ABSTRACT. DNA isolation from some fungal organisms of agronomic importance is difficult because they have cell walls or capsules that are relatively unsusceptible to lysis. We have developed a fast DNA isolation protocol for Fusarium oxysporum, which causes fusarium wilt disease in more than 100 plant species, and for Pyrenochaeta terrestris, which causes pink root in onions. This protocol was based on the sodium dodecyl sulfate/ phe nol method, without β-mercaptoethanol and without maceration in liquid nitrogen; it uses phenol/chloroform extraction to remove proteins and co-precipitated polysaccharides. The A 260/280 absorbance ratios of isolated DNA were around 1.9, suggesting that the DNA fraction was pure and may be used for further analysis. Additionally, the A 260/230 values were higher than 1.8, suggesting negligible contamination by polysaccharides. The DNA isolated by this protocol is of sufficient quality for molecular applications; this tech nique could be applied to other organisms that have similar substances that hinder DNA extraction.

In the present study the characterization and properties of silver nanoparticles from Prosopis glandulosa leaf extract (AgNPs) were investigated using UV-Vis spectroscopic techniques, energy dispersive X-ray spectrometers (EDS), zeta potential and dynamic light scattering. The UV-Vis spectroscopic analysis showed the absorbance peaked at 487 nm, which indicated the synthesis of silver nanoparticles. The experimental results showed silver nanoparticles had Z-average diameter of 421 nm with higher stability (−200 mV). The EDS analysis also exhibited presentation of silver element. Additionally, the different concentrations of AgNPs (25, 50, 75 and 100 mg/mL) showed antibacterial activity against Acinetobacter calcoaceticus and Bacillus cereus. Finally, AgNPs from leaf extracts of P. glandulosa may be used as an agent of biocontrol of microorganism of importance medical. However, further studies will be needed to fully understand the antimicrobial activity of silver nanoparticles obtain from P. glandulosa.

The effect of heavy metal stress on the cell viability and leakage of electrolytes of Avicennia germinans leaf discs was investigated by the tissue tolerance test. Foliar discs were incubated with different Cd2+ or Cu2+ concentrations for 24 h; thereafter, the cell membrane stability of the tissue was assayed by the cell viability Evans blue and leakage electrolytes methods. The results indicated that electrolyte leakage of the leaf discs increased 24 h after exposure to heavy metal stress, as shown by a reduction of the cell viability by 30% in discs exposed to higher doses of Cd2+ (0.546 M) and Cu2+ (0.7 M), respectively. Additionally, the histological analysis of the leaf discs exposed to heavy metal stress revealed that at higher Cd2+ and/or Cu2+ concentrations an increase in the intercellular spaces and destruction of mesophyll cells was observed 24 h after exposure. In summary, the biochemical and structural changes observed in foliar tissues of A. germinans suggest that higher cadmium and copper concentrations may result in structural changes and altered physiological characters in leaves

The effect of phenolic and carotenoid extracts from chiltepin fruits on mycelial growth and the inhibition of conidial germination of Alternaria alternata and Fusarium oxysporum were investigated in the present work. Phenolic extracts inhibited mycelial growth of A.alternata by 38.46%, and significantly reduced conidial germination on the fifth day after treatment to 92% in relation to control. No significant changes were observed in the inhibition of mycelial growth in Fusarium oxysporum; however, the number of germinated conidia was reduced, showing 85% inhibition five days after treatment in relation to control. Moreover, carotenoid extracts showed 38.5% inhibition of mycelial growth and 85.3% inhibition of conidial germination of A.alternata, five days after treatment. Carotenoid extracts showed less inhibition of mycelial growth (20.3%) in F.oxysporum, with respect to A.alternata; while there was greater inhibition of conidial germination (96%) on the fifth day after treatment. Phenolic and carotenoid extracts from chiltepin may be a promising alternative as a natural fungicide against fungi of agricultural importance.

The aim of the present work is to evaluate the changes on the phenylalanine ammonia lyase (PAL) activity, phenolic compounds accumulation and photochemical efficiency in leaves of P. glandulosa treated with Cd2+ (0.001 M) and Cu2+ (0.52 M) concentrations for 96 h under hydroponic conditions. The results showed that only leaves treated with copper had a decrease in photochemical efficiency and leaf epidermal polyphenols in P. glandulosa leaves after 96 h of exposure. On the other hand the reverse-phase HPLC analysis revealed higher levels of phenolic compound (gallic, vanillic and caffeic acids) and flavonoids (rutin and kaempferol-3-O-glucosides) in plant leaves from Cu and Cd-treatments with respect to control plants. Finally, highest increments in PAL activity was observed in extracts of leaves treated with Cu and Cd (about 205 and 284%), respectively, with respect to control plants after 96 h treatment. These suggest that activation of phenylpropanoid pathway represent a source of nonenzymatic antioxidants that protect at P. glandulosa against oxidative stress when exposed to cadmium and copper. Hence future studies are necessary to elucidate the participation of phenylpropanoid pathway in the reduction of metal toxicity in Prosopis species.

In the present study the production of phenolic acids and fl avonoid compounds by E. gracilis exposed to two cadmium concentrations (0.02 and 0.14 mM) was evaluated using high-performance liquid chromatography (HPLC). The results showed that E. gracilis exposed to 0.02 mM Cd +2 increased signifi cantly the total content of phenolic compounds (798.46 ± 12.61 μg GA/g) and total fl avonoids (241.34 ± 47.63 μg QE/g) with respect to the control (137.34 ± 19.80 μg QE/g DW and 549.00 ± 8.57 μg GA/g DW, respectively). However, no signifi cant increase in the total content of phenolic compounds (568.54 ± 17.42 μg GA/g DW) and total fl avonoids (141.11 ± 9.36 μg QE/g DW) were observed in E. gracilis exposed to 0.14 mM Cd +2. Further research is necessary to determine the specifi c role of fl avonoids in E. gracilis exposed to high concentrations of Cd +2 .

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