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    Öğe
    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.
  • [ X ]
    Öğe
    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.
  • Küçük Resim
    Öğe
    Assessment of Thinopyrum ponticum (Podp.) Barkworth & D. R. Dewey accessions using universal rice primers and molecular cytogenetics
    (Springer, 2021) Tiryaki, İskender; Baytekin Karaoğlu, Gülhan; Yücel, Gülru; Tuna, Metin
    The aim of this study was to make morphological and molecular characterization of tall wheatgrass (Thinopyrum ponticum) accessions naturally found in Canakkale flora, Turkey. The seeds collected from 24 different locations in Canakkale vicinity were planted in the nursery field to determine thirteen morphological parameters. Twelve universal rice primers (URPs) were used to reveal genetic relationship among the accessions while ploidy analysis was done based on the nuclear DNA content of plants determined by Flow cytometer. Fluorescence in situ hybridization (FISH) method was used to determine 5S and 45S rDNA distributions on mitotic chromosomes. The agro-morphological data showed significant variation among the accessions for all parameters measured, except the number of nodes per plant. Twelve URP primers produced 73 alleles in total and 63 of those were polymorphic. The highest polymorphism information content value was obtained from URP 17R with 0.82. The first three components of Eigen values in PCA analysis explained 41.1% of total variation. The 2C nuclear DNA contents of the accessions ranged from 41.17 to 45.49 pg. All the accessions were determined as decaploid with 2n = 70 chromosomes. FISH analysis provided 18-20 interstial 5S rDNA and 12-14 terminal 45S rDNA loci. The results concluded that tall wheatgrass accessions used in this study contain a significant variation in morphological traits and PCR-based DNA polymorphism which could be used as a new genetic resource in breeding programs of tall wheatgrass and wheat improvement for intra- and intergeneric crosses.
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    Öğe
    Genetic relationship and nuclear dna content variation in Tef [Eragrostis tef (Zucc.) Trotter] accessions
    (Springer, 2020) Kaya, Caglar; Tiryaki, Iskender; Sari, Ugur; Tuna, Metin
    This study was initiated to reveal genetic relationship of 25 tef (Eragrostis tef (Zucc.) accessions by using 10 SSR markers and to determine DNA content variation by using flow cytometer analysis. Ten markers produced a total of 18 alleles and 11 of those were polymorphic. The mean polymorphism rate was 66.6%. The highest polymorphism information content value was obtained from marker CNLTs370 with 0.69 while markers CNTLs11 and CNTLs133 produced monomorphic bands only. UPGMA analysis divided 25 tef genotypes into three main clades. The accessions PI193511 and PI195934 were distinctly separated from the others. No ploidy differences were determined among the 25 tef accessions. 2C mean nuclear DNA content ranged from 1.406 pg to 1.510 with mean of 1.460 pg. The results of this study indicated that SSR markers successfully determined genetic relationship of 25 tef accession although they had a low rate of polymorphism. This study also revealed that available tef related SSR markers should be optimized before use and their efficiency may vary based on tef genotypes or accessions used.
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    Öğe
    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.
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    Öğe
    In vitro Üretilen Digitalis Trojana Ivanina Bitkilerinin Genetik Kararlılığının Flow Sitometri ve Sitolojik Analizler ile Değerlendirilmesi
    (2017) Çördük, Nurşen; Yücel, Gülru; Akıncı, Nihan; Tuna, Metin
    Bu çalışmada, Digitalis trojana Ivanina türüne ait dolaylı sürgün organogenezis ile üretilen bitkilerin genetik kararlılığı flow sitometri ve sitolojik analizler ile değerlendirilmiştir. İlk aşamada yaprak eksplantları 3 mg/L BA + 0,1 mg/L NAA içeren MS besin ortamında kültüre alınmıştır. Bu ortamda dolaylı organogenez yoluyla oluşan sürgünler 2 haftada bir altkültüre alınarak çoğaltılmıştır. Bitkiler % 0.1 aktif karbon içeren MS besin ortamında köklendirilmiştir. Üretilen bitkilerin genetik kararlılığı flow sitometri ve sitolojik analizler ile tohumdan yetiştirilen D. trojana bitki örnekleri ile karşılaştırılarak değerlendirilmiştir. Flow sitometri analizinde, in vitro üretilen bitki örneklerinin DNA içeriği 2.80±0.03 piko gram, tohumdan yetiştirilen bitki örneklerinin DNA içeriği ise 2.80±0.1 piko gram belirlenmiştir. Bitkinin mikroskop ile yapılan mitoz kromozomu sayımlarında kromozom sayısının 56 olduğu ve in vitro üretilen bitkilerin kromozom sayısında bir değişiklik olmadığı belirlenmiştir. Yapılan analizler sonucunda 3 mg/L BA + 0,1 mg/L NAA içeren MS besin ortamında üretilen bitkilerde genetik farklılık görülmemiştir. Yapılan bu çalışma ile D. trojana türünün ilk kez DNA içeriği ve kromozom sayısı belirlenmiştir. Elde edilen sonuçlar, türün in vitro kültüre alınması sırasında kültür koşullarından dolayı meydana gelebilecek olan genetik varyasyonların analizinde flow sitometrinin kullanılabileceğini göstermektedir.