Plant growth promoting activities of endophytic bacteria from Melia azedarach (Meliaceae) and their influence on plant growth under gnotobiotic conditions.

Bacteria that live asymptomatically within plant tissues are known as endophytes. Because of the close relation with the plant host, they have been a matter of interest for application as plant growth promoters. Melia azedarach is a widely distributed medicinal tree with proven insecticidal, antimicrobial, and antiviral activity. The aim of this study was to isolate and characterize endophytic bacteria from M. azedarach and analyze their plant growth promoting activities for the potential application as biological products. Bacteria were isolated from roots and leaves of trees growing in two locations of Northeastern Argentina. The isolates were characterized by repetitive extragenic palindromic sequence PCR and 16S rDNA sequence analysis. The plant growth-promoting activities were assayed in vitro, improvement of plant growth of selected isolates was tested on M. azedarach plantlets, and the effect of selected ACC deaminase producing isolates was tested on tomato seedlings under salt-stress conditions. The highest endophytic bacterial abundance and diversity were obtained from the roots. All isolates had at least one of the assayed plant growth-promoting activities and 80 % of them had antagonistic activity. The most efficient bacteria were Pseudomonas monteilii, Pseudomonas farsensis, Burkholderia sp. and Cupriavidus sp. for phosphate solubilization (2064 µg P ml-1), IAA production (94.7 µg ml-1), siderophore production index (5.5) and ACC deaminase activity (1294 nmol α-ketobutyrate mg-1 h-1). M. azedarach inoculation assays revealed the bacterial growth promotion potential, with Pseudomonas monteilii, Pseudomonas farsensis and Cupriavidus sp. standing out for their effect on leaf area, leaf dry weight, specific leaf area, and total Chl, Mg and N content, with increases of up to 149 %, 58 %, 65 %, 178 %, 76 % and 97.7 %, respectively, compared to NI plants. Efficient ACC deaminase-producing isolates increased stress tolerance of tomato plants under saline condition. Overall, these findings indicate the potential of the endophytic isolates as biostimulant and biocontrol agents.

Complete mitochondrial genome of Melia azedarach L., reveals two conformations generated by the repeat sequence mediated recombination.

Melia azedarach is a species of enormous value of pharmaceutical industries. Although the chloroplast genome of M. azedarach has been explored, the information of mitochondrial genome (Mt genome) remains surprisingly limited. In this study, we used a hybrid assembly strategy of BGI short-reads and Nanopore long-reads to assemble the Mt genome of M. azedarach. The Mt genome of M. azedarach is characterized by two circular chromosomes with 350,142 bp and 290,387 bp in length, respectively, which encodes 35 protein-coding genes (PCGs), 23 tRNA genes, and 3 rRNA genes. A pair of direct repeats (R1 and R2) were associated with genome recombination, resulting in two conformations based on the Sanger sequencing and Oxford Nanopore sequencing. Comparative analysis identified 19 homologous fragments between Mt and chloroplast genome, with the longest fragment of 12,142 bp. The phylogenetic analysis based on PCGs were consist with the latest classification of the Angiosperm Phylogeny Group. Notably, a total of 356 potential RNA editing sites were predicted based on 35 PCGs, and the editing events lead to the formation of the stop codon in the rps10 gene and the start codons in the nad4L and atp9 genes, which were verified by PCR amplification and Sanger sequencing. Taken together, the exploration of M. azedarach gap-free Mt genome provides a new insight into the evolution research and complex mitogenome architecture.

Transcriptome analysis reveals the molecular mechanisms underlying the enhancement of salt-tolerance in Melia azedarach under salinity stress.

Melia azedarach demonstrates strong salt tolerance and thrives in harsh saline soil conditions, but the underlying mechanisms are poorly understood. In this study, we analyzed gene expression under low, medium, and high salinity conditions to gain a deeper understanding of adaptation mechanisms of M. azedarach under salt stress. The GO (gene ontology) analysis unveiled a prominent trend: as salt stress intensified, a greater number of differentially expressed genes (DEGs) became enriched in categories related to metabolic processes, catalytic activities, and membrane components. Through the analysis of the category GO:0009651 (response to salt stress), we identified four key candidate genes (CBL7, SAPK10, EDL3, and AKT1) that play a pivotal role in salt stress responses. Furthermore, the KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis revealed that DEGs were significantly enriched in the plant hormone signaling pathways and starch and sucrose metabolism under both medium and high salt exposure in comparison to low salt conditions. Notably, genes involved in JAZ and MYC2 in the jasmonic acid (JA) metabolic pathway were markedly upregulated in response to high salt stress. This study offers valuable insights into the molecular mechanisms underlying M. azedarach salt tolerance and identifies potential candidate genes for enhancing salt tolerance in M. azedarach.

New anti-inflammatory limonoids from the fruits of Melia azedarach.

The phytochemical study of the fruits of Melia azedarach (Meliaceae) led to the isolation and characterisation of two novel natural limonoids1-deoxy- 3, 20-dicinnamoyl-11-methoxy-meliacarpinin (1) and 12ß- O- methyl nimbolinin A (2), along with twelve known limonoids. Its structure was identified by 1D- and 2D-NMR, HR-ESI-MS and comparison with published data. The anti-inflammatory effect of the compounds was measured in vitro in RAW 264.7 cells by evaluating the production of NO stimulated by LPS. Compounds 1, 8 and 14 indicated significant anti-inflammatory effect with inhibition rate of 11.76, 8.45 and 6.59 µM, respectively. Limonoid 1 significantly inhibited the production of NO, TNF-α and IL-1ß in RAW 264.7 cells. Therefore, limonoid derivative may be a promising source of bioactive metabolite for inflammatory diseases.

Occurrence of 'Candidatus Phytoplasma asteris'-Related Strains infecting Chinaberry (Melia azedarach) Showing Chlorotic Leaf Symptoms on Hainan Island of China.

Chinaberry (Melia azedarach), belonging to the family of Meliaceae, is an ornamental tree distributes across southern of China. In the autumn of 2021, In an area of 400 acres located in Wanning city of Hainan Province, a tropical island in China, with coordinates of 110°28'42.72″E, 19°2'9.96″N, about 20 % (100) of the chinaberry trees showed disease symptoms included chlorotic leaves. The disease symptoms were consistent with infections by a phloem-limited prokaryotic pathogen phytoplasma. The samples of six symptomatic and three asymptomatic were collected for pathogen detection. To identify the pathogen, total nucleic acids were extracted from 0.10 g fresh leaf tissues from the diseased and healthy plant using CTAB DNA extraction method based on Doyle and Doyle. Three primer pairs of R16mF2/R16mR1, secAfor1/secArev3 and fTuf1/rTuf1 were used for specific identification of phytoplasma conserved gene fragments of 16S rDNA, secA and tuf, PCR amplification. Target PCR bands were amplified from the DNA of six diseased chinaberry samples, but not from the DNA of the healthy samples. The products of amplified were cloned and sequenced by Biotechnology (Shanghai) Co., Ltd. (Guangzhou, China). The phytoplasma gene sequences of 16S rRNA, secA and tuf were obtained and all the sequences were identical with the length of 1336 bp, 710 bp and 955 bp, respectively. Representative sequence data for strain MaCL-hn were deposited in Genbank under accession Nos. OR438638 (16S rDNA), OR513089 (secA) and OR860415 (tuf). The phytoplasma strain identified in the study was described as chinaberry chlorotic leaf (MaCL) phytoplasma, MaCL-hn strain. BLAST search based on 16S rRNA genes showed that 43 strains in 16SrI group 'Candidatus Phytoplasma asteris' showed 100% similarity with the 16SRNA sequence of MaCL-hn. BLAST search based on secA genes showed that 9 strains in the phytoplasma group showed 100% similarity with the 16SRNA sequence of MaCL-hn. BLAST search based on tuf genes showed that 21 strains in the phytoplasma group showed 100% similarity with the 16SRNA sequence of MaCL-hn. RFLP analysis based on iPhyClassifier indicated that the MaCL-hn strain was a member of 16SrI-B subgroup with a similarity coefficient 1.00 to the reference phytoplasma strain (AP006628). Phylogenetic tree was constructed based on 16S rRNA by MEGA 11.0 using neighbor-joining (NJ) method with 1000 bootstrap value. The results showed that the MaCL-hn strains were clustered into one clade with 16SrI group 'Ca. Phytoplasma asteris' related strains with 99 % bootstrap value. Multilocus sequence analysis (MLSA) based on the concatenated sequences with the length of 3001 bp including the sequences of 16S rRNA, secA and tuf showed that the MaCL-hn strains were clustered into one clade with the phytoplasma strains in the group with 100 % bootstrap value. To our knowledge, this is the first report that chinaberry can be infected by 'Ca. Phytoplasma asteris'-related strains belonging to 16SrI-B subgroup on Hainan Island of China. This finding in the study will contribute to the epidemic monitoring and the preventive management of the phytoplasmas and their related diseases.

Limonoids from the roots of Melia azedarach and their anti-inflammatory activity.

Twelve undescribed limonoids, meliazedarines J-U (1-12), along with a known one, were isolated from the roots of Melia azedarach. Their structures were elucidated by extensive spectroscopic investigations, X-ray diffraction analyses, and ECD calculations. Compounds 1-8 were identified as ring intact limonoids, while compounds 9-12 were established as ring C-seco ones. The anti-inflammatory potential of compounds 1-4, 6, 8, 9, and 11-13 was evaluated on macrophages. Compounds 1, 3, 4, 6, and 9 significantly suppressed nitric oxide production in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages, among them compound 3 showed the best inhibitory effect with an IC50 value of 7.07 ± 0.48 µΜ. Furthermore, compound 3 effectively reduced interleukin-1ß secretion in LPS plus nigericin-induced THP-1 macrophages by inhibiting NLRP3 inflammasome activation. The results strongly suggested that limonoids from the roots of M. azedarach might be candidates for treating inflammation-related diseases.

Production of biochar from Melia azedarach seeds for the crystal violet dye removal from water: combining of hydrothermal carbonization and pyrolysis.

Biochar has shown large potential in water treatment because of its low cost, good textural properties, and high reusability. In this study, two porous biochars were developed from the Melia azedarach seeds via direct pyrolysis process (B-700) and through hydrothermal carbonization followed with pyrolysis (HB-700). They were characterized by morphology, structural characteristics, and surface features and used to adsorb the crystal violet (CV) dye in water environment. Results of the isotherm approaches demonstrated that the removal capacity of these biochars reached 119.4 mg/g for B-700, and 209 mg/g for HB-700 (at 45°C). Also, the Avrami model best fitted the kinetic data. The electrostatic attraction was regarded as one of the adsorptions mechanisms of CV dye. The regeneration tests reveal that both B-700 and HB-700 are good reusable adsorbents. Finally, findings of the study showed that the hydrothermal carbonization method that precede the pyrolysis process can improve significantly the adsorption capacity of the produced biochar.

Along with cyclic electron flow and non-photochemical quenching, water-to-water cycle is involved uniquely in alleviating Zn stress-caused photodamage in Melia azedarach.

Zinc (Zn) is a widespread industrial pollutant that has detrimental effects on plant growth and development. Photoprotective properties ensure plant survival during stress by protecting the photosynthetic apparatus. This occurs via numerous mechanisms, including non-photochemical quenching (NPQ), cyclic electron flow (CEF) and the water-to-water cycle (WWC). However, whether and how Zn stress affects the photoprotective properties of plants to enhance the tolerance of Zn toxicity remains unknown. In this study, we treated Melia azedarach plants with different Zn concentrations ranging from 200 to 1000 mg kg-1. We then analyzed the activities of two leaf photosynthetic pigment components-photosystems I and II (PSI and PSII)-and the relative expression levels of their subunit genes. As expected, we found that Zn treatment decreases photosynthesis and increases photodamage in M. azedarach leaves. The Zn treatments exacerbated a variety of photodamage phenotypes in photosystem activities and altered the expression levels of key photosystem complex genes and proteins. Furthermore, our results demonstrated that PSI was more seriously damaged than PSII under Zn stress. Subsequently, we compared differences in photodamage in the NPQ, CEF and WWC photoprotection pathways under Zn stress and found that each exerted a protective function again photodamage under 200 mg kg-1 Zn stress. The NPQ and CEF may also play major protective roles in the avoidance of irreversible photodamage and helping to ensure survival under higher (i.e., 500 and 1000 mg kg-1) levels of Zn stress. Thus, our study revealed that NPQ- and CEF-based photoprotection mechanisms are more effective than WWC in M. azedarach upon Zn stress.

Climate-Driven Adaptive Differentiation in Melia azedarach: Evidence from a Common Garden Experiment.

Studies of local adaptation in populations of chinaberry (Melia azedarach L.) are important for clarifying patterns in the population differentiation of this species across its natural range. M. azedarach is an economically important timber species, and its phenotype is highly variable across its range in China. Here, we collected M. azedarach seeds from 31 populations across its range and conducted a common garden experiment. We studied patterns of genetic differentiation among populations using molecular markers (simple sequence repeats) and data on phenotypic variation in six traits collected over five years. Our sampled populations could be subdivided into two groups based on genetic analyses, as well as patterns of isolation by distance and isolation by environment. Significant differentiation in growth traits was observed among provenances and families within provenances. Geographic distance was significantly correlated with the quantitative genetic differentiation (QST) in height (HEIT) and crown breadth. Climate factors were significantly correlated with the QST for each trait. A total of 23 climatic factors were examined. There was a significant effect of temperature on all traits, and minimum relative humidity had a significant effect on the survival rate over four years. By comparing the neutral genetic differentiation (FST) with the QST, the mode of selection acting on survival rate varied, whereas HEIT and the straightness of the main trunk were subject to the same mode of selection. The variation in survival rate was consistent with the variation in genetic differentiation among populations, which was indicative of local adaptation. Overall, our findings provide new insights into the responses of the phenological traits of M. azedarach to changes in the climate conditions of China.

Phytochemical and Antioxidant Profile of the Medicinal Plant Melia azedarach Subjected to Water Deficit Conditions.

Environmental stress triggered by climate change can alter the plant's metabolite profile, which affects its physiology and performance. This is particularly important in medicinal species because their economic value depends on the richness of their phytocompounds. We aimed to characterize how water deficit modulated the medicinal species Melia azedarach's lipophilic profile and antioxidant status. Young plants were exposed to water deficit for 20 days, and lipophilic metabolite profile and the antioxidant capacity were evaluated. Leaves of M. azedarach are rich in important fatty acids and oleamide. Water deficit increased the radical scavenging capacity, total phenol, flavonoids, and catechol pools, and the accumulation of ß-sitosterol, myo-inositol, succinic acid, sucrose, d-glucose and derivatives, d-psicofuranose, d-(+)-fructofuranose, and the fatty acids stearic, α-linolenic, linoleic and palmitic acids. These responses are relevant to protecting the plant against climate change-related stress and also increase the nutritional and antioxidant quality of M. azedarach leaves.

Role of quercetin and rutin in enhancing the therapeutic potential of mesenchymal stem cells for cold induced burn wound.

Introduction: Cold burn wounds differ in their pathophysiological spectrum as compared to other types of burn wounds. These wounds have prolonged devastating effects on the body including hypertrophic scars, contracture, and necrosis. Mesenchymal stem cells (MSCs) are considered promising candidates for the complete regeneration of burn wounds. However, transplanted MSCs face the challenge to survive under the harsh tissue conditions. Preconditioning of MSCs with bioactive compounds may enhance their survival and regenerative potential for use in clinical applications. Bioactive compounds of Melia azedarach are well known for their potential role in treating different types of skin wounds due to their anti-inflammatory, anti-viral, anti-cytotoxic, and anti-oxidative properties. This study aims to evaluate the synergistic effects of human umbilical cord derived MSCs (hUC-MSCs) after preconditioning them with bioactive compounds of M. azedarach (quercetin and rutin) for cold induced burn wounds. Method: Human umbilical cord MSCs (hUC-MSCs) were characterized based on their specific cell surface markers and treated with 20 µM of quercetin or rutin. In vitro scratch assay was performed to measure cell migration and wound closure. In vivo cold burn wound model was developed via direct exposure of the dorsal rat skin to liquid nitrogen. hUC-MSCs were subcutaneously transplanted next day of burn wound induction and wound was examined at different time points corresponding to the wound healing phases (days 3, 7, and 14). The regenerative potential of preconditioned hUC-MSCs was assessed in different groups; control (treated only with hUC-MSCs), and treated groups (quercetin or rutin treated hUC-MSCs). Healing potential and wound closure were evaluated by histological, gene expression, and immunohistochemical analyses of the wound tissues before and after treatment. Results: Scratch assay exhibited enhanced cell migration towards wound closure in the treated groups as compared to the control. Macroscopic examination of the wound revealed scab formation at day 14 in control, whereas scab was detached and the wound tissue was remarkably remodeled in the treated groups. Comparison between the treated groups showed that burn wound treated with quercetin significantly increased healing potential than the rutin treated MSCs. Histological findings showed enhanced regeneration of skin layers along with hair follicles in the quercetin group, while increased neovascularization was noted in both treatment groups. Gene profile of wound healing mediators illustrated significant upregulation of IL-5, IL-4, GPX-7, TXNRD-2, PRDX, VEGF, and FGF and downregulation of inflammatory cytokines IL-1ß and IL-6. Conclusion: In conclusion, synergistic effect of hUC-MSCs and bioactive compounds of M. azedarach enhances wound healing by reducing the inflammation, mitigating oxidative stress and enhancing neovascularization. The study findings will aid in designing more effective treatment options for cold burn wounds.

Triterpenoids from the fruits of Melia azedarach L. and their cytotoxic activities.

Eleven undescribed tetracyclic triterpenoids, meliazedarachins A-K, along with twenty-six known compounds were isolated from the fruits of Melia azedarach L.. Their structures were determined by HRESIMS, UV, IR, NMR, X-ray diffraction, electronic circular dichroism (ECD) spectra, and the modified Mosher's method. The cytotoxic activities of all the isolates were measured. Meliazedarachin K and mesendanin N showed cytotoxicity against five human cancer cell lines with IC50 values ranging from 9.02 to 31.31 µM. Meliazedarachin K showed significant cytotoxicity against HCT116 cell line with IC50 value of 9.02 ± 0.84 µM. 21α-methylmelianodiol showed significant cytotoxicity against HCT116 and RKO cell lines with IC50 values of 10.16 ± 1.22 and 8.57 ± 0.80 µM, respectively.

The complete genome sequence of Melia azedarach Linn. (Meliaceae): a multi-purpose pesticide species.

The plant genus Melia has two or four species in the modern world, and is a natural source of a traditional pesticide, Toosendanin. In this study, we report the complete chloroplast genome of Melia azedarach, assembled from whole-genome high-throughput sequencing data, as a resource for future studies on the taxonomy and evolution of Melia. The chloroplast genome was 160,393 bp in length, with a large single-copy region of 87,598 bp, a small single-copy region of 18,709 bp, separated by two inverted repeat regions of 27,043 bp each. It was predicted to contain a total of 133 genes, with an overall GC content of 37.37%. Phylogenetic analysis placed M. azedarach closest to Azadirachta sp. in Meliaceae.

Acaricidal Efficacy of Biosynthesized Zinc Oxide Nanoparticles Against Hyalomma dromedarii (Acari: Ixodidae) and Their Toxic Effects on Swiss Albino Mice.

PURPOSE: The current study aimed to investigate the efficacy of zinc oxide nanoparticles (ZnO NPs) synthesized by Melia azedarach aqueous extract to control Hyalomma dromedarii tick, and to evaluate their toxic effects on Swiss albino mice. METHODS: ZnO NPs were synthesized using M. azedarach aqueous extract. UV-visible spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive spectroscopy were used to characterize the synthesized NPs. Egg, nymph, larva, and adult immersion tests were used for bioassay of tick stages with the synthesized ZnO NP. A toxicity study was performed on Swiss albino mice after treatment with 1/10 of the oral LD50 of ZnO NPs (8437 mg/kg) for 5 successive days by oral gavage. RESULTS: The LC50 of ZnO NPs on the eggs, larvae, and nymphs was 11.6, 8.03, and 3.9 mg/ml, respectively. The reproductive performance of females treated with ZnO NPs was lower than that of untreated females. The hematological results showed an insignificant increase in the level of white blood cells with normal red blood cells, hemoglobin, in addition to normal platelet count. The biochemical analysis showed an insignificant increased level (P > 0.05) of alkaline phosphatase and alanine aminotransferase. The liver and kidney suffered few histopathological changes after oral administration of ZnO NPs. CONCLUSION: These results suggest that ZnO NPs have good acaricidal activity against eggs, larvae, and engorged nymphs of H. dromedarii. ZnO NPs minimized the number of eggs laid by engorged females and the hatchability of their eggs. ZnO NPs did not affect unfed adults. The toxicity results of the mice revealed insignificant changes in the hemogram, biochemistry, with liver and kidney suffering few histopathological changes. Future studies are needed to assess application routes (topical vs oral). Based on these findings, ZnO NPs may be incorporated in the control of camel tick H. dromedarii.

Sterilization protocols and the effect of plant growth regulators on callus induction and secondary metabolites production in in vitro cultures Melia azedarach L.

Melia azedarach L. is a valuable source of antioxidants and secondary metabolites. This study is a first extensive report about the effect of different serialization protocols and plant growth regulators (PGRs) on explant disinfection efficiency, callus induction and secondary metabolites production and accumulation in callus cultures of M. azedarach L. In this regard, the effect of plant growth regulators on callus induction and secondary metabolites production were examined. In addition, different sterilization agents were evaluated for disinfection of chinaberry leaf explants. The results showed that the lowest percentage of explant contamination and browning with the highest percentage of callus induction and callus growth obtained with explants pretreated with benomyl (2 g/L) for 2 h and sterilized with 7% H2O2 for 10 min and NaOCl 2% (without pH adjustment) for 12 min. Although adjusting the pH of NaOCl to pH = 7 and 10 significantly reduced the microbial contamination and increased the percentage of contamination-free cultures of M. azedarach L., adversely influenced the explant viability and callus induction and growth. The highest percentage of callus induction obtained on the MS medium containing 3 mg/L NAA/2,4-D and 1 or 3 mg/L Kin/BAP, and the highest callus yield (1804.833 mg/explant) belonged to the MS medium supplemented with 5 mg/L 2,4-D and 5 mg/L Kin. The callus cultures grown on the MS medium supplemented with 3 mg/L NAA and 1 mg/L Kin produced the highest amount of Quercetin (2.06 mg/g fresh weight), Rutin (5.56 mg/g fresh weight) and Kaempferol (1.84 mg/g fresh weight).

The Development of a Green Innovative Bioactive Film for Industrial Application as a New Emerging Technology to Protect the Quality of Fruits.

Today, the most significant challenge encountered by food manufacturers is degradation in the food quality during storage, which is countered by expensive packing, which causes enormous monetary and environmental costs. Edible packaging is a potential alternative for protecting food quality and improving shelf life by delaying microbial growth and providing moisture and gas barrier properties. For the first time, the current article reports the preparation of the new films from Ditriterpenoids and Secomeliacins isolated from Melia azedarach (Dharek) Azadirachta indica plants to protect the quality of fruits. After evaluating these films, their mechanical, specific respirational, coating crystal elongation, elastic, water vapor transmission rate (WVTR), film thickness, and nanoindentation test properties are applied to apple fruit for several storage periods: 0, 3, 6, 9 days. The fruits were evaluated for postharvest quality by screening several essential phytochemical, physiological responses under film coating and storage conditions. It was observed that prepared films were highly active during storage periods. Coated fruits showed improved quality due to the protection of the film, which lowered the transmission rate and enhanced the diffusion rate, followed by an increase in the shelf life. The coating crystals were higher in Film-5 and lower activity in untreated films. It was observed that the application of films through dipping was a simple technique at a laboratory scale, whereas extrusion and spraying were preferred on a commercial scale. The phytochemicals screening of treated fruits during the storage period showed that a maximum of eight important bioactive compounds were present in fruits after the treatment of films. It was resolved that new active films (1-5) were helpful in the effective maintenance of fruit quality and all essential compounds during storage periods. It was concluded that these films could be helpful for fruits growers and the processing industry to maintain fruit quality during the storage period as a new emerging technology.

Essential oils from Melia azedarach L. (Meliaceae) leaves: chemical variability upon environmental factors.

The chemical composition of the essential oils extracted from the leaves of Melia azedarach L. collected monthly from July 2019 to June 2020 was examined via gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) techniques. Analysis of the essential oils identified about 17 compounds representing more than 85% of the oil. Oil yields were higher in the months of June and August, and the primary compounds identified were ß-caryophyllene (3.50-63.41%), benzaldehyde (3.50-55.98%), and azulene (1.27-19.05%). A correlation analysis was performed to determine the relationship between yields and climatic conditions, and between constituent concentration and temperature and precipitation values during the study period. As per our findings, although not significant, a positive correlation was determined between yield and climatic parameters. However, the oil components were categorized into four groups based on their correlation with temperature and precipitation indices. Among the major components of the essential oils, only azulene and ß-caryophyllene exhibited a negative correlation with both precipitation and temperature. The results show substantial differences in the chemical composition of M. azedarach essential oils and provide further insight into the phytochemical constituents that are sensitive to climate fluctuations. Furthermore, it provides an indication of the optimal time that the plant produces the important mono- and sesquiterpene components and the biological significance of their regulation.

Enhancing plant growth and biomass production by overexpression of GA20ox gene under control of a root preferential promoter.

Overexpression of GA20 oxidase gene has been a recent trend for improving plant growth and biomass. Constitutive expression of GA20ox has successfully improved plant growth and biomass in several plant species. However, the constitutive expression of this gene causes side-effects, such as reduced leaf size and stem diameter, etc. To avoid these effects, we identified and employed different tissue-specific promoters for GA20ox overexpression. In this study, we examined the utility of At1g promoter to drive the expression of GUS (ß-glucuronidase) reporter and AtGA20ox genes in tobacco and Melia azedarach. Histochemical GUS assays and quantitative real-time-PCR results in tobacco showed that At1g was a root-preferential promoter whose expression was particularly strong in root tips. The ectopic expression of AtGA20ox gene under the control of At1g promoter showed improved plant growth and biomass of both tobacco and M. azedarach transgenic plants. Stem length as well as stem and root fresh weight increased by up to 1.5-3 folds in transgenic tobacco and 2 folds in transgenic M. azedarach. Both tobacco and M. azedarach transgenic plants showed increases in root xylem width with xylem to phloem ratio over 150-200% as compared to WT plants. Importantly, no significant difference in leaf shape and size was observed between At1g::AtGA20ox transgenic and WT plants. These results demonstrate the great utility of At1g promoter, when driving AtGA20ox gene, for growth and biomass improvements in woody plants and potentially some other plant species.

Phytochemical composition of almond oil from Melia azedarach L. and its larvicidal, ovicidal, repellent and enzyme activities in Culex pipiens L

@#Melia azedarach L. (Meliaceae) is a botanical species with focal point of global research for its biological properties. The Melia azedarach tree is distinguished by its rapid growth, its adaptation to different temperate zones, as well as its insecticidal properties. All this made us think of exploiting it in biological control against different stages of mosquitoes. To this end, we aim, through the present work, to evaluate the effectiveness of Melia azedarach extracts against Culex pipiens mosquito. More specifically, our study focuses on determining the chemical composition of Melia almond oil, as well as the larvicidal, ovicidal and repellent activities on Culex pipiens L. mosquito as well as the activities of acetylcholinesterase (AChE) and glutathione-S-transferase (GST). Almond oil was extracted by a Soxhlet and subjected to gas chromatography–mass spectrometry (GC/MS). The yield was found to be 35.17%. The chemical composition revealed the presence of various phytoconstituents. A total of 7 compounds were identified, the main ones being 9,11-Octadecadienoic acid, methyl ester, (E,E)- (79.32%), 9-octadecenoic acid (Z)-, methyl ester (13.24%), hexadecanoic acid and methyl ester (3.69%). The larvicidal bioassays were performed according to the protocol recommended by the World Health Organization with concentrations varying from 20 to 80 mg/L depending on the exposure time (24, 48 and 72 hours). The almond oil exhibited remarkable larvicidal activity against fourth instar larvae and the lethal concentrations were determined (LC25= 23.70 mg/L, LC50=35.49 mg/L, LC90=79.61 mg/L). The results also showed that the oil caused an ovicidal activity with a significant effect on egg hatch. The recorded hatching percentages were respectively 88.79% and 72.40% for the LC25 and LC50, and this compared to the control series. Moreover, this oil exhibited significant repellency against adult mosquitoes. Furthermore, the enzymatic measurements performed on LC50 and LC90 treated larvae revealed a neurotoxic activity and a stimulation of the detoxification system as evidenced, respectively, by an inhibition of AChE and induction in GST activity. Overall, our data proved that Melia azedarach almond oil could be considered as a potent biorational alternative to synthetic insecticides for mosquito control.

In silico estrogen receptor alpha antagonist studies and toxicity prediction of Melia azedarach leaves bioactive ethyl acetate fraction.

The estrogen hormone dependent accounts for a major cause in the incidence of women breast cancer. Thus, their receptor, especially the estrogen receptor α (ER-α), is becoming a target in endocrine treatment. These ligand-inducible nuclear functions are regulated by an array of phytochemical and synthetic compounds, such as 17 ß-estradiol and tamoxifen (4-hydroxytamoxifen [4OHT]). The Chinaberry (Melia azedarach) leaves are known naturally for relieving internal and external diseases. Previous studies revealed the potency of Melia's ethanolic extract and ethyl acetate fractions as anticancer; furthermore, this study aimed to resolve possible ER-α antagonist's mechanism and safety from M. azedarach leaves ethyl acetate fraction contents. Melia's phytochemical content was analyzed with electrospray ionization liquid chromatography-mass spectrometry, while its ER-α antagonist's potency was investigated by in silico. The computational docking was used to 3ERT (a human ER-α-4OHT binding domain complex) with Autodock Vina and related programs. The results presented Energy binding (ΔG) of Melia's quercetin 3-O-(2'',6''-digalloyl)-ß-D-galactopyranoside was similar to 4OHT, and lower than its agonist 17 ß-estradiol. Furthermore, the toxicity prediction of these compounds were revealed safer than 4OHT. The Melia's leaves ethyl acetate fraction, therefore, is a potential pharmacological material for further studies.