Fungi Rhizopus Stolonifer Research Articles

Exploring the biotechnological prospects of a recently discovered fungus isolated from marine mucilage

AbstractThis study aimed to determine the usability of the identified fungus isolated from the marine-mucilage formation in the biotechnological processes. For this purpose, the antimicrobial and antioxidant activities of the extract obtained from the fungus were examined, and the fatty acid methyl ester composition was determined. The molecular identification of the pure fungal culture was made using LSU regions. In addition, fungal extracts were prepared using different solvents, and the antimicrobial activity of these extracts was investigated by disk diffusion and minimum inhibition concentration methods. At the same time, the antioxidant properties of these extracts were analyzed using the DPPH and ABTS free radical removal methods. Also, FAME analysis was performed to determine the fatty acid content of the fungal extract. According to the study results, the new isolate was identified as the fungus Rhizopus stolonifer. Although fungal extracts have no significant antimicrobial activity, it has been determined that they performed successful DPPH and ABTS scavenging activity without needing additional reactions. FAME results indicate that the mucilage-originated fungus R. stolonifer is a valuable provider of fatty acids that, when purified at large bioreactors, can be good and cheap sources of next-generation biologicals for wide-ranging biotechnological applications.

Metabolic engineering of omega-3 long chain polyunsaturated fatty acids in plants using different ∆6- and ∆5-desaturases co-expressed with LPCAT from the marine diatom Phaeodactylum tricornutum

Continuous research on obtaining an even more efficient production of very long-chain polyunsaturated fatty acids (VLC-PUFAs) in plants remains one of the main challenges of scientists working on plant lipids. Since crops are not able to produce these fatty acids due to the lack of necessary enzymes, genes encoding them must be introduced exogenously from native organisms producing VLC-PUFAs. In this study we reported, in tobacco leaves, the characterization of three distinct ∆6-desaturases from diatom Phaeodactylum tricornutum, fungi Rhizopus stolonifer and microalge Osterococcus tauri and two different ∆5-desaturases from P. tricornutum and single-celled saprotrophic eukaryotes Thraustochytrium sp. The in planta agroinfiltration of essential ∆6-desaturases, ∆6-elongases and ∆5-desaturases allowed for successful introduction of eicosapentaenoic acid (20:5∆5,8,11,14,17) biosynthesis pathway. However, despite the desired, targeted production of ω3-fatty acids we detected the presence of ω6-fatty acids, indicating and confirming previous results that all tested desaturases are not specifically restricted to neither ω3- nor ω6-pathway. Nevertheless, the additional co-expression of acyl-CoA:lysophosphatidylcholine acyltransferase (LPCAT) from Phaeodactylum tricornutum boosted the proportion of ω3-fatty acids in newly synthesized fatty acid pools. For the most promising genes combinations the EPA content reached at maximum 1.4% of total lipid content and 4.5% of all fatty acids accumulated in the TAG pool. Our results for the first time describe the role of LPCAT enzyme and its effectiveness in alleviating a bottleneck called ‘substrate dichotomy’ for improving the transgenic production of VLC-PUFAs in plants.

Biocontrol Potential of Ocimum Gratissimum on Some Fungal Isolates from Arachias Hypogea

Isolation of fungi was carried out from groundnut (arachais hypogae) seeds, and two fungi was isolated, Aspergillus spp and Rhizopus stolonifer. Proximate analysis of Ocimum gratissimum was carried out with moisture content of 9.32 , ash content of 10.90, nitrogen content of 3.78 and protein content of 22.03. Aqueous and Ethanolic extract of scent leaf ( Ocimum Gratissimum) was obtained and tested on the fungi isolates and Rhizopus stolonifer associated with groundnut was inhibited by aqueous and Ethanolic extracts of Ocimum Gratissimum while both aqueous and Ethanolic extracts of the plant did not show any activity against Aspergillus Niger associated with groundnut.

Biocontrol mechanisms of the Antarctic yeast Debaryomyces hansenii UFT8244 against post-harvest phytopathogenic fungi of strawberries.

The use of yeasts has been explored as an efficient alternative to fungicide application in the treatment and prevention of post-harvest fruit deterioration. Here, we evaluated the biocontrol abilities of the Antarctic yeast strain Debaryomyces hansenii UFT8244 against the post-harvest phytopathogenic fungi Botrytis cinerea and Rhizopus stolonifer for the protection and preservation of strawberry fruit. The strongest inhibition of germination of B. cinerea (57%) was observed at 0 °C, followed by 40% at 25 °C. In addition, germ tubes and hyphae of B. cinerea were strongly surrounded and colonized by D. hansenii. Production of the enzymes β-1,3-glucanase, chitinase and protease by D. hansenii was detected in the presence of phytopathogenic fungus cell walls. The activity of β-1,3-glucanase was highest on day 12 of incubation and remained high until day 15. Chitinase and protease activities reached their highest levels on the day 15 of incubation. D. hansenii additionally demonstrated the ability to resist oxidative stress. Our data demonstrated that the main biocontrol mechanisms displayed by D. hansenii were the control of phytopathogenic fungal spore germination, production of antifungal enzymes and resistance to oxidative stress. We conclude that isolate D. hansenii UFT8422 should be further investigated for use at commercial scales at low temperatures.

Complete genome sequence of a novel double-stranded RNA virus infecting the phytopathogenic fungus Rhizopus stolonifer.

In this study, we report the complete genome sequence of a novel toti-like virus, tentatively named "Rhizopus stolonifer double-stranded RNA virus 1" (RsDSV1), identified from a phytopathogenic fungal agent of apple fruit rot disease, Rhizopus stolonifer strain A2-1. RsDSV1 has a double-stranded RNA genome. The complete RsDSV1 genome is 5178 nucleotides (nt) in length and contains two open reading frames (ORFs) encoding a putative coat protein (CP) and an RNA-dependent RNA polymerase (RdRp). Phylogenetic analysis based on RdRp and CP amino acid sequences revealed that RsDSV1 is closely related to unclassified members of the family Totiviridae. In stress-inducing Vogel's minimal and sodium dodecyl sulfate-containing media, hyphal growth of A2-1 was suppressed, but the accumulation of RsDSV1 RNA increased, indicating that stresses promote RsDSV1 replication. To our knowledge, this is the first report of a mycovirus found in R. stolonifer.

Extracellular self-DNA induced a PTI-related local defence against Rhizopus rot in postharvest peach fruit

The plant immune response against various pathogens relies on a convoluted recognition system based on perceiving exogenous or endogenous stimuli, such as pathogen- or damage-associated molecular patterns (PAMPs or DAMPs). Extracellular self-DNA (sDNA) is an established DAMP source, but the plant intracellular signaling mechanism after sDNA perception remains largely unclear. Here, we observed that peach fruit was more resistant to the fungus Rhizopus stolonifer after sDNA perception. We identified a complete MAPK cascade (MAPKKK1, MAPKK2 and MAPK1) that functions downstream of the FLS2-BAK1 receptor complex following the perception of sDNA-derived DAMPs by means of transcriptome sequencing. The results of yeast two-hybrid (Y2H) and glutathione S-transferase (GST) pull-down assays confirmed the protein interaction of FLS2 and BAK1; meanwhile, the interaction of MAPKKK1 and MAPKK2 and its downstream interaction of MAPKK2 and MAPK1 were both exhibited following the another Y2H results. Accordingly, MAPK1 interacted with 1-aminocyclopropane-1-carboxylate oxidase (ACO1), which was directly associated with increased ethylene production and upregulation of the transcription profile of ethylene synthesis-related genes (PpSAMDC, PpACS1 and PpACO1), ethylene receptors (ETRs and ERS1), ethylene-responsive transcription factors (ERFs) and three ethylene-inducible marker genes, PLANT DEFENSIN1.2 (PDF1.2), CHITINASE (CHI) and HOOKLESS1 (HLS1). Accordingly, the overexpression of MAPK1 enhanced the disease resistance of transgenic tomato against the fungus R. stolonifer and increased ethylene biosynthesis and ethylene signaling-related gene transcription. Taken together, our data provide insight into the putative mechanism underlying the recognition of sDNA components by the FLS2-BAK1 complex and its downstream events in the transduction of ethylene signaling by the MAPK cascade to mediate local resistance in postharvest peach fruit.

Pseudomonas aeruginosa isolate PM1 effectively controls virus infection and promotes growth in plants.

A bacterial isolate PM1 obtained from the rhizosphere of healthy plants was identified as Pseudomonas aeruginosa by biochemical characteristics and 16S rRNA gene sequence (GenBank ID OL321133.1). It induced resistance in Nicotiana tabacum cv. Xanthi-nc and Cyamopsis tetragonoloba, against Tobacco mosaic virus (TMV) and Sunn-hemp rosette virus (SRV), respectively. Foliar treatment with isolate PM1 curbed TMV accumulation in susceptible N. tabacum cv. White Burley. PM1 was more effective as a foliar than a root/soil drench treatment, evident through a comparative decrease in ELISA values, and reduced viral RNA accumulation. Foliar and soil drench treatment with PM1 resulted in a disease index of 48 and 86 per cent, and a control rate of 48.9 and 8.5 per cent, respectively. PM1 exhibited phosphate solubilization, produced siderophores, auxins, HCN, and ammonia, all important plant growth-promoting traits. Foliar treatment with PM1 enhanced growth in tobacco, while its volatiles significantly promoted seedling growth in C. tetragonoloba. Of the several metabolites produced by the isolate, many are known contributors to induction of systemic resistance, antibiosis, and growth promotion in plants. Soluble metabolites of PM1 were less effective in inducing antiviral resistance in N. tabacum cv. Xanthi-nc in comparison with its broth culture. PM1 and its metabolites were antagonistic to Gram-positive Bacillus spizizenii and Staphylococcus aureus, and fungi Fusarium oxysporum, Aspergillus niger, and Rhizopus stolonifer. Its volatiles were inhibitory to F. oxysporum and R. stolonifer. Thus, PM1 exhibited considerable potential for further evaluation in plant virus control and production of diverse metabolites of use in agriculture and medicine.

Antifungal activity of Mentha × Piperita L. essential oil

The objective of this study was to investigate the chemical composition and in vitro antifungal activity of Mentha × Piperita L. (peppermint) essential oil (EO) against some plant pathogenic fungi (Alternaria alternata, Penicillium expansum, Rhizoctonia solani, and Rhizopus stolonifer). Antifungal activity of EO against the selected fungi was conducted using the agar diffusion method by adding peppermint EO concentrations (0, 250, 500, 750, 1000, and 2000 ppm). The gas chromatography-mass spectrometry (GC-MS) analysis of peppermint EO showed that the main constituent was menthol (36.4%), followed by menthone (27.7%) and menthyl acetate (11.2%). The mycelium growth of the selected fungi was significantly inhibited by peppermint EO. Light and electron microscopy studies showed that mycelium morphology was seriously changed after treatment with peppermint essential oil. The level of malondialdehyde illustrated that peppermint EO led to lipid peroxidation in the fungal pathogens. Therefore, due to its antifungal properties, peppermint EO can be used as an additive in the food industry and as an active substance in pharmaceuticals.

Bioproduction of Silver Nanoparticles by Myrtus Communis Leaf Extract and Their Effect on Plant Pathogenic Fungi in Vitro

In this study, Myrtus communis was used for the bioproduction of colloidal silver nanoparticles (AgNPs). The resultant AgNPs were characterized by X-ray diffraction (XRD), UV-Visible spectroscopy and field-emission scanning electron microscopy (FESEM). The hump-like peaks were recorded near the wavenumber 422 nm and such peaks originated due to the electronic structures of silver tiny particles and silver nanoparticles. The four intensive peaks of XRD patterns indicated the crystalline nature and the face-centered cubic structure of the AgNPs. The average crystallite size of the AgNPs ranged from 19[Formula: see text]nm to 25[Formula: see text]nm. The FESEM image illustrates the good dispersion of the AgNPs and the spherical shape of the nanoparticles. The AgNPs were prepared to study antifungal activity against plant pathogenic fungi Aspergillus niger, Rhizopus stolonifer and Neurospora crassain vitro. Results showed that the AgNPs demonstrated high antifungal activity against these fungi with significant differences between treatments when compared with the control (without nanoparticles).

Restricted Nitrogen and Water Applications in the Orchard Modify the Carbohydrate and Amino Acid Composition of Nonpareil and Carmel Almond Hulls.

Hull rot disease of almond (Prunus dulcis), caused by the fungus Rhizopus stolonifer, is prevalent in well maintained orchards where trees are provided plenty of water and nitrogen to increase the growth and yield. The predominantly grown variety Nonpareil is considered very susceptible to hull rot, while the pollinator variety Carmel is more resistant. Reduced nitrogen rates and restricted irrigation scheduling decreased the incidence and severity of hull rot in Californian orchards. As a part of our research, the hull composition of Australian almond fruits of Nonpareil and Carmel varieties, grown under two levels of irrigation (high and low) and two levels of nitrogen (high and low), were analysed using 1H NMR-based metabolomics. Both Nonpareil and Carmel hulls contained sugars such as glucose, sucrose, fructose and xylose, and amino acids, particularly asparagine. Variety was the major factor with Nonpareil hulls significantly higher in sugars and asparagine than Carmel. Within varieties, nitrogen influenced the relative concentrations of glucose, sucrose and asparagine. In Nonpareil, high nitrogen high water (the control) had relatively high glucose and asparagine content. High nitrogen low water increased the sucrose component, low nitrogen high water increased the glucose component and low nitrogen low water increased the sucrose and asparagine components. In Carmel, however, high nitrogen low water and low nitrogen high water increased sucrose and asparagine, and low nitrogen low water increased sucrose and glucose. Hull rot symptoms are caused by fumaric acid production by R. stolonifer growing within the hull. These changes in the hull composition under different nitrogen and water scenarios have the potential to affect the growth of R. stolonifer and its metabolite production in hull rot disease.

Activation of the BABA-induced priming defence through redox homeostasis and the modules of TGA1 and MAPKK5 in postharvest peach fruit.

The priming of defence responses in pathogen‐challenged model plants undergoes a preparation phase and an expression phase for defence function. However, the priming response in postharvest fruits has not been elucidated. Here, we found that 50 mM β‐aminobutyric acid (BABA) treatment could induce two distinct pathways linked with TGA1‐related systemic acquired resistance (SAR), resulting in the alleviation of Rhizopus rot in postharvest peach fruit. The first priming phase was elicited by BABA alone, leading to the enhanced transcription of redox‐regulated genes and posttranslational modification of PpTGA1. The second phase was activated by an H2O2 burst via up‐regulation of PpRBOH genes and stimulation of the MAPK cascade on pathogen invasion, resulting in a robust defence. In the MAPK cascade, PpMAPKK5 was identified as a shortcut interacting protein of PpTGA1 and increased the DNA binding activity of PpTGA1 for the activation of salicylic acid (SA)‐responsive PR genes. The overexpression of PpMAPKK5 in Arabidopsis caused the constitutive transcription of SA‐dependent PR genes and as a result conferred resistance against the fungus Rhizopus stolonifer. Hence, we suggest that the BABA‐induced priming defence in peaches is activated by redox homeostasis with an elicitor‐induced reductive signalling and a pathogen‐stimulated H2O2 burst, which is accompanied by the possible phosphorylation of PpTGA1 by PpMAPKK5 for signal amplification.

Identification of some Pathogenic Organisms Involved in Rotting of the Libyan Tirmania Truffles

Truffles are ectomycorrhizal fungi that grow symbiotically with several trees. Truffles are highly valued edible mycorrhizae fungi in economic and gastronomic terms. Desert truffles have been traditionally used as food in Libya. The rotting of truffle due to environmental factors is very common in Libya. The mechanism of truffle rot is complex and attributed mainly due to the infection with fungi, followed by bacteria. In order to document and highlight the rotting of Libyan Desert truffle phenomenon, the existence of microorganisms in the infected fruit body of Tirmania truffles has been investigated. The investigations showed changes in physical characteristics such as color, hardness, and humidity of the studied truffle samples. Also, the results showed a diversity of pathogens involved in the deterioration of truffle includes bacteria (Bacillus pumilus, Citrobacteryoungea), saprophytic fungi (Fusarium solani, Alternariachlamydosporia, and Rhizopusstolonifer). This study concluded the importance of studying microbial flora living in truffle and their environment. Keywords: Libyan Desert truffle; Tirmania; Rotting; Pathogens

PpWRKY22 physically interacts with PpHOS1/PpTGA1 and positively regulates several SA-responsive PR genes to modulate disease resistance in BABA-primed peach fruit

WRKY transcription factors (TFs) are essential for the regulation of disease resistance in plants, and ubiquitination is one of the major post-translational modification types for a suite of TFs, while limited information is available about the involvement of E3 ubiquitin ligase-mediated regulation of WRKY proteins in β-amino-butyric acid (BABA)-primed defense resistance in peach fruit. Here, we observed that the treatment of peach fruit with BABA elicited disease resistance against the necrotrophic fungus Rhizopus stolonifer, resulting in an obvious inhibition of Rhizopus rot, which was accompanied by accumulation of total phenolics and lignins and potentiated the activity of several defensive enzymes, including phenylalanine ammonia-lyase (PAL), chitinase (CHI) and β-1,3-glucanase (GNS). Additionally, the expression of PpHOS1 that encodes a C3HC4-type Really Interesting New Gene (RING) finger E3 ligase was induced in BABA-treated peach fruit in response to R. stolonifer attack, following which the specific PpWRKY22 protein was exclusively targeted by PpHOS1 for possible degradation. Notably, PpWRKY22 functions as a positive regulator in disease resistance, as determined by overexpression of PpWRKY22 in Arabidopsis thaliana, which conferred enhanced resistance to the fungus R. stolonifer. Moreover, the physical interaction between PpWRKY22 and PpTGA1 increased the transactivation of PpNPR1, PpPR1, PpPR2, and PpPR5 by PpWRKY22 alone, leading to elevated resistance of peach fruit against the fungus R. stolonifer. Taken together, our results indicated that PpWRKY22 might be a positive participant in BABA-primed resistance in peach fruit, which could involve the post-translational ubiquitination of PpWRKY22 by PpHOS1 and physical interaction between PpWRKY22 and PpTGA1 for modification and activation.

Use of an exopolysaccharide-based edible coating and lactic acid bacteria with antifungal activity to preserve the postharvest quality of cherry tomato

One of the main causes of cherry tomato postharvest loss is diseases caused by food-contaminating fungi. Edible coatings (ECs) can preserve the quality of this crop and can serve as carriers of lactic acid bacteria (LAB), which have demonstrated inhibitory potential against phytopathogenic fungi. In the present study, the effectiveness of the LAB strain Lactiplantibacillus plantarum A6 incorporated into an EC based on exopolysaccharide from Weissella confusa JCA4 on the physicochemical and microbiological quality of cherry tomato was evaluated. The fruit were artificially inoculated with the fungi Aspergillus niger, Fusarium sp., and Rhizopus stolonifer to test the antifungal potential of the coating. The physicochemical and microbiological quality of cherry tomato was studied at two storage temperatures to calculate its shelf life. L. plantarum A6 remained viable both in the solution and on the surface of the fruit after coating, protecting the fruit against two of the three evaluated fungi (Fusarium sp. and Rhizopus stolonifer). The EC controlled weight loss, maintained firmness, and slowed the respiration rate of cherry tomato; the other physicochemical properties and the appearance of the fruit were not negatively affected. Based on these results, prolongation of shelf life was achieved by the application of the exopolysaccharide coating.

Monitoring the infection process of Rhizopus stolonifer on strawberry fruit during storage using films based on chitosan/polyvinyl alcohol/polyvinylpyrrolidone and plant extracts

Different formulations based on nanoparticles of chitosan-plant extracts were evaluated to detect the infection process from the earliest stage of the fungus Rhizopus stolonifer on strawberry fruit during storage. Chitosan/polyvinyl alcohol (Ch/PVA) and chitosan/polyvinylpyrrolidone (Ch/PVP) films enriched with nanoparticles (NPs) of chitosan blended with plant extracts were prepared. They were placed inside a plastic package containing inoculated fruits and stored at 25 °C for 72 h. The thickness values of the films were in the range of 0.10 to 0.25 mm. All samples showed a maximum absorbance peak of about 300–320 nm; however, the Ch/PVP films enriched with NPs of chitosan and 10% of radish extract had an evident decrease in the optical absorbance as the fungal infection progressed. Additionally, as observed by scanning electron microscopy, the cross-section and surface morphology of films were not modified during storage, and the growth of R. stolonifer was evident after 48 h. Therefore, the Ch/PVP films enriched with chitosan NPs blended with 10% radish extract could be a reliable indicator of this fungus's growth.

Antagonistic Activity of Lactic Acid Bacteria Against Phytopathogenic Fungi Isolated from Cherry Tomato (Solanum lycopersicum var. cerasiforme).

The postharvest deterioration of cherry tomatoes due to diseases caused by fungi is one of the main causes of the loss of this product. The objective of this study was to determine the antagonistic capacity by evaluating the antifungal power of nine strains of lactic acid bacteria (LAB) in vitro against the phytopathogenic fungi Aspergillus niger, Fusarium sp., and Rhizopus stolonifer isolated from cherry tomatoes (Solanum lycopersicum var. cerasiforme) and to measure the biosurfactant production capacity, its antagonism in vivo, and the production of organic acids. The results showed that seven of the nine strains were able to inhibit at least one of the three fungi isolated in the in vitro assay. In eight of nine strains, biosurfactant production was identified, and the strains Weissella confusa and Lactiplantibacillus plantarum A6 showed the highest antifungal activity in vitro and in vivo against the fungi evaluated, with the identification of organic acid production in both strains. LAB demonstrated the ability to inhibit cherry tomato fungi, thus emerging as an alternative to the use of chemical preservatives in the production of this fruit and being projected as a preservation technology for this type of product through the use of strains or their metabolites.

Antifungal potential of essential oils from two varieties of Citrus sinensis (lima orange and bahia navel orange) in postharvest control of Rhizopus stolonifer (Ehrenb.: Fr.) Vuill.

Abstract Brazil, the world’s largest citros producer, yields around 19 million tons per year and exports most part of its production as orange juice. Essential oils (EOs) extracted from leaves, fruit and flowers of many species of the genus Citrus have been widely used as the result of their promising bioactivities. The fungus Rhizopus stolonifer (Ehrenb.) Vuill., an agent which causes soft rot in fruit, has been considered one of the main factors that cause postharvest diseases, thus, leading to major economic losses in agribusiness. This research aimed at evaluating the chemical composition and in vitro antifungal effect of EOs from two varieties of fresh Citrus sinensis (lima orange and bahia navel orange) peel on mycelial growth of R. stolonifer. EOs were obtained by hydrodistillation, which was carried out by a Clevenger-type apparatus, while their chemical composition was analyzed by gas chromatography-flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Limonene was the major monoterpene identified in EOs from lima orange (95.2%) and bahia navel orange (93.2%) peel. EOs from lima orange and bahia navel orange inhibited 91.95% and 80.05% mycelial growth of R. stolonifer, respectively, at the highest dose under evaluation (100 µL). This study revealed the biotechnological potential of EOs extracted from fruit peel of two varieties of citros which may applied to biofilm preparation, so as to coat and preserve different types of fruit.

Essential oils from Eucalyptus staigeriana F. Muell. ex Bailey and Eucalyptus urograndis W. Hill ex Maiden associated to carboxymethylcellulose coating for the control of Botrytis cinerea Pers. Fr. and Rhizopus stolonifer (Ehrenb.:Fr.) Vuill. in strawberries

The fungi Rhizopus stolonifer (Ehrenb.:Fr.) Vuill. (R. stolonifer (Ehrenb.:Fr.) Vuill.) and Botrytis cinerea Pers. Fr. (B. cinerea Pers. Fr.) are causal agents of important post-harvest diseases in strawberries. For their control, the essential oil (EO) extracted from the leaves of Eucalyptus staigeriana Muell. ex Bailey (E. staigeriana F. Muell. ex Bailey) and Eucalyptus urograndis W. Hill ex Maiden (E. urograndis W. Hill ex Maiden) can be a good alternative to the synthetic fungicides. Thus, the antifungal activity of the EOs from E. staigeriana F. Muell. ex Bailey and from E. urograndis W. Hill ex Maiden was evaluated in vitro on the fungi B. cinerea Pers. Fr. and R. stolonifer (Ehrenb.:Fr.) Vuill.. The effects on pathogen morphology and the in vivo antifungal activity of the EO that presented the highest activity in vitro, associated with carboxymethylcellulose (CMC), were also verified. The chemical composition of the essential oils evaluated was determined. The EO from E. staigeriana F. Muell. ex Bailey was the most efficient in the in vitro assays on the two fungi. This EO, whose major compound is limonene, caused dehydration and rupture of the fungal hyphae, suggesting its action is on the cell wall and membrane. The incorporation of this EO to CMC in the curative way reduced the severity of soft rot, caused by R. stolonifer (Ehrenb.:Fr.) Vuill.; nevertheless, it did not present an effect on the inhibition of B. cinerea Pers. Fr.. These results suggest that E. staigeriana F. Muell. ex Bailey EO, associated to CMC coating, can be an alternative to the use of synthetic fungicides for the control of R. stolonifer (Ehrenb.:Fr.) Vuill., mainly.

Effects of Fermentation on the Antioxidant and Antinutritional Compositions of Green Pea

Green peas are known to contain anti-nutritional factors like enzymes inhibitors, phytates, oxalates, saponins and polyphenolic compounds, all of which limit their utilization hence, the study evaluate the effect of fermentation on the antioxidant and antinutrients content of green pea. Fermentation of green pea was done using both submerged and solid state fermentation for 7days. Isolation and identification of microorganism from the fermented sample was done on daily basis using standard microbiological and molecular techniques. The type of organism isolated from the submerged fermentation of Green pea included the bacteria (Bacillus subtilis, Lactobacillus Plantarum, Micrococcus roseus, Lactobacillus lactis, and Lactobacillus fermentum) and the fungi Rhizopus oryzae, Penicillium chrysogenum and Rhizopus stolonifer. While the type of organism isolated from the solid state fermentation of Green pea included some bacteria (Bacillus subtilis, Lactobacillus Plantarum and Lactobacillus lactis) and fungi (Penicillium notatum, Rhizopus oryzae, Penicillium chrysogenum, Candida albicans, Alternaria alternate and Rhizopus stolonifer). Fermentation reduced the antinutritional content of the fermented sample with submerged fermentation resulting in the highest reduction from 32.18 mg/g, 4.14 mg/g, 1.62 mg/g, 51.08 mg/g and 36.37 mg/g in the raw sample to 26.27 mg/g, 0.48 mg/g, 0.27 mg/g, 7.82 mg/g and 24.07 mg/g in submerged fermented green pea for saponin, tannin, oxalate, phytate and alkaloid respectively. However, Fermentation significantly p ≤ 0.05 increased the phenol, flavonoid and FRAP content of the fermented green pea with the solid state fermentation resulting in the highest increase from 3.50, 0.03 and 1.41 in the raw sample to 9.32, 0.12 and 9.66 in the solid state fermented green pea for phenol, flavonoid and FRAP content respectively. This study revealed that fermentation had significant effect on the antioxidant and antinutritional compositions of Green pea thereby reducing the antinutrient composition of Green pea in which will improve the nutrient value of Green pea.

Tricomponent composite containing copper-hydroxyapatite/chitosan/polyvinyl pyrrolidone for bone tissue engineering.

The tricomponent composite chitosan/polyvinyl pyrrolidone (CS/PVP) with different weight ratios of (0, 20, 40, 60, and 80 wt%) of copper-hydroxyapatite (Cu-HAP) were prepared by solvent casting technique, which was characterized by X-ray diffraction, Fourier-transform infrared, and scanning electron microscopy with energy-dispersive X-ray to confirm the formation of Cu-HAP/CS/PVP composite. The Cu-HAP/CS/PVP composite with 80 wt% Cu-HAP showed 98.73 ± 1.14% of porosity with highest tensile strength (101.45 ± 0.98 MPa) and less swelling percentage (19.51 ± 1.03%) compared to others. In vitro antimicrobial activity was investigated against bacteria (Staphylococcus aureus, Bacillus subtilis, and Escherichia coli) and fungus (Candida albicans, Penicillium notatum, and Rhizopus stolonifer). In vitro hemocompatibility study proves that the Cu-HAP/CS/PVP composites are blood compatible with the hemolytic ratio of less than 2%. In vitro bioactivity study revealed the formation of apatite on the optimized Cu-HAP/CS/PVP composite (80 wt% of Cu-HAP) in simulated body fluid (SBF) solution. ICP-OES (Inductively Coupled Plasma-Optical Emission Spectroscopy) analysis was used to find out the leaching of Ca, P, and Cu ions from the SBF. In vitro biocompatibility was studied against human osteosarcoma cell line by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-DiphenyltetrazoliumBromide) assay.