophytes from the xerophytic cucurbit Citrullus colocynthis L. (Cucurbitaceae), collected in a semi-arid environment in central Morocco. Using culture-based isolation, phenotypic characterization, and whole-genome sequencing, we analyzed three representative isolates from leaf and root tissues. Genome-based taxonomy combined with polyphasic analyses identified two novel species, Microbacterium xerophyticum sp. nov. and Microbacterium umsixpiens sp. nov., with genome sizes of approximately 4.0 Mb and 3.9 Mb, respectively. Functional annotation revealed traits consistent with endophytism in water-limited ecosystems, including oxidative and osmotic stress responses, metal homeostasis, and high-affinity phosphate uptake. Differences in siderophore acquisition and nitrogen metabolism suggest niche partitioning between the two species. These findings document two novel bacterial species from a medicinal plant native to arid ecosystems, broaden the known diversity of plant-associated Microbacterium, and provide region-specific genomic references with adaptive traits relevant to host resilience under arid conditions.

Members of the genus Rhizobium are best known for nitrogen-fixing symbioses with legumes, yet their diversity and evolutionary roles in non-legume hosts remain poorly explored, particularly in arid ecosystems. We report the isolation and characterization of strain AGC32, an endophytic bacterium obtained from surface-sterilized roots of the desert medicinal plant Peganum harmala collected in Moroccan drylands. Phylogenomic analyses placed AGC32 within the genus Rhizobium but clearly distinct from described species, with average nucleotide identity values below 96% and digital DNA–DNA hybridization values below 70%, supporting its designation as a novel species for which the name Rhizobium moroccans sp. nov. is proposed. Comparative genomics revealed extensive structural genome rearrangements relative to its closest sequenced relative, Rhizobium deserti, indicating a divergent evolutionary trajectory. The high-quality draft genome encodes metabolic pathways associated with adaptation to nutrient limitation and environmental stress, including complete allantoin utilization, polyphosphate metabolism, organic acid assimilation, and multiple systems involved in oxidative and osmotic stress tolerance. Phenotypic assays corroborated these genomic predictions, demonstrating the ability to metabolize diverse organic acids and carbohydrates and to express multiple plant growth–promoting traits, including nitrogen fixation and the solubilization of phosphorus, potassium, and silicon. Collectively, these findings expand the ecological and evolutionary diversity of Rhizobium, demonstrate its capacity to associate with non-legume medicinal plants in extreme environments, and highlight desert ecosystems as reservoirs of previously unrecognized microbial diversity with potential applications in sustainable agriculture in arid regions.

Ziziphus lotus (L.) Lam. (Rhamnaceae), a key shrub species native to North Africa, is commonly found in arid and semi-arid regions. Renowned for its resilience under harsh conditions, it forms vegetation clusters that influence the surrounding environment. These clusters create microhabitats that promote biodiversity, reduce soil erosion, and improve soil fertility. However, in agricultural fields, Z. lotus is often regarded as an undesirable species. This study investigated the bacterial diversity and community composition along spatial gradients around Z. lotus patches in barley-planted and non-planted fields. Using 16S rRNA gene sequencing, 84 soil samples were analyzed from distances of 0, 3, and 6 m from Z. lotus patches. MiSeq sequencing generated 143,424 reads, representing 505 bacterial ASVs across 22 phyla. Alpha-diversity was highest at intermediate distances (3 m), while beta-diversity analyses revealed significant differences in community composition across distances (p = 0.035). Pseudomonadota dominated close to the shrub (44% at 0 m) but decreased at greater distances, whereas Bacillota and Actinobacteriota displayed distinct spatial patterns. A core microbiome comprising 44 ASVs (8.7%) was shared across all distances, with the greatest number of unique ASVs identified at 3 m. Random forest analysis highlighted Skermanella and Rubrobacter as key discriminatory taxa. These findings emphasize the spatial structuring of bacterial communities around Z. lotus patches, demonstrating the shrub’s substantial influence on bacterial dynamics in arid ecosystems.

Endophytic bacteria from plants adapted to arid and semi-arid environments represent an underexplored reservoir of microbial diversity with potential agricultural applications. Here, we report a polyphasic taxonomic and genome-based characterization of Achromobacter sp. isolates recovered from root and foliar tissues of Citrullus colocynthis and Peganum harmala, two medicinal plants thriving under harsh environmental conditions. Whole-genome sequencing, phylogenomic analyses, average nucleotide identity (ANI), digital DNA–DNA hybridization (dDDH), multilocus sequence typing, and detailed phenotypic profiling revealed three previously undescribed species, for which we propose the names Achromobacter colocynthi sp. nov., Achromobacter maghribensis sp. nov., and Achromobacter semiaridum sp. nov. Genome assemblies were highly complete (98.7–99.2%) with minimal contamination (<1%), supporting robust taxonomic inference. All three species displayed ANI and dDDH values below accepted thresholds relative to their closest phylogenetic neighbors, despite partial inconsistencies in 16S rRNA similarity for one isolate, highlighting the value of genome-wide metrics for species delineation. Phylogenomic analyses placed the novel taxa within Achromobacter sp. as distinct evolutionary lineages. Phenotypic characterization indicated broad metabolic versatility, including utilization of carbohydrates, organic acids, and amino acids, tolerance to moderate salinity and acidic pH, and resistance to multiple antimicrobial compounds, traits likely linked to adaptation to endophytic lifestyles under semi-arid conditions. Beyond their taxonomic novelty, the isolates exhibited in vitro traits associated with plant adaptation and stress tolerance, including IAA production, ACC deaminase activity, and tolerance to Zn, Cu, and Cd. Genomic analyses further indicated functions related to phosphate acquisition and stress response. These findings expand the taxonomic framework of Achromobacter sp., establish C. colocynthis and P. harmala as reservoirs of novel endophytic bacteria, and highlight their potential relevance for agricultural biotechnology in stress-prone environments.

Ziziphus lotus (L.) Lam., (Rhamnaceae) a resilient shrub native to Moroccan’s arid regions, functions as a keystone species by creating microhabitats that buffer temperature extremes, retain soil moisture, and accumulate organic matter. However, its role in structuring soil fungal diversity and community composition in these environments remains largely unexplored. This study investigated the spatial distribution of fungal communities associated with Z. lotus in barley-planted and non-planted fields. Soil samples were collected at 0, 3, and 6 m from shrub clusters during the barley harvest. The fungal community was dominated by Ascomycota (93.5%). Alpha diversity indices (Shannon–Wiener and Simpson) were significantly higher near shrub bases (0 and 3 m) compared to more distant soils (6 m), indicating a clear decline in diversity with distance (0 m vs. 6 m: p = 0.0012; 3 m vs. 6 m: p = 0.0007). Soil physicochemical parameters, including calcium carbonate content, nitrate, and salinity, significantly influenced fungal diversity (p ≤ 0.05). Beta diversity analysis revealed significant spatial differentiation in fungal community composition (PERMANOVA: p = 0.001). Overall, fungal richness and diversity were highest near shrub. Genera such as Cladosporium, Fusarium, and Alternaria were more abundant near shrub bases, while taxa like Didymellaceae and Alfaria were specially restricted. Functional predictions indicated dominance of fungi with mixed trophic modes (pathotroph–saprotroph–symbiotroph), suggesting ecological plasticity. Despite barley cultivation, the fungal community structure remained largely similar between the planted and non-planted fields. Overall, our findings underscore the ecological importance of Z. lotus as a reservoir of stress-tolerant fungi and as a potential keystone species for restoring degraded arid ecosystems.