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Determination of Nuclear DNA Content and Chromosome Number of Verbascum scamandri Murb. (Scrophulariaceae)

Yıl 2023, Cilt: 38 Sayı: 2, 397 - 406, 04.07.2023
https://doi.org/10.7161/omuanajas.1286961

Öz

The study aimed to determine the chromosome number and nuclear DNA content of an endemic plant, Verbascum scamandri, using 14-week-old plants germinated from seed and to determine the genetic stability based on flow cytometry analyses in callus tissue induced MS medium containing 1 mg/L Kin + 1 mg/L 2,4-D and 9-week-old propagated plants, induced and developed on MS medium containing 2 mg/L Kin + 0.1 mg/L 2,4-D. In the mitotic chromosome counts showed that V. scamandri had 2n = 32 chromosome number. Flow cytometric analysis revealed that 14-week-old in vitro grown plants have 0.73 pg/2C ± 0.01, callus tissue has 0.76 pg/2C ± 0.02 and propagated plantlets have 0.79 pg/2C ± 0.01. The results proved that propagated plants had similar DNA content to the seed-derived plants which showed analysed plants were genetically stable.

Destekleyen Kurum

Çanakkale Onsekiz Mart University The Scientific Research Coordination Unit, Çanakkale, Türkiye

Proje Numarası

FYL-2021- 3732

Kaynakça

  • Akdemir, Z., Kahraman, Ç., Tatlı, I. I., Akkol, E. K., Süntar, I., Keles, H., 2011. Bioassay-guided isolation of anti-inflammatory, antinociceptive and wound healer glycosides from the flowers of Verbascum mucronatum Lam. Journal of Ethnopharmacology, 136(3): 436-443. https://doi.org/10.1016/j.jep.2010.05.059
  • Araújo, L.G., Prabhu, A.S., Filippi, M.C., Chaves, L.J., 2001. RAPD analysis of blast resistant somaclones from upland rice cultivar IAC 47 for genetic divergence. Plant Cell Tissue Organ Culture, 67: 165–172. https://doi.org/10.1023/A:1011960225472.
  • Babaoğlu, M., Yorgancılar, M. Akbulak, A.M., 2001. Doku Kültürü: Temel Laboratuar Teknikleri. In: Babaoğlu, M., Gürel, E., Özcan, S., (Eds). Bitki Biyoteknolojisi I Doku Kültürü ve Uygulamaları. Selçuk Üniversitesi Vakfı Yayınları, Konya. pp 1-35.
  • Benedi, C., Rico, E., Guemes, J., Herrero, A., 2009. Flora Iberica Vol. 13. Madrid: Real Jardin Botanico. Bennett, M.D., Leitch, I.J., 1995. Nuclear DNA Amounts in Angiosperms. Annals of Botany, 76(2): 113–176. https://doi.org/10.1006/anbo.1995.1085
  • Bhatia, S., Sharma, K., 2015. Technical Glitches in Micropropagation. In Bhatia S, Sharma K, Dahiya R, Bera T., (Eds) Modern Applications of Plant Biotechnology in Pharmaceutical Sciences. Academic Press, pp 393-404.
  • Çalışkan, T., Hatipoğlu, R., Kırıcı, S., 2019. Production of plant secondary metabolites from cell and organ cultures under in vitro conditions. Turkish Journal of Agriculture-Food Science and Technology, 7(7): 971-980. https://doi.org/10.24925/turjaf.v7i7.971-980.2447
  • Cambaz, E., 2022. Verbascum scamandri Murb. Türünün Kallus Kültürü ile Sekonder Metabolit Üretimi. Master Thesis. Çanakkale Onsekiz Mart University, School of Graduate Studies, p 94, Çanakkale.
  • Castro, M., Castro, S., Loureiro, J., 2012. Genome size variation and incidence of polyploidy in Scrophulariaceae sensu lato from the Iberian Peninsula, AoB PLANTS, pls037. https://doi.org/10.1093/aobpla/pls037
  • Chinnusamy, V., Zhu, J.K., 2009. Epigenetic regulation of stress responses in plants. Curr Opin Plant Biol, 12: 133–39.
  • Çıngay, B., Demir, O., Cabi, E., 2018. Verbascum faik-karaveliogullarii (Scrophulariaceae), a new species from southeastern Anatolia, Turkey. Phytotaxa, 372(4): 263-272. https://doi.org/10.11646/phytotaxa.372.4.3.
  • Çıngay, B., Karavelioğulları, F.A., 2016. A new species of Verbascum, V. nihatgoekyigitii (Scrophulariaceae), from southeastern Anatolia, Turkey. Phytotaxa, 269(4): 287-293. https://doi.org/10.11646/phytotaxa.269.4.4
  • Çördük, N., Yücel, G., Akıncı, N., Tuna, M., 2017. Assessment of the genetic stability of indirect shoot organogenesis-derived plantlets of Digitalis trojana Ivanina by flow cytometry and cytological analyses. Journal of Tekirdag Agricultural Faculty, 14 (1): 70-76.
  • Çördük, N., Yücel, G., Akıncı, N., Tuna, M., Esen, O., 2018. In vitro propagation of Silene bolanthoides Quézel, Contandr. & Pamukç. and assessment of genetic stability by flow cytometry. Archives of Biological Sciences, 70(1): 141-148. https://doi.org/10.2298/ABS170410033C.
  • Dolezel, J., Binarova, P., Lucretti, S., 1989. Analysis of nuclear DNA content in plant cells by flow cytometry. Biology Plantarum, 31(2): 113–120. https://doi.org/10.1007/BF02907241.
  • Dolezel, J., Greilhuber, J., Suda, J., 2007. Estimation of nuclear DNA content in plants using flow cytometry. Nat Protoc, 2: 2233–2244. https://doi.org/10.1038/nprot.2007.310.
  • Duman, H., Uzunhisarcıklı, M.E., Tan, K., 2017. Verbascum mughlaeum (Scrophulariaceae), a new species from SW Anatolia, Turkey. Phytotaxa, 291(3): 231-236. https://doi.org/10.11646/phytotaxa.291.3.8
  • Ekim, T., Koyuncu, M., Vural, M., Duman, H., Aytac, Z., Adıguzel, N., 2000. Red Data Book of Turkish Plants (Pteridophyta and Spermatophyta). Ankara: Foundation for Turkish Nature Conservation and Van Centinential University Press. pp. 141.
  • Escobedo-Gracia-Medrano, R.M., Burgos-Tan, M.J., Ku-Cauich, J.R., Quiroz-Moreno, A., 2018. Using Flow Cytometry Analysis in Plant Tissue Culture Derived Plants. Methods in Molecular Biology, 1815: 317-332. https://doi.org/10.1007/978-1-4939-8594-4_22.
  • Esposito, E., Dal Toso, R., Pressi, G. Bramanti, P., Meli, R., Cuzzocrea, S., 2010. Protective effect of verbascoside in activated C6 glioma cells: possible molecular mechanisms. Naunyn-Schmiedeberg's Archives of Pharmacology, 381: 93-105. https://doi.org/10.1007/s00210-009-0466-0
  • Galbraith, D.W., 2009. Simultaneous flow cytometric quantification of plant nuclear DNA contents over the full range of described angiosperm 2C values. Cytometry A., 75(8): 692-8. https://doi.org/10.1002/cyto.a.20760.
  • Hacıoğlu Doğru, N., Demir, N., Yılmaz, Ö., 2021. Three species of Verbascum L. from Northwest Anatolia of Turkey as a source of biological activities. Turk J Anal Chem, 3(1): 19-26. https://doi.org/10.51435/turkjac.886692
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  • Karavelioğulları, F.A., 2015a. Verbascum ibrahim-belenlii (Scrophulariaceae), a new species from East Anatolia, Turkey. Phytotaxa, 212(3): 246-248. http://doi.org/10.11646/phytotaxa.212.3.8
  • Karavelioğulları, F.A., 2015b. Verbascum misirdalianum (Scrophulariaceae), a new species from central Anatolia, Turkey. Phytotaxa, 217(1): 96-99. https://doi.org/10.11646/phytotaxa.217.1.10
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  • Kubalaková, M., Dolezel, J., Lebeda, A., 1996. Ploidy instability of embryogenic cucumber (Cucumis sativus L.) callus culture. Biologia Plantarum, 38(3): 475–480. https://doi.org/10.1007/BF02896685.
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  • Lee, Y.I., Tseng, Y-F., Lee, Y.C., Chung, M.C., 2020. Chromosome constitution and nuclear DNA content of Phalaenopsis hybrids. Scientia Horticulturae, 262. 109089.https://doi.org/10.1016/j.scienta.2019.109089.
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Verbascum scamandri Murb. (Scrophulariaceae) Türünün Çekirdek DNA İçeriğinin ve Kromozom Sayısının Belirlemesi

Yıl 2023, Cilt: 38 Sayı: 2, 397 - 406, 04.07.2023
https://doi.org/10.7161/omuanajas.1286961

Öz

Çalışmanın amacı, endemik bir bitki olan Verbascum scamandri türünün in vitro büyütülen 14 haftalık bitkiler kullanılarak kromozom sayısı ve çekirdek DNA içeriğinin belirlenmesi, 2 mg/L Kin + 0.1 mg/L 2,4-D içeren MS ortamında çoğaltılan 9 haftalık in vitro bitkilerde ve 1 mg/L Kin + 1 mg/L 2,4-D içeren MS ortamında indüklenen kallus dokularında flow sitometri analizleri ile genetik kararlılığın tespit edilmesidir. Mitoz kromozom sayımlarında türün kromozom sayısının 2n = 32 olduğu belirlenmiştir. Flow sitometri analizinde, in vitro yetiştirilen bitki örneklerinin 0.73 pg/2C ± 0.01, kallus örneklerinin 0.76 pg/2C ± 0.02, in vitro çoğaltılmış bitkiciklerin DNA içeriği ise 0.79 pg/2C ± 0.01 belirlenmiştir. Yapılan flow sitometri analizleri tohumdan gelişen bitkiler ile in vitro çoğaltılan bitkilerin benzer çekirdek DNA içeriklerine sahip olduklarını göstermiş ve dolayısıyla genetik stabil oldukları anlaşılmıştır.

Proje Numarası

FYL-2021- 3732

Kaynakça

  • Akdemir, Z., Kahraman, Ç., Tatlı, I. I., Akkol, E. K., Süntar, I., Keles, H., 2011. Bioassay-guided isolation of anti-inflammatory, antinociceptive and wound healer glycosides from the flowers of Verbascum mucronatum Lam. Journal of Ethnopharmacology, 136(3): 436-443. https://doi.org/10.1016/j.jep.2010.05.059
  • Araújo, L.G., Prabhu, A.S., Filippi, M.C., Chaves, L.J., 2001. RAPD analysis of blast resistant somaclones from upland rice cultivar IAC 47 for genetic divergence. Plant Cell Tissue Organ Culture, 67: 165–172. https://doi.org/10.1023/A:1011960225472.
  • Babaoğlu, M., Yorgancılar, M. Akbulak, A.M., 2001. Doku Kültürü: Temel Laboratuar Teknikleri. In: Babaoğlu, M., Gürel, E., Özcan, S., (Eds). Bitki Biyoteknolojisi I Doku Kültürü ve Uygulamaları. Selçuk Üniversitesi Vakfı Yayınları, Konya. pp 1-35.
  • Benedi, C., Rico, E., Guemes, J., Herrero, A., 2009. Flora Iberica Vol. 13. Madrid: Real Jardin Botanico. Bennett, M.D., Leitch, I.J., 1995. Nuclear DNA Amounts in Angiosperms. Annals of Botany, 76(2): 113–176. https://doi.org/10.1006/anbo.1995.1085
  • Bhatia, S., Sharma, K., 2015. Technical Glitches in Micropropagation. In Bhatia S, Sharma K, Dahiya R, Bera T., (Eds) Modern Applications of Plant Biotechnology in Pharmaceutical Sciences. Academic Press, pp 393-404.
  • Çalışkan, T., Hatipoğlu, R., Kırıcı, S., 2019. Production of plant secondary metabolites from cell and organ cultures under in vitro conditions. Turkish Journal of Agriculture-Food Science and Technology, 7(7): 971-980. https://doi.org/10.24925/turjaf.v7i7.971-980.2447
  • Cambaz, E., 2022. Verbascum scamandri Murb. Türünün Kallus Kültürü ile Sekonder Metabolit Üretimi. Master Thesis. Çanakkale Onsekiz Mart University, School of Graduate Studies, p 94, Çanakkale.
  • Castro, M., Castro, S., Loureiro, J., 2012. Genome size variation and incidence of polyploidy in Scrophulariaceae sensu lato from the Iberian Peninsula, AoB PLANTS, pls037. https://doi.org/10.1093/aobpla/pls037
  • Chinnusamy, V., Zhu, J.K., 2009. Epigenetic regulation of stress responses in plants. Curr Opin Plant Biol, 12: 133–39.
  • Çıngay, B., Demir, O., Cabi, E., 2018. Verbascum faik-karaveliogullarii (Scrophulariaceae), a new species from southeastern Anatolia, Turkey. Phytotaxa, 372(4): 263-272. https://doi.org/10.11646/phytotaxa.372.4.3.
  • Çıngay, B., Karavelioğulları, F.A., 2016. A new species of Verbascum, V. nihatgoekyigitii (Scrophulariaceae), from southeastern Anatolia, Turkey. Phytotaxa, 269(4): 287-293. https://doi.org/10.11646/phytotaxa.269.4.4
  • Çördük, N., Yücel, G., Akıncı, N., Tuna, M., 2017. Assessment of the genetic stability of indirect shoot organogenesis-derived plantlets of Digitalis trojana Ivanina by flow cytometry and cytological analyses. Journal of Tekirdag Agricultural Faculty, 14 (1): 70-76.
  • Çördük, N., Yücel, G., Akıncı, N., Tuna, M., Esen, O., 2018. In vitro propagation of Silene bolanthoides Quézel, Contandr. & Pamukç. and assessment of genetic stability by flow cytometry. Archives of Biological Sciences, 70(1): 141-148. https://doi.org/10.2298/ABS170410033C.
  • Dolezel, J., Binarova, P., Lucretti, S., 1989. Analysis of nuclear DNA content in plant cells by flow cytometry. Biology Plantarum, 31(2): 113–120. https://doi.org/10.1007/BF02907241.
  • Dolezel, J., Greilhuber, J., Suda, J., 2007. Estimation of nuclear DNA content in plants using flow cytometry. Nat Protoc, 2: 2233–2244. https://doi.org/10.1038/nprot.2007.310.
  • Duman, H., Uzunhisarcıklı, M.E., Tan, K., 2017. Verbascum mughlaeum (Scrophulariaceae), a new species from SW Anatolia, Turkey. Phytotaxa, 291(3): 231-236. https://doi.org/10.11646/phytotaxa.291.3.8
  • Ekim, T., Koyuncu, M., Vural, M., Duman, H., Aytac, Z., Adıguzel, N., 2000. Red Data Book of Turkish Plants (Pteridophyta and Spermatophyta). Ankara: Foundation for Turkish Nature Conservation and Van Centinential University Press. pp. 141.
  • Escobedo-Gracia-Medrano, R.M., Burgos-Tan, M.J., Ku-Cauich, J.R., Quiroz-Moreno, A., 2018. Using Flow Cytometry Analysis in Plant Tissue Culture Derived Plants. Methods in Molecular Biology, 1815: 317-332. https://doi.org/10.1007/978-1-4939-8594-4_22.
  • Esposito, E., Dal Toso, R., Pressi, G. Bramanti, P., Meli, R., Cuzzocrea, S., 2010. Protective effect of verbascoside in activated C6 glioma cells: possible molecular mechanisms. Naunyn-Schmiedeberg's Archives of Pharmacology, 381: 93-105. https://doi.org/10.1007/s00210-009-0466-0
  • Galbraith, D.W., 2009. Simultaneous flow cytometric quantification of plant nuclear DNA contents over the full range of described angiosperm 2C values. Cytometry A., 75(8): 692-8. https://doi.org/10.1002/cyto.a.20760.
  • Hacıoğlu Doğru, N., Demir, N., Yılmaz, Ö., 2021. Three species of Verbascum L. from Northwest Anatolia of Turkey as a source of biological activities. Turk J Anal Chem, 3(1): 19-26. https://doi.org/10.51435/turkjac.886692
  • Heywood, V.H., 1993. Flowering Plants of the World. Oxford University Press.
  • Huber-Morath, A., 1978. Verbascum L In: Davis PH (ed) Flora of Turkey and the East Aegean Islands. Vol 6. Edinburgh University Press Edinburgh. pp 461-603.
  • Hung, J. Y., Yang, C. J., Tsai, Y. M., Huang, H. W., Huang, M. S., 2008. Antiproliferative activity of aucubin is through cell cycle arrest and apoptosis in human non-small cell lung cancer A549 cells. Clinical and Experimental Pharmacology and Physiology, 35(9): 995-1001. https://doi.org/10.1111/j.1440-1681.2008.04935.x
  • IUCN, International Union for Conservation of Nature. 2012. IUCN Red List Categories and Criteria: Version 3.1. Gland, Switzerland.
  • Karavelioğulları, F.A., 2015a. Verbascum ibrahim-belenlii (Scrophulariaceae), a new species from East Anatolia, Turkey. Phytotaxa, 212(3): 246-248. http://doi.org/10.11646/phytotaxa.212.3.8
  • Karavelioğulları, F.A., 2015b. Verbascum misirdalianum (Scrophulariaceae), a new species from central Anatolia, Turkey. Phytotaxa, 217(1): 96-99. https://doi.org/10.11646/phytotaxa.217.1.10
  • Kevers, C., Greimers, R., Franck, T., Bisbis, T., Dommes, J., Gaspar, T., 1999. Flow cytometry estimation of nuclear size and ploidy level of habituated calli of sugar beet. Biologia Plantarum, 42(3): 321–332. https://doi.org/10.1023/A:1002469331895
  • Kozan, E., Çankaya, I.T., Kahraman, C., Akkol, E.K., Akdemir, Z., 2011. The in vivo anthelmintic efficacy of some Verbascum species growing in Turkey. Experimental Parasitology, 129(2): 211-214. https://doi.org/10.1016/j.exppara.2011.06.005
  • Kubalaková, M., Dolezel, J., Lebeda, A., 1996. Ploidy instability of embryogenic cucumber (Cucumis sativus L.) callus culture. Biologia Plantarum, 38(3): 475–480. https://doi.org/10.1007/BF02896685.
  • Kupeli, E., Tatli, I.I., Akdemir, Z.S., Yesilada, E. 2007. Bioas- say-guided isolation of anti-inflammatory and antinociceptive glycoterpenoids from the flowers of Verbascum lasianthum Boiss. ex Bentham, J. Ethnopharmacol., 110, 444-450., 2007. https://doi.org/10.1016/j.jep.2006.10.004
  • Lee, Y.I., Tseng, Y-F., Lee, Y.C., Chung, M.C., 2020. Chromosome constitution and nuclear DNA content of Phalaenopsis hybrids. Scientia Horticulturae, 262. 109089.https://doi.org/10.1016/j.scienta.2019.109089.
  • Leljak-Levanic, D., Bauer, N., Mihaljevic, S., Jelaska, S., 2004. Changes in DNA methylation during somatic embryogenesis in Cucurbita pepo L. Plant Cell Rep, 23: 120–27. https://doi.org/10.1007/s00299-004-0819-6
  • Lira-Medeiros, C.F., Parisod, C., Fernandes, R.A., Mata, CS, Cardoso, M.A., Ferreira, P.C.G., 2010. Epigenetic variation in mangrove plants occurring in contrasting natural environment. PLOS One, 5(4): e10326. https://doi.org/10.1371/journal.pone.0010326
  • Lysak, M.A., Weiss-Schneeweiss, H., 2021. Editorial: Chromosomal Evolution in Plants. Front Plant Sci. 2021 Jul 29;12:726330. doi: 10.3389/fpls.2021.726330.
  • Makowczyńska, J., Andrzejewska-Golec, E. and Sliwinska, E., 2008. Nuclear DNA content in different plant materials of Plantago asiatica L. cultured in vitro. Plant Cell Tiss Organ Cult, 94: 65–71. https://doi.org/10.1007/s11240-008-9387-8.
  • Mihailović, V., Kreft, S., Benković, E.T., Ivanović, N., Stanković, M.S., 2016. Chemical profile, antioxidant activity and stability in stimulated gastrointestinal tract model system of three Verbascum species. Industrial Crops and Products, 89, 141-151. https://doi.org/10.1016/j.indcrop.2016.04.075
  • Mikulík, J., 1999. Propagation of endangered plant species by tissue cultures. Acta Universitatis Palackianae Olomucensis, Biologica, 37, 27-33.
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  • Wu, Q.Q., Xiao, Y., Duan, M.X., Yuan, Y., Jiang, X.H., Yang, Z., Liao, H.H., Deng, W., Tang, Q.Z., 2018. Aucubin protects against pressure overload‐induced cardiac remodelling via the β3‐adrenoceptor–neuronal NOS cascades. British Journal of Pharmacology, 175(9): 1548-1566. https://doi.org/10.1111/bph.14164
  • Xue, H.Y., Lu, Y.N., Fang, X.M., Xu, Y.P., Gao, G.Z., Jin, L.J., 2012. Neuroprotective properties of aucubin in diabetic rats and diabetic encephalopathy rats. Molecular Biology Reports, 39(10): 9311-9318. https://doi.org/10.1007/s11033-012-1730-9
  • Young, I.C., Chuang, S.T., Hsu, C.H., Sun, Y.J., Liu, H.C., Chen, Y.S., Lin, F.H., 2017. Protective effects of aucubin on osteoarthritic chondrocyte model induced by hydrogen peroxide and mechanical stimulus. BMC complementary and Alternative Medicine, 17(91): 1-11. https://doi.org/10.1186/s12906-017-1581-y
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Toplam 56 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Nükleik Asitlerin ve Proteinlerin Tarımsal Moleküler Mühendisliği
Bölüm Anadolu Tarım Bilimleri Dergisi
Yazarlar

Gülru Yücel 0000-0001-9785-929X

Ebru Cambaz 0000-0001-7301-9288

Nurşen Çördük 0000-0001-8499-4847

Proje Numarası FYL-2021- 3732
Erken Görünüm Tarihi 30 Haziran 2023
Yayımlanma Tarihi 4 Temmuz 2023
Kabul Tarihi 23 Mayıs 2023
Yayımlandığı Sayı Yıl 2023 Cilt: 38 Sayı: 2

Kaynak Göster

APA Yücel, G., Cambaz, E., & Çördük, N. (2023). Determination of Nuclear DNA Content and Chromosome Number of Verbascum scamandri Murb. (Scrophulariaceae). Anadolu Tarım Bilimleri Dergisi, 38(2), 397-406. https://doi.org/10.7161/omuanajas.1286961
Online ISSN: 1308-8769