Mohamed El-Esawi

Blue-light dependent reactive oxygen species formation by Arabidopsis cryptochrome may define a novel evolutionarily conserved signaling mechanism

Cryptochromes are widespread blue-light absorbing flavoproteins with important signaling roles. In plants they mediate de-etiolation, developmental and stress responses resulting from interaction with downstream signaling partners such as transcription factors and components of the proteasome. Recently, it has been shown that Arabidopsis cry1 activation by blue light also results in direct enzymatic conversion of molecular oxygen (O2 ) to reactive oxygen species (ROS) and hydrogen peroxide (H2 O2 ) in vitro. Here we explored whether direct enzymatic synthesis of ROS by Arabidopsis cry1 can play a physiological role in vivo. ROS formation resulting from cry1 expression was measured by fluorescence assay in insect cell cultures and in Arabidopsis protoplasts from cryptochrome mutant seedlings. Cell death was determined by colorimetric assay. We found that ROS formation results from cry1 activation and induces cell death in insect cell cultures. In plant protoplasts, cryptochrome activation results in rapid increase in ROS formation and cell death. We conclude that ROS formation by cryptochromes may indeed be of physiological relevance and could represent a novel paradigm for cryptochrome signaling.

Cellular metabolites modulate in vivo signaling of Arabidopsis cryptochrome-1

Cryptochromes are blue-light absorbing flavoproteins with multiple signaling roles. In plants, cryptochrome (cry1, cry2) biological activity has been linked to flavin photoreduction via an electron transport chain to the protein surface comprising 3 evolutionarily conserved tryptophan residues known as the ‘Trp triad.’ Mutation of any of the Trp triad residues abolishes photoreduction in isolated cryptochrome protein in vitro and therefore had been suggested as essential for electron transfer to the flavin. However, photoreduction of the flavin in Arabidopsis cry2 proteins occurs in vivo even with mutations in the Trp triad, indicating the existence of alternative electron transfer pathways to the flavin. These pathways are potentiated by metabolites in the intracellular environment including ATP, ADP, AMP, and NADH. In the present work we extend these observations to Arabidopsis cryptochrome 1 and demonstrate that Trp triad substitution mutants at W400F and W324F positions which are not photoreduced in vitro can be photoreduced in whole cell extracts, albeit with reduced efficiency. We further show that the flavin signaling state (FADH°) is stabilized in an in vivo context. These data illustrate that in vivo modulation by metabolites in the cellular environment may play an important role in cryptochrome signaling, and are discussed with respect to possible effects on the conformation of the C-terminal domain to generate the biologically active conformational state.

Blue-light dependent ROS formation by Arabidopsis cryptochrome-2 may contribute toward its signaling role

Cryptochromes are blue-light absorbing flavoproteins with many important signaling roles in plants, including in de-etiolation, development, and stress response. They interact with downstream signaling partners such as transcription factors and components of the proteasome, and thereby alter regulation of nuclear gene expression in a light dependent manner. In a prior study, it has also been shown that Arabidopsis cry1 activation by blue light results in direct enzymatic conversion of molecular oxygen (O2) to ROS (reactive oxygen species) in vivo leading to cell death in overexpressing lines. Here we extend these observations to show that Atcry2 is translocated from the cytosol to the nucleus in response to blue light illumination, resulting in nuclear accumulation of ROS in expressing insect cell cultures. These observations suggest that ROS formation may represent a novel means of signaling by Atcry2 distinct from, and perhaps complementary to, the currently known mechanism of light-mediated conformational change.

Genetic diversity and evolution of Brassica genetic resources: from morphology to novel genomic technologies – a review

Brassica species have an economic and medicinal importance. Estimation of the amount and distribution of genetic diversity within Brassica species is essential for establishing efficient management, conservation and breeding practices. This review discusses the taxonomy, gene pool, and Brassica -derived phytochemicals and their nutraceutical importance. It also surveys the recently advanced studies of the genetic diversity and phylogenetic studies of Brassica species at the level of morphological, cytological, biochemical and molecular markers that have proven to be useful for evaluating the genetic variation, taxonomic relationships and species identity, and could be useful for improving Brassica crops through future promising breeding programmes.

Blue-light induced biosynthesis of ROS contributes to the signaling mechanism of Arabidopsis cryptochrome

Cryptochromes are evolutionarily conserved blue light receptors with many roles throughout plant growth and development. They undergo conformational changes in response to light enabling interaction with multiple downstream signaling partners. Recently, it has been shown that cryptochromes also synthesize reactive oxygen species (ROS) in response to light, suggesting the possibility of an alternate signaling mechanism. Here we show by fluorescence imaging and microscopy that H202 and ROS accumulate in the plant nucleus after cryptochrome activation. They induce ROS-regulated transcripts including for genes implicated in pathogen defense, biotic and abiotic stress. Mutant cryptochrome alleles that are non-functional in photomorphogenesis retain the capacity to induce ROS-responsive phenotypes. We conclude that nuclear biosynthesis of ROS by cryptochromes represents a new signaling paradigm that complements currently known mechanisms. This may lead to novel applications using blue light induced oxidative bursts to prime crop plants against the deleterious effects of environmental stresses and toxins.

Salicylic Acid-Regulated Antioxidant Mechanisms and Gene Expression Enhance Rosemary Performance under Saline Conditions

Salinity stress as a major agricultural limiting factor may influence the chemical composition and bioactivity of Rosmarinus officinallis L. essential oils and leaf extracts. The application of salicylic acid (SA) hormone may alleviate salinity stress by modifying the chemical composition, gene expression and bioactivity of plant secondary metabolites. In this study, SA was applied to enhance salinity tolerance in R. officinallis. R. officinallis plants were subjected to saline water every 2 days (640, 2,000, and 4,000 ppm NaCl) and 4 biweekly sprays of SA at 0, 100, 200, and 300 ppm for 8 weeks. Simulated salinity reduced all vegetative growth parameters such as plant height, plant branches and fresh and dry weights. However, SA treatments significantly enhanced these plant growth and morphological traits under salinity stress. Salinity affected specific major essential oils components causing reductions in α-pinene, β-pinene, and cineole along with sharp increases in linalool, camphor, borneol, and verbenone. SA applications at 100–300 ppm largely reversed the effects of salinity. Interestingly, SA treatments mitigated salinity stress effects by increasing the total phenolic, chlorophyll, carbohydrates, and proline contents of leaves along with decline in sodium and chloride. Importantly, this study also proved that SA may stimulate the antioxidant enzymatic mechanism pathway including catalase (CAT), superoxide dismutase (SOD), and ascorbate peroxidase (APX) as well as increasing the non-enzymatic antioxidants such as free and total ascorbate in plants subjected to salinity. Quantitative real-time PCR analysis revealed that APX and 3 SOD genes showed higher levels in SA-treated rosemary under salinity stress, when compared to non-sprayed plants. Moreover, the expression level of selected genes conferring tolerance to salinity (bZIP62, DREB2, ERF3, and OLPb) were enhanced in SA-treated rosemary under salt stress, indicating that SA treatment resulted in the modulation of such genes expression which in turn enhanced rosemary tolerance to salinity stress.

Effect of lipid-free microalgal biomass and waste glycerol on growth and lipid production of Scenedesmus obliquus: Innovative waste recycling for extraordinary lipid production

In the present work, a novel approach of using growth medium with different substitutions of lipid-free algal hydrolysate (LFAH, 0, 5, 10 and 15%) and/or waste glycerol (WG, 0, 5, 10 and 20 g L-1) for enhanced biodiesel production from Scenedesmus obliquus was studied. Combination of different concentrations of WG with 15% LFAH showed the maximum significant biomass productivity, which represented 27.4, 30.5 and 28.9% over the control at combined 5, 10 and 20 g L-1 WG, respectively. The combinations of different LFAH with 20 g L-1 WG showed the maximum significant lipid accumulation, where lipid productivity showed its maximum significant value of 59.66 mg L-1 d-1 using LFAH15-WG10. In addition, LFAH15-WG10 significantly enhanced total FAMEs yield by 21.2% over the control. Moreover, it reduced polyunsaturated fatty acids (PUFAs) ratio from 52.1% to 47.8% of total FAMEs, and increased monounsaturated fatty acids (MUFAs) ratio from 26.6% to 31.3% of total FAMEs.

Comparative Physiological, Biochemical, and Genetic Responses to Prolonged Waterlogging Stress in Okra and Maize Given Exogenous Ethylene Priming

Waterlogging is an environmental challenge affecting crops worldwide. Ethylene induces the expression of genes linked to important agronomic traits under waterlogged conditions. The ability of okra (Abelmoschus esculentus L. Moench.) and maize (Zea mays L.) given exogenous ethylene priming to tolerate prolonged waterlogged conditions was investigated in this study. The investigation was carried out as field experiments using 3 week-old plants grouped into four treatments; control, waterlogged plants, ethylene priming of plants before waterlogging, and ethylene priming of plants after waterlogging. Different growth parameters were recorded. Soil chemical and bacterial analyses were performed. The activity and gene expression of antioxidant enzymes were studied. The ethylene biosynthetic genes expression analysis and root anatomy of surviving okra plants were also carried out. Results revealed that okra and maize plants showed increase in their height under waterlogged conditions. Ethylene priming and waterlogged conditions induced early production of adventitious roots in okra and maize. Maize survival lasted between 5 and 9 weeks under waterlogging without reaching the flowering stage. However, okra survived up to 15 weeks under waterlogging producing flower buds and fruits in all treatments. Variable changes were also recorded for total soluble phenolics of soil. Cross sections of waterlogged okra roots showed the formation of a dark peripheral layer and numerous large aerenchyma cells which may have assisted in trapping oxygen required for survival. The activity and gene expression levels of antioxidant enzymes were studied and showed higher increases in the root and leaf tissues of okra and maize subjected to both waterlogging and ethylene priming, as compared to control or waterlogged condition. Quantitative RT-PCR analysis also showed that the ethylene biosynthetic gene expression levels in all okra and maize tissues were up-regulated and showed much higher levels under ethylene-treated waterlogged conditions than those expressed under control or waterlogged conditions at all time points. These results indicate that okra and maize tissues respond to the conditions of waterlogging and exogenous ethylene priming by inducing their ethylene biosynthetic genes expression in order to enhance ethylene production and tolerate the prolonged waterlogging stress. In conclusion, this study revealed that exogenously generated ethylene gas as a priming treatment before or after waterlogging could enhance waterlogging tolerance in maize and okra crops.

Analysis of the Genetic Diversity and Population Structure of Austrian and Belgian Wheat Germplasm within a Regional Context Based on DArT Markers

Analysis of crop genetic diversity and structure provides valuable information needed to broaden the narrow genetic base as well as to enhance the breeding and conservation strategies of crops. In this study, 95 Austrian and Belgian wheat cultivars maintained at the Centre for Genetic Resources (CGN) in the Netherlands were characterised using 1052 diversity array technology (DArT) markers to evaluate their genetic diversity, relationships and population structure. The rarefacted allelic richness recorded in the Austrian and Belgian breeding pools (A25 = 1.396 and 1.341, respectively) indicated that the Austrian germplasm contained a higher genetic diversity than the Belgian pool. The expected heterozygosity (HE) values of the Austrian and Belgian pools were 0.411 and 0.375, respectively. Moreover, the values of the polymorphic information content (PIC) of the Austrian and Belgian pools were 0.337 and 0.298, respectively. Neighbour-joining tree divided each of the Austrian and Belgian germplasm pools into two genetically distinct groups. The structure analyses of the Austrian and Belgian pools were in a complete concordance with their neighbour-joining trees. Furthermore, the 95 cultivars were compared to 618 wheat genotypes from nine European countries based on a total of 141 common DArT markers in order to place the Austrian and Belgian wheat germplasm in a wider European context. The rarefacted allelic richness (A10) varied from 1.224 (Denmark) to 1.397 (Austria). Cluster and principal coordinates (PCoA) analyses divided the wheat genotypes of the nine European countries into two main clusters. The first cluster comprised the Northern and Western European wheat genotypes, whereas the second included the Central European cultivars. The structure analysis of the 618 European wheat genotypes was in a complete concordance with the results of cluster and PCoA analyses. Interestingly, a highly significant difference was recorded between regions (26.53%). In conclusion, this is the first study to reveal the high diversity levels and structure of the uncharacterised Austrian and Belgian wheat germplasm maintained at the CGN as well as place them in a wider European context. The results should help plant breeders to utilise the most promising wheat genotypes of this study in future breeding programmes for enhancing wheat cultivars.

Bioactivities of Traditional Medicinal Plants in Alexandria

In traditional folklore, medicinal herbs play a vital role in the prevention and treatment of microbial diseases. In the present study, the phenolic profiles of the medicinal plants Asparagus aethiopicus L., Citrullus colocynthis L., Senna alexandrina L., Kalanchoe delagoensis L., Gasteria pillansii L., Cymbopogon citratus, Brassica juncea, and Curcuma longa L. were determined by high-performance liquid chromatography with a diode-array detector method. The results revealed rich sources of important compounds such as robinin in the fruits and leaves of A. aethiopicus; caffeic acid in the tubers of A. aethiopicus and quercitrin in the leaves of G. pillansii. Further, relatively high antioxidant, antibacterial, and antifungal activities were observed in C. colocynthis fruit coat, S. alexandrina pods, and A. aethiopicus leaves, respectively. The relatively higher the bioactivities of plants extracts associated with the phenols in these plants, in particular, the more abundant the phenols. Therefore, it was concluded that the fruit coat of C. colocynthis, pods of S. alexandrina, and leaves of A. aethiopicus might be excellent sources of natural products. These plant extracts also have a wide spectrum of antimicrobial activities that could be used in the pharmaceutical industries and to control diseases.