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    Assessment of The Genetic Stability of Indirect Shoot Organogenesis-Derived Plantlets of Digitalis Trojana Ivanina by Flow Cytometry and Cytological Analyses
    (Univ Namik Kemal, 2017) Corduk, Nursen; Yucel, Gulru; Akinci, Nihan; Tuna, Metin
    In this study, flow cytometry and cytological analysis was used to evaluate the genetic stability of Digitalis trojana Ivanina plants regenerated via indirect shoot organogenesis. For in vitro propagation, leaf explants were excised from seedlings grown in sterile conditions and cultured MS medium supplemented with 3.0 mg/L BA + 0.1 mg/L NAA. Shoots and calli were subcultured for a period of 2 weeks for shoot multiplication. For rooting, shoots were separated individually and transferred to MS medium containing 0.1% activated charcoal. Genetic stability of the regenerated plants was assessed by flow cytometry and cytological analyses. Flow cytometric analysis revealed that regenerated plantlets has as 2.80 +/- 0.03 pg nuclear DNA (2C) and seed-derived plants has on average 2.80 +/- 0.1 pg/2C. Cytological analysis showed that regenerated plantlets have the same number of chromosome with seed-derived plantlets of D. trojana (2n=56). Our results have showed that the plantlets propagated in MS medium with 3 mg/L BA + 0.1 mg/L NAA did not differ genetically from donor plants. Therefore, this system can be effective and suitable for clonal propagation of D. trojana. Our results also confirmed that flow cytometry is fast, easy, accurate and relatively cheap method to determine ploidy of in vitro propagated D. trojana plantlets.
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    Assessment of the genetic stability of indirect shoot organogenesis-derived plantlets of digitalis trojana ivanina by flow cytometry and cytological analyses
    (Namik Kemal University - Agricultural Faculty, 2017) Çördük, Nurşen; Yücel, Gülru; Akinci, Nihan; Tuna, Metin
    In this study, flow cytometry and cytological analysis was used to evaluate the genetic stability of Digitalis trojana Ivanina plants regenerated via indirect shoot organogenesis. For in vitro propagation, leaf explants were excised from seedlings grown in sterile conditions and cultured MS medium supplemented with 3.0 mg/L BA + 0.1 mg/L NAA. Shoots and calli were subcultured for a period of 2 weeks for shoot multiplication. For rooting, shoots were separated individually and transferred to MS medium containing 0.1% activated charcoal. Genetic stability of the regenerated plants was assessed by flow cytometry and cytological analyses. Flow cytometric analysis revealed that regenerated plantlets has as 2.80±0.03 pg nuclear DNA (2C) and seed-derived plants has on average 2.80±0.1 pg/2C. Cytological analysis showed that regenerated plantlets have the same number of chromosome with seed-derived plantlets of D. trojana (2n=56). Our results have showed that the plantlets propagated in MS medium with 3 mg/L BA + 0.1 mg/L NAA did not differ genetically from donor plants. Therefore, this system can be effective and suitable for clonal propagation of D. trojana. Our results also confirmed that flow cytometry is fast, easy, accurate and relatively cheap method to determine ploidy of in vitro propagated D. trojana plantlets. © 2017 Namik Kemal University - Agricultural Faculty. All Rights Reserved.
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    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.
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    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.
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    Genotoxic effects of chlorophenoxy herbicide diclofop-methyl in mice in vivo and in human lymphocytes in vitro
    (Taylor & Francis Ltd, 2011) Unal, Fatma; Yuzbasioglu, Deniz; Yilmaz, Serkan; Akinci, Nihan; Aksoy, Huseyin
    Diclofop-methyl (DM) is a chlorophenoxy derivative used in large quantities for the control of annual grasses in grain and vegetable crops. In this study, the genotoxic effects of DM were investigated by measuring chromosomal aberrations (CAs) in mouse bone-marrow cells and CA and the comet assay in human peripheral lymphocytes. Mice were treated with 15.63, 31.25, 62.5, and 125 mg/kg body weight of DM intraperitoneally for 24 hours, and 15.63-, 31.25-, 62.5-, 125-, and 250-mu g/mL concentrations were applied to human lymphocytes for both 24 and 48 hours. In in vivo treatments, DM significantly, but not dose dependently, increased the total chromosome aberrations, compared to both negative and solvent controls. Cell proliferation was significantly, but not dose dependently, affected by all doses. In in vitro treatments, DM (except 15.63 mu g/mL) significantly and dose dependently increased the frequency of chromosome aberrations. Also, 250 mu g/mL of 48-hour treatment was found to be toxic. Cell proliferation was significantly and dose dependently affected by DM applications, when compared to negative control. In in vitro treatments, DM significantly decreased the mitotic index only at the highest concentration for 24 hours, and 62.5- and 125-mu g/mL concentrations for 48 hours. In the comet assay, a significant and dose-dependent increase in comet-tail intensity was observed at 62.5-, 125-, and 250-mu g/mL concentrations. The mean comet-tail length was significantly increased in all concentrations. Our results demonstrate that DM is genotoxic in mammalian cells in vivo and in vitro.
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    In vitro propagation of Silene bolanthoides Quezel, Contandr. & Pamukc. and assessment of genetic stability by flow cytometry
    (Inst Bioloska Istrazivanja Sinisa Stankovic, 2018) Corduk, Nursen; Yucel, Gulru; Akinci, Nihan; Tuna, Metin; Esen, Onur
    Silene bolanthoides Quezel, Contandr. & Pamukc. is an endemic species from Kazdagi (Mt. Ida), Canakkale-Balikesir, Turkey. In order to develop an efficient shoot regeneration protocol, the leaf, nodal and internodal explants of S. bolanthoides were cultured on Murashige and Skoog (MS) medium containing benzyladenine (BA) alone or in combination with alpha-naphthaleneacetic acid (NAA). The highest number of regenerated shoots (5.75 +/- 0.1) was obtained from nodal explants that were cultured on MS medium with 8.8 mu M BA+0.54 mu M NAA. Regenerated shoots were rooted on MS medium without plant growth regulators (PGRs). Rooted plants (2-3 cm) were separately transferred to pots containing a mixture of peat and perlite (3:1 v/v) and acclimatized successfully in a growth chamber. Genetic stability of the propagated plants was assessed by flow cytometry and cytological analysis. Flow cytometry analysis demonstrated that regenerated plants had 2.61 +/- 0.01 pg nuclear DNA (2C) and seed-derived plants had on average 2.58 +/- 0.02 pg/2C. Cytological analysis showed that the regenerated plants had the same chromosome number as seed-derived plants of S. bolanthoides (2n=24). It was determined that regenerated plants were uniform in chromosome number and had a similar DNA content to the seed-derived ones, indicating that the described efficient shoot regeneration protocol can be applied for ex situ conservation of this species.