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Öğe 24-Epibrassinolide promotes arsenic tolerance in Arabidopsis thaliana L. by altering stress responses at biochemical and molecular level(Elsevier Gmbh, 2019) Surgun-Acar, Yonca; Zemheri-Navruz, FahriyeIn this study, the effect of 24-Epibrassinolide (EBL) on antioxidant system in Arabidopsis thaliana were investigated under arsenate [As(V)] stress. The enzyme activity of superoxide dismutase (SOD) and catalase (CAT), total antioxidant status, malondialdehyde (MDA) level and free proline content, as well as the expression levels of SOD isoforms (Cu-ZnSODs, FeSODs and MnSOD), CAT isoforms (CAT1, CAT2 and CAT3), some heat shock proteins (Hsp70-4 and Hsp90-1) and proline biosynthesis (P5CS1 and P5CS2) genes were determined in rosette leaves of eight-week old plants under exposure of 100 and 200 mu M As(V) and/or 1 mu M EBL treatments for 24 h. Total SOD and CAT enzyme activities increased as a result of 100 mu M As(V) + EBL treatments compared to 100 As(V) treatment. Total antioxidant and proline levels increased in plants subjected to As(V), and the treatment of EBL together with stress caused further increase. As the MDA level increased in As-treated plants, 100 mu M As(V) + EBL treatment decreased MDA level. Transcript levels of CSD1, CSD2, FSD1, FSD2, MSD1 and CAT2 genes increased as a result of combined treatment of EBL and As(V) compared to control and alone stress treatments (except CSD1 gene). Expression level of CSD3, CAT1 and CAT3 genes were downregulated in response to As(V) and/or EBL treatments. EBL application alone and in combination with As(V) elevated the expression level of P5CS1 gene dramatically. Treatment with 100 mu M As(V) and EBL increased the transcript level of Hsp70-4 and Hsp90-1 genes in leaves compared to 100 mu M As(V) treatment. To our best knowledge, this is the first detailed study to evaluate the improving effect of EBL on antioxidant defense system at biochemical and transcriptional level in A. thaliana plants under As(V) stress.Öğe Determination of heavy metal-induced dna damage in pisum sativum L. At the molecular and population level(Pakistan Agricultural Scientists Forum, 2018) Surgun-Acar, YoncaIn the current study, the effects of heavy metals on pea (Pisum sativum L.), which were exposed to 150 mg L-1 and 300 mg L-1 Cd, Pb and Cu treatments for 12 days, were investigated by using population parameters and Random Amplified Polymorphic DNA (RAPD) assay in the r oots and leaves. In RAPD assay, 17 RAPD primers generated polymorphic band profiles, and a total of 110 bands were produced in the leaves and roots of the control seedlings. Changes were detected in the RAPD patterns of the roots and leaves of the pea plants that subjected to heavy metal treatment in terms of band intensities, appearance of new bands and disappearance of bands. The changes in the RAPD patterns were found to be more in roots when compared leaves. The results of germination, shoot and primary root length showed a correlation with the genomic template stability (GTS) values. Determination of the DNA damage by using the RAPD assay together with different biomarkers may be a practical eco-genotoxicological tool in the biomonitoring of chemical contaminants. © 2018, Pakistan Agricultural Scientists Forum. All rights reserved.Öğe DETERMINATION OF HEAVY METAL-INDUCED DNA DAMAGE IN PISUM SATIVUM L. AT THE MOLECULAR AND POPULATION LEVEL(Pakistan Agricultural Scientists Forum, 2018) Surgun-Acar, YoncaIn the current study, the effects of heavy metals on pea (Pisum sativum L.), which were exposed to 150 mg L-1 and 300 mg L-1 Cd, Pb and Cu treatments for 12 days, were investigated by using population parameters and Random Amplified Polymorphic DNA (RAPD) assay in the roots and leaves. In RAPD assay, 17 RAPD primers generated polymorphic band profiles, and a total of 110 bands were produced in the leaves and roots of the control seedlings. Changes were detected in the RAPD patterns of the roots and leaves of the pea plants that subjected to heavy metal treatment in terms of band intensities, appearance of new bands and disappearance of bands. The changes in the RAPD patterns were found to be more in roots when compared leaves. The results of germination, shoot and primary root length showed a correlation with the genomic template stability (GTS) values. Determination of the DNA damage by using the RAPD assay together with different biomarkers may be a practical eco-genotoxicological tool in the biomonitoring of chemical contaminants.Öğe Estimation of arsenic-induced genotoxicity in melon (Cucumis melo) by using RAPD-PCR and comet assays(Institute of Botany and Botanical Garden "Jevremovac", University of Belgrade, 2021) Surgun-Acar, YoncaIn this study, arsenic (As)-induced genotoxicity in the roots and shoots of Cucumis melo (melon) seedlings were investigated by using the random amplified polymorphic DNA - polymerase chain reaction (RAPD-PCR) and comet assays. For this purpose, melon seedlings were exposed to arsenate [As(V)] at 0, 100, 200, 300, and 400 μM concentrations in the hydroponic system for 14 days to examine the level of As accumulation, alterations in growth parameters, and DNA damage. A reduction in growth with increasing As(V) concentration was observed in the melon seedlings. Total As accumulations in the shoot and root tissue increased in a dose-dependent manner; however, the level was higher in the roots than the shoots. In RAPD-PCR analysis, 26 primers gave reproducible and scorable results and produced a total of 128 bands in the control seedlings. Alterations in RAPD profiles, including the loss or appearance of new bands, were determined in the As-treated seedlings when compared to the control. The values of genomic template stability (GTS) were decreased by increasing the concentration of the As(V) in both tissue types. DNA strand breaks were observed in all the tested As(V) concentrations in the alkaline comet assay; furthermore, the loss of DNA integrity was higher with 300 and 400 μM As(V) treatments. The results clearly indicate that the combination of DNA-based molecular and cytogenetic techniques (e.g. the comet assay) may be proposed as a reliable evaluation of genotoxicity in plants after exposure to heavy metal pollution.Öğe Exogenous Application of 24-Epibrassinolide Improves Manganese Tolerance in Arabidopsis thaliana L. via the Modulation of Antioxidant System(Springer, 2022) Surgun-Acar, Yonca; Zemheri-Navruz, FahriyeThe goal of the present study was to investigate the role of 24-Epibrassinolide (EBL) hormone in supporting adaptation of plant to manganese (Mn) stress. For this purpose, changes in antioxidant system and stress-related gene expression were determined in rosette leaves of Arabidopsis thaliana following 24-h exposure to Mn (0.5 and 1.0 mM) and/or EBL (1 µM). Decreased chlorophyll level in the rosette leaves of seedlings due to Mn stress increased consequent to exogeneous EBL application. Superoxide dismutase (SOD) and catalase (CAT) enzyme activities alongside transcript level of copper/zinc superoxide dismutase 1 (CSD1), copper/zinc superoxide dismutase 2 (CSD2), copper/zinc superoxide dismutase 3 (CSD3), Fe superoxide dismutase 2 (FSD2), Fe superoxide dismutase 3 (FSD3), and catalase 2 (CAT2) genes were found to increase depending on Mn concentration; however, co-exposure of EBL and Mn led to a further improvement in enzyme activities and gene expressions (except CSD1 and CSD2 genes). The mRNA level of Fe superoxide dismutase 1 (FSD1) gene was downregulated following Mn and/or EBL treatments. Exogenous treatment of EBL improved the total antioxidant and proline level, while it decreased the lipid peroxidation under Mn stress. EBL treatment upregulated transcript level of delta 1-pyrroline-5-carboxylate synthase 1 (P5CS1) and delta 1-pyrroline-5-carboxylate synthase 2 (P5CS2) genes in seedlings treated with Mn. Co-treatment of Mn and EBL enhanced expression level of A. thaliana detoxification 1 (AtDTX1) and A. thaliana detoxification 3 (AtDTX3) genes when compared to the seedlings treated with Mn. Biochemical and molecular results from the current study revealed that EBL regulates antioxidant system in a coordinated manner under A. thaliana Mn stress.Öğe Expression analysis of cell wall assembly and remodelling-related genes in Arabidopsis roots subjected to boron stress and brassinosteroid at different developmental stages(Soc Botanica Brasil, 2018) Iskil, Rabia; Surgun-Acar, YoncaPlant cell walls are affected by many biotic and abiotic stress conditions. The aim of this study is to determine the effects of 24-Epibrassinolide (EBL) on some cell wall-related genes in root tissue of five- and ten-week-old Arabidopsis thaliana plants exposed to boron (B) deficiency (0 mu M) or toxicity (3000( )mu M) at the transcriptional level. Expressions of the genes that encode cellulose synthase (CESA1, CESA4, CESA6 and CESA8), cellulose synthase-like (CSLBS), expansin (EXPAS, EXPA8 and EXPA14) and cell wall protein (SEB1) decreased under conditions of B deficiency and toxicity. EBL treatments, in general, led the expressions of these genes to reduce significantly. Expressions of xyloglucan endotransglucosylase/hydrolase genes (XTH21 and XTH23) changed only under conditions of B toxicity. Boron stress and/or EBL treatments caused different responses in expression of pectin methylesterase (PME2 and PME41) genes. As a result of B stress, the expression levels of investigated genes changed more in roots of five-weekold plants than in roots of ten-week-old plants. Results of the present study include new findings that support the ability of BRs to increase molecular aspects of tolerance to stress in plants.Öğe GENOTOXICITY ASSESSMENT OF HEAVY METALS (Zn, Cr, Pb) ON STRAWBERRY PLANTS USING RAPD ASSAY(Parlar Scientific Publications (P S P), 2018) Surgun-Acar, Yonca; Iskil, Rabia; Ceylan, Keyser Betul; Ceylan, YusufThe aim of the present study is to assess DNA damage in leaves of strawberry (Fragaria x ananassa Duch.) seedlings treated with different heavy metals (zinc, chromium, lead) using random amplified polymorphic DNA (RAPD) assay and analysis of total soluble protein content. For this purpose, strawberry seedlings were treated with 400 and 800 mu M Zn, Pb and Cr for 7 days. Thirty four RAPD primers produced 218 bands at molecular weight ranging from 183 bp to 5180 bp. Compared with control, RAPD patterns of heavy metal exposed groups showed differences in band loss, gain of new bands and increase and decrease of band intensity. The highest polymorphism rate (32.11%) was observed in 800 mu M Pb applied strawberry seedlings. To evaluate the alterations in RAPD profiles qualitatively, genomic template stability (GTS) was performed and the values were 75.08% and 69.59% for 400 and 800 mu M Zn treatments, 74.52% and 68.60% for 400 and 800 mu M Cr treatments, 70.11% and 63.78% for 400 and 800 mu M Pb treatments, respectively. Total soluble protein content in heavy metal-treated groups also showed a similar correlation to GTS values. RAPD analyses are useful biomarker assays to determine the genotoxicity induced by environmental pollutants such as heavy metals in plant model systems.Öğe Mercury toxicity affects oxidative metabolism and induces stress responsive mechanisms in wheat (Triticum aestivum L.)(Springer, 2022) Iskil, Rabia; Surgun-Acar, Yonca; Catav, Sukru Serter; Zemheri-Navruz, Fahriye; Erden, YavuzMercury (Hg) toxicity is an increasing problem worldwide, with a negative impact on the environment and living organisms including both animals and plants. In this study, we analyzed molecular and biochemical changes related to Hg toxicity in wheat (Triticum aestivum L.) seedlings. Seven-day-old seedlings were exposed to various concentrations (5, 10, and 20 mu M) of HgCl2 for 24 and 48 h. Our results showed that HgCl2 treatments led to an increase in the Hg content of wheat leaves in a time- and concentration-dependent manner. Furthermore, significant increases were observed in hydrogen peroxide, malondialdehyde, and proline contents in response to Hg toxicity. While all HgCl2 treatments decreased the level of superoxide dismutase (SOD), the level of catalase (CAT) was reduced only in seedlings exposed to 5 mu M of HgCl2. Mercury stress caused a decline in the expression of Cu/Zn-SOD, Fe-SOD, TaWRKY19, and TaDREB1 genes at both exposure times. On the other hand, 10 and 20 mu M HgCl2 treatments caused significant induction (1.9 to 6.1-fold) in the expression of the CAT gene in wheat leaves. The mRNA level of the Mn-SOD and TaWRKY2 genes showed different patterns depending on the concentration and exposure period of HgCl2. In conclusion, the findings of this work demonstrate that Hg toxicity causes oxidative damage in wheat seedlings and changes the expression of some genes encoding WRKY and DREB transcription factor families, which have important functions in abiotic stress response.Öğe Physiological, biochemical, and molecular responses of wheat seedlings to salinity and plant-derived smoke(Elsevier, 2021) Çatav, Şükrü Serter; Surgun-Acar, Yonca; Zemheri-Navruz, FahriyeThere is great interest in developing strategies to reduce the detrimental effects of salinity on plant growth and crop yield. Plant-derived smoke containing many promotive and inhibitory compounds is known to influence seed germination and plant growth in a concentration-dependent manner. It has been suggested that smoke can also alleviate abiotic stress-induced growth inhibition in several plants. However, there is still a lack of comprehensive knowledge regarding smoke-mediated biochemical and molecular changes in plants grown under stress conditions. Here we show that smoke regulates the expression of some genes encoding transcription factors and antioxidant enzymes in wheat seedlings subjected to salt stress (150 mM NaCl). We found that smoke-water (0.1%) increased the expression levels of TaDREB1, TaWRKY2, TaWRKY19, CAT, and Cu/Zn-SOD in salt-treated seedlings. Smoke-water also enhanced the transcription of TaWRKY19 and Cu/Zn- SOD genes under normal growth conditions. In addition, smoke was found to modulate physiological responses of wheat seedlings to salinity. The results demonstrate that smoke-water increases total phenolic content and free radical scavenging activity under salt stress but it leads to a significant decrease in proline accumulation. In conclusion, the current study reveals that smoke, although not having a remarkable effect on growth at the concentration tested, can affect physiological, biochemical, and molecular processes in wheat seedlings exposed to short-term salt stress. Our data also indicate the need for long-term field experi- ments for a better understanding of the effect of smoke on plant growth in the presence of salinity.Öğe Response of soybean (Glycine max L.) seedlings to polystyrene nanoplastics: Physiological, biochemical, and molecular perspectives(Elsevier Sci Ltd, 2022) Surgun-Acar, YoncaMicro and nanoplastics are new generation contaminants of global concern. It is important to evaluate the effects on edible products due to the presence of micro-and nano-sized plastics in the treated wastewater. A hydroponic experiment was carried out to explore the effect of polsytrene nanoplastics (PS-NPs; 20 nm) at different con-centrations (0, 12.5, 25, and 50 mg L-1) on Glycine max L. (soybean) seedlings for 7-days. In the current study, firstly the uptake of PS-NPs by Glycine max L. (soybean) roots were confirmed by laser confocal scanning mi-croscope. Exposure to PS-NPs, negatively affected growth parameters and increased Fe, Zn and Mn contents in roots and leaves of soybean seedlings. PS-NPs treatments caused oxidative stress in soybean seedlings. The hydrogen peroxide and malondialdehyde contents, showed similar increase pattern in seedlings exposed to PS -NPs. Response to PS-NPs, the level of antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase, and guaiacol peroxidase) and proline content were generally enhanced in roots and leaves of soybean. The expression level of stress-related genes examined in the study included CSD5, FSD3, APX1, and POD up-regulated in PS-NPs treated-soybean seedlings in a tissue specific manner. The results of the present study showed the adverse effects of PS-NPs on soybean seedlings, which may have important implications for the risk assessment of NPs on crop production and environmental safety.Öğe The effect of 24-Epibrassinolide on gene expression related to cell walls under boron deficiency and toxicity in the leaves of Arabidopsis thaliana(Univ Belgrade, Inst Botany & Botanical Garden, 2022) Iskil, Rabia; Surgun-Acar, YoncaThe changing composition of plant cell walls allows for the continuation of the existing structure under normal conditions and also the protection of physical integrity under altering environmental conditions. In this study, the possible effects of the 24-Epibrassinolide (EBL) hormone under boron (B) deficiency and toxicity conditions on the expression of cell wall-related genes [cellulose synthase (CESA), expansin (EXP), xyloglucan endotransglucosylase/hydrolase (XTH) and pectin methylesterase (PME)] were investigated in the rosette leaves of Arabidopsis thaliana. For this purpose, 0 or 3000 mu M of boric acid (BA) and/or 1 mu M of EBL were applied to the plants which were grown in a hydroponic medium for five and ten weeks. While B-toxicity elevated the mRNA levels of the CESA4 and CESA8 genes in the leaves of the five-week-old plants, B-stress (B-deficiency and-toxicity) caused an increase in the expression of the CESA4, CESA6, and CESA8 genes in the ten -weekold plants. The transcript levels of the EXPA5 gene increased under B-stress in the ten-week-old plants whereas the expression of the EXPA8 gene decreased when compared to the control at two developmental stages. Co-treatment of EBL and B stress strongly elevated the transcript level of the EXPA5 gene in the ten-week-old plants and the EXPA8 gene at both developmental stages. The EXPA14 and XTH23 genes exhibited distinct expression profiles under B-deficiency and-toxicity in both the five-and ten-week-old plants. The transcript level of the XTH21 gene was upregulated in the leaves of the plants exposed to B-stress. The mRNA level of the PME2 and PME41 genes was generally upregulated in response to B-stress in both the five-and ten-week-old plants. 24-Epibrassinolide alone and in combination with B-stress led to a remarkable increase in the expression of the XTH and PME genes compared to the control. These results demonstrate that cell wall genes generally show a similar pattern of expression at both developmental stages and the EBL hormone induces changes in the expression levels of cell wall-related genes under B-stress.
