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Bitki büyüme düzenleyicilerinin Calendula officinalis L. ve C. arvensis L. türlerinin hücre süspansiyon kültürlerinde belirli sekonder metabolitler üzerine etkileri
(Çanakkale Onsekiz Mart Üniversitesi, Lisansüstü Eğitim Enstitüsü, 2016) Kaya, Nergis; Akı, Cüneyt
Araştırmamızın ilk aşamasında tıbbi ve ekonomik öneme sahip olan Calendula officinalis ve Calendula arvensis fideleri embriyodan itibaren in vivo ve in vitro olarak yetiştirilmiştir. İkinci aşamada, in vitro olarak yetiştirilen iki Calendula fidesinin yaprak eksplantları ile başlatılan ve MS0, MS1, MS4, MS6 besin ortamlarında yapılan biyokütle ölçümleri ile kallus kültürü optimize edilmiştir. Üçüncü aşamada, MS1, MS3, MS4, MS6 besin ortamlarının bulunduğu kallus kültürlerinden hücre süspansiyon kültürlerine geçiş yapılarak hücre sayımı, taze ve kuru ağırlık ölçümleri yapılmıştır. Dördüncü aşamada, in vivo ve in vitro olarak yetiştirilen bitkilerin yapraklarında; kallus kültüründe ve hücre süspansiyon kültüründe biriken sekonder metabolitlerin (kuersetin, kaemferol, kafeik asit, beta karoten) miktarları araştırılmıştır. En fazla kafeik asit ve beta karoten miktarları, her iki Calendula türünün MS1 besin ortamındaki hücre süspansiyon kültürlerinin 120. gününde saptanmıştır. Kafeik asit miktarının C. officinalis ve C. arvensis türünde sırası ile 268,585 µg/g ve 235,865 µg/g; beta karoten miktarının ise C. officinalis ve C. arvensis türünde sırası ile 523,685 µg/g ve 385,115 µg/g olduğu ortaya konmuştur. Tamamlanan araştırmada C. officinalis ve C.arvensis türünün hücre süspansiyon kültürlerinde biriken kafeik asit ve beta karoten miktarları ilk defa belirlenmiştir.
Catharanthus roseus (L.) G. Don. türünde bitki büyüme düzenleyicilerinin in vitro somaklonal varyasyon üzerine etkilerinin belirlenmesi
(Çanakkale Onsekiz Mart Üniversitesi, 2012) Kaya, Nergis; Akı, Cüneyt
Bu araştırmada, in vivo olarak yetiştirilen Catharanthus roseus (L.) G. Don bitkisinden yaprak eksplantları alınarak 5:2; 8:2; 10:2 mg/L NAA:BAP içeren MS ortamında in vitro kültüre alınmıştır. Eksplantlar dört haftada bir alt kültüre alınmış ve uygulamaların beş alt kültür boyunca kallus üzerindeki biyokütle değişimleri ve somaklonal varyasyon üzerine etkileri takip edilmiştir. Tüm denemeler üç tekrarlı olarak gerçekleştirilmiştir. Biyokütle değişimlerinde alt kültürler boyunca ilk üç alt kültür boyunca tüm serilerde genel bir artış gözlenmiştir. Kallus biyokütlesi ekimin başlangıcından 1. alt kültür sonuna kadar 5:2; 8:2; 10:2 mg/L NAA:BAP içeren MS ortamında sırası ile %168, %139 ve %142 arttığı belirlenmiştir. Kallustan ezme preparat yöntemi ile mikroskopta yapılan incelemelerde, somaklonal varyasyon ile ilgili poliploidi, metafaz kromozom dağınıklılığı gibi anormalliklere rastlanmıştır. Sonuç olarak, biyokütle eldesinde MS ortamına eklenen bitki büyüme düzenleyicilerinin en uygun kombinasyonun 5:2 mg/L NAA:BAP olduğu ve bitki büyüme düzenleyicilerinin farklı kombinasyon ve konsantrasyonlarının kallus biyokütlesinin de farklı şekilde artış gösterdiği saptanmış ve ölçümler istatistiksel olarak verilmiştir.
Effect of Ferrous Gluconate on Chromosomal Abnormality Index of Allium Cepa Root Tip
(2021) Kaya, Nergis
In completed research, ferrous gluconate -a food additive- used to preserve black color to prevent discoloration during storage in ripe black olives, and Allium cepa L. species. A. cepa L. roots were treated with different doses of ferrous gluconate. The effective concentration EC50 (0.068 g/l) was determined. A. cepa root tips were treated with EC50/2 (0.034 g/l), EC50 (0.068 g/l), 2XEC50 (0.136 g/l) dose for 24, 48, 72 hours, and afterward, the root tips were prepared for observation under the light microscope according to the method of preparing mitotic preparation. Chromosomal abnormality index (CAI) and genotoxic effect of ferrous gluconate in A. cepa root tip cells were determined. Repeated measurement ANOVA and TUKEY multiple comparison tests were used to investigate the effect of time and dose together on genotoxicity. C-mitosis, polyploidy, polar shifting in anaphase, polar shifting in telophase, equatorial plate shifting, laggard chromosome was observed by microscope. The highest CAI (70.16±4.85) was observed at 72h for 2XEC50 dose. Chromosomal aberration is also observed in control group. While the most common chromosomal aberration is determined as C-mitosis; The least observed chromosomal aberration is determined as polyploidy. Research results revealed that ferrous gluconate has a genotoxic effect on the root tip of A. cepa.
Effects of dodine on total protein content and peroxidase activity in Vicia faba L.
(2016) Çördük, Nurşen; Akıncı, Nihan; Kaya, Nergis; Yücel, Gülru; Akı, Cüneyt
Bu araştırmada dodin fungusitinin Vicia faba L. fizyolojisi üzerindeki etkisi, total protein miktarı ve peroksidaz (POX) aktivitesi aracılığıyla değerlendirilmiştir. Bu doğrultuda, dört haftalık V. faba fideleri 0.04 mL/L, 0.08 mL/L ve 0.16 mL/L'lik dodin konsantrasyonları ile muamele edilmiştir. Bu işlemlerden sonra bitki örneklerinin yaprakları 24, 48 ve 72 saat ara ile hasat edilmiştir. Yapılan analizler sonucunda, artan dodin konsantrasyonu ve muamele süresiyle total protein miktarı kontrole göre önemli ölçüde azalırken, peroksidaz aktivitesi yükselmiştir. Elde edilen sonuçlar dodinin bitki savunma sistemini uyardığını göstermiştir.
Evaluation of Anti-Cytotoxicity and Anti-Genotoxicity of Raphanus sativus L. Tuber on Allium cepa L. Root Tips
(Pleiades Publishing, 2023) Kaya, Nergis
Abstract: Raphanus sativus (radish) tuber extracts stock solutions were prepared by dissolving radish extracts prepared with certain solvents (distilled water, ethanol and methanol) in dimethyl sulfoxide. Negative control (dimethyl sulfoxide, DMSO), positive control mutagen (ethyl methanesulfonate) and R. sativus extracts (10 and 20 mg/mL doses) were applied to Allium cepa root tips at for 24 and 48 h. For examination under the light microscope, the root tips were placed in fixative, then hydrolyzed and treated with acetocarmine. Chromosomal aberration index (CAI%) was increased as compared with the negative control, but decreased as compared with the positive control after the application of three different extracts. It was put forward that mitotix index decreased with increasing extract concentration applications. In this way, it has been determined that these extracts have an inhibitory effect on cell division at high doses. It has been found that CAI% was an increase with increasing extract concentration. It has been revealed that distilled water, ethanol and methanol extracts have more inhibitory effect on cell division and an increasing frequency of chromosomal anomalies, respectively. As a result of this research, R. sativus tuber might cause damage to cells genetic material and spindle formation when used incorrectly.
Evaluation of cytotoxic and genotoxic effects of hosap stream gumuscay biga canakkale on Allium cepa L.
(Parlar Scientific Publications, 2021) Kaya, Nergis
Since the Hosap Stream is located near the urban settlement area, water pollution might occur. In this study, the cytotoxic and genotoxic effect of Hosap Stream, which is also used in irrigation of agricultural lands, was investigated at the root tip cells of Allium cepa. The EC50 value in A. cepa roots was determined by exposure to different concentrations of the water sample taken from Hosap Stream. Then, the root tips were treated with EC50 values for 24 h, 48 h and 72 h. The root tips were prepared according to the method of preparing the mitotic preparation and examined under a microscope. It was determined that the water sample taken from Hosap Stream had a cytotoxic and genotoxic effect on A. cepa root tip cells. It was evaluated that the chromosomal aberration index increased statistically significantly depending on the dose in all application periods except for the 24 h application. It was determined that the mitotix index and chromosomal aberration index were higher than the control groups. The lowest mitotix index (2,217±0,625) and the highest chromosomal aberration index (53,153±1,642) was observed with EC50X2 dose at 72h. The prevalent chromosome aberration observed in A. cepa root tip cells were -mitosis, laggard chromosome, polar shifting in anaphase and telophase, equatorial plate shifting, chromosome fragment. According to the results of this study, it can be said that Hosap tea has cytotoxic and genotoxic effects on other living organism.
Evaluation of cytotoxic effect of ferrous gluconate on Allium cepa root tip
(2021) Kaya, Nergis
It was used Allium cepa L and ferrous gluconate (E579). A. cepa roots were treated with ferrous gluconate of different concentrations. A. cepa root lengths were measured. So, the EC50 value was determined as 0.068 g/l. Then, A. cepa L. roots were treated with EC50 / 2 (0.034 g/l), EC50 (0.068 g/l), 2XEC50 (0.136 g/l) for 24, 48, 72 hours. The root tips were cut and prepared for observation in the light microscope. For each group, at least 5000 cell counts were made. Mitotic index was used in the determination of cytotoxicity. TUKEY multiple comparison test and Repeated measurement ANOVA was used. It was determined that the difference between ferrous gluconate concentrations varied as far as the treatment duration. In the way of mitotic index, it was stated that mean of control doses at all treatment duration were significantly higher than the EC50/2, EC50, EC50X2 doses and decreased with the increasing dose. After 24, 48 and 72 hours treatment of EC50X2 dose, it was found that mitotic index decreased due to increasing in application duration. It was detected that ferrous gluconate reduced mitotic division at the root of A. cepa in comparison to the control group. In this way, it was established that ferrous gluconate can be cytotoxic. In addition to this, it was determined that the cytotoxic effect of ferrous gluconate increased and mitotic index decreased due to increasing of treatment duration and dose. Results of this research are parallel with the literatüre that food additives might cause cytotoxicity.
Evaluation of Genotoxic Effect of Phloxine by Allium Test
(2022) Kaya, Nergis
Phloxine is used as a food dye. In this study, genotoxicity of phloxine at the root tip of Allium cepa\rL. was investigated. A. cepa L. meristematic root tip cells were treated with ten different doses of\rphloxine. In this way, the EC50 value was determined. Then, phloxine was applied to root tips at\rEC50/2, EC50 and EC50×2 doses. Treatment time was determined as 24, 48 and 72 hours. As a result,\rit was revealed that phloxine caused chromosomal aberrations in cells in mitotic cycle at the root tip\rof A. cepa. There are equatorial plate shifting in metaphase, laggard chromosome, disturbed spindle,\rchromosome stickiness, C-mitosis, polar shifting among the observed chromosomal aberrations. It\rwas stated that the % chromosomal aberration index (CAI) increased depending on concentration\rincrease. It has been demonstrated that the highest % chromosomal aberration index occurred at the\rEC50×2 dose for 72 hours. According to the research, it was revealed that phloxine has a genotoxic\reffect on the root cells of A. cepa. For this reason, it can be emphasized that care should be taken in\rits use in foods
Evaluation of genotoxicity and cytotoxicity of dodine (1-dodecylguanidium acetate) by Allium test
(Parlar Scientific Publications, 2015) Çördük, Nursen; Akinci, Nihan; Yücel, Gülru; Kaya, Nergis; Aki, Cüneyt
In this study, we evaluated the genotoxic and cytotoxic effects of dodine, a fungicide extensively used to control scab on apples, pears and pecans, brown rot on peaches and several foliar diseases of cherries, strawberries, peaches and black walnuts. For this purpose the Allium cepa test was carried out exposing roots to dodine for 24,48 and 72 h at the concentrations of EC50/2, EC50 and 2×EC50. The mitotic index was calculated as the number of dividing cells per number of 3000-4000 observed cells and the mitotic aberrations also were scored at each concentration. The results showed that dodine induced significant increases of mitotic aberrations such as C-mitosis, polar shifting, laggard chromosome and chromosome fragments. In addition, mitotic index decreased significantly with increasing of concentration and the exposure time as compared to their controls. Hence dodine should be used under control in agricultural fields due to its possible toxic effects. © by PSP.
EVALUATION OF GENOTOXICITY AND CYTOTOXICITY OF DODINE (1-dodecylguanidium acetate) BY Allium TEST
(Parlar Scientific Publications (P S P), 2015) Corduk, Nursen; Akinci, Nihan; Yucel, Gulru; Kaya, Nergis; Akı, Cüneyt
In this study, we evaluated the genotoxic and cytotoxic effects of dodine, a fungicide extensively used to control scab on apples, pears and pecans, brown rot on peaches and several foliar diseases of cherries, strawberries, peaches and black walnuts. For this purpose the Allium cepa test was carried out exposing roots to dodine for 24, 48 and 72 h at the concentrations of EC50/2, EC50 and 2xEC(50). The mitotic index was calculated as the number of dividing cells per number of 3000-4000 observed cells and the mitotic aberrations also were scored at each concentration. The results showed that dodine induced significant increases of mitotic aberrations such as C-mitosis, polar shifting, laggard chromosome and chromosome fragments. In addition, mitotic index decreased significantly with increasing of concentration and the exposure time as compared to their controls. Hence dodine should be used under control in agricultural fields due to its possible toxic effects.
Evaluation of the Effects of Some Raphanus sativus Tuber Extracts on Certain Antioxidant Enzyme Activity and Lipid Peroxidation Level Against Gray Mold (Botrytis Cinerea) in Vicia Faba Leaves
(2023) Kaya, Nergis; Kaya, Tayfun; Yiğit, Soner
The antioxidant defense system of the medicinal plant Raphanus sativus L. tuber root in terms of certain antioxidant enzymes (ascorbat peroxidase, glutathion reductase, catalase, superoxide dismutase) and lipid peroxidation against gray mold disease caused by Botrytis cinerea fungus on Vicia faba L. (bean) leaves was analyzed spectrophotometrically. V. faba plants were grown from seed under controlled conditions. A spore suspension of 105 spores/ml of B. cinerea was prepared. The leaves of V. faba were treated with 1-) only distilled water, 2-)only DMSO, 3-) only B. cinerea (positive control), 4-) only extract applications (distilled water, ethanol and methanol extract),5-) B. cinerea treatment after extract applications (extract:fungus). The leaves of V. faba were harvested 24 and 48 hours after the applications. Analyzes of the supernatants obtained from the extracts were performed spectrophotometrically. It has been revealed that the activity of all investigated enzymes generally increased in the extract:fungus application groups compared to the fungus application group. It was determined that the application of fungus alone caused an increase in the activity of enzymes compared to application groups other than the extract:fungus application groups. It was observed that there was an increase in lipid peroxidation (MDA) only in the fungus application group compared to all extract:fungus application groups except 10mg/ml distilled water extract:fungus treatment. As a result of the research, it was determined that the extract:fungus applications generally increased the antioxidant enzyme activity levels and decreased the MDA content when compared to the negative control and positive control groups. In extract:fungus applications, enzyme activities were obtained from 10mg/ml distilled water extract:fungus, 10mg/ml EtOH extract:fungus, 10mg/ml MeOH extract:fungus applications. From this point of view, it can be stated that R. sativus distilled water, ethanol and methanol extracts generally give an increased antioxidant defense response in V. faba leaves compared to B. cinerea inoculation alone.
Evaluation of the Effects of Specific Extracts of Raphanus Sativus Tuber on Total Protein Content and Peroxidase Activity Against Botrytis cinerea in Vicia Faba
(2022) Kaya, Nergis; Kaya, Tayfun; Yiğit, Soner
The antioxidant defense system of the medicinal plant Raphanus sativus L. tuber in terms of total protein content and peroxidase (POX) activity against gray mold disease caused by Botrytis cinerea fungus on Vicia faba L. (bean) leaves was analyzed spectrophotometrically. Stock solutions of distilled water, ethanol and methanol extracts of R. sativus tuber were prepared with dimethyl sulfoxide (DMSO). A spore suspension of 105 spores/mL of B. cinerea was prepared. The leaves of V. faba were treated with distilled water (negative control), only DMSO, only B. cinerea (positive control), only extract treatment (distilled water, ethanol and methanol extract) and B. cinerea applications after extract treatments. The leaves were extracted in accordance with the methods to be analyzed. Analyzes of the supernatants obtained from the extracts were performed spectrophotometrically. As a result of the research, it was determined that the total protein content increased in the extract:fungus treatment groups compared to the group treated with distilled water (negative control). However, it was determined that it decreased in generally compared to the group that was only treated with fungus (positive control). POX activity was found to be increased compared to the negative control group. Total protein content and POX activity increased in only the extract groups compared to the negative control group. The highest POX activity was obtained from the treatment of 10 mgmL distilled water:fungus. In this respect, it can be stated that the best antioxidant activity among the extract:fungus treated groups is 10 mg/mL distilled water:fungus treatment. From this point of view, it can be stated that R. sativus distilled water, ethanol and methanol extracts give an antioxidan defense response in V. faba leaves.
Hplc Analysis of Certain Phenolic Compounds and Carotenoid in Two Calendula Species
(2020) Kaya, Nergis; Akı, Cüneyt
Calendula officinalis (pot marigold), a medicinal ornamental plant belonging to the Asteraceaefamily, has various medicinal activities such as anticancer, antimicrobial, antioxidant and antitumor.The retention time of quercetin (6.445 min), kaempferol (11.246 min), caffeic acid (2.333 min), betacarotene (9.614 min) standards, which were planned to be analyzed by HPLC, was determined. TheHPLC conditions that are most suitable for these metabolites have been determined. The amounts ofquercetin, kaempferol, caffeic acid and beta carotene in the seedling leaves of these two Calendulaspecies on the 0., 15., 30., 45., 60. days were compared. Caffeic acid and beta carotene have beendetected in the leaves. However, quercetin and kaempferol compounds were not detected. It wasdetermined that the caffeic acid and beta carotene amounts of C. officinalis species were higher thanthat of C. arvensis. The highest caffeic acid in both C. officinalis and C. arvensis seedlings(0.4042±0.0123 µg/g dry weight for C. officinalis, 0.1918±0.0437 µg / g dry weight for C. arvensis)and beta carotene amounts (0.8520±0.0692 µg/g dry weight for C. officinalis, 0.6389±0.0189 µg/gdry weight for C. arvensis) were determined on 60. day. In addition, it was determined that the amountof these metabolites differed according to the seedling development period.

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