Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics 75(3): 13-18, 2022 Firenze University Press www.fupress.com/caryologia ISSN 0008-7114 (print) | ISSN 2165-5391 (online) | DOI: 10.36253/caryologia-1660 Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Citation: Esra Kavcı, Esra Mar- tin, Halil Erhan Eroğlu, Fatih Serdar Yıldırım (2022). Chromosome number of some Satureja species from Turkey. Caryologia 75(3): 13-18. doi: 10.36253/ caryologia-1660 Received: May 17, 2022 Accepted: February 12, 2022 Published: April 5, 2023 Copyright: © 2022 Esra Kavcı, Esra Mar- tin, Halil Erhan Eroğlu, Fatih Serdar Yıldırım. This is an open access, peer-reviewed article published by Firenze University Press (http://www. fupress.com/caryologia) and distrib- uted under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, pro- vided the original author and source are credited. Data Availability Statement: All rel- evant data are within the paper and its Supporting Information files. Competing Interests: The Author(s) declare(s) no conflict of interest. ORCID EK: 0000-0002-5235-6979 EM: 0000-0002-5484-0676 HEE: 0000-0002-4509-4712 FSY: 0000-0003-4080-8488 Chromosome number of some Satureja species from Turkey Esra Kavcı1, Esra Martin1, Halil Erhan Eroğlu2,*, Fatih Serdar Yıldırım3 1 Necmettin Erbakan University, Faculty of Science, Department of Biotechnology, Konya, Turkey 2 Yozgat Bozok University, Faculty of Science and Arts, Department of Biology, Yozgat, Turkey 3 Akdeniz University, Faculty of Education, Department of Science Education, Antalya, Turkey *Corresponding author: herhan.eroglu@bozok.edu.tr Abstract. The genus Satureja belonging to the Lamiaceae family includes about 200 species, generally aromatic, distributed in the Mediterranean basin. In genus Satureja, the chromosomal data were reported from only 26 species. In this study, it is aimed to eliminate the deficiencies in the chromosomal data of Satureja species, which are dis- tributed in Turkey, which is center of origin and diversity of the genus Satureja. It was reported only one chromosome number (2n = 30), the first report for chromosome numbers of three taxa, the same chromosome count (excluding B-chromosomes) with previous report in only one species, and the new chromosome number in only one species. In conclusion, this study presented new data into the chromosomal records of genus Satureja that might be useful for interpreting or understanding relationships among the species. In addition, dysploidy and polyploidy variations might probably have played an important role in speciation. In this regard, the results contributed to some missing data in Satureja cytotaxonomy. Keywords: Satureja, chromosome, dysploidy, polyploidy, Turkey. INTRODUCTION The genus Satureja L. belonging to the Lamiaceae family includes about 200 species, generally aromatic, distributed in the Mediterranean basin. Satureja species are distributed in Morocco, Libya, Saudi Arabia, Cauca- sus, Iran, Iraq and Turkey, which are mostly Mediterranean countries from Europe, Asia and North Africa. Turkey is center of origin and diversity of the genus Satureja (Harley et al. 2004; Dirmenci et al. 2019). According to Flora of Turkey records, the genus Satureja is represented by a total of 15 species, five of which are endemic. Satureja species, popularly known as “pointed thyme” or “stone thyme”, have a wide range of uses in the food, pharmaceutical and cosmetic industries due to high amounts of thymol 14 Esra Kavcı, Esra Martin, Halil Erhan Eroğlu, Fatih Serdar Yıldırım and carvacrol (Momtaz and Abdollahi 2010; Dirmenci et al. 2019). The dried leaves of Satureja species are used as spice, food additive and herbal tea (Babajafari et al. 2014). The genus Satureja has been the focus of many studies due to its biological activities such as antimicro- bial, antioxidant and anti-HIV-1. It has been stated that the rich essential oil (eg mono- and sesquiterpenes) and phenolic content (eg phenolic acids, catechins, and flavo- noids) of Satureja species are responsible for these activi- ties (Eminağaoğlu et al. 2007; Bektaş 2020). Cytotaxonomy is a branch of taxonomy that uses karyological parameters to classify organisms. In cyto- ta xonomy, chromosomal conf iguration is the most widely used parameter to understand the relationship between organisms. Inference of species relationships is based on the assumption that closely related species share similar features in their chromosomal arrange- ment. By analyzing similarities and differences in chromosomes, karyotype evolution and species evolu- tion can be reconstructed. The number, structure and behaviour of chromosomes are very valuable in tax- onomy and the chromosome numbers (x, 2n) are the most commonly used characters (Guerra 2008; Eroğlu et al. 2020; Martin et al. 2020; Eroğlu et al. 2021). In genus Satureja, the chromosomal data were reported from only 26 species. Eighteen species were only dip- loid; however, they revealed two different basic num- bers: x = 13 (2n = 26) and x = 15 (2n = 30). Seven spe- cies were polyploid and revealed two different poly- ploidy levels: tetraploidy (2n = 4x = 24, 28, 44, and 60) and hexaploidy (2n = 6x = 48). The polyploid species revealed four different basic numbers such as x = 6, 7, 11, and 15. Satureja sahendica Bornm. had diploid and polyploid records. In addition, S. hortensis L. presented dysploidy, which is an alteration in basic number, gen- erally by fusion, without the significant loss or gain of genetic material (Shariat et al. 2013; Irani et al. 2014; Bordbar et al. 2021; Chromosome Counts Database 2022; Vozhdehnazari et al. 2022). In Turkish Satureja, the chromosomal data were reported from eight species, which were S. macran- tha C.A. Mey. (2n = 24), S. coerulea Janka, S. cuneifolia Ten., S. pilosa Velen. S. thymbra L. (2n = 30), S. spinosa L. (2n = 30 + 2B), S. spicigera Boiss. (2n = 44, 60), and S. hortensis (2n = 45-48) (Shariat et al. 2013; Irani et al. 2014; Chromosome Counts Database 2022; Vozhdehnaz- ari et al. 2022). There was no record of the chromosome number of seven species, which were S. aintabensis P.H.Davis, S. amani P.H.Davis, S. boissieri Hausskn. ex Boiss., S. cilicica P.H.Davis, S. icarica P.H.Davis, S. par- nassica Heldr. & Sartori ex Boiss., and S. wiedemanniana Lall. ex Velen. Therefore, the lack of some chromosomal reports from Turkey, which is located in the Mediter- ranean Basin, may lead to some uncertainties in the cytotaxonomy. In this study, it is aimed to eliminate the deficiencies in the chromosomal data of Satureja species, which are distributed in Turkey, which is center of ori- gin and diversity of the genus. MATERIALS AND METHODS Plant material Within the scope of this study, S. aintabensis, S. boissieri, S. icarica, S. macrantha and S. spinosa distrib- uted in different localities of Turkey were examined by chromosome numbers. The examined plant samples were collected from their natural habitats and identi- fied by Prof. Dr. Tuncay Dirmenci et al (Figure 1). The collected plant samples were preserved in Balıkesir Uni- versity, Necatibey Faculty of Education, Department of Biology Education. The distribution regions and collec- tion information are given in Figure 2 and Table 1. Cytogenetic procedure The seeds of the collected plant samples were kept at -40°C for 1 month and then planted in petri dishes. The samples were kept in the dark at 4°C for 21 days, and at the end of 21 days, the samples placed in the climate cabinet were kept until the root tip tis- sue reached a few centimeters in length. Germinated root tips were kept in α-monobromonaphthalene for 16 hours at 4°C for the first treatment. Afterwards, root tips were fixed in 3:1 absolute alcohol:glacial acetic acid and stored in 70% alcohol in the refrigerator. The root tips were removed from the refrigerator, hydro- lyzed in 1N HCl at room temperature for 10 minutes and stained with 2% aceto-orcein for 2 hours at room temperature. Then, squash preparations were prepared with 45% acetic acid. After the preparations were fro- zen in liquid nitrogen, they were dried at room tem- perature and stabilized with Depex medium (Martin et al. 2018; Eroğlu et al. 2021). Ten metaphase plates were used for counting the somatic chromosomes of each species. After the mitotic chromosomes, which were well distributed, had good morphology, and were on the same plane, were detect- ed, their photographs were taken at 1000× magnifica- tion with a camera attached to the microscope (Olym- pus BX51). The chromosome photographs were analyzed using the Image Analysis System (Bs200ProP). 15Chromosome number of some Satureja species from Turkey Figure 1. Habitat (1) and flowers (2) of Satureja species. (A) S. aintabensis; (B) S. boissieri; (C) S. icarica; (D) S. macrantha; and (E) S. spi- nosa. Scale bar, 10 cm for habitat (1) and 5 mm for flowers (2). Figure 2. Distribution map of the studied species in Turkey. (A) S. aintabensis; (B) S. boissieri; (C) S. icarica; (D) S. macrantha; and (E) S. spinosa. 16 Esra Kavcı, Esra Martin, Halil Erhan Eroğlu, Fatih Serdar Yıldırım RESULTS Figure 3 presented the mitotic metaphase chromo- somes of five Satureja species. The chromosome num- bers of studied Satureja species were given in Table 2. In all species, the diploid number was 2n = 30, three of which were reported for the first time and the basic number was x = 15. In genus Satureja, because the chromosomes were very small and the centromere region is unclear, detailed chromosomal measurements were not made. Table 1. The collection information and localities of studied Satureja species. Species Collection cite Altitude Date Voucher number S. aintabensis Gaziantep, Samköy, Behind Dülükbaba promenade. 1000 m 03.09.2018 Dirmenci 5210 & Arabacı S. boissieri Adıyaman, Çelikhan, between Yazıbaşı village and Ulubaba mountain, 8th km. 1700 m 02.09.2018 Dirmenci 5207 & Arabacı S. icarica Çanakkale, between Gökçeada and Aydıncık, 2nd-3rd km. 300 m 15.08.2018 Dirmenci 5166 S. macrantha Ardahan, between Göle and Şenkaya, 10th km. 1800 m 31.08.2018 Dirmenci 5196 & Arabacı S. spinosa Muğla, Fethiye, Babadağ, above telpher. 1900 m 11.09.2018 Dirmenci 5224 & Yıldız Figure 3. Metaphase chromosomes of Satureja species. (A) S. aintabensis; (B) S. boissieri; (C) S. icarica; (D) S. macrantha; and (E) S. spinosa. Scale bar 10 µm. Table 2. The chromosome numbers of studied Satureja species. Species x = basic number, 2n (ploidy level) S. aintabensis x = 15, 2n = 30 (diploid) S. boissieri x = 15, 2n = 30 (diploid) S. icarica x = 15, 2n = 30 (diploid) S. macrantha x = 15, 2n = 30 (diploid) S. spinosa x = 15, 2n = 30 (diploid) 17Chromosome number of some Satureja species from Turkey DISCUSSION The genus Satureja was represented by a total of 15 species in Turkey. Eleven Turkish Satureja whose chro- mosome numbers had been reported with present and previous studies were given in the Table 3 for compari- son. In the present study, the diploid number of all spe- cies was 2n = 30, three of which were reported for the first time: S. aintabensis, S. boissieri, and S. icarica. The chromosome number represented new cytotype in only one species, which was S. macrantha (2n = 30). Vozh- dehnazari et al. (2022) reported that the chromosome number of S. macrantha was 2n = 24. The chromosome number of S. spinosa agreed with the previous report excluding B-chromosomes. In Turk ish Satureja, f ive different chromosome numbers were recorded such as 2n = 24, 30, 44, 48, 60 and 2n = 30 was the most common diploid number. S. spinosa was the only species to have B-chromosomes. Montmollin (1986) reported that the karyotype of S. spinosa was 2n = 30 + 2B, which were small super- numerary chromosomes other than A-chromosomes. B-chromosomes originated from the A-chromosomes and were a basic source of intraspecific variations of nuclear DNA (Heneida k et a l. 2019). Although we obtained the same diploid number, we did not observe B-chromosomes. This was probably due to the locality difference. In Table 3, Satureja was a polybasic genus by x = 11, 12, 15 with ploidy levels of 2x and 4x. Nine species were diploid with 2n = 2x = 30. S. hortensis and S. spinosa were polyploid, which revealed only one polyploidy level of tetraploidy (2n = 4x = 44, 48, and 60). A basic number of x = 15 dominated in reported Satureja species (all species excluding S. hortensis in Table 3), which were S. aintabensis, S. boissieri, S. coer- ulea, S. cuneifolia, S. icarica, S. macrantha (only in this study), S. pilosa, S. spicigera (only in this study), S. spi- nosa, and S. thymbra. In addition, the basic numbers of x = 11 and 12 were recorded. However, different basic numbers were reported, such as x = 6 for S. multiflora Briq. and x = 7 for S. sahendica (Krogulevich 1978; Irani et al. 2014). In detecting karyotype evolution and spe- ciation processes, basic chromosome number is one of the most important parameter. In genus Satureja, basic number variations were probably caused by descending or ascending dysploidy and dibasic polyploidy. In Table 3, nine species were diploid with 2x = 30 (81.82% of the species) and two species were polyploid with 4x = 44, 48, and 60 (18.18% of the species). Poly- ploidy was probably one of the important mechanisms in the karyotype evolution of the genus, as it occurred at a non-negligible rate (Chromosome Counts Database 2022) in the genus Satureja. In the present study, it was reported only one chro- mosome number (2n = 30), the first report for chromo- some numbers of three taxa, the same chromosome count (excluding B-chromosomes) with previous report in only one species, and the new chromosome number in only one species. In conclusion, this study presented new data into the chromosomal records of genus Satureja that might be useful for interpreting or understanding relationships among the species. In addition, dysploidy and polyploidy variations might probably have played an important role in speciation. In this regard, the results contributed to some missing data in Satureja cytotaxon- Table 3. The chromosome numbers of Turkish Satureja in present and previous studies. Species (alphabetically) x = basic number, 2n (ploidy level) References Observation S. aintabensis x = 15, 2n = 30 (diploid) Present study First report S. boissieri x = 15, 2n = 30 (diploid) Present study First report S. coerulea x = 15, 2n = 30 (diploid) Chromosome Count Database 2022 Previous report S. cuneifolia x = 15, 2n = 30 (diploid) Chromosome Count Database 2022 Previous report S. hortensis x = 12, 2n = 48 (tetraploid) 45-47 (probably dysploidy) Chromosome Count Database 2022 Previous report S. icarica x = 15, 2n = 30 (diploid) Present study First report S. macrantha x = 15, 2n = 30 (diploid) x = 12, 2n = 24 (diploid) Present study Vozhdehnazari et al. 2022 New count S. pilosa x = 15, 2n = 30 (diploid) Chromosome Count Database 2022 Previous report S. spicigera x = 15, 2n = 60 (tetraploid) x = 11, 2n = 44 (tetraploid) Shariat et al. 2013 Irani et al. 2014 Previous report S. spinosa x = 15, 2n = 30 (diploid) x = 15, 2n = 30 + 2B (diploid) Present study Chromosome Count Database 2022 B-chromosomes not observed S. thymbra x = 15, 2n = 30 (diploid) Chromosome Count Database 2022 Previous report 18 Esra Kavcı, Esra Martin, Halil Erhan Eroğlu, Fatih Serdar Yıldırım omy. 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