Potential fungicidal and antiaflatoxigenic effects of cinnamon essential oils on Aspergillus flavus inhabiting the stored wheat grains.

Wheat is one of the essential crops for the human and animal nutrition, however, contamination with aflatoxigenic fungi, due to the improper storage conditions and high humidity, was the main global threats. So, preventing the growth of aflatoxigenic fungi in stored wheat grains, by using different essential oils was the main objective of this work. Aspergillus flavus EFBL-MU12 PP087400, EFBL-MU23 PP087401 and EFBL-MU36 PP087403 isolates were the most potent aflatoxins producers inhabiting wheat grains. The effect of storage conditions of wheat grains "humidity, temperature, incubation period, and pH" on growth of A. flavus, was assessed by the response surface methodology using Plackett-Burman design and FCCD. The highest yield of aflatoxins EFBL-MU12 B1 and B2 by A. flavus grown on wheat grains were 145.3 and 7.6 µg/kg, respectively, at incubation temperature 35°C, 16% moisture contents, initial pH 5.0, and incubated for 14 days. The tested oils had a powerful antifungal activity for the growth and aflatoxins production by A. flavus in a concentration-dependent manner. Among these oils, cinnamon oil had the highest fungicidal activity for A. flavus at 0.125%, with about 85-90 % reduction to the aflatoxins B1 and B2, conidial pigmentation and chitin contents on wheat grains. From the SEM analysis, cinnamon oils had the most deleterious effect on A. flavus with morphological aberrations to the conidial heads, vegetative mycelia, alteration in conidiophores identity, hyphae shrank, and winding. To emphasize the effect of the essential oils on the aflatoxins producing potency of A. flavus, the molecular expression of the aflatoxins biosynthetic genes was estimated by RT-qPCR. The molecular expression of nor-1, afLR, pKsA and afLJ genes was suppressed by 94-96%, due to cinnamon oil at 0.062% compared to the control. Conclusively, from the results, cinnamon oils followed by the peppermint oils displayed the most fungicidal activity for the growth and aflatoxins production by A. flavus grown on wheat grains.

Proteomics and metabolomics analyses of camptothecin-producing Aspergillus terreus reveal the integration of PH domain-containing proteins and peptidylprolyl cis/trans isomerase in restoring the camptothecin biosynthesis.

IMPORTANCE: Decreasing the camptothecin productivity by fungi with storage and subculturing is the challenge that halts their further implementation to be an industrial platform for camptothecin (CPT) production. The highest differentially abundant proteins were Pleckstrin homology (PH) domain-containing proteins and Peptidyl-prolyl cis/trans isomerase that fluctuated with the subculturing of A. terreus with a remarkable relation to CPT biosynthesis and restored with addition of F. elastica microbiome.

The effects of microbial fertilizers application on growth, yield and some biochemical changes in the leaves and seeds of guar (Cyamopsis tetragonoloba L.).

Guar (Cyamopsis tetragonoloba L.) is a summer legume that is becoming a crucial industrial crop because of its high gum and protein content. Thus far, the combined effects of arbuscular mycorrhizal fungi (AMF) and Bradyrhizobium on the yield and chemical composition of guar plants are not well studied. Therefore, the current investigation was designed to estimate the individual as well as the combined effects of AMF and Bradyrhizobium on plant growth, yield and nutritional quality of seeds and leaves of guar. AMF and/or Bradyrhizobium inoculation improved chemical composition of guar seeds and its morpho-physiological (plant height, fresh weight, dry weight, and yield production) traits. In addition to increased guar growth and yield production, the inoculation of AMF and/or Bradyrhizobium increased guar leaf and seed minerals, fiber, lipids, crude protein and ash contents. At primary metabolites, there were increases in sugar levels including raffinose stachyose, verbascose and galactomannan. These increases in sugar provided a route for organic acids, amino acids and fatty acids production. Interestingly, there was an increase in essential amino acids and unsaturated fatty acids. At the bioactive secondary metabolite levels, biofertilizers improved phenols and flavonoids levels and anthocyanin and polyamines biosynthesis. In line with these increases, precursors of anthocyanin (phenylalanine, p-coumaric acid, and cinnamic acid) and the levels of polyamines (diaminopropane, putrescine, cadaverine, spermidine, spermine, and agmatine) were increased. Overall, for the first time, our study shed the light on how AMF and Bradyrhizobium improved guar yield and metabolism. Our findings suggested that the combined inoculation of AMF and Bradyrhizobium is an innovative approach to improve guar growth, yield production and yield quality.

Production, bioprocessing and antiproliferative activity of camptothecin from Aspergillus terreus, endophyte of Cinnamomum camphora: restoring their biosynthesis by indigenous microbiome of C. camphora.

Fungal producing potency of camptothecin (CPT) raise the hope for their usage to be a platform for industrial production of CPT, nevertheless, attenuation of their productivity of CPT with the subculturing and preservation is the challenge. So, screening for novel endophytic fungal isolates with a reliable CPT-biosynthetic stability was the objective. Among the isolated endophytic fungi from the tested medicinal plants, Aspergillus terreus OQ642314.1, endophyte of Cinnamomum camphora, exhibits the highest yield of CPT (89.4 µg/l). From the NMR, FT-IR and LC-MS/MS analyses, the extracted CPT from A. terreus gave the same structure and molecular mass fragmentation pattern of authentic CPT (349 m/z). The putative CPT had a significant activity against MCF7 (0.27 µM) and HEPG-2 (0.8 µM), with a strong affinity to inhibits the human Topoisomerase 1 activity (IC50 0.362 µg/ml) as revealed from the Gel-based DNA relaxation assay. The purified CPT displayed a strong antimicrobial activity for various bacterial (E. coli and B. cereus) and fungal (A. flavus and A. parasiticus) isolates, ensuring the unique tertiary, and stereo-structure of A. terreus for penetrating the microbial cell walls and targeting the topoisomerase I. The higher dual activity of the purified CPT as antimicrobial and antitumor, emphasize their therapeutic efficiency, especially with growth of the opportunistic microorganisms due to the suppression of human immune system with the CPT uses in vivo. The putative CPT had an obvious activity against the tumor cell (MCF7) metastasis, and migration as revealed from the wound healing assay. The overall yield of A. terreus CPT was maximized with the Blackett-Burman design by twofolds increment (164.8 µg/l). The CPT yield by A. terreus was successively diminished with the multiple fungal subculturing, otherwise, the CPT productivity of A. terreus was restored, and increased over the zero culture upon coculturing with C. camphora microbiome (1.5% w/v), ensuring the restoring of CPT biosynthetic potency of A. terreus by the plant microbiome-derived chemical signals "microbial communication". This is the first report exploring the feasibility of A. terreus "endophyte of C. camphora" to be a preliminary platform for commercial production of CPT with a reliable sustainability upon uses of indigenous C. camphora microbiome.

Funneliformis constrictum modulates polyamine metabolism to enhance tolerance of Zea mays L. to salinity.

The mechanisms underlie increased stress tolerance in plants of salinity stress in plants by arbuscular mycorrhizal fungi (AMF) are poorly understood, particularly the role of polyamine metabolism. The current study was conducted to investigate how inoculation with the AMF, Funneliformis constrictum, affects maize plant tolerance to salt stress. To this end, we investigated the changes in photosynthesis, redox status, primary metabolites (amino acids) and secondary metabolism (phenolic and polyamine metabolism). Control and inoculated maize plants were grown using different concentrations of diluted seawater (0%, 10%, 20% and 40%). Results revealed that treatment with 10% seawater had a beneficial effect on AMF and its host growth. However, irrigation with 20% and 40% significantly reduced plant growth and biomass. As seawater concentration increased, the plants' reliance on mycorrhizal fungi increased resulting in enhanced growth and photosynthetic pigments contents. Under higher seawater concentrations, inoculation with AMF reduced salinity induced oxidative stress and supported redox homeostasis by reducing H2O2 and MDA levels as well as increasing antioxidant-related enzymes activities (e.g., CAT, SOD, APX, GPX, POX, GR, and GSH). AMF inoculation increased amino acid contents in shoots and roots under control and stress conditions. Amino acids availability provides a route for polyamines biosynthesis, where AMF increased polyamines contents (Put, Spd, Spm, total Pas) and their metabolic enzymes associated (ADC, SAMDC, Spd synthase, and Spm synthase), particularly under 40% seawater irrigation. Consistently, the transcription of genes, involved in polyamine metabolism was also up regulated in salinity-stressed plants. AMF further increased the expression in genes involved in polyamine biosynthesis (ODC, SAMDC, SPDS2 and decreased expression of those in catabolic biosynthesis (ADC and PAO). Overall, inoculation with Funneliformis constrictum could be adopted as a practical strategy to alleviate salinity stress.