Statistical optimization as a powerful tool for indole acetic acid production by Fusarium oxysporum

Crop production is challenged in our world by increasing food demands, decrease natural resource bases and climatic change. Nowadays plant growth regulators works like fertilizers in increasing plant growth production efficiency and needed to produce in large industrial scale. Fermentation condition and medium constituents can significantly affect on the product production and designing an acceptable fermentation medium is critical importance. In this paper Fusarium sp. could be considered as promising indole-3-acetic acid producers with the ability to improve the production using statistical methods. The results showed that fermentation type, incubation temperature and L-tryptophan were the most influencing parameters on the production. Maximum IAA production by Fusarium oxysporum was 300.4 mg/l obtained under the fermentation conditions: temperature at 25oC, incubation period 5 days, pH 7, inoculums size 2%, shaking rate at 150 rpm and medium constituents: Glucose 40 g/l, yeast extract 3 g/l, L-tryptophan 1 g/l, KH2PO4 2 g/l, NaNO3 4 g/l, MgSO4·7H2O 0.1 g/l with regression analysis (R2) 99.67% and 2.12-fold increase in comparison to the production of the original level (142 mg/l). DOI: http://dx.doi.org/10.5281/zenodo.101234

Optimization of kojic acid production conditions from cane molasses using Plackett-Burman design

Fungal synthesis of kojic acid has gained more interest in these days as an alternative way to chemical synthetic. The aspect of the microbial fermentation process is to develop a suitable culture medium to obtain the maximum amount of kojic acid using statistical methods. In this study; different selected three isolates of Aspergillus flavus (No 1, 2 and 3) were screened for their ability to produced kojic acid and the isolate No 3 was the highest kojic acid producer one. The capability of A. flavus No 3 to produce kojic acid was improved using Plackett-Burman design. From ten different agro-industrial wastes cane molasses recorded the highest kojic acid productivity with 2.24 g/l-1 day-1 and was the most effective parameter plays a crucial role in Plackett-Burman design. Maximum kojic acid production (24.65 g/l) by A. flavus (No. 3) obtained under the fermentation conditions: incubation temperature at 25oC, incubation time 9 days, pH 3, inoculum size 0.5%, shaking rate at 150 rpm and medium constituents: Cane molasses 60 g/l, yeast extract 7 g/l, KH2PO4 2 g/l, ZnSO4·7H2O 100 µg/l and MgSO4·7H2O 1 g/l with regression analysis (R2) 99.45% and 2.33-fold increase in comparison to the production of the original level (10.6 g/l). DOI: http://dx.doi.org/10.5281/zenodo.121151

Bioconversion of plant wastes to β-carotene by Rhodotorula glutinis KU550702

Microbial synthesis of β-carotene has gained more interest as an alternative to synthetic β-carotene due to easy extraction and high yield. The vitamin microbial production is mainly dependent on culture conditions and the medium compositions. In this study, the β-carotene production by the Rhodo-torula glutinis ASU6 (KU550702) was evaluated under different growth conditions and nutrient composition. Different agro-renewable wastes were tested as carbon source for R. glutinis to obtain maximum amount of β-carotene. Meanwhile, it is clear that R. glutinis could grow well on acid extract of onion peels and produced large amount of β-carotene. Initial statistical screening using a Plackett-Burman design showed temperature, incubation time, fermentation type, non-treated onion waste, KH2PO4 and L-asparagine as significantly, influencing β-carotene production. Response surface methodology was applied to determine the mutual interactions between these parameters and optimal levels for β-carotene production. The maximum value of β-carotene production was 204.29 mg/l (7.5-fold) of value observed as central point of the central composite design. All the experimental data are in good agreement with predicted ones, confirming the responsibility of the proposed empirical model in describing β-carotene production by R. glutinis. In the whole, the outcomes of this study support the exploitation of onion peels through microbial fermentation for β-carotene production

Induction of resilience strategies against biochemical deteriorations prompted by severe cadmium stress in sunflower plant when Trichoderma and bacterial inoculation were used as biofertilizers

BackgroundCadmium (Cd) is a highly toxic heavy metal. Its emission is suspected to be further increased due to the dramatic application of ash to agricultural soils and newly reclaimed ones. Thereby, Cd stress encountered by plants will exacerbate. Acute and chronic exposure to Cd can upset plant growth and development and ultimately causes plant death. Microorganisms as agriculturally important biofertilizers have constantly been arising as eco-friendly practices owing to their ability to built-in durability and adaptability mechanisms of plants. However, applying microbes as a biofertilizer agent necessitates the elucidation of the different mechanisms of microbe protection and stabilization of plants against toxic elements in the soil. A greenhouse experiment was performed using Trichoderma harzianum and plant growth-promoting (PGP) bacteria (Azotobacter chroococcum and Bacillus subtilis) individually and integrally to differentiate their potentiality in underpinning various resilience mechanisms versus various Cd levels (0, 50, 100, and 150 mg/kg of soil). Microorganisms were analyzed for Cd tolerance and biosorption capacity, indoleacetic acid production, and phosphate and potassium solubilization in vitro. Plant growth parameters, water relations, physiological and biochemical analysis, stress markers and membrane damage traits, and nutritional composition were estimated.ResultsUnequivocal inversion from a state of downregulation to upregulation was distinct under microbial inoculations. Inoculating soil with T. harzianum and PGPB markedly enhanced the plant parameters under Cd stress (150 mg/kg) compared with control plants by 4.9% and 13.9%, 5.6% and 11.1%, 55.6% and 5.7%, and 9.1% and 4.6% for plant fresh weight, dry weight, net assimilation rate, and transpiration rate, respectively; by 2.3% and 34.9%, 26.3% and 69.0%, 26.3% and 232.4%, 135.3% and 446.2%, 500% and 95.6%, and 60% and 300% for some metabolites such as starch, amino acids, phenolics, flavonoids, anthocyanin, and proline, respectively; by 134.0% and 604.6% for antioxidants including reduced glutathione; and by 64.8% and 91.2%, 21.9% and 72.7%, and 76.7% and 166.7% for enzymes activity including ascorbate peroxidase, glutathione peroxidase, and phenylalanine ammonia-lyase, respectively. Whereas a hampering effect mediated by PGP bacterial inoculation was registered on levels of superoxide anion, hydroxyl radical, electrolyte leakage, and polyphenol oxidase activity, with a decrease of 0.53%, 14.12%, 2.70%, and 5.70%, respectively, under a highest Cd level (150 mg/kg) compared with control plants. The available soil and plant Cd concentrations were decreased by 11.5% and 47.5%, and 3.8% and 45.0% with T. harzianum and PGP bacterial inoculation, respectively, compared with non-inoculated Cd-stressed plants. Whereas, non-significant alternation in antioxidant capacity of sunflower mediated by T. harzianum action even with elevated soil Cd concentrations indicates stable oxidative status. The uptake of nutrients, viz., K, Ca, Mg, Fe, nitrate, and phosphorus, was interestingly increased (34.0, 4.4, 3.3, 9.2, 30.0, and 1.0 mg/g dry weight, respectively) owing to the synergic inoculation in the presence of 150 mg of Cd/kg.ConclusionsHowever, strategies of microbe-induced resilience are largely exclusive and divergent. Biofertilizing potential of T. harzianum showed that, owing to its Cd biosorption capability, a resilience strategy was induced via reducing Cd bioavailability to be in the range that turned its effect from toxicity to essentiality posing well-known low-dose stimulation phenomena (hormetic effect), whereas using Azotobacter chroococcum and Bacillus subtilis, owing to their PGP traits, manifested a resilience strategy by neutralizing the potential side effects of Cd toxicity. The synergistic use of fungi and bacteria proved the highest efficiency in imparting sunflower adaptability under Cd stress

Statistical optimization as a powerful tool for indole acetic acid production by Fusarium oxysporum

Crop production is challenged in our world by increasing food demands, decrease natural resource bases and climatic change. Nowadays plant growth regulators works like fertilizers in increasing plant growth production efficiency and needed to produce in large industrial scale. Fermentation condition and medium constituents can significantly affect on the product production and designing an acceptable fermentation medium is critical importance. In this paper Fusarium sp. could be considered as promising indole-3-acetic acid producers with the ability to improve the production using statistical methods. The results showed that fermentation type, incubation temperature and L-tryptophan were the most influencing parameters on the production. Maximum IAA production by Fusarium oxysporum was 300.4 mg/l obtained under the fermentation conditions: temperature at 25oC, incubation period 5 days, pH 7, inoculums size 2%, shaking rate at 150 rpm and medium constituents: Glucose 40 g/l, yeast extract 3 g/l, L-tryptophan 1 g/l, KH2PO4 2 g/l, NaNO3 4 g/l, MgSO4·7H2O 0.1 g/l with regression analysis (R2) 99.67% and 2.12-fold increase in comparison to the production of the original level (142 mg/l). DOI: http://dx.doi.org/10.5281/zenodo.101234

Toxicity Evaluation and Genetic Improvement of Bacillus thuringiensis Isolated from Different Regions in Assiut, Egypt against Mosquito Larvae

This study aims to evaluate the toxicity and genetic improvement of Bacillus thuringiensis isolates. Isolates were obtained from soil, insect and water samples from different regions of Assiut, Egypt for biological control of mosquito larvae. B. thuringiensis colonies were identified based on morphological and then by PCR which detect the Cry toxic genes in the isolates. Bioassays were performed to evaluate the toxicity of different strains of B. thuringiensis against mosquito larvae such as (Culex spp). In general, 36 B. thuringiensis isolates were obtained (31 from soil, 4 from insects, and 1 from water). And they were all toxic to mosquito larvae with different mortality percentages from 7 to 97% after 48 hours. Isolate Am2 recorded the highest mortality percentage 97% and Mn3 lowest mortality percentage 7%. PCR revealed that Am2 isolate which caused the highest mortality encodes three different types of Cry toxins, Cyt1AA, Cry1Ac and Cry2Aa. This isolate Am2 was examined by scanning electron microscopy to observe the shape of the Cry proteins. The results showed that the Am2 isolate contained of spherical and cuboidal toxic proteins. Then UV-mutagenesis was performed on the Am2 isolate to improve its toxicity. Out of 30 obtained UV-mutants, only one mutant showed improvement in the mortality of mosquito larvae since it caused a mortality rate of 100%. The results of the present study revealed the larvicidal efficacy of B. thuringiensis (Am2) isolate found in the soil of Assiut, could be used in biological control program of mosquito larvae

Controlling Alternaria cerealis MT808477 Tomato Phytopathogen by Trichoderma harzianum and Tracking the Plant Physiological Changes

Plant responses during the pathogen infection and the pathogen control reflect its strategies to protect its cells. This work represents the Alternaria cerealis MT808477 as a phytopathogen causing leaf spot disease in tomatoes. A. cerealis was identified morphologically and genetically by 18SrRNA, and its pathogenicity was confirmed by light and scanning electron microscopy. Trichoderma harzianum has the ability to control A. cerealis MT808477 by stimulating various cell responses during the controlling process. The cell behavior during the biological control process was observed by analyses of total phenol, flavonoids, terpenoids, antioxidant, malondialdehyde and antioxidant enzymes (catalase and peroxidase). The extracts of infected tomato leaves were tested against plant and human pathogenic microorganisms. Results showed that the biological control process activates the defense cell strategies by increasing the plant tolerance, and activation of plant defense systems. The total phenol, flavonoids, terpenoids, antioxidant and malondialdehyde were increased after 48 h. Catalase and peroxidase were increased in infected tomato plants and decreased during the biological control process, reflecting the decrease of cell stress. Leaves extract inhibited the growth of nine plant and human pathogenic microorganisms. Biological control represents a safe and effective solution to phytopathogens that decreases plant cell stress by stimulating various defensive agents

Pb (II) and Hg (II) thiosemicarbazones for inhibiting the broad-spectrum pathogen <i>Cladosporium sphaerospermum</i> ASU18 (MK387875) and altering its antioxidant system

Pb (II) and Hg (II) ions were reacted in aqueous solution with two tridentate thiosemicarbazones, resulting in four complexes {[Pb(L1)2] 1, [Pb(L2)2] 2, [Hg(L1)2] 3 and [Hg(L2)2] 4; HL1 = 4-(4-nitrophenyl)-1-((pyridin-2-yl)methylene)thiosemicarbazide and HL2 = 4-(2-chlorophenyl)-1-((pyridin-2-yl)methylene)thiosemicarbazide}. The solid-state structures of complexes 2 and 3 were determined by X-ray crystallography, which also confirmed the monobasic tridentate NNS nature of the ligands, and the octahedral coordination geometry around the metal centers. Complexes 1-4 exhibited high antifungal activity against the pathogen Cladosporium sphaerospermum; all four complexes giving rise to a reduction in the fungal dry mass and sugar consumption. Complexes 2 and 4 that inhibited Cladosporium sphaerospermum growth at the concentration of 1.8 mM are more efficient inhibitors than the free ligand and metal ions. Further, the complexes gave varied values of the fungal total antioxidants (TA), superoxide dismutase (SOD) and catalase (CAT) activities dependent on their concentrations