Mehmet Cengiz Baloglu

Genome-wide analysis of the bZIP transcription factors in cucumber.

bZIP proteins are one of the largest transcriptional regulators playing crucial roles in plant development, physiological processes, and biotic/abiotic stress responses. Despite the availability of recently published draft genome sequence of Cucumis sativus, no comprehensive investigation of these family members has been presented for cucumber. We have identified 64 bZIP transcription factor-encoding genes in the cucumber genome. Based on structural features of their encoded proteins, CsbZIP genes could be classified into 6 groups. Cucumber bZIP genes were expanded mainly by segmental duplication rather than tandem duplication. Although segmental duplication rate of the CsbZIP genes was lower than that of Arabidopsis, rice and sorghum, it was observed as a common expansion mechanism. Some orthologous relationships and chromosomal rearrangements were observed according to comparative mapping analysis with other species. Genome-wide expression analysis of bZIP genes indicated that 64 CsbZIP genes were differentially expressed in at least one of the ten sampled tissues. A total of 4 CsbZIP genes displayed higher expression values in leaf, flowers and root tissues. The in silico micro-RNA (miRNA) and target transcript analyses identified that a total of 21 CsbZIP genes were targeted by 38 plant miRNAs. CsbZIP20 and CsbZIP22 are the most targeted by miR165 and miR166 family members, respectively. We also analyzed the expression of ten CsbZIP genes in the root and leaf tissues of drought-stressed cucumber using quantitative RT-PCR. All of the selected CsbZIP genes were measured as increased in root tissue at 24th h upon PEG treatment. Contrarily, the down-regulation was observed in leaf tissues of all analyzed CsbZIP genes. CsbZIP12 and CsbZIP44 genes showed gradual induction of expression in root tissues during time points. This genome-wide identification and expression profiling provides new opportunities for cloning and functional analyses, which may be used in further studies for improving stress tolerance in plants.

Diverse expression pattern of wheat transcription factors against abiotic stresses in wheat species

Abiotic stress including drought and salinity affects quality and yield of wheat varieties used for the production of both bread and pasta flour. bZIP, MBF1, WRKY, MYB and NAC transcription factor (TF) genes are the largest transcriptional regulators which are involved in growth, development, physiological processes, and biotic/abiotic stress responses in plants. Identification of expression profiling of these TFs plays a crucial role to understand the response of different wheat species against severe environmental changes. In the current study, expression analysis of TaWLIP19 (wheat version of bZIP), TaMBF1, TaWRKY10, TaMYB33 and TaNAC69 genes was examined under drought and salinity stress conditions in Triticum aestivum cv. (Yuregir-89), Triticum turgidum cv. (Kiziltan-91), and Triticum monococcum (Siyez). After drought stress application, all five selected genes in Kiziltan-91 were induced. However, TaMBF1 and TaWLIP19 were the only downregulated genes in Yuregir-89 and Siyez, respectively. Except TaMYB33 in Siyez, expression level of the remaining genes increased under salt stress condition in all Triticum species. For determination of drought response to selected TF members, publicly available RNA-seq data were also analyzed in this study. TaMBF1, TaWLIP19 and TaNAC69 transcripts were detected through in silico analysis. This comprehensive gene expression analysis provides valuable information for understanding the roles of these TFs under abiotic stresses in modern wheat cultivars and ancient einkorn wheat. In addition, selected TFs might be used for determination of drought or salinity-tolerant and susceptible cultivars for molecular breeding studies.

Genome-wide characterization and expression analysis of common bean bHLH transcription factors in response to excess salt concentration

Members of basic helix-loop-helix (bHLH) gene family found in all eukaryotes play crucial roles in response to stress. Though, most eukaryotes carry the proteins of this family, biological functions of the most bHLH family members are not deeply evaluated in plants. In this study, we conducted a comprehensive genome-wide analysis of bHLH transcription factors in salt tolerant common bean. We identified 155 bHLH protein-encoding genes (PvbHLH) by using in silico comparative genomics tools. Based on the phylogenetic tree, PvbHLH genes were classified into 8 main groups with 21 subfamilies. Exon–intron analysis indicated that proteins belonging to same main groups exhibited a closely related gene structure. While, the PvbHLH gene family has been mainly expanded through segmental duplications, a total of 11 tandem duplication were detected. Genome-wide expression analysis of bHLH genes showed that 63 PvbHLH genes were differentially expressed in at least one tissue. Three of them displayed higher expression values in both leaf and root tissues. The in silico micro-RNA target transcript analyses revealed that totally 100 PvHLH genes targeted by 86 plant miRNAs. The most abundant transcripts, which were targeted by all 18 plant miRNA, were belonging to PvHLH-22 and PvHLH-44 genes. The expression of 16 PvbHLH genes in the root and leaf tissues of salt-stressed common bean was evaluated using qRT-PCR. Among them, two of PvbHLHs, PvbHLH-54, PvbHLH-148, were found to be up-regulated in both tissues in correlation with RNA-seq measurements. The results of this study could help improve understanding of biological functions of common bean bHLH family under salt stress. Additionally, it may provide basic resources for analyzing bHLH protein function for improving economic, agronomic and ecological benefit in common bean and other species.

Differential Gene Expression in Liver Tissues of Streptozotocin-Induced Diabetic Rats in Response to Resveratrol Treatment

This study was conducted to elucidate the genome-wide gene expression profile in streptozotocin induced diabetic rat liver tissues in response to resveratrol treatment and to establish differentially expressed transcription regulation networks with microarray technology. In addition to measure the expression levels of several antioxidant and detoxification genes, real-time quantitative polymerase chain reaction (qRT-PCR) was also used to verify the microarray results. Moreover, gene and protein expressions as well as enzymatic activities of main antioxidant enzymes; superoxide dismutase (SOD-1 and SOD-2) and glutathione S-transferase (GST-Mu) were analyzed. Diabetes altered 273 genes significantly and 90 of which were categorized functionally which suggested that genes in cellular catalytic activities, oxidation-reduction reactions, co-enzyme binding and terpenoid biosynthesis were dominated by up-regulated expression in diabetes. Whereas; genes responsible from cellular carbohydrate metabolism, regulation of transcription, cell signal transduction, calcium independent cell-to-cell adhesion and lipid catabolism were down-regulated. Resveratrol increased the expression of 186 and decreased the expression of 494 genes in control groups. While cellular and extracellular components, positive regulation of biological processes, biological response to stress and biotic stimulants, and immune response genes were up-regulated, genes responsible from proteins present in nucleus and nucleolus were mainly down-regulated. The enzyme assays showed a significant decrease in diabetic SOD-1 and GST-Mu activities. The qRT-PCR and Western-blot results demonstrated that decrease in activity is regulated at gene expression level as both mRNA and protein expressions were also suppressed. Resveratrol treatment normalized the GST activities towards the control values reflecting a post-translational effect. As a conclusion, global gene expression in the liver tissues is affected by streptozotocin induced diabetes in several specific pathways. The present data suggest the presence of several processes which contribute and possibly interact to impair liver functions in type 1 diabetes, several of which are potentially amenable to therapeutic interventions with resveratrol.

Drought-Responsive Hsp70 Gene Analysis in Populus at Genome-Wide Level

The heat shock protein 70 (Hsp70) family members are known as molecular chaperones. They play a crucial role in protecting plant cells and tissues from thermal or abiotic stress through protein folding and in assembly, stabilization, activation, and degradation processes. Although many studies have been performed to identify molecular functions of individual family members, there is a limited study on genome-wide identification and characterizations of Hsps in the Populus model tree genus. We have identified 34 poplar Hsp70 genes, which were phylogenetically clustered into three major groups. Gene structure and motif composition are relatively conserved in each group. Mainly tandem and infrequently segmental duplications have a significant role in poplar Hsp70 gene expansion. The in silico microRNA (miRNA) and target transcript analyses identified that a total of 19 PtHsp70 genes were targeted by 27 plant miRNAs. PtHSP70-14 and PtHSP70-33 are the most targeted by miR390 and miR414 family members, respectively. For determination of drought response to Hsp70 genes, publicly available RNA-seq data were analyzed. Poplar Hsp70s are differentially expressed upon exposure to different drought stress conditions. Expression analysis of PtHsp70 genes was also examined under drought stress in drought-sensitive and drought-resistant Populus clones with quantitative real-time PCR (qRT-PCR). PtHsp70-16 and PtHsp70-26 genes might provide adaptation to drought stress for both clones. Because of high expression responses to drought in only resistant Populus clone, PtHsp70-25 and PtHsp70-33 genes might be used for determination of drought-tolerant clones for molecular breeding studies. This research provides a fundamental clue for contribution of PtHsp70s to drought tolerance in poplar.

A Multidirectional Perspective for Novel Functional Products: In vitro Pharmacological Activities and In silico Studies on Ononis natrix subsp. hispanica

The genus Ononis has important value as traditional drugs and foods. In the present work, we aimed to assess the chemical profiles and biological effects of Ononis natrix subsp. hispanica extracts (ethyl acetate, methanol, and water). For chemical profile, total and individual phenolic components were detected. For biological effects, antioxidant (DPPH, ABTS, CUPRAC, FRAP, phosphomolybdenum, and metal chelating assays), enzyme inhibitory (against cholinesterase, tyrosinase, α-amylase and α-glucosidase), antimicrobial, DNA protection and cytotoxic abilities were tested. The predominant phenolics were apigenin, luteolin, and quercetin in the tested extracts. Generally, the ethyl acetate and methanol extracts were noted as the most active in the antioxidant and enzyme inhibitory assays. Water extract with different concentrations indicated high level of DNA protection activity. Methanol and ethyl acetate extracts showed antibacterial effect against to Staphylococcus aureus and Staphylococcus epidermidis strains. The cytotoxic effects of O. natrix subsp. hispanica extracts on the survival of HeLa and PC3 cells were determined by MTT cell viability assay. Water and methanol extracts caused initiation of apoptosis for PC3 cell line. Furthermore, molecular docking was performed to better understand interactions between dominant phenolic compounds and selected enzymes. Our results clearly indicate that O. natrix subsp. hispanica could be considered a potential candidate for designing novel pharmaceuticals, cosmeceuticals and nutraceuticals.

Novel in vitro and in silico insights of the multi-biological activities and chemical composition of Bidens tripartita L.

Bidens tripartita L. is a traditional phyto-remedy used in several countries, yet there is still a paucity of data on its biological potential. We aimed to provide new insights on the pharmacological potential of extracts prepared from B. tripartita via highlighting its antioxidant, key enzymes inhibitory potency, and DNA protecting effects. Phytochemical profile was established using High-Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD) and bioactive compound(s) docked against target enzymes using in silico methods. Cytotoxicity against three cancer cell lines was assessed using the methylthiazolyldiphenyl-tetrazolium bromide (MTT) cell viability test. The main compounds were luteolin-7-glucoside (cynaroside), chlorogenic acid, and epicatechin in the extracts. The methanol extract exhibited the highest radical scavenging activity. Ethyl acetate extract showed strongest α-amylase inhibitory activity, while the best α-glucosidase inhibitory effect recorded for the methanol extract. Molecular docking showed that cynaroside strongly interact to α-glucosidase cavity by establishing six hydrogen bonds. B. tripartita extracts were found to protect supercoiled form of pUC19 plasmid (>70%) and also showed anti-proliferative properties. Results amassed in the present study add on to a growing body of literature on the multi-pharmacological potency of B. tripartita which can be applied to bio-products development geared towards management of common diseases.

DNA protection, antioxidant, antibacterial and enzyme inhibition activities of heartwood and sapwood extracts from juniper and olive woods

In this study, DNA protective, antioxidant, antibacterial and enzyme inhibiting properties of methanol extracts obtained from juniper and olive heartwood and sapwood were determined. These extracts were tested by five antioxidant methods (DPPH scavenging, FRAP, CUPRAC, metal chelating and phosphomolybdenum). Generally, heartwood extracts of both species are more efficient for DPPH radical scavenging activity, cupric ion reducing activity, ferric reducing antioxidant power and metal chelating activity than sapwood extracts. When compared to heartwood extracts, sapwood extracts have larger inhibition zone in disk diffusion test. In addition, all extracts showed high antibacterial activity against Staphylococcus aureus. DNA protection of both extracts had a capacity to inhibit the DNA damage arisen from Fentons reagent. The highest DNA protective activity was observed in juniper sapwood extract with 84%. Furthermore, other extracts also indicated more than 60% of DNA protective activity. Olive wood extracts displayed the strongest enzyme inhibition activities against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Although juniper heartwood extracts showed highest anti-amylase, anti-glucosidase and anti-tyrosinase effects, they had no ability for inhibition BChE. The methanol extracts of olive samples demonstrated the most antioxidant activity (DPPH, CUPRAC and FRAP). In addition, juniper samples showed the highest anti-amylase, anti-tyrosinase, metal chelating and DNA protective activity. According to these results, the extracts of juniper and olive wood can be considered as a source of natural bio active agents for dietary, pharmacological and medicinal applications. This research will also serve as a base for future studies about biological activities of wood extracts.

Molecular Characterization, 3D Model Analysis, and Expression Pattern of the CmUBC Gene Encoding the Melon Ubiquitin-Conjugating Enzyme Under Drought and Salt Stress Conditions

Ubiquitin-conjugating (UBC) enzyme is a key enzyme in ubiquitination. Here, we describe the cloning, characterization, and expression pattern of a novel gene, CmUBC, from a melon. Comparison of the deduced amino acid sequences allowed the identification of highly conserved motifs. Synteny analysis between Cucumis sativus L. and Arabidopsis demonstrated that homologs of several Cucumis UBC genes were found in corresponding syntenic blocks of Arabidopsis. The homology structure model of the CmUBC protein was constructed. UBCs from melon, yeast, and Arabidopsis were highly conserved in their three-dimensional folding. CmUBC was ubiquitously expressed in all melon tissues. Increased transcript levels of CmUBC were observed during drought and salinity stresses, which suggested that the expression of the CmUBC gene in melon plants is responsive to physiological water stress. These results suggested that the CmUBC gene might play an important role in the modulation of the ubiquitination pathway.

Identification, molecular characterization and expression analysis of RPL24 genes in three Cucurbitaceae family members: cucumber, melon and watermelon

Ribosomal proteins are crucial for the proper growth and development of any organism, including plants. The ribosomal protein L24 (RPL24) is found in the large subunit of the ribosome and is responsible for the stabilization of the peptidyl transferase activity. Although RPL24 gene has been individually identified in different organisms, little is known about the genome-wide survey and expression patterns of R24 genes in Cucurbitaceae family members. We identified seven Cucurbitaceae RPL24 genes from cucumber, melon and watermelon. They were phylogenetically clustered into seven major groups. Gene structure and motif composition are relatively conserved in each group. Three-dimensional homology modelling of RPL24 proteins was performed with higher confidence level. CmRPL24-01 was isolated from melon and characterized at a molecular level. The regulation of ribosomal proteins in melon under drought stress conditions was also studied. The expression of CmRPL24-01 gene increased in melon leaf tissue at 3 h upon polyethylene glycol treatment and showed a gradual induction after 12 h. Our study provided a very useful reference for identification and functional analysis of RPL24 protein members in different plants. In addition, this research indicated a potential usage of ribosomal proteins in response to drought stress.