Impact of two Erwinia sp. on the response of diverse Pisum sativum genotypes under salt stress.

Currently, salinization is impacting more than 50% of arable land, posing a significant challenge to agriculture globally. Salt causes osmotic and ionic stress, determining cell dehydration, ion homeostasis, and metabolic process alteration, thus negatively influencing plant development. A promising sustainable approach to improve plant tolerance to salinity is the use of plant growth-promoting bacteria (PGPB). This work aimed to characterize two bacterial strains, that have been isolated from pea root nodules, initially called PG1 and PG2, and assess their impact on growth, physiological, biochemical, and molecular parameters in three pea genotypes (Merveille de Kelvedon, Lincoln, Meraviglia d'Italia) under salinity. Bacterial strains were molecularly identified, and characterized by in vitro assays to evaluate the plant growth promoting abilities. Both strains were identified as Erwinia sp., demonstrating in vitro biosynthesis of IAA, ACC deaminase activity, as well as the capacity to grow in presence of NaCl and PEG. Considering the inoculation of plants, pea biometric parameters were unaffected by the presence of the bacteria, independently by the considered genotype. Conversely, the three pea genotypes differed in the regulation of antioxidant genes coding for catalase (PsCAT) and superoxide dismutase (PsSOD). The highest proline levels (212.88 µmol g-1) were detected in salt-stressed Lincoln plants inoculated with PG1, along with the up-regulation of PsSOD and PsCAT. Conversely, PG2 inoculation resulted in the lowest proline levels that were observed in Lincoln and Meraviglia d'Italia (35.39 and 23.67 µmol g-1, respectively). Overall, this study highlights the potential of these two strains as beneficial plant growth-promoting bacteria in saline environments, showing that their inoculation modulates responses in pea plants, affecting antioxidant gene expression and proline accumulation. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01419-8.

Mechanistic insight of Staphylococcus aureus associated skin cancer in humans by Santalum album derived phytochemicals: an extensive computational and experimental approaches.

An excessive amount of multidrug-resistant Staphylococcus aureus is commonly associated with actinic keratosis (AK) and squamous cell carcinoma (SCC) by secreted virulence products that induced the chronic inflammation leading to skin cancer which is regulated by staphylococcal accessory regulator (SarA). It is worth noting that there is currently no existing published study that reports on the inhibitory activity of phytochemicals derived from Santalum album on the SarA protein through in silico approach. Therefore, our study has been designed to find the potential inhibitors of S. aureus SarA protein from S. album-derived phytochemicals. The molecular docking study was performed targeting the SarA protein of S. aureus, and CID:5280441, CID:162350, and CID: 5281675 compounds showed the highest binding energy with -9.4 kcal/mol, -9.0 kcal/mol, and -8.6 kcal/mol respectively. Further, molecular dynamics simulation revealed that the docked complexes were relatively stable during the 100 ns simulation period whereas the MMPBSA binding free energy proposed that the ligands were sustained with their binding site. All three complexes were found to be similar in distribution with the apoprotein through PCA analysis indicating conformational stability throughout the MD simulation. Moreover, all three compounds' ADMET profiles revealed positive results, and the AMES test did not show any toxicity whereas the pharmacophore study also indicates a closer match between the pharmacophore model and the compounds. After comprehensive in silico studies we evolved three best compounds, namely, Vitexin, Isovitexin, and Orientin, which were conducted in vitro assay for further confirmation of their inhibitory activity and results exhibited all of these compounds showed strong inhibitory activity against S. aureus. The overall result suggests that these compounds could be used as a natural lead to inhibit the pathogenesis of S. aureus and antibiotic therapy for S. aureus-associated skin cancer in humans as well.

Novel computational and drug design strategies for the inhibition of human T-cell leukemia virus 1-associated lymphoma by Astilbin derivatives.

Human T-cell leukemia virus 1 (HTLV-1) associated lymphoma is a devastating malignancy triggered by HTLV-1 infections. We employeda comprehensive drug design and computational strategy in this work to explore the inhibitory activitiesof Astilbin derivatives against HTLV-1-associated lymphoma. We evaluated the stability, binding affinities, and various computational analysis of Astilbin derivatives against target proteins, such as HTLV-1 main protease and HTLV-1 capsid protein. The root mean square deviation (RMSD), root mean square fluctuation, radius of gyration, hydrogen bond analysis, principal component analysis (PCA) and dynamic cross-correlation matrix (DCCM) were applied to characterize these protein-ligand interactions further. Ligand-03 and ligand-04 exhibited notable binding affinity to HTLV-1 capsid protein, while ligand-05 displayed high binding affinity to HTLV-1 protease. MD simulation analysis revealed that ligand-03, bound to HTLV-1 capsid protein, demonstrated enhanced stability with lower RMSD values and fewer conformational changes, suggesting a promising binding orientation. Ligand-04, despite stable binding, exhibited increased structural deviations, making it less suitable. Ligand-05 demonstrated stable binding to HTLV-1 protease throughout the simulation period at 100 nanoseconds. Hydrogen bond analysis indicated that ligand-05 formed persistent hydrogen bonds with significantresidues, contributing to its stability. PCA highlighted ligand-03's more remarkable conformational changes, while DCCM showed ligand-05's distinct dynamics, indicating its different behavior in the complex. Furthermore, binding free energy calculations supported the favorable interactions of ligand-03 and ligand-04 with HTLV-1 capsid protein, while ligand-05 showed weaker interactions with HTLV-1 protease. Molecular electrostatic potential and frontier molecular orbital analyses provided insights into these compounds' charge distribution and stability. In conclusion, this research found Astilbin derivatives as potential inhibitors of HTLV-1-associated lymphoma. Future attempts at drug development will benefit from the steady interaction landscape provided by Ligand-03, Ligand-04 and Ligand-05, which showed the most attractive binding profile with the target protein. These results open up new opportunities for innovative drug development, and more experimental testing should be done between Astilbin derivatives and HTLV-1-associated lymphoma.Communicated by Ramaswamy H. Sarma.

Napthyridine-derived compounds as promising inhibitors for Staphylococcus aureus CrtM: a primer for the discovery of potential anti-Staphylococcus aureus agents.

The disease-free existence of humans is constantly under attack by a variety of infections caused by a variety of organisms including bacteria. Notable among the bacteria is Staphylococcus aureus which is an etiological organism for infections including impetigo, folliculitis, and furuncles. The response of the human immune system against this disease is often neutralized by the production of a pigment called Staphyloxanthin (STX) via a series of reactions mediated by several enzymes. Among these enzymes, dehydrosqualene synthase, also known as CrtM, has emerged as a viable drug target due to its role in mediating the first step of the pathway. Consequently, this study employs molecular modeling approaches including molecular docking, quantum mechanical calculations, and molecular dynamics (MD) simulations among others to investigate the potential of napthyridine derivatives to serve as inhibitors of the CrtM. The results of the study revealed the high binding affinities of the compounds for the target as demonstrated by their docking scores, while further subjection to screening pipeline aimed at determining their fitness for development into drugs revealed just one compound namely 6-[[1-[(2-fluorophenyl) methyl]triazol-4-yl]methoxy]-4-oxo-1H-1,5-naphthyridine-3-carboxylic acid as the compound with good drug-like, pharmacokinetics, and toxicity properties profiles. A 100 ns-long MD simulation of the complexes formed after molecular docking revealed the stable interaction of the compound with the target. Ultimately, this study can be a promising outlet to discover a weapon to fight against clinically resistant bacteria, however, further experimental studies are suggested to carry out in the wet lab, pre-clinical, and clinical levels.

Insight into biological activities of chemically characterized extract from Marrubium vulgare L. in vitro, in vivo and in silico approaches.

Aqueous extracts of Marrubium vulgare L. (M. vulgare) are widely used in traditional medicine for their therapeutic effects. Hence, this study aims to evaluate in vitro, in vivo, and in silico the biological activities of M. vulgare aqueous extract to further support their traditional use. Qualitative phytochemical tests of M. vulgare extracts showed the presence of primary and secondary metabolites, while quantitative analyses recorded revealed the contents of total phenols, flavonoids, and tannins, with values of 488.432 ± 7.825 mg/EAG gallic acid extract/g, 25.5326 ± 1.317 mg/EQ Quercetin extract/g and 23.966 ± 0.187 mg/EC catechin extract/g, respectively. Characterization of the phytochemical constituents of the extract revealed the presence of catechin and maleic acid as the most abundant while the evaluation of the antioxidant power revealed that the extract possesses significant antioxidant capacity, antimitotic potential, and antimicrobial properties against Streptococcus agalactiae and Staphylococcus epidermidis among many others. The antidiabetic activity of the extract showed a potent antihyperglycemic effect and a significant modulation of the pancreatic α-amylase activity as revealed by both in vitro and in vivo analysis, while an in silico evaluation showed that chemicals in the studied extract exhibited the aforementioned activities by targeting 1XO2 antimitotic protein, W93 antidiabetic protein and 1AJ6 antimicrobial protein, which revealed them as worthy of exploration in drug discovery odyssey. Conclusively, the result of this study demonstrates the numerous biological activities of M. vulgare and gives credence to their folkloric and traditional usage.

Volatile Constituents in Essential Oil from Leaves of Withania adpressa Coss. Ex Exhibit Potent Antioxidant and Antimicrobial Properties against Clinically-Relevant Pathogens.

Withania adpressa Coss. ex is a plant used in traditional medications. Antioxidant, antibacterial, and antifungal properties of the essential oil from leaves of Withania adpressa Coss ex. (EOW) were investigated. EOW was extracted using a Clevenger apparatus, and its volatile compounds were characterized by GC-MS. Antioxidant potency was determined using DPPH, FRAP, and TAC assays. Antibacterial effects were determined vs. Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, and Streptococcus pneumonia; while its antifungal efficacy was determined vs. Candida albicans, Aspergillus flavus, Aspergillus niger, and Fusarium oxysporum using the disc diffusion and minimum inhibitory concentration bioassays. A chromatographic analysis showed that EOW contained eight phytochemical compounds constituting 99.14% of the total mass of oil. Caryophyllene (24.74%), Longifolene (21.37%), δ-Cadinene (19.08%), and Carene (14.86%) were predominant compounds in EOW. The concentrations required to inhibit 50% of free radical (IC50) values of antioxidant activities of EOW were 0.031 ± 0.006 mg/mL (DPPH), 0.011 ± 0.003 mg/mL (FRAP), and 846.25 ± 1.07 mg AAE/g (TAC). Inhibition zone diameters of EOW vs. bacteria were 18.11 ± 0.5 mm (E. coli), 17.10 ± 0.42 mm (S. aureus), 12.13 ± 0.31 mm (K. pneumoniae), and 11.09 ± 0.47 mm (S. pneumoniae), while MIC values were 51 ± 3, 47 ± 5, 46 ± 3 and 31 ± 1 µg/mL, respectively. Inhibition zone diameters of EOW vs. fungi were 31.32 ± 1.32, 29.00 ± 1.5, 27.63 ± 2.10, and 24.51 ± s1.07 mm for A. flavus, C. albicans, F. oxysporum, and A. niger, respectively. MIC values were 8.41 ± 0.40, 28.04 ± 0.26, 9.05 ± 0.76, and 22.26 ± 0.55 µg/mL, respectively. Importantly, the highest dose of EOW (1 mg/mL) showed negligible (~5%) cytotoxicity against MCF-12, a normal human epithelial cell line derived from the mammary gland, thus underscoring its wide safety and selectivity against tested microbes. To sum it up, EOW has exhibited promising antioxidant and antimicrobial properties, which suggests potential to abrogate antibiotic resistance.

Phytochemical Analysis and Antioxidant, Antibacterial, and Antifungal Effects of Essential Oil of Black Caraway (Nigella sativa L.) Seeds against Drug-Resistant Clinically Pathogenic Microorganisms.

Nigella sativa (NS) is a plant that has long been utilized in traditional medicine as a treatment for certain diseases. The aim of this work was to valorize the essential oil (EO) of this species by phytochemical analysis and antimicrobial and antioxidant evaluation. EO was extracted by hydrodistillation from the seeds of Nigella sativa (EO-NS). Phytochemical content of EO-NS was evaluated by use of gas chromatography coupled to mass spectrometry (GC-MS/MS). Antioxidant ability was in vitro determined by use of three assays: 2.2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing power (FRAP), and total antioxidant capacity (TAC) relative to two synthetic antioxidants: BHT and quercetin. Antimicrobial effect was evaluated against four clinically important bacterial strains (Staphylococcus aureus, ATCC 6633; Escherichia coli, K12; Bacillus subtilis, DSM 6333; and Proteus mirabilis, ATCC 29906) and against four fungal strains (Candida albicans, ATCC 10231; Aspergillus niger, MTCC 282; Aspergillus flavus, MTCC 9606; and Fusarium oxysporum, MTCC 9913). Fifteen constituents that accounted for the majority of the mass of the EO-NS were identified and quantified by use of GC-MSMS. The main component was O-cymene (37.82%), followed by carvacrol (17.68%), α-pinene (10.09%), trans-sabinene hydrate (9.90%), and 4-terpineol (7.15%). EO-NS exhibited significant antioxidant activity with IC50, EC50, and total antioxidant capacity (TAC) of 0.017 ± 0.0002, 0.1196 ± 0.012, and 114.059 ± 0.97 mg EAA/g, respectively. Additionally, EO-NS exhibited promising antibacterial activity on all strains under investigation, especially on E. coli K12 resulting in inhibition diameter of 38.67 ± 0.58 mm and a minimum inhibitory concentration (MIC) of 1.34 ± 0.00 µg/mL. Also, EO-NS had significant antifungal efficacy, with a percentage of inhibition of 67.45 ± 2.31% and MIC of 2.69 ± 0.00 µg/mL against F. oxysporum, MTCC 9913 and with a diameter of inhibition 42 ± 0.00 mm and MIC of 0.67 ± 0.00 µg/mL against C. albicans. To minimize development of antibiotic-resistant bacteria, EO-NS can be utilized as a natural, alternative to synthetic antibiotics and antioxidants to treat free radicals implicated in microbial infection-related inflammatory reactions.

Chemical Composition and Antimicrobial Activity of Essential Oils from Three Mediterranean Plants against Eighteen Pathogenic Bacteria and Fungi.

The chemical composition and antimicrobial activity of essential oils (EOs) obtained from three medicinal plants of the Moroccan flora were evaluated. The chemical composition of EOs of Thymus leptobotrys, Laurus nobilis and Syzygium aromaticum was determined using a gas chromatograph coupled with mass spectrometry. Carvacrol (75.05%) was the main constituent of T. leptobotrys EOs, while 1,8-cineole (31.48%) and eugenol (82.16%) were the predominant components of L. nobilis and S. aromaticum EOs, respectively. The antimicrobial activity of the EOs was evaluated qualitatively and quantitatively against 18 microbial strains pathogenic to humans by using the disc diffusion method, and by measuring the minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC). The EOs of T. leptobotrys were the most active against the strains tested, with inhibitory zone values ranging from 7.00 to 45.00 mm, and MIC and MMC values ranging from 0.312 to 80.00 mg/mL. In many cases, these EOs exhibited higher antibacterial and antifungal activities than the chemical compounds ciprofloxacin and fluconazole, respectively. This high antimicrobial activity can be ascribed to their richness in carvacrol. The EOs of T. leptobotrys, L. nobilis, and S. aromaticum could be considered a promising alternative to replace chemical antimicrobials, and a readily available natural source of bioactive compounds.

Genome-wide footprints in the carob tree (Ceratonia siliqua) unveil a new domestication pattern of a fruit tree in the Mediterranean.

Intense research efforts over the last two decades have renewed our understanding of plant phylogeography and domestication in the Mediterranean basin. Here we aim to investigate the evolutionary history and the origin of domestication of the carob tree (Ceratonia siliqua), which has been cultivated for millennia for food and fodder. We used >1000 microsatellite genotypes to delimit seven carob evolutionary units (CEUs). We investigated genome-wide diversity and evolutionary patterns of the CEUs with 3557 single nucleotide polymorphisms generated by restriction-site associated DNA sequencing (RADseq). To address the complex wild vs. cultivated status of sampled trees, we classified 56 sampled populations across the Mediterranean basin as wild, seminatural or cultivated. Nuclear and cytoplasmic loci were identified from RADseq data and separated for analyses. Phylogenetic analyses of these genomic-wide data allowed us to resolve west-to-east expansions from a single long-term refugium probably located in the foothills of the High Atlas Mountains near the Atlantic coast. Our findings support multiple origins of domestication with a low impact on the genetic diversity at range-wide level. The carob was mostly domesticated from locally selected wild genotypes and scattered long-distance westward dispersals of domesticated varieties by humans, concomitant with major historical migrations by Romans, Greeks and Arabs. Ex situ efforts to preserve carob genetic resources should prioritize accessions from both western and eastern populations, with emphasis on the most differentiated CEUs situated in southwest Morocco, south Spain and eastern Mediterranean. Our study highlights the relevance of wild and seminatural habitats in the conservation of genetic resources for cultivated trees.

The first use of morphologically isolated arbuscular mycorrhizal fungi single-species from Moroccan ecosystems to improve growth, nutrients uptake and photosynthesis in Ceratonia siliqua seedlings under nursery conditions.

The carob tree (Ceratonia siliqua L.) is an important component in semi-arid Mediterranean ecosystems, particularly in Morocco where it plays a considerable socio-economic role. This species is widely used in the reforestation programmes and in the rehabilitation of degraded soils serving both environmental and socio-economic objectives. In spite of these assets, this species is suffering the particular climatic conditions, rare and irregular rains, long hot and dry summers, generally, leading to desertification processes. To withstand these contrasting conditions, selected arbuscular mycorrhizal fungi (AMF) were tested for their contribution to the growth, nutrient uptake and photosynthesis improvement of the carob tree C. siliqua under nursery conditions. The objective of this study was, to evaluate the effects of some arbuscular mycorrhizal fungi complexes isolated in different Mediterranean ecosystems compared to single-species isolates selected using morphological tools on the growth, mineral nutrition, and chlorophyll content of C. siliqua seedlings. The results indicate that all the used AMF inocula stimulated significantly the height of C. siliqua seedlings after eight months under nursery conditions. An increase in plant height between 33% and 70% compared to a control without inoculation was recorded. Similarly, the aerial dry weight recorded an increase of 62% to 124% comparing inoculated and non-inoculated seedlings. The root dry weight has shown an increase rate of 24% to 86% compared to the control. The analysis of mineral contents in plant tissues, showed a highly significant increase in P. N. K. Ca and Mg levels of the aerial parts compared to the control. A significant increase in chlorophyll contents was noticed when inoculated seedlings were compared to non-inoculated ones. This study had confirmed the importance of AMF improving the growth of C. siliqua seedlings; the AMF complexes remain to have the important growth and mineral nutrition responses. However some single- species have shown similar magnitude to the complexes for all analysed parameters. A large biofertilizer potential of the single-species isolates in the inoculation of C. siliqua is demonstrated for the first time.

Chemical Composition and Antifungal, Insecticidal and Repellent Activity of Essential Oils From Origanum compactum Benth. Used in the Mediterranean Diet.

Essential oils (EO) of Origanum compactum Benth. (O. compactum) are well known for their biological and pharmacological activities. This study aimed to assess the chemical composition, antifungal, insecticidal and repellent activities of EO of O. compactum used in the Mediterranean diet. Phytochemical screening was conducted using gas chromatography-mass spectrometry (GC/MS). Antifungal activity was tested by the disc diffusion method followed by a minimal inhibitory concentration (MIC) assay against Candida albicans (C. albicans), Aspergillus flavus (A. flavus), Aspergillus niger (A. nige), and Fusarium oxysporum (F. oxysporum). Repellent potential and toxicity of EO by contact and inhalation were tested against Callosobruchus maculatus (C. maculatus). The yield of essential oil obtained by hydrodistillation of O. compactum was 4.41 ± 0.35%, mainly composed of Carvacrol (38%) and Thymol (31.46%). Regarding antifungal activity, the results revealed a wide antifungal spectrum of the studied EO against the tested strains, which reached 100% growth inhibition, especially against A. niger and C. albicans even at the lowest MIC values (3.125 µg/mL). Concerning insecticidal activity, the EO caused total mortality of C. maculatus adults at a dose of 20 µL/L air with LC50 value of 5.3 µL/L air. A significant reduction in the number of eggs and emergence was proportionally recorded with increasing doses up to 100% at 20 µL/L air. For repellent activity, the studied EO showed a moderate repellent activity with an average percentage of 39.16%. The outcome of this work revealed that O. Compactum EO could be a sustainable and environmentally friendly alternative bioinsecticide and bio-fungicide to replace the chemically synthesized forms.

Ethnobotanical Study of Medicinal Plants Used as Therapeutic Agents to Manage Diseases of Humans.

OBJECTIVE: This work aimed to survey medicinal plants used in phytotherapy in the Fez-Boulemane region, Morocco. METHODS: A comprehensive ethnobotanical survey was conducted using a questionnaire to collect data from thirty herbalists on medicinal plants used for therapeutic purposes in the Fez-Boulemane region, Morocco. RESULTS: The mean age of herbalists interviewed was 52.13 years. Forty percent of the herbalists were illiterate, and 73% referred to the experiences of their parents as knowledge of the properties and uses of medicinal plants. One hundred and eight medicinal plants belonging to 51 botanical families were recommended by herbalists in the region of Fez-Boulemane, Morocco, for treatment. According to the respondents, Lawsonia inermis L., Rosmarinus officinalis L., and Lavandula coronopifolia L. were the most used plants with the percentage of 13%, 12%, and 11%, respectively. Most plants had been involved in the treatment of digestive disorders (25%) and osteoarticular diseases (24%). Some (7.4%) of the plants mentioned in our survey were potentially toxic. CONCLUSION: It was learned that irrational use of toxic plants and unknown compositions of recipes are recommended to consumers. As a result, particular attention should be paid to risks related to plants used in traditional treatment without scientific validation. It is envisaged that increasing awareness, by conducting educational campaigns and transferring evidence-based scientific knowledge, on traditional treatments among the local population is expected to have beneficial impacts on health and disease management.

Lavandula dentata L.: Phytochemical Analysis, Antioxidant, Antifungal and Insecticidal Activities of Its Essential Oil.

Antioxidant, antifungal and insecticidal activities of essential oil (EO) extracted from the Moroccan lavender (Lavandula dentata) were investigated and their chemical constituents determined. Gas chromatography with flame ionization detection (GC-FID) and gas chromatography-mass spectrometry analyses (GC-MS) were used to examine the phytochemical composition of EO. Antioxidant potential was examined in vitro by use of three tests: DPPH inhibition, reducing power (FRAP) and total antioxidant capacity (TAC). Antifungal activity was assessed by calculating inhibition of growth of Alternaria alternata, Botrytis cinerea and Fusarium oxysporum. Repellent potential and toxicity of EO by contact and inhalation were performed against Callosobruchus maculatus. Sixteen constituents were detected in the EO of Lavandula dentata. The major component was linalool (45.06%) followed by camphor (15.62%) and borneol (8.28%). EO exhibited a significant antioxidant activity, as measured by DPPH and FRAP assays, with IC50 and EC50 values of 12.95 ± 1.300 mg/mL and 11.88 ± 0.23 mg/mL, respectively. EO of lavender exhibited total antioxidant capacity of 81.28 ± 2.28 mg AAE/g EO. EO of lavender showed an inhibitory effect on mycelial growth against tested fungi and was 100% in the case of B. cinerea. EO caused total mortality of adult C. maculatus from 5 µL/L air with LC50 value of 4.01 µL/L air. Significant reduction in numbers of eggs laid (99.2%) and emergence (100%) was observed in a dose-dependent manner up to maxima of 100% and 99.2%, respectively. EO of lavender also showed a moderate potency to repel insects with a mean of 34.44%. EO of Moroccan Lavandula dentata has potential to be an effective natural agent against free radical damage and could be an environmentally friendly alternative bio-fungicide and bio-insecticide.

Insecticidal and Antifungal Activities of Chemically-Characterized Essential Oils from the Leaves of Withania frutescens L.

The current study was conducted to investigate antifungal and insecticidal activities of essential oil extracted from the Moroccan Withania frutescens L. (EOW), and their chemical composition was profiled. To achieve this goal, EOW was extracted by the hydro-distillation method and their phytochemical constituents were characterized by gas chromatography-mass spectrometry analyses (GC-MS). Insecticidal activity was evaluated by use of four tests: contact toxicity, inhalation toxicity, and repellent tests. Antifungal activity was evaluated on Fusarium oxysporum f. sp. Ciceris (F. oxysporum) using different concentrations of EOW. GC/MS analysis revealed that EOW was rich in carvacrol (31.87%), thymol (30.08%), and camphor (9.13%). At a 1-µL/L dose, EOW exhibited mortality rates of 23.13 ± 1.07% and 24.41 ± 1.21% against Callosobruchus maculatus (C.maculatus) by inhalation and contact, respectively. Notably, EOW dose of 20 µL/L caused significant mortality rates of 95.1 ± 3.5% and 76.69 ± 1.71% by inhalation and contact, respectively. EOW exhibited an inhibitory effect on mycelial growth against the tested fungi F. oxysporum of 100% and 93.5 ± 1.1% for the 9 and 4.5 mg/mL doses, respectively. The reduced mycelial growth rate for F. oxysporum was recorded to be 0.3 ± 0.1 and 0.6 ± 0.1 mm/h for the EOW doses of 2.25 and 4.5 mg/mL, respectively. The outcome of the present work showed that EOW has a promising antifungal and insecticidal activity, and it can therefore be employed as a natural alternative insecticidal and mycocidal agent to replace the chemically-synthesized ones.

Application of Native or Exotic Arbuscular Mycorrhizal Fungi Complexes and Monospecific Isolates from Saline Semi-Arid Mediterranean Ecosystems Improved Phoenix dactylifera's Growth and Mitigated Salt Stress Negative Effects.

The date, the palm tree (Phoenix dactylifera L.) is an important component of arid and semi-arid Mediterranean ecosystems, particularly in Morocco where it plays a considerable socio-economic and ecological role. This species is largely affected by desertification, global warming, and anthropic pressure. Salinity is a very worrying problem that negatively affects the growth and the physiological and biochemical activities of the date palm. In these arid zones, the main challenge is to develop new environmentally friendly technologies that improve crop tolerance to abiotic restraints including salinity. In this sense, Arbuscular mycorrhizal fungi (AMF) have received much attention due to their capability in promoting plant growth and tolerance to abiotic and biotic stresses. It is thus fitting that the current research work was undertaken to evaluate and compare the effects of native AMF on the development of the growth and tolerance of date palm to salt stress along with testing their role as biofertilizers. To achieve this goal, two complexes and two monospecific isolates of native and non-native AMF were used to inoculate date palm seedlings under saline stress (0 g·L-1 Na Cl, 10 g·L-1, and 20 g·L-1 Na Cl). The obtained results showed that salinity drastically affected the physiological parameters and growth of date palm seedlings, whilst the application of selected AMF significantly improved growth parameters and promoted the activities of antioxidant enzymes as a protective strategy. Inoculation with non-native AMF complex and monospecific isolates showed higher responses for all analyzed parameters when compared with the native complex and isolate. It therefore becomes necessary to glamorize the fungal communities associated with date palm for their use in the inoculation of Phoenix dactylifera L. seedlings.

Phylogenetic study of rhizobia nodulating pea (Pisum sativum) isolated from different geographic locations in Tunisia.

Nodulated Pisum sativum plants showed the presence of native rhizobia in 16 out of 23 soil samples collected especially in northern and central Tunisia. A total of 130 bacterial strains were selected and three different ribotypes were revealed after PCR-RFLP analysis. Sequence analyses of rrs and four housekeeping genes (recA, atpD, dnaK and glnII) assigned 35 isolates to Rhizobium laguerreae, R. ruizarguesonis, Agrobacterium radiobacter, Ensifer meliloti and two putative genospecies. R. laguerreae was the most dominant species nodulating P. sativum with 63%. The isolates 21PS7 and 21PS15 were assigned to R. ruizarguesonis, and this is the first report of this species in Tunisia. Two putative new lineages were identified, since strains 25PS6, 10PS4 and 12PS15 clustered distinctly from known rhizobia species but within the R. leguminosarum complex (Rlc) with the most closely related species being R. indicum with 96.4% sequence identity. Similarly, strains 16PS2, 3PS9 and 3PS18 showed 97.4% and 97.6% similarity with R. sophorae and R. laguerreae, respectively. Based on 16S-23S intergenic spacer (IGS) fingerprinting, there was no clear association between the strains and their geographic locations. According to nodC and nodA phylogenies, strains of Rlc species and, interestingly, strain 8PS18 identified as E. meliloti, harbored the symbiotic genes of symbiovar viciae and clustered in two different clades showing heterogeneity within the symbiovar. All these strains nodulated and fixed nitrogen with pea plants. However, the strains belonging to A. radiobacter and the two remaining strains of E. meliloti were unable to nodulate P. sativum, suggesting that they were non-symbiotic strains. The results of this study further suggest that the Tunisian Rhizobium community is more diverse than previously reported.

Inoculation with selected indigenous mycorrhizal complex improves Ceratonia siliqua's growth and response to drought stress.

In the current study, we investigated the impact of inoculation with a selected indigenous arbuscular mycorrhizal fungi (AMF) complex on the growth and physiology of carob plants at increasing levels of watering (25, 50, 75 and 100% field capacity). The following growth and stress parameters were monitored in carob seedlings after 6 months of growth and 2 months of applied drought stress: fresh and dry weight, root and shoot lengths, leaf surface area, relative water content, stomatal conductance and membrane stability. Chlorophyll a and b, total soluble sugars, proline and protein contents were also determined along with the activities of stress enzymes: Catalase, Peroxidase and Superoxide dismutase. The obtained results indicate that inoculation with the indigenous AMF complex has a positive impact on the plant's growth as all the assessed parameters were significantly improved in the mycorrhizal plants. Additionally, our results show that mycorrhization contributes to the minimization of the impact of drought stress on the carob plants and allows a better adaptation to dry conditions.

Advances in genotyping microsatellite markers through sequencing and consequences of scoring methods for Ceratonia siliqua (Leguminosae).

PREMISE OF THE STUDY: Simple sequence repeat (SSR) or microsatellite markers have been used in a broad range of studies mostly scoring alleles on the basis of amplicon size as a proxy for the number of repeat units of an SSR motif. However, additional sources of variation within the SSR or in the flanking regions have largely remained undetected. METHODS: In this study, we implemented a next-generation sequencing-based genotyping approach in a newly characterized set of 18 nuclear SSR markers for the carob tree, Ceratonia siliqua. Our aim was to evaluate the effect of three different methods of scoring molecular variation present within microsatellite markers on the genetic diversity and structure results. RESULTS: The analysis of the sequences of 77 multilocus genotypes from four populations revealed SSR variation and additional sources of polymorphism in 87% of the loci analyzed (42 single-nucleotide polymorphisms and five insertion/deletion polymorphisms), as well as divergent paralog copies in two loci. Ignoring sequence variation under standard amplicon size genotyping resulted in incorrect identification of 69% of the alleles, with important effects on the genetic diversity and structure estimates. DISCUSSION: Next-generation sequencing allows the detection and scoring of SSRs, single-nucleotide polymorphisms, and insertion/deletion polymorphisms to increase the resolution of population genetic studies.

Use of arbuscular mycorrhizal fungi to improve the drought tolerance of Cupressus atlantica G.

In this study, we investigated whether indigenous arbuscular mycorrhizal (AM) fungi could improve the tolerance of Cupressus atlantica against water deficit. We tested a gradient of watering regime spanning from 90% to 25% of soil retention capacity of water on mycorhized and non-mycorhized seedlings in pot cultures with sterilized and non-sterilized soils. Our result showed a positive impact of AM fungi on shoot height, stem diameter and biomass as well as on the growth rate. We also observed that inoculation with AM fungi significantly improved uptake of minerals by C. atlantica in both sterilized and non-sterilized soils independently of water regimes. We found that mycorhized plants maintained higher relative water content (RWC) and water potential compared with non-mycorhized plants that were subjected to drought-stress regimes (50% and 25% of soil retention capacity). The contents of proline and of soluble sugars showed that their concentrations decreased in non-mycorhized plants subjected to DS. Superoxide dismutase (SOD) and catalase (CAT) activities also decreased in non-mycorhized plants submitted to DS compared to mycorhized plants. The same pattern was observed by measuring peroxidase (POD) enzyme activity. The results demonstrated that AM fungal inoculation promoted the growth and tolerance of C. atlantica against DS in pot cultures. Therefore, mycorrhizal inoculation could be a potential solution for the conservation and reestablishment of C. atlantica in its natural ecosystem.