Functional analysis and comparative genomics of Rahnella perminowiae S11P1 and Variovorax sp. S12S4, two plant growth-promoting rhizobacteria isolated from Crocus sativus L. (saffron) rhizosphere.

BACKGROUND: Rahnella perminowiae S11P1 and Variovorax sp. S12S4 are two plant growth-promoting rhizobacteria that were previously isolated from the rhizosphere of Crocus sativus L. (saffron), and have demonstrated interesting PGP activities and promising results when used as inoculants in field trials. To further elucidate the molecular mechanisms underlying their beneficial effects on plant growth, comprehensive genome mining of S11P1 and S12S4 and comparative genomic analysis with closely related strains were conducted. RESULTS: Functional annotation of the two strains predicted a large number of genes involved in auxin and siderophore production, nitrogen fixation, sulfur metabolism, organic acid biosynthesis, pyrroloquinoline quinone production, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, volatile organic compounds production, and polyamine biosynthesis. In addition, numerous genes implicated in plant-bacteria interactions, such as those involved in chemotaxis and quorum sensing, were predicted. Moreover, the two strains carried genes involved in bacterial fitness under abiotic stress conditions. Comparative genomic analysis revealed an open pan-genomic structure for the two strains. COG annotation showed that higher fractions of core and accessory genes were involved in the metabolism and transport of carbohydrates and amino acids, suggesting the metabolic versatility of the two strains as effective rhizosphere colonizers. Furthermore, this study reports the first comparison of Multilocus sequence analysis (MLSA) and core-based phylogenies of the Rahnella and Variovorax genera. CONCLUSIONS: The present study unveils the molecular mechanisms underlying plant growth promotion and biocontrol activity of S11P1 and S12S4, and provides a basis for their further biotechnological application in agriculture.

Genomic analysis of Paenibacillus sp. MDMC362 from the Merzouga desert leads to the identification of a potentially thermostable catalase.

Microorganisms in hot deserts face heat and other environmental conditions, such as desiccation, UV radiation, or low nutrient availability. Therefore, this hostile environment harbour microorganisms with acquired characteristics related to survival in their habitat, which can be exploited in biotechnology. In this work, the genome of Paenibacillus sp. MDMC362 isolated from the Merzouga desert in Morocco was sequenced to understand its survival strategy's genetic basis; and to evaluate the thermostability of a catalase extracted from genomic annotation files using molecular dynamics. Paenibacillus sp. MDMC362 genome was rich in genetic elements involved in the fight against different stresses, notably temperature stress, UV radiations, osmotic stress, carbon starvation, and oxidative stress. Indeed, we could identify genes of the operons groES-groEL and hrcA-grpE-dnaK and those involved in the different stages of sporulation, which can help the bacteria to survive the high temperatures imposed by a desertic environment. We also observed the genetic components of the UvrABC system and additional mechanisms involved in DNA repair, which help overcome UV radiation damage. Other genes have been identified in the genome, like those coding for ectoine and proline, that aids fight osmotic stress and desiccation. Catalase thermostability investigation using molecular dynamics showed that the protein reached stability and conserved its compactness at temperatures up to 373.15 K. These results suggest a potential thermostability of the enzyme. Since the studied protein is a core protein, thermostability could be conserved among Paenibacillus sp. MDMC362 closely related strains; however, bacteria from harsh environments may have a slight advantage regarding protein stability.

Genomic analysis of two Bacillus safensis isolated from Merzouga desert reveals desert adaptive and potential plant growth-promoting traits.

Deserts represent extreme environments for microorganisms, and conditions such as high soil salinity, nutrient deficiency, and increased levels of UV radiation make desert soil communities of high biotechnological potential. In this study, we isolated, sequenced, and assembled the genomes of Bacillus safensis strains BcP62 and Bcs93, to which we performed comparative genome analyses. Using the DDH and ANI of both strains with the available B. safensis genomes, we identified three potential subspecies within this group. Intra-species core genome phylogenetic analysis did not result in clustering genomes by niche type, with some exceptions. This study also revealed that the genomes of the analyzed strains possessed plant growth-promoting characteristics, most of which were conserved in all B. safensis strains. Furthermore, we highlight the genetic features of B. safensis BcP62 and Bcs93 related to survival in the Merzouga desert in Morocco. These strains could be potentially used in agriculture as PGPB in extreme environments, given their high tolerability to unfavorable conditions.

Draft Genome Sequences of Four Plant Growth-Promoting Rhizobacteria Isolated from Saffron (Crocus sativus L.) Rhizosphere in Morocco.

We report the draft genome sequences of plant growth-promoting Rahnella perminowiae strain S11P1, Variovorax sp. strain S12S4, and Pseudomonas sp. strains S11A4 and S11P7, which were isolated from saffron (Crocus sativus L.) rhizosphere. Several genes were predicted to be involved in auxin production, phosphate solubilization, and other specialized functions in plant growth and defense.

Draft Genome Sequences of Three Pseudomonas chengduensis Strains Isolated from Desert Soil in Morocco.

We report the draft genome sequences of three Pseudomonas chengduensis strains isolated from the sand dunes of the Merzouga (MDMC17 strain) and Erg Lihoudi (MDMC216 and MDMC224 strains) regions in the Moroccan desert. These bacteria are able to tolerate the harsh environmental conditions of the desert ecosystem.

Draft Genome Sequences of Two Enterobacter hormaechei subsp. xiangfangensis Strains Isolated from the Moroccan Sahara.

We report the draft genome sequences of Enterobacter hormaechei subsp. xiangfangensis strains MDMC82 and MDMC76, which were isolated from the sand dunes of the Merzouga desert in the Moroccan Sahara. These bacteria are able to tolerate the harsh environmental conditions of the Moroccan desert.

Lipidomic Profiling of Argania spinosa L. (Skeels) Following Drought Stress.

Argan tree is frequently constrained by environmental stresses, especially drought. Lipids play crucial roles in stress adaptation, but lipidomic profiles in Argania spinosa under drought stress is largely unknown. The aim of this study was to identify lipid components potentially responsive to drought stress from leaves in the four argan ecotypes. For this, non-polar metabolite profiling was carried out using gas chromatography-mass spectrometry. We identified 228 components, the majority of which belonged to fatty acids and prenol lipids classes. The principal component analysis and partial least-squares discriminant analysis were applied to the lipidomics data to determine the component changes between all ecotypes. Based on the common components present in all ecotypes under contrast conditions, 21 metabolites belong fatty acids were identified with significant change. It was concluded that the majority of these components show up-accumulation in their content and involve in different pathways, especially in alpha-linolenic acid metabolism. The findings of this study provided new insights into the lipidomic study of argan leaves under drought stress and may be eventually contribute to overcoming drought; in addition, this could serve as a base for future studies on transcriptomic and proteomic to enhance the drought tolerance of forest trees.

Native Heavy Metal-Tolerant Plant Growth Promoting Rhizobacteria Improves Sulla spinosissima (L.) Growth in Post-Mining Contaminated Soils.

The potential of rhizobacteria in assisting plants used in the phytostabilization or re-vegetation of soils contaminated by heavy metals is gaining interest all around the world. In this context, six rhizobacterial strains isolated from highly heavy metal-contaminated soils situated in abandoned mining sites around the Oujda region (Morocco) were tested with Sulla spinosissima (L.), a native leguminous plant expanding in this area. The strains used were multi-resistant to heavy metals and possessed multiple plant growth-promoting traits. Potential beneficial effects of the strains were also evaluated in planta by measuring various growth and physiological parameters of inoculated Sulla plants grown in sterilized sand. Inoculation with the Rhodococcus qingshengii strain LMR340 boosted plant biomass (39% to 83% increase compared to uninoculated plants), chlorophyll and carotenoid content (up to 29%), and antioxidant enzyme activities (15% to 80% increase). Based on these interesting findings, selected strains were inoculated into plants growing in a heavy metal, multi-polluted, and poor soil. Under these conditions, non-inoculated plants and those inoculated with the strain LMR250 were unable to grow, while the other five bacterial inoculants restored plant growth. The best performing strain, Pseudarthrobacter oxydans LMR291, could be considered as a good biofertilizer and/or biostimulant candidate to be used for promoting the growth of selected plants in re-vegetation and/or phytostabilization programs of degraded and contaminated soils.

Characterization of plant growth promoting activities of indigenous bacteria of phosphate mine wastes, a first step toward revegetation.

Morocco holds the vast majority of the world's phosphate reserves, but due to the processes involved in extracting and commercializing these reserves, large quantities of de-structured, nutritionally deficient mine phosphate wastes are produced each year. In a semi-arid climate, these wastes severely hamper plant growth and development leading to huge unvegetated areas. Soil indigenous Plant Growth-Promoting Bacteria (PGPB) play a pivotal role in restauration of these phosphate mining wastes by revegetation, by increasing plants development, soil functioning, and nutrient cycling. The development of a vegetative cover above the degraded phosphate wastes, could stabilize and reintegrate these wastes in the surrounding environment. The current study's objectives were to isolate, characterize, and identify indigenous bacterial strains, and test their PGP activity in vitro and, for the best-performing strains in planta, in order to assess their potential for acting as biofertilizers. A quantitative test for the synthesis of auxin and the production of siderophores as well as a qualitative test for the solubilization of phosphate were performed on all isolated bacterial strains. The production of hydrogen cyanide (HCN), exopolysaccharides (EPS), and enzymes were also examined. Three bacteria, selected among the best PGPB of this study, were tested in planta to determine whether such indigenous bacteria could aid plant growth in this de-structured and nutrient-poor mining soil. Using 16S rRNA gene sequencing, 41 bacterial strains were isolated and 11 genera were identified: Acinetobacter, Agrococcus, Bacillus, Brevibacterium, Microbacterium, Neobacillus, Paenibacillus, Peribacillus, Pseudarthrobacter, Stenotrophomonas, and Raoultella. Among the three best performing bacteria (related to Bacillus paramycoides, Brevibacterium anseongense, and Stenotrophomonas rhizophila), only Stenotrophomonas rhizophila and Brevibacterium anseongense were able to significantly enhance Lupinus albus L. growth. The best inoculation results were obtained using the strain related to Stenotrophomonas rhizophila, improving the plant's root dry weight and chlorophyll content. This is also, to our knowledge, the first study to show a PGP activity of Brevibacterium anseongense.

Past mastering of metal transformation enabled physicians to increase their therapeutic potential.

BACKGROUND: Metals are trace elements, vital in some instances or toxic in others. Due to this toxicity, they have been used since ancient time as antimicrobials, and prescribed when plant-only remedies were not efficient enough. These remedies could still contain secrets that may lead to the discovery of new therapeutically interesting combinations. The objective of this study was to give a proof of concept that such remedies combining metals and plants are worth studying again. METHODS: We exploited 4 medical formularies (aqrabadhin), from three Arab authors from the 9-12th century. We reproduced a remedy, and analyzed the role of each ingredient. We further looked for the minimum inhibitory concentration against three pathogenic bacteria, and we analyzed toxic and inflammatory effects of this remedy on macrophages. RESULTS: Even if plants were extensively used (almost 80 % of all ingredients), more than 36 different minerals have been found in these 4 aqrabadhin. When it came to remedies against infections that could be applied externally, the use of metals grew to 70 %. We focused on a remedy, containing mainly metals. We have been able to attribute a role for each ingredient, to show that this skin remedy helped to combat the infection and to resorb the wound, and to highlight the mastering of metal transformation by these physicians. CONCLUSIONS: With a very simple recipe, mainly composed of metals, these past physicians designed a complete and synergistic remedy to combat abscesses, while restricting the toxic effect of metals to the site of infection. It is a first example showing that different metal manufactures were evolved to improve their therapeutic potentials. The knowledge acquired by these physician should deserve more attention, and unexpected features, original organo-metallic compounds or therapeutic synergy could still be found from such research.

Evaluation of microalgae polysaccharides as biostimulants of tomato plant defense using metabolomics and biochemical approaches.

Microalgal polysaccharides (PSs) may be an effective elicitor agent that can efficiently protect plants against biotic stresses. In this study, wee investigates, the effect of PS obtained from microalgae and cyanobacteria (D. salina MS002, P. tricorontum MS023, Porphyridium sp. MS081, Desmodesmus sp., D. salina MS067 and A. platensis MS001) on the biochemical and metabolomics markers linked to defense pathways in tomato plants. The phenylalanine ammonia lyase (PAL), chitinase, 1,3-beta-glucanase and peroxidase (POX) activities have been improved in tomato plants leaves treated by polysaccharides extracted from P. triocnutum (238.26%); Desmodesmus sp. (19.95%); P. triocnutum (137.50%) and Porphyridium sp. (47.28%) respectively. For proteins, polyphenols and H2O2, the maximum effect was induced by D. salina 067 (55.01%), Porphyridium sp. (3.97%) and A. platensis (35.08%) respectively. On the other hand, Gas Chromatography-mass spectrometry (GC-MS) metabolomics analysis showed that PSs induced the modification of metabolite profile involved in the wax construction of tomato leaves, such as fatty acids, alkanes, alkenes and phytosterol. PS treatments improved the accumulation of fatty acids C16:3, C18:2 and C18:3 released from the membrane lipids as precursors of oxylipin biosynthesis which are signaling molecules of plant defense. In addition, PS treatment induced the accumulation of C18:0 and Azelaic acid which is a regulator of salicylic acid-dependent systemic acquired resistance. However, molecular and metabolic studies can determine more precisely the mode of action of microalgal polysaccharides as biostimulants/elicitors plant defense.

Description of Microbial Communities of Phosphate Mine Wastes in Morocco, a Semi-Arid Climate, Using High-Throughput Sequencing and Functional Prediction.

Soil microbiota are vital for successful revegetation, as they play a critical role in nutrient cycles, soil functions, and plant growth and health. A rehabilitation scenario of the abandoned Kettara mine (Morocco) includes covering acidic tailings with alkaline phosphate mine wastes to limit water infiltration and hence acid mine drainage. Revegetation of phosphate wastes is the final step to this rehabilitation plan. However, revegetation is hard on this type of waste in semi-arid areas and only a few plants managed to grow naturally after 5 years on the store-and-release cover. As we know that belowground biodiversity is a key component for aboveground functioning, we sought to know if any structural problem in phosphate waste communities could explain the almost absence of plants. To test this hypothesis, bacterial and archaeal communities present in these wastes were assessed by 16S rRNA metabarcoding. Exploration of taxonomic composition revealed a quite diversified community assigned to 19 Bacterial and two Archaeal phyla, similar to other studies, that do not appear to raise any particular issues of structural problems. The dominant sequences belonged to Proteobacteria, Bacteroidetes, Actinobacteria, and Gemmatimonadetes and to the genera Massilia, Sphingomonas, and Adhaeribacter. LEfSe analysis identified 19 key genera, and metagenomic functional prediction revealed a broader phylogenetic range of taxa than expected, with all identified genera possessing at least one plant growth-promoting trait. Around 47% of the sequences were also related to genera possessing strains that facilitate plant development under biotic and environmental stress conditions, such as drought and heat.

Molecular analysis of SCARECROW genes expressed in white lupin cluster roots.

The Scarecrow (SCR) transcription factor plays a crucial role in root cell radial patterning and is required for maintenance of the quiescent centre and differentiation of the endodermis. In response to phosphorus (P) deficiency, white lupin (Lupinus albus L.) root surface area increases some 50-fold to 70-fold due to the development of cluster (proteoid) roots. Previously it was reported that SCR-like expressed sequence tags (ESTs) were expressed during early cluster root development. Here the cloning of two white lupin SCR genes, LaSCR1 and LaSCR2, is reported. The predicted amino acid sequences of both LaSCR gene products are highly similar to AtSCR and contain C-terminal conserved GRAS family domains. LaSCR1 and LaSCR2 transcript accumulation localized to the endodermis of both normal and cluster roots as shown by in situ hybridization and gene promoter::reporter staining. Transcript analysis as evaluated by quantitative real-time-PCR (qRT-PCR) and RNA gel hybridization indicated that the two LaSCR genes are expressed predominantly in roots. Expression of LaSCR genes was not directly responsive to the P status of the plant but was a function of cluster root development. Suppression of LaSCR1 in transformed roots of lupin and Medicago via RNAi (RNA interference) delivered through Agrobacterium rhizogenes resulted in decreased root numbers, reflecting the potential role of LaSCR1 in maintaining root growth in these species. The results suggest that the functional orthologues of AtSCR have been characterized.

Microalgae polysaccharides bio-stimulating effect on tomato plants: Growth and metabolic distribution.

Microalgae polysaccharides represent a potentially bioressource for the enhancement and the protection of agricultural crops. We investigate the possibility to use microalgae polysaccharides as a plant biostimulant. The crude polysaccharides extract (PS) from three microalgae strains were applied to Solanum lycopersicum plants by irrigation and compared basing on their effects on shoot and root length, nodes number and shoot and root dry weight. The application of 1 mg mL-1 PS from A. platensis, D. salina and Porphorydium sp. on tomato plants improved significantly the nodes number (NN), shoot dry weight (SDW), and shoot length (SS) by75 %, 46,6 %, 25,26 % compared to control respectively. Furthermore, crude PS treatment showed an improvement of carotenoid, chlorophyll and proteins content, and Nitrate Reductase (NR), NAD-Glutamate Dehydrogenase (NAD-GDH) activities in plants leaves compared to control. 1 mg mL-1 of Porphorydium sp. enhanced significantly the carotenoid content and NAD-GDH activity by 400 %, 200 % compared to control respectively. In the same way, A. platensis PS improved chl a, chl b and NR activity by 90.1 %, 102.7 % and 88.34 compared to control respectively. In addition, it is found that a PS treatment has affected the protein content, which reaches 88.3 % under 0.5 mg mL-1 of D. salina PS treatment. GC-MS metabolomics analysis also showed a change in lipids, sterol and alkanes profiles. Some sterols precursors were increased such as Cholesta-6,22,24-triene, which may indicate an enhancement of the biosynthesis of sterols and/or steroidal glycoalkaloids in treated plants. Therefore, this is an evidence to use microalgae polysaccharides as a plant biostimulant.

Draft Genome Sequence of Stenotrophomonas maltophilia MDMC339, Isolated from Soil of Merzouga Desert in Morocco.

Here, we report the draft genome sequence of Stenotrophomonas maltophilia MDMC339, a strain able to survive in the difficult conditions imposed by the Merzouga desert. The analyzed genome contains 4,788,525 bp with 4,262 genes coding for proteins, including several genes related to stress.

Genome Sequences of Six SARS-CoV-2 Strains Isolated in Morocco, Obtained Using Oxford Nanopore MinION Technology.

Here, we report the draft genome sequences of six severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains. SARS-CoV-2 is responsible for the COVID-19 pandemic, which started at the end of 2019 in Wuhan, China. The isolates were obtained from nasopharyngeal swabs from Moroccan patients with COVID-19. Mutation analysis revealed the presence of the spike D614G mutation in all six genomes, which is widely present in several genomes around the world.

Genomic Diversity and Hotspot Mutations in 30,983 SARS-CoV-2 Genomes: Moving Toward a Universal Vaccine for the "Confined Virus"?

The COVID-19 pandemic has been ongoing since its onset in late November 2019 in Wuhan, China. Understanding and monitoring the genetic evolution of the virus, its geographical characteristics, and its stability are particularly important for controlling the spread of the disease and especially for the development of a universal vaccine covering all circulating strains. From this perspective, we analyzed 30,983 complete SARS-CoV-2 genomes from 79 countries located in the six continents and collected from 24 December 2019, to 13 May 2020, according to the GISAID database. Our analysis revealed the presence of 3206 variant sites, with a uniform distribution of mutation types in different geographic areas. Remarkably, a low frequency of recurrent mutations has been observed; only 169 mutations (5.27%) had a prevalence greater than 1% of genomes. Nevertheless, fourteen non-synonymous hotspot mutations (>10%) have been identified at different locations along the viral genome; eight in ORF1ab polyprotein (in nsp2, nsp3, transmembrane domain, RdRp, helicase, exonuclease, and endoribonuclease), three in nucleocapsid protein, and one in each of three proteins: Spike, ORF3a, and ORF8. Moreover, 36 non-synonymous mutations were identified in the receptor-binding domain (RBD) of the spike protein with a low prevalence (<1%) across all genomes, of which only four could potentially enhance the binding of the SARS-CoV-2 spike protein to the human ACE2 receptor. These results along with intra-genomic divergence of SARS-CoV-2 could indicate that unlike the influenza virus or HIV viruses, SARS-CoV-2 has a low mutation rate which makes the development of an effective global vaccine very likely.

Local genetic structure and worldwide phylogenetic position of symbiotic Rhizobium leguminosarum strains associated with a traditional cultivated crop, Vicia ervilia, from Northern Morocco.

A total of 212 symbiotic bacteria were isolated from nodules of Vicia ervilia, a traditional crop cultivated in Northern Morocco. The isolates were recovered from 10 different sites, trapped each time with the local cultivar grown in the same field. Four loci were sequenced in order to characterize the isolates, including two housekeeping genes (recA and glnII), one plasmidic symbiotic gene (nodC) and one locus from another plasmid (prL11). In several isolates, two different copies of glnII were detected and sequenced, suggesting a unique duplication event, which has never been reported previously. There was no correlation between the genetic differentiation among cultivars and among bacteria, showing that the evolution of the bacterial population was independent, at least partially, from the host plant. By placing the haplotypes in a wide-ranging phylogenetic reconstruction, it was shown that the diversity detected in Morocco was spread throughout the different clades detected worldwide. The differentiation between areas relied on frequency variations of haplotypes rather than a presence/absence pattern. This finding raises new questions concerning bacterial genetic resource preservation, and confirms the old tenet "everything is everywhere but the environment selects".

Recovery of symbiotic nitrogen fixing acacia rhizobia from Merzouga Desert sand dunes in South East Morocco--Identification of a probable new species of Ensifer adapted to stressed environments.

Bacteria capable of nodulating Acacia tortilis and A. gummifera could be recovered from sand dunes collected in the Moroccan Merzouga desert. The trapping approach enabled the recovery of 17 desert rhizobia that all clustered within the Ensifer (Sinorhizobium) genus. Four isolates of the dominant genotype comprising 15 strains as well as 2 divergent strains were further characterized by MLSA. Phylogenetic analyzes indicated that the dominant genetic type was belonging to a new and yet undefined species within the Ensifer genus. Interestingly, housekeeping gene phylogenies showed that this possibly new species is also present in another desert but in India. Phylogenetic analyses of nifH and nodC sequences showed high sequence conservation among the Moroccan strains belonging to the dominant genotype but high divergence with sequences from Indian isolates suggesting acquisition of symbiotic genes through Horizontal Gene Transfer. These desert rhizobia were capable of growing in media containing high salt concentrations, under high pH and most of the strains showed growth at 45°C. Only recovered from desert type of Biome, yet, this new taxon appears particularly adapted to such harsh environment.