Unveiling resilience: coelomic fluid bacteria's impact on plant metabolism and abiotic stress tolerance.

Earthworms' coelomic fluid (CF) has been discovered to possess properties that promote plant development. In particular, the earthworm's coelomic fluid-associated bacteria (CFB) are the primary factor influencing the plants' response. To investigate this, we used bacteria isolated from the CF and selected based on different plant growth-promoting traits, in a mesocosm ecosystem that includes plants. This experiment aimed to assess their impact on the metabolism of plants growing under abiotic stress environments (alkaline soil and nitrogen (N), phosphate (P), and potassium (K) deficit) and compare the lipid profiles of plants under the various treatments. We used seven different bacterial species isolated from the CF of Aporrectodea molleri and as a plant model Zea mays L. For the metabolomic analysis method, we used gas chromatography-mass spectrometry lipidomic. After observing the metabolomic profiles, we found that a few molecular pathways are involved in how plants react to bacterial biostimulants. The bacterial isolates belonging to Pantoea vagans, Pseudomonas aeruginosa, Bacillus paramycoides, and Bacillus thuringiensis have led to a significant increase in synthesizing several metabolites belonging to various chemical categories. Contrary to predictions, abiotic stress did not cause a drop in the composition and concentration of lipids in plants treated with the CFB, demonstrating the rigidity of the protective mechanisms. The statistical analysis based on the Pearson method revealed a positive significant correlation between plant growth parameters (length of the aerial part, surface of the leaves, and biomass) and some metabolites belonging to fatty acids, carboxylic acids, benzene derivatives, and alkanes. Moreover, the standard metabolic components of all treatments in much higher concentrations during bacterial treatments than the control treatment suggests that the bacteria have stimulated the overexpression of these metabolic components. According to these results, we could assume that plants treated with CFB exhibit an adaptability of abiotic stress defense mechanisms, which may be attributed to the upregulation of genes involved in lipid biosynthesis pathways.

Agronomic advantage of bacterial biological nitrogen fixation on wheat plant growth under contrasting nitrogen and phosphorus regimes.

Introduction: Given their remarkable capacity to convert atmospheric nitrogen into plant-accessible ammonia, nitrogen-fixing microbial species hold promise as a sustainable alternative to chemical nitrogen fertilizers, particularly in economically significant crops like wheat. This study aimed to identify strains with optimal attributes for promoting wheat growth sustainably, with a primary emphasis on reducing reliance on chemical nitrogen fertilizers. Methods: We isolated free nitrogen-fixing strains from diverse rhizospheric soils across Morocco. Subsequently, we conducted a rigorous screening process to evaluate their plant growth-promoting traits, including nitrogen fixation, phosphate solubilization, phytohormone production and their ability to enhance wheat plant growth under controlled conditions. Two specific strains, Rhodotorula mucilaginosa NF 516 and Arthrobacter sp. NF 528, were selected for in-depth evaluation, with the focus on their ability to reduce the need for chemical nitrogen supply, particularly when used in conjunction with TSP fertilizer and natural rock phosphate. These two sources of phosphate were chosen to assess their agricultural effectiveness on wheat plants. Results and discussion: Twenty-two nitrogen-fixing strains (nif-H+) were isolated from various Moroccan rhizospheric soils, representing Bacillus sp., Pseudomonas sp., Arthrobacter sp., Burkholderia sp. and a yeast-like microorganism. These strains were carefully selected based on their potential to promote plant growth. The findings revealed that the application of Rhodotorula mucilaginosa NF 516 and Arthrobacter sp. NF 528 individually or in combination, significantly improved wheat plant growth and enhanced nutrients (N and P) uptake under reduced nitrogen regimes. Notably, their effectiveness was evident in response to both natural rock phosphate and TSP, demonstrating their important role in wheat production under conditions of low nitrogen and complex phosphorus inputs. This research underscores the significant role of nitrogen-fixing microorganisms, particularly Rhodotorula mucilaginosa NF 516 and Arthrobacter sp. NF 528, in wheat production under conditions of low nitrogen and complex phosphorus inputs. It showcases their potential to reduce chemical nitrogen fertilization requirements by up to 50% without compromising wheat plant yields. Our study emphasizes the importance of bacterial biological nitrogen fixation in meeting the remaining nitrogen requirements beyond this reduction. This underscores the vital role of microbial contributions in providing essential nitrogen for optimal plant growth and highlights the significance of biological nitrogen fixation in sustainable agriculture practices.

Isolation, identification and pathogenicity of local entomopathogenic bacteria as biological control agents against the wild cochineal Dactylopius opuntiae (Cockerell) on cactus pear in Morocco.

The Opuntia ficus-indica (L.) cactus, a crucial crop in Morocco, is threatened by the wild cochineal, Dactylopius opuntiae (Cockerell). The aim of this research was to investigate the efficacy of nine bacterial strains against both D. opuntiae nymphs and adults females applied individually or after black soap in the laboratory, greenhouse, and field conditions. Using the partial 16S ribosomal DNA, the bacterial isolates were identified as Pseudomonas koreensis, Pseudomonas sp., Burkholderia sp. and Bacillus sp. Under laboratory conditions, the insecticidal activity of P. koreensis strain 66Ms.04 showed the level mortality (88%) of adult females' at 108 CFU/mL, 7 days after application. At a concentration of 108 CFU/mL, P. koreensis strain 66Ms.04 and Pseudomonas sp. (strains 37 and 5) caused 100% nymphs mortality rate three days after application. Under greenhouse conditions, the use of P. koreensis strain 66Ms.04 at 108 CFU/mL following the application of black soap (60 g/L) demonstrated the maximum levels of females and nymphs' mortalities with 80 and 91.25%, respectively, after 8 days of treatment. In field conditions, the combined application of the P. koreensis strain 66Ms.04 at 108 CFU/mL with black soap at 60 g/L, for an interval of 7 days, significantly increased the mortality of adult females to 93.33% at 7 days after the second application. These findings showed that the combined treatment of P. koreensis strain 66Ms.04 with black soap can be a potent and eco-friendly pesticide against D. opuntiae.

Aphanothece sp. as promising biostimulant to alleviate heavy metals stress in Solanum lycopersicum L. by enhancing physiological, biochemical, and metabolic responses.

Heavy metals (H.M) are a major environmental concern around the world. They have harmful impact on plant productivity and pose a serious risk to humans and animals health. In the present study, we investigated the effect of Aphanothece crude extract (ACE) on physiological, biochemical, and metabolic responses of tomato plant exposed to 2 mM Pb and Cd. The results showed a significant reduction of tomato plant weights and perturbation in nutrients absorption under 2 mM Pb and Cd conditions. Moreover, ACE treatment showed a significant enhancement of plant biomass compared to plants under Pb and Cd. On the other hand, ACE application favoured H.M accumulation in root and inhibited their translocation to shoot. In addition, ACE treatment significantly enhanced several stress responses in plant under Pb and Cd stress such as scavenging enzymes and molecules: POD, CAT, SOD, proline, and polyphenols etc. Furthermore, ACE treatment showed remodulation of metabolic pathways related to plant tolerance such as wax construction mechanism, particularly SFA, UFA, VLFA, alkanes, alkenes, and sterols biosynthesis to enhance tolerance and resistance to H.M stress. In the present study, we emphasized that ACE alleviates H.M stress by minimizing metal translocation to above-part of plant and enhancing plant growth, nutrients absorption, and biochemical responses.

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.

Investigation of bacterial diversity using 16S rRNA sequencing and prediction of its functionalities in Moroccan phosphate mine ecosystem.

Native plants in extreme environments may harbor some unique microbial communities with particular functions to sustain their growth and tolerance to harsh conditions. The aim of this study was to investigate the bacterial communities profiles in some native plants and samples of the Moroccan phosphate mine ecosystem by assessing the percentages of taxonomic identification using six hypervariable regions of the 16S rRNA. The rhizosphere of the three wild plants in the Moroccan phosphate mine is characterized by interesting bacterial diversity including Proteobacteria (62.24%, 71.15% and 65.61%), Actinobacteria (22.53%, 15.24%, 22.30%), Bacteroidetes (7.57%; 4.23%; 7.63%), and Firmicutes (5.82%; 1.17%; 2.83%). The bulk phosphate mine samples were dominated by Actinobacteria with average relative abundance of 97.73% that are different from those inferred in the rhizosphere samples of the native plants. The regions V3, V4 and V67 performed better in the taxonomic profiling at different taxonomic levels. Results indicated that both plant genotype and mainly soil conditions may be involved in the shaping of bacterial diversity. Such indication was also confirmed by the prediction of functional profiles that showed enrichment of many functions related to biological nitrogen fixation in the rhizosphere of native plants and the stress related functions in the bulk phosphate mine in comparison with the wheat rhizosphere samples.

Chemical Composition and Insecticidal Potential of Six Essential Oils from Morocco against Dactylopius opuntiae (Cockerell) under Field and Laboratory Conditions.

The carmine cochineal Dactylopius opuntiae (Cockerell) is the major insect pest of the prickly-pear cactus Opuntia ficus-indica (L.) in Morocco. The present study investigated the insecticidal activities of six essential oils (EOs) against nymphs and adult females of D. opuntiae applied singly or in combination with a detergent under laboratory and field conditions. Under laboratory conditions, M. pulegium and O. vulgare L. essential oils showed a high level of insecticidal activity at 5%, with 98% and 92% females' mortality, respectively, 5 days after treatments. The M. pulegium and O. vulgaris oils at 5% applied in combination with black soap at (60 g/L) induced the highest toxic activity on adult females, 100% and 96% at 5 days after treatments, respectively. Under field conditions, M. pulegium and O. vulgare oils at 5% in combination with black soap (60 g/L) showed the highest adult female mortalities with 96.33 and 92.56%, respectively, 7 days after the first application. The double application of M. pulegium oil at 5% significantly increased the mortality of adult females up to 91%, 5 days after the second spray. GC-MS analysis revealed that the most abundant constituent of M. pulegium and O. vulgare oils was pulegone (84.69%) and durenol (76.53%), respectively. These findings showed that the use of M. pulegium and O. vulgare in combination with black soap or in double sprays could be incorporated in the management package for the control of the wild cochineal D. opuntiae, as a safe and natural alternative to chemical insecticides.

Nitrogen Fixing Azotobacter Species as Potential Soil Biological Enhancers for Crop Nutrition and Yield Stability.

Biological nitrogen fixation (BNF) refers to a microbial mediated process based upon an enzymatic "Nitrogenase" conversion of atmospheric nitrogen (N2) into ammonium readily absorbable by roots. N2-fixing microorganisms collectively termed as "diazotrophs" are able to fix biologically N2 in association with plant roots. Specifically, the symbiotic rhizobacteria induce structural and physiological modifications of bacterial cells and plant roots into specialized structures called nodules. Other N2-fixing bacteria are free-living fixers that are highly diverse and globally widespread in cropland. They represent key natural source of nitrogen (N) in natural and agricultural ecosystems lacking symbiotic N fixation (SNF). In this review, the importance of Azotobacter species was highlighted as both important free-living N2-fixing bacteria and potential bacterial biofertilizer with proven efficacy for plant nutrition and biological soil fertility. In addition, we described Azotobacter beneficial plant promoting traits (e.g., nutrient use efficiency, protection against phytopathogens, phytohormone biosynthesis, etc.). We shed light also on the agronomic features of Azotobacter that are likely an effective component of integrated plant nutrition strategy, which contributes positively to sustainable agricultural production. We pointed out Azotobacter based-biofertilizers, which possess unique characteristics such as cyst formation conferring resistance to environmental stresses. Such beneficial traits can be explored profoundly for the utmost aim to research and develop specific formulations based on inoculant Azotobacter cysts. Furthermore, Azotobacter species still need to be wisely exploited in order to address specific agricultural challenges (e.g., nutrient deficiencies, biotic and abiotic constraints) taking into consideration several variables including their biological functions, synergies and multi-trophic interactions, and biogeography and abundance distribution.

Role of Inorganic Phosphate Solubilizing Bacilli Isolated from Moroccan Phosphate Rock Mine and Rhizosphere Soils in Wheat (Triticum aestivum L) Phosphorus Uptake.

This work aimed to assess the ability of plant growth-promoting Bacilli isolated from wheat rhizosphere and rock phosphate mine soils to convert inorganic phosphate (Pi) from Moroccan natural phosphate (NP) to soluble forms. The effect of these bacteria on wheat plants in order to increase their phosphorus (P) uptake in vitro was also investigated. Bacteria were isolated from wheat rhizosphere and natural rock phosphate soils and screened for their ability to solubilize Tri-Calcium Phosphate (TCP) and Natural Rock Phosphate (NP), to produce indole-3-acetic acid (IAA), siderophores and 1-aminocyclopropane-1-carboxylate (ACC) deaminase. Isolates were identified by 16S rRNA sequencing and tested for their capacity to increase wheat plants growth and their phosphorus uptake.Twenty-four strains belonging to Bacillus genus isolated from both biotopes were screened for their ability to solubilize Pi. The highest NP solubilization was showed by strains isolated from wheat rhizosphere. Solubilization of Pi was accompanied by organic acid production. Strains produce IAA, siderophore and ACC deaminase. Inoculation assays using efficient NP-solubilizing bacilli strains from both sources showed the ability of these isolates to increase wheat growth and the phosphorus uptake under in vitro conditions. Bacilli strains isolated from rhizosphere soil and natural rock phosphorus soil showed effective solubilization of Pi from rock phosphate. Phosphate solubilizing Bacilli were evaluated for their plant growth promotion under in vitro conditions. Results revealed the positive effect of all strains on biometric parameters and P content of wheat seedlings.