Functional characterization and expression profiling of glyoxalase III genes in date palm grown under abiotic stresses.

Methylglyoxal (MG), a by-product of various metabolic processes, including glycolysis, is a highly reactive cytotoxic metabolite. The level of MG in the cell is maintained at a non-toxic level via MG detoxification pathways such as the universal glyoxalase system, including glyoxalase I/II/III enzymes. Glyoxalase III (DJ-1) can breakdown MG to d-lactate in a single step without reducing glutathione (GSH). Elucidating the function of the DJ-1 gene family may provide further knowledge about its role in plants under abiotic stresses. Here, we characterize four glyoxalase III genes (PdDJ-1B1, PdDJ-1B2, PdDJ-1C, and PdDJ-1D) encoding the conserved DJ-1 domain in the genome of the date palm, a crop with high drought and salinity tolerance. The expression level of the PdDJ-1 genes increased in date palm leaves upon salinity treatment. In addition, overexpression of PdDJ-1 genes in Escherichia coli and the complementation in yeast hsp31Δ knockout mutant cells enhanced their growth rate and reduced the accumulation of reactive oxygen species (ROS) under MG and oxidative stress conditions as shown by the flow cytometry assay. Subcellular localization using confocal microscopy revealed the accumulation of PdDJ-1B1, PdDJ-1C, and PdDJ-1D in the chloroplast, whereas PdDJ-1B2 was localized to the cytosol. Remarkably, constitutive expression of the PdDJ-1C gene in Arabidopsis thaliana Columbia (Col-0) resulted in the generation of non-viable albino plants implying that PdDJ-1C plays a critical function in chloroplast development. These findings suggest that PdDJ-1 protein has an important function in MG-detoxification and maintaining the redox balance in date palm plants under abiotic stress conditions.

Impact of Pretreatments and Drying Temperatures on Quality of Siwi and Sakkoti Dates.

BACKGROUND AND OBJECTIVE: Drying of date helps in preserving it to be consumed outside the harvest season and removes some moisture from dates and also slows down the action of date endogenous enzymes. This study was carried out to investigate pretreatments and drying temperature on the physical and chemical properties of 2 date varieties (Siwi and Sakkoti) at the khalal stage. MATERIALS AND METHODS: The date fruits at khalal stage were dipped in ascorbic acid solution, sodium metabisulfite solution and sulfur dioxide before cut into pieces, halves and as whole. Then dates were dried at 50, 55, 60 and 65°C, respectively till ~20% moisture content and examined the physical and chemical properties of dried dates. RESULTS: Moisture content of Siwi and Sakkoti at the khalal stage was 56.90 and 51.72%, respectively, while total sugars were 79.76 and 75.74%, respectively on dry weight bases. The color of dates Hunter (L and b) were the highest of treated with meta-bisulfate solution or sulfur dioxide and the lowest of color date observed (Hunter, a) comparing with control and ascorbic acid. CONCLUSION: The pretreatments indicated that the dipping dates in sodium meta-bisulfate solution or sulfur dioxide then, dried at 60˚C produce high quality parameters of semi-dry dates comparing control and ascorbic acid.

Purification and characterization of catechol oxidase from Tadela (Phoenix dactylifera L.) date fruit.

Catechol oxidase (PPO) was extracted and purified from Tadela (Phoenix dactylifera L.) date fruit, by a procedure that included (NH4)2SO4 precipitation followed by dialysis, Q-Sepharose bb ion-exchange chromatography and HPLC gel filtration chromatography. Some of its biochemical characteristics were studied. The purification rate and the yield were 80% and 20%, respectively. The Tadela date fruit catechol oxidase exhibited a molecular weight of 90 kDa using SDS-PAGE. The catechol oxidase showed only o­diphenolase and triphenolase activities while no monophenolase activity was detected. A better affinity was observed using catechol as substrate (Km = 35 mM) with thus, a higher Vmax/Km ratio (80 U/mM·mL). This enzyme is thermostable in the temperature range (30-60 °C) with optimum activity in acidic range of pH. Four inhibitors were used for the control of enzymatic browning, of which sodium metabisulfite was the most potent (IC50 = 0, 11 mM). The values of KI and mechanism of inhibition were also determined. No significant change on enzyme activity was noticed in the presence of metal ion and detergents. Therefore, thermal inactivation was studied in the temperature range between 60 and 80 °C using catechol as substrate. Their kinetic (K, D, t1/2, Zt, Ea) and thermodynamic (ΔH, ΔG and ΔS) parameters were also estimated.

The phytochelatin synthase gene in date palm (Phoenix dactylifera L.): Phylogeny, evolution and expression.

We studied date palm phytochelatin synthase type I (PdPCS1), which catalyzes the cytosolic synthesis of phytochelatins (PCs), a heavy metal binding protein, in plant cells. The gene encoding PdPCS1 (Pdpcs) consists of 8 exons and 7 introns and encodes a protein of 528 amino acids. PCs gene history was studied using Notung phylogeny. During evolution, gene loss from several lineages was predicted including Proteobacteria, Bilateria and Brassicaceae. In addition, eleven gene duplication events appeared toward interior nodes of the reconciled tree and four gene duplication events appeared toward the external nodes. These latter sequences belong to species with a second copy of PCs suggesting that this gene evolved through subfunctionalization. Pdpcs1 gene expression was measured in seedling hypocotyls exposed to Cd, Cu and Cr using quantitative real-time polymerase chain reaction (qPCR). A Pdpcs1 overexpression was evidenced in P. dactylifera seedlings exposed to metals suggesting that 1-the Pdpcs1 gene is functional, 2-there is an implication of the enzyme in metal detoxification mechanisms. Additionally, the structure of PdPCS1 was predicted using its homologue from Nostoc (cyanobacterium, NsPCS) as a template in Discovery studio and PyMol software. These analyses allowed us to identify the phytochelatin synthase type I enzyme in date palm (PdPCS1) via recognition of key consensus amino acids involved in the catalytic mechanism, and to propose a hypothetical binding and catalytic site for an additional substrate binding cavity.

Extraction, partial purification and characterization of amylase from parthenocarpic date (Phoenix dactylifera): effect on cake quality.

BACKGROUND: Phoenix dactylifera L. plays an important role in social, economic and ecological Tunisian sectors. Some date palms produce parthenocarpic fruit named Sish. The objective of the present study was to extract biomolecules from parthenocarpic fruit by producing value-added products from the fruits. RESULTS: The extraction of amylolytic activity from parthenocarpic fruit (AmyPF) was optimized using Box-Behnken design (BBD). Partial purification of about 250-fold with an activity yield of 47% was achieved. The amylase exhibited a specific activity of 80 U mg-1 protein. The optimum pH and temperature for enzyme activity were 5 and 55 °C respectively. The enzyme was highly active over a wide range of pH (5-10), and significant stabilization was observed at 60 °C. The purified enzyme belongs to the exo type of amylases. Given the economic and industrial relevance of amylases used in the food industry, three different concentrations of AmyPF (0.007, 0.014 and 0.018 U g-1 ) were incorporated into a cake formulation, resulting in a decrease in density, moisture retention and water activity and an increase in hardness. CONCLUSION: The beneficial effect of AmyPF on the technological characteristics of cakes was confirmed by sensory evaluation. © 2017 Society of Chemical Industry.

Exogenous proline mediates alleviation of cadmium stress by promoting photosynthetic activity, water status and antioxidative enzymes activities of young date palm (Phoenix dactylifera L.).

The ability of exogenous compatible solutes, such as proline, to counteract cadmium (Cd) inhibitory effects in young date palm plants (Phoenix dactylifera L. cv Deglet Nour) was investigated. Two-year-old date palm plants were subjected for five months at different Cd stress levels (0, 10 and 30 mg CdCl2 kg(-1) soil) whether supplied or not with exogenous proline (20mM) added through the irrigation water. Different levels of Cd stress altered plant growth, gas exchanges and chlorophyll content as well as water status, but at different extent among them. In contrast, an increase of antioxidant enzymes activities of Cd-treated plants in association with high amounts of proline content, hydrogen peroxide (H2O2), thiobarbituric acid reactive substances (TBARS) and electrolyte leakage (EL) were observed. Interestingly, exogenous proline mitigated the adverse effects of Cd on young date palm. Indeed, it alleviated the oxidative damage induced by Cd accumulation and established better levels of plant growth, water status and photosynthetic activity. Moreover, proline-treated plants showed high antioxidant enzymes activities (superoxide dismutase, catalase and glutathione peroxydase) in roots and leaves as compared to Cd-treated plants.