Abstract
A detailed study of the Turkish endemic Gypsophila pilulifera were presented by both morphological and molecular investigations of several sites (each one divided in four biotopes). The characteristics were investigated using statistic analyses, i.e. t-Test and One-way ANOVA tests, as well as Discriminant analysis (á=0.05). Some differences were detected among the individuals in the locations stufied where the species was distributed in terms of morphological characteristics and obvious similarities/differences were determined among the biotopes as a result of the discriminant analysis. Morphological characterization was supported by molecular data. Four individuals from two different locations were analyzed by PCR using twenty-four ISSR primers. Analysis of PCR products on agarose gel yielded three primary reproductive band profiles, and the similarity frequencies calculated with the Jaccard coefficient were found to be among 0.229 and 0.769. In this context, according to the results, there was no difference in species, subspecies or variety level between individuals between the two locations. But the obtained molecular differences support the morphological differences. It is predicted that this may change towards speciation as ecological changes increase in the locations where the species is distributed.
References
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<p>Boissier, E. (1867) <em>Flora Orientalis: sive Enumeratio Plantarum in Oriente a Graecia et Aegypto ad Indiae fines hucusque observatarum</em>, vol. 1. Primum, A.Ascher & Co., Genevӕ, 548 pp. https://doi.org/10.5962/bhl.title.20323</p>
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<p>Davis, P.H., Mill, R.R. & Tan, K. (Eds.) (1988) <em>Gypsophila </em>L. <em>Flora of Turkey and the East Aegean Islands</em>, vol. 10 (Supplement). Edinburgh University Press, Edinburgh, pp. 73–75.</p>
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<p>Doyle, J.J. & Doyle, J.L. (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. <em>Phytochemical Bulletin</em> 19: 11–15.</p>
<p>Düşen, O., Göktürk, R.S., Kaya, E., Sarpkaya, U. & Gürcan, B. (2018) Morphological and molecular determination of a new <em>Viola</em> species (Violaceae) from Turkey. <em>Phytotaxa</em> 369 (1): 37–46. https://doi.org/10.11646/phytotaxa.369.1.4</p>
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<p>Hernández-Ledesma, P., Berendsohn, W.G., Borsch, T., Mering, S.V., Akhani, H., Arias, S., CastañedaNoa, I., Eggli, U., Eriksson, R., Flores-Olvera, H., Fuentes-Bazán, S., Kadereit, G., Klak, C., Korotkova, N., Nyffeler, R., Ocampo, G., Ochoterena, H., Oxelman, B., Rabeler, R.K., Sanchez, A., Schlumpberger, B.O. & Uotila, P. (2015) A taxonomic backbone for the global synthesis of species diversity in the angiosperm order Caryophyllales. <em>Willdenowia</em> 45: 281–383. https://doi.org/10.3372/wi.45.45301</p>
<p>Huber-Morath, A. (1967) <em>Gypsophila</em> L. <em>In</em>: Davis, P.H. (Ed.) <em>Flora of Turkey and the East Aegean Islands, vol. 2.</em> Edinburgh University Press, Edinburgh, pp. 149–171.</p>
<p>IUCN Standards and Petitions Committee (2019) <em>Guidelines for Using the IUCN Red List Categories and Criteria. Version 14. Prepared by the Standards and Petitions Committee</em>. Gland, Switzerland and Cambridge, UK,16 pp.</p>
<p>Kaya, E. (2015) ISSR Analysis for determination of genetic diversity and relationship in some Turkish olive (<em>Olea europaea</em> L.) Cultivars. <em>Notulae Botanicae Horti Agrobotanici Cluj-Napoca </em>43 (1): 96–99. https://doi.org/10.15835/nbha4319818</p>
<p>Kaya, E. & Souza, F.V.D. (2017) Comparison of two PVS2-based procedures for cryopreservation of commercial sugarcane (<em>Saccharum</em> spp.) germplasm and confirmation of genetic stability after cryopreservation using ISSR markers. <em>In Vitro Cellular and Developmental Biology – Plant</em> 53: 410–417. https://doi.org/10.1007/s11627-017-9837-2</p>
<p>Kaya, E. & Yılmaz Gökdoğan, E. (2016) Using two retrotransposon based marker systems (IRAP and REMAP) for Molecular Characterization of olive (<em>Olea europaea</em> L.) cultivars. <em>Notulae Botanicae Horti Agrobotanici Cluj-Napoca</em> 44 (1): 167–174. https://doi.org/10.15835/nbha44110158</p>
<p>Kołodziej, B., Okoñ, S., Nucia, A., Ociepa, T., Luchowska, K., Sugier, D., Gevrenova, R. & Henry, M. (2018) Morphological, chemical, and genetic diversity of <em>Gypsophila</em> L. (Caryophyllaceae) species and their potential use in the pharmaceutical industry. <em>Turkish Journal of Botany</em> 42 (3): 257–270. https://doi.org/10.3906/bot-1707-13</p>
<p>Korkmaz, M. (2007) <em>Türkiye’de yetişen tek yıllık Gypsophila L. (Caryophyllaceae) taksonları üzerinde biyosistematik çalışmalar (in Turkish).</em> PhD thesis. Süleyman Demirel University, Isparta, 260 pp.</p>
<p>Korkmaz, M., Özçelik, H. & İlhan, V. (2012) Türkiye’nin bazı <em>Gypsophila</em> L. (Caryophyllaceae) taksonlarının habitat özellikleri (in Turkish). <em>Biyoloji Bilimleri Araştırma Dergisi</em> 5 (2): 111–125.</p>
<p>Linnaeus, C. (1753) <em>Species Plantarum</em>, <em>vol. 1.</em> Laurentii Salvii, Stockholm, 560 pp.</p>
<p>Mabberley, D.J. (2018) <em>Mabberley’s Plant-Book: A portable dictionary of plants, their classification and uses, fourth edition</em>. Cambridge University Press, UK, 1102 pp. https://doi.org/10.1017/9781316335581</p>
<p>Markus, S.D. & Joachim W.K. (2022) The distinction between <em>Sagina apetala</em> and <em>S. micropetala</em> (Caryophyllaceae: Sagineae), their phylogenetic relationships, and a note on the coastal origin of some widespread ruderals. <em>Willdenowia</em> 52 (1): 5–23. https://doi.org/10.3372/wi.52.52101</p>
<p>Muca, B. (2017) <em>Türkiye Gypsophila L. (Caryophyllaceae) cinsine ait Capituliformes Williams seksiyonunun revizyonu (in Turkish).</em> PhD thesis. Süleyman Demirel University, Isparta, 338 pp.</p>
<p>Özbek Yazıcı, S. & Özmen, İ. (2018) Effect of the crude saponin extract from <em>Gypsophila pilulifera</em> Boiss. & Heldr. on protease from <em>Bacillus subtilis</em> ATCC 6633 and antioxidant properties of the extract<em>. Iranian Journal of Science and Technology</em> 42 (4): 1707–1713. https://doi.org/10.1007/s40995-017-0366-y</p>
<p>Özçelik, H. & Muca, B. (2010) <em>Ankyropetalum </em>Fenzl (Caryophyllaceae) cinsine ait türlerin Türkiye’deki yayılışı ve habitat özellikleri (in Turkish). <em>BIBAD (Research Journal of Biology Sciences) </em>3 (2): 47–56.</p>
<p>Özçelik, H. & Özgökçe, F. (2021) Taxonomic contributions to genus <em>Gypsophila</em> in Turkey and a new taxon from Erzurum: <em>G. venusta</em> subsp. <em>staminea</em>. <em>Anatolian Journal of Botany</em> 5 (1): 6–18. https://doi.org/10.30616/ajb.791872</p>
<p>Özhatay, N. & Kültür, Ş. (2006) Check-list of additional taxa to the supplement Flora of Turkey III. <em>Turkish Journal of Botany</em> 30: 281–316.</p>
<p>Özüdoğru, E.A., Kaya, E., Kırdök, E. & İşsever-Öztürk, S. (2011) In vitro propagation from young and mature explants of thyme (<em>Thymus vulgaris</em> and <em>Thymus longicaulis</em>) resulting in genetically stable shoots. <em>In Vitro Cell Dev Biol-Plant</em> 47: 309–320. https://doi.org/10.1007/s11627-011-9347-6</p>
<p>POWO (2020) International Plant Names Index, Available from: https://powo.science.kew.org/results?q=Gypsophila%20pilulifera (accessed 1 January 2022)</p>
<p>Rohlf, F.J. (1998) <em>NTSYS-pc numerical taxonomy and multivariate analysis system version 2.0</em>. Exeter Publications, New York, 43 pp.</p>
<p>Stenoien, H., Fenster, C., Tonteri, A. & Savolainen, O. (2005) Genetic variability in natural populations of <em>Arabidopsis thaliana</em> in northern Europe. <em>Molecular Ecology</em> 14: 137–148. https://doi.org/10.1111/j.1365-294X.2004.02359.x</p>
<p>Sumaira, S., Mir, A.K., Akbar, A.M. & Asma, J. (2008) Polen Morphology of the Genus <em>Silene</em> (<em>Silene</em>-Caryopyllaceae) from Pakistan. <em>Biological Diversity and Conservation (BioDiCon)</em> 1 (2): 74–85.</p>
<p>Washitani, I., Ishikama, F., Matsumura, C., Nagal, M. & Nishihiro, M.A. (2005) Conservation ecology of <em>Primula sieboldii</em>: syntesisis of information toward the prediction of the genetic /demographic fate of a population<em>. Plant Species Biology</em> 20: 3–15. https://doi.org/10.1111/j.1442-1984.2005.00127.x</p>
<p>Williams, F.N. (1989) Revision of the forms of the genus <em>Gypsophila </em>L. <em>Jown Botanica of London</em> 27: 321–329.</p>
<p>Zhang, M.L., Zeng, X.Q., Li, C., Sanderson, S.C., Byalt, V.V. & Lei, Y. (2017) Molecular phylogenetic analysis and character evolution in Pseudostellaria (Caryophyllaceae) and description of a new genus, Hartmaniella, in North America, <em>Botanical Journal of the Linnean Society</em> 184 (4): 444–456. https://doi.org/10.1093/botlinnean/box036</p>
<p>Arslan, I., Çelik, A. & Chol, J.H.A. (2012) Cytotoxic triterpenoid saponin from under-ground parts of <em>Gypsophila pilulifera</em> Boiss. & Heldr. <em>Fitoterapia</em> 83: 699–703. https://doi.org/10.1016/j.fitote.2012.02.005</p>
<p>Ataşlar, E. (2000) <em>Gypsophila </em>L. <em>In</em>: Güner, A., Özhatay, N., Ekim, T. & Başer, K.H.C. (Eds.) <em>Flora of Turkey and the East Aegean Islands, </em>vol. 11 (Supplement). Edinburgh Univ. Press, Edinburgh, pp. 49–50.</p>
<p>Barkoudah, Y.I. (1962) <em>A Revision of Gypsophila, Bolanthus, Ankyropetalum and Phryna</em>. Wentia, Amsterdam, North-Holland, 203 pp. https://doi.org/10.1111/j.1438-8677.1962.tb00012.x</p>
<p>Boissier, E. (1842) <em>Diagnoses Plantarum Orientalium Novarum, vol. 2(8).</em> A.Ascher & Co., Leipzig, 128 pp.</p>
<p>Boissier, E. (1867) <em>Flora Orientalis: sive Enumeratio Plantarum in Oriente a Graecia et Aegypto ad Indiae fines hucusque observatarum</em>, vol. 1. Primum, A.Ascher & Co., Genevӕ, 548 pp. https://doi.org/10.5962/bhl.title.20323</p>
<p>Cain, M., Milligan, B. & Strand, A. (2000) Long-distance seed dispersal in plant populations. <em>American Journal of Botany</em> 87: 1217–1227. https://doi.org/10.2307/2656714</p>
<p>Chima, N.K., Nahar, L., Majinda, R.R.T., Çelik, S. & Sarker, S.D. (2014) Assessment of free-radical scavenging activity of <em>Gypsophila pilulifera</em>: Assay-Guided isolation of verbascoside as the main active component<em>. Revista Brasileira de Farmacognosia </em>24 (1): 38–43. https://doi.org/10.1590/0102-695X20142413391</p>
<p>Davis, P.H. (1965) <em>Flora of Turkey and the East Aegean Islands</em>, vol. 1<em>. </em>Edinburgh University Press, Edinburgh, 567 pp.</p>
<p>Davis, P.H., Mill, R.R. & Tan, K. (Eds.) (1988) <em>Gypsophila </em>L. <em>Flora of Turkey and the East Aegean Islands</em>, vol. 10 (Supplement). Edinburgh University Press, Edinburgh, pp. 73–75.</p>
<p>Dittbrenner, A., Hensen, I. & Wesche, K. (2005) Genetic structure and random amplified polymorphic DNA diversity of the rapidly declining <em>Angelica palustris (Apiaceae)</em> in Eastern Germany in relation to population size and seed production. <em>Plant Species Biology</em> 20: 191–200. https://doi.org/10.1111/j.1442-1984.2005.00138.x</p>
<p>Dole, J.M. & Wilkins, H.F. (2005) <em>Floriculture: Principles and species</em>. 2nd Edition. Pearson/Prentice Hall, Upper Saddle River, 1023 pp.</p>
<p>Doyle, J.J. & Doyle, J.L. (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. <em>Phytochemical Bulletin</em> 19: 11–15.</p>
<p>Düşen, O., Göktürk, R.S., Kaya, E., Sarpkaya, U. & Gürcan, B. (2018) Morphological and molecular determination of a new <em>Viola</em> species (Violaceae) from Turkey. <em>Phytotaxa</em> 369 (1): 37–46. https://doi.org/10.11646/phytotaxa.369.1.4</p>
<p>Ekim, T., Koyuncu, M., Vural, M., Duman, H., Aytaç, Z. & Adıgüzel, N. (2000) <em>Red Data Book of Turkish Plants</em>. Barışcan Press, Ankara, 246 pp.</p>
<p>Göktürk, R.S., Düşen, O., Kaya, E., Gürcan, B. & Sarpkaya, U. (2019) A new variety of <em>Plocama calabrica</em> (Rubiaceae) from Denizli (Turkey) confirmed by morphological and molecular ISSR markers. <em>Acta Botanica Croatica</em> 78 (2): 142–146. https://doi.org/10.2478/botcro-2019-0016</p>
<p>Hernández-Ledesma, P., Berendsohn, W.G., Borsch, T., Mering, S.V., Akhani, H., Arias, S., CastañedaNoa, I., Eggli, U., Eriksson, R., Flores-Olvera, H., Fuentes-Bazán, S., Kadereit, G., Klak, C., Korotkova, N., Nyffeler, R., Ocampo, G., Ochoterena, H., Oxelman, B., Rabeler, R.K., Sanchez, A., Schlumpberger, B.O. & Uotila, P. (2015) A taxonomic backbone for the global synthesis of species diversity in the angiosperm order Caryophyllales. <em>Willdenowia</em> 45: 281–383. https://doi.org/10.3372/wi.45.45301</p>
<p>Huber-Morath, A. (1967) <em>Gypsophila</em> L. <em>In</em>: Davis, P.H. (Ed.) <em>Flora of Turkey and the East Aegean Islands, vol. 2.</em> Edinburgh University Press, Edinburgh, pp. 149–171.</p>
<p>IUCN Standards and Petitions Committee (2019) <em>Guidelines for Using the IUCN Red List Categories and Criteria. Version 14. Prepared by the Standards and Petitions Committee</em>. Gland, Switzerland and Cambridge, UK,16 pp.</p>
<p>Kaya, E. (2015) ISSR Analysis for determination of genetic diversity and relationship in some Turkish olive (<em>Olea europaea</em> L.) Cultivars. <em>Notulae Botanicae Horti Agrobotanici Cluj-Napoca </em>43 (1): 96–99. https://doi.org/10.15835/nbha4319818</p>
<p>Kaya, E. & Souza, F.V.D. (2017) Comparison of two PVS2-based procedures for cryopreservation of commercial sugarcane (<em>Saccharum</em> spp.) germplasm and confirmation of genetic stability after cryopreservation using ISSR markers. <em>In Vitro Cellular and Developmental Biology – Plant</em> 53: 410–417. https://doi.org/10.1007/s11627-017-9837-2</p>
<p>Kaya, E. & Yılmaz Gökdoğan, E. (2016) Using two retrotransposon based marker systems (IRAP and REMAP) for Molecular Characterization of olive (<em>Olea europaea</em> L.) cultivars. <em>Notulae Botanicae Horti Agrobotanici Cluj-Napoca</em> 44 (1): 167–174. https://doi.org/10.15835/nbha44110158</p>
<p>Kołodziej, B., Okoñ, S., Nucia, A., Ociepa, T., Luchowska, K., Sugier, D., Gevrenova, R. & Henry, M. (2018) Morphological, chemical, and genetic diversity of <em>Gypsophila</em> L. (Caryophyllaceae) species and their potential use in the pharmaceutical industry. <em>Turkish Journal of Botany</em> 42 (3): 257–270. https://doi.org/10.3906/bot-1707-13</p>
<p>Korkmaz, M. (2007) <em>Türkiye’de yetişen tek yıllık Gypsophila L. (Caryophyllaceae) taksonları üzerinde biyosistematik çalışmalar (in Turkish).</em> PhD thesis. Süleyman Demirel University, Isparta, 260 pp.</p>
<p>Korkmaz, M., Özçelik, H. & İlhan, V. (2012) Türkiye’nin bazı <em>Gypsophila</em> L. (Caryophyllaceae) taksonlarının habitat özellikleri (in Turkish). <em>Biyoloji Bilimleri Araştırma Dergisi</em> 5 (2): 111–125.</p>
<p>Linnaeus, C. (1753) <em>Species Plantarum</em>, <em>vol. 1.</em> Laurentii Salvii, Stockholm, 560 pp.</p>
<p>Mabberley, D.J. (2018) <em>Mabberley’s Plant-Book: A portable dictionary of plants, their classification and uses, fourth edition</em>. Cambridge University Press, UK, 1102 pp. https://doi.org/10.1017/9781316335581</p>
<p>Markus, S.D. & Joachim W.K. (2022) The distinction between <em>Sagina apetala</em> and <em>S. micropetala</em> (Caryophyllaceae: Sagineae), their phylogenetic relationships, and a note on the coastal origin of some widespread ruderals. <em>Willdenowia</em> 52 (1): 5–23. https://doi.org/10.3372/wi.52.52101</p>
<p>Muca, B. (2017) <em>Türkiye Gypsophila L. (Caryophyllaceae) cinsine ait Capituliformes Williams seksiyonunun revizyonu (in Turkish).</em> PhD thesis. Süleyman Demirel University, Isparta, 338 pp.</p>
<p>Özbek Yazıcı, S. & Özmen, İ. (2018) Effect of the crude saponin extract from <em>Gypsophila pilulifera</em> Boiss. & Heldr. on protease from <em>Bacillus subtilis</em> ATCC 6633 and antioxidant properties of the extract<em>. Iranian Journal of Science and Technology</em> 42 (4): 1707–1713. https://doi.org/10.1007/s40995-017-0366-y</p>
<p>Özçelik, H. & Muca, B. (2010) <em>Ankyropetalum </em>Fenzl (Caryophyllaceae) cinsine ait türlerin Türkiye’deki yayılışı ve habitat özellikleri (in Turkish). <em>BIBAD (Research Journal of Biology Sciences) </em>3 (2): 47–56.</p>
<p>Özçelik, H. & Özgökçe, F. (2021) Taxonomic contributions to genus <em>Gypsophila</em> in Turkey and a new taxon from Erzurum: <em>G. venusta</em> subsp. <em>staminea</em>. <em>Anatolian Journal of Botany</em> 5 (1): 6–18. https://doi.org/10.30616/ajb.791872</p>
<p>Özhatay, N. & Kültür, Ş. (2006) Check-list of additional taxa to the supplement Flora of Turkey III. <em>Turkish Journal of Botany</em> 30: 281–316.</p>
<p>Özüdoğru, E.A., Kaya, E., Kırdök, E. & İşsever-Öztürk, S. (2011) In vitro propagation from young and mature explants of thyme (<em>Thymus vulgaris</em> and <em>Thymus longicaulis</em>) resulting in genetically stable shoots. <em>In Vitro Cell Dev Biol-Plant</em> 47: 309–320. https://doi.org/10.1007/s11627-011-9347-6</p>
<p>POWO (2020) International Plant Names Index, Available from: https://powo.science.kew.org/results?q=Gypsophila%20pilulifera (accessed 1 January 2022)</p>
<p>Rohlf, F.J. (1998) <em>NTSYS-pc numerical taxonomy and multivariate analysis system version 2.0</em>. Exeter Publications, New York, 43 pp.</p>
<p>Stenoien, H., Fenster, C., Tonteri, A. & Savolainen, O. (2005) Genetic variability in natural populations of <em>Arabidopsis thaliana</em> in northern Europe. <em>Molecular Ecology</em> 14: 137–148. https://doi.org/10.1111/j.1365-294X.2004.02359.x</p>
<p>Sumaira, S., Mir, A.K., Akbar, A.M. & Asma, J. (2008) Polen Morphology of the Genus <em>Silene</em> (<em>Silene</em>-Caryopyllaceae) from Pakistan. <em>Biological Diversity and Conservation (BioDiCon)</em> 1 (2): 74–85.</p>
<p>Washitani, I., Ishikama, F., Matsumura, C., Nagal, M. & Nishihiro, M.A. (2005) Conservation ecology of <em>Primula sieboldii</em>: syntesisis of information toward the prediction of the genetic /demographic fate of a population<em>. Plant Species Biology</em> 20: 3–15. https://doi.org/10.1111/j.1442-1984.2005.00127.x</p>
<p>Williams, F.N. (1989) Revision of the forms of the genus <em>Gypsophila </em>L. <em>Jown Botanica of London</em> 27: 321–329.</p>
<p>Zhang, M.L., Zeng, X.Q., Li, C., Sanderson, S.C., Byalt, V.V. & Lei, Y. (2017) Molecular phylogenetic analysis and character evolution in Pseudostellaria (Caryophyllaceae) and description of a new genus, Hartmaniella, in North America, <em>Botanical Journal of the Linnean Society</em> 184 (4): 444–456. https://doi.org/10.1093/botlinnean/box036</p>