Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics 76(1): 67-75, 2023 Firenze University Press www.fupress.com/caryologiaCaryologia International Journal of Cytology, Cytosystematics and Cytogenetics ISSN 0008-7114 (print) | ISSN 2165-5391 (online) | DOI: 10.36253/caryologia-2084 Citation: Meryem Nassar, Nora Sakhraoui, Gianniantonio Domina (2023). Karyomorphology of two subspecies of Anthemis maritima (Asteraceae) from Algeria. Caryologia 76(1): 67-75. doi: 10.36253/caryologia-2084 Received: March 10, 2023 Accepted: July 4, 2023 Published: September, 19, 2023 Copyright: © 2023 Meryem Nassar, Nora Sakhraoui, Gianniantonio Domina. This is an open access, peer-reviewed article published by Firenze University Press (http://www.fupress.com/caryo- logia) and distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided 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 MN: 0000-0002-1161-6957 NS: 0000-0002-9853-5702 GD: 0000-0002-9125-764X Karyomorphology of two subspecies of Anthemis maritima (Asteraceae) from Algeria Meryem Nassar1,4,*, Nora Sakhraoui2,4, Gianniantonio Domina3 1Department of Natural Sciences and Life BP 26, University of 20 August 1955, Route d’El Hadaiek-Skikda 21000, Algeria 2Department of Ecology and Environment BP 26, University of 20 August 1955, Route d’El Hadaiek-Skikda 21000, Algeria 3Department of Agricultural, Food and Forest Sciences University of Palermo, Viale delle Scienze, Bldg 4. 90128, Palermo, Italy 4Laboratory of Research in Biodiversity Interaction, Ecosystem and Biotechnology ‘LRIBEB’, University of 20 August 1955 BP 26, Route d’El Hadaiek-Skikda 21000, Algeria *Corresponding author. E-mail: meryem4321@yahoo.fr; m.nassar@univ-skikda.dz Abstract. The chromosome number and karyomorphology of two subspecies of the Anthemis maritima complex collected from two different coastal localities in the Skik- da region (northeastern Algeria) are reported in this study: Anthemis maritima subsp. maritima, which is common throughout the Mediterranean, and A. maritima subsp. bolosii, a strict Algerian endemic recently rediscovered after 100 years of disappear- ance. The Feulgen staining method indicated that Anthemis maritima subsp. maritima is a tetraploid with 2n=4x=36 chromosomes (2n=11m+3sm+4st), and Anthemis mar- itima subsp. bolosii is a diploid with 2n=2x=18 chromosomes (2n=4m+3sm+2st). Both taxa have symmetrical karyotypes, 2A and 1A, respectively, according to Stebbins clas- sification. These findings are novel for both the subspecies Anthemis maritima subsp. bolosii and the Algerian population of Anthemis maritima subsp. maritima. Keywords: Anthemis maritima subsp. bolosii, Anthemis maritima subsp. maritima, karyotype, chromosome, symmetrical, Skikda. INTRODUCTION The Asteraceae is an extremely diverse family of flowering plants, repre- senting a large part of the world’s flora with approximately 1,900 genera and 32,000 species (Mandela et al. 2019) distributed across 40 tribes (Funk et al. 2009). The tribe Anthemideae, which is a member of this family, stands out with 111 genera and nearly 1,800 species, making it one of the most impor- tant tribes of the Asteraceae family (Oberprieler et al. 2006; Oberprieler et al. 2007). Following a thorough revision based on molecular phylogenetic analyses, six new subtribes of the Asteraceae -Anthemideae family were iden- tified, which are: the Anthemidinae, Artemisiinae, Glebionidinae, Handelii- nae, Leucantheminae, and Leucanthemopsidinae. Five other monotypic sub- https://doi.org/10.36253/caryologia-2084 https://doi.org/10.36253/caryologia-2084 https://orcid.org/0000-0002-1161-6957 https://orcid.org/0000-0002-9853-5702 https://orcid.org/0000-0002-9125-764X 68 Meryem Nassar, Nora Sakhraoui, Gianniantonio Domina tribes were also added. As a consequence, this impor- tant tribe currently contains 19 subtribes (Oberprieler et al. 2022a). One of the most important genera in the Anthemideae tribe and a member of the Anthemidinae subtribe is Anthemis, which has 175 species distributed throughout the Mediterranean and Southwest Asia. However, due to the huge micromorphological differ- entiation of its species and those of related genera, the taxonomy of this genus is often challenging. Hybridi- zation and polyploidy may have played crucial roles in the evolution of this genus, further complicating its tax- onomy (Oberprieler 1998; Oberprieler 2001; Lo Presti et al. 2010). Phylogenetic analyses based on ITS sequences have improved our understanding of the evolution- ary relationships among Anthemis species, revealing that some species previously included in the subgenus Anthemis were actually more closely related to species in adjacent genera (Tripleurospermum and Nananthea) than to those in the subgenus Cota. This finding resulted to the reclassification of several species, as well as the exclusion of the subgenus Cota as a distinct genus (Greu- ter et al. 2003; Oberprieler 2001; Oberprieler et al. 2006; Oberprieler et al. 2009). Anthemis appears to be one of the few genera with an ancestral base number of x=9, evolved from x=10 fairly early in the evolutionary process (Bremer and Humphries 1993). In spite the fact that cytogenetic events such as ascending dysploidy (Leptinella, Artemi- sia) and descending dysploidy (Cotula, Artemisia, Atha- nasia, and Ursinia) are common in the Anthemideae tribe, resulting in base number alterations ranging from x=5 to x=11, 13, and 17, and driving their evolution. Indeed, all chromosomal counts in Anthemis sec- tions indicated a majority of diploid species (2n=2x=18) with an x=9 base number. Thus, polyploidization is pri- marily responsible for the evolution in some of its spe- cies (Oberprieler 1998; Inceer and Hayirlioglu-Ayaz 2007; Pellicer et al. 2007; Javadi et al. 2013; Shariat et al. 2021). Hiorthia is regarded as one of the six sections of the genus that have undergone true evolution due to the occurrence of polyploid species (2n=4x=36) (Oberprieler 2001). One of these is the North African A. maritima, specifically the subspecies A. maritima subsp. maritima (Oberprieler 1998). Despite the revision of (Oberprieler 1998) sev- eral taxa of the genus in North Africa are still poorly understood. This concerns not only the morphologi- cal aspects but above all the karyological and molecu- lar ones. The study of ploidy levels within the various subspecies allows us to better clarify the phylogenetic relationships between the investigated taxa. Although Algeria holds 20 taxa (12 species and 8 subspecies) among North African species (Dobignard and Chat- elain 2011), but unfortunately, karyological information on these taxa is scarce. The Anthemis maritima complex is present with three subspecies: A. maritima subsp. maritima and two strictly endemic subspecies to Algeria, A. maritima subsp. bolosii Benedí et Molero and A. maritima subsp. Pseudopunctata Oberpr. (Dobignard and Chatelain 2011; Tison and De Foucaul 2014). A. maritima  subsp. bolosii was first distinguished by Benedi Gonzalez and Molero Briones (1990) from A. maritima subsp. maritima by its more erect stem. Oberprieler (1998) reported the total absence of hairs on the peduncle and involucre of A. maritima subsp. bolosii, and Sakhraoui et al. (2021) add- ed other salient features, namely color and leaf appear- ance, mode of reproduction, habitat, and aromaticity. A. maritima subsp. maritima occurs on coastal dunes throughout the Mediterranean region (Dobignard and Chatelain 2010–2011; Tison and De Foucaul 2014). In Algeria, the subspecies is given as quite common on the littoral of the small and great Kabylia (Quézel and Santa 1963; Boulemtafes et al. 2018). A. maritima subsp. bolosii is considered rare, particularly since its distri- bution is very restricted. According to Sakhraoui et  al. (2021), the plant has been reported only at two coastal localities (Annaba and Skikda) in northeastern Algeria, and the presence of the subspecies has not been recorded for more than a century. The oldest record dates back to 6/23/1920 (specimen: P 03697947). The same author reported the rediscovery of the subspecies in the Stora locality (Skikda region). A. maritima subsp. bolosii is a perennial plant with a woody stump, a hairless stem and pinnate leaves that are sessile, or petiolate. The flowers with ligules and yellow centers are arranged into capitula that range in diameter from 14 to 35 mm and are carried by glabrous pedun- cles, the plant grows on cliffs and sea rocks (Sakhraoui et al. 2021) unlike A. maritima subsp. maritima, which prefers coastal dunes and sandy beachs (Boulemtafes et al. 2018; Sakhraoui et al. 2021). This taxon includes aro- matic plants of 20 to 70 cm in size with creeping stems rising during flowering and achenes with smooth ribs. The spangles of the receptacle are oblong-lanceolate, and the pinnate leaves are more or less fleshy, weakly to strongly puberulent (Quézel and Santa 1963; Tison and De Foucaul 2014; Sakhraoui et al. 2021). The aim of this contribution is to investigate and characterize, from a karyological point of view, A. mar- itima subsp. bolosii in comparison with the geographi- cally closest populations of A. maritima subsp. maritima. In fact, no cytogenetic investigation of these subspecies from Algeria has been conducted. This study is therefore 69Karyomorphology of two subspecies of Anthemis maritima (Asteraceae) from Algeria the first to report on the chromosome number and kary- ological features of A. maritima subsp. bolosii as well. MATERIALS AND METHODS The seeds used in this study were collected from wild plants growing in two separate locations in Skik- da’s region (Table 1). Chromosome observation and detection were achieved by the standard Feulgen stain- ing. Roots of 0.5-1.5 cm in length were obtained from the germinated seeds, then pretreated with 0.03% 8-hydroxyquinoline for 4 h at 4°C before being fixed in a 3 ethanol/1 acetic acid solution for 24 h. Hydroly- sis was performed in 1M hydrochloric acid for 5 min at 60°C. Root tips were stained with Schiff’s reagent for 2 h before being crushed in a drop of 45% acetic acid between the slide and coverslip. Five metaphase plates for each subspecies were analyzed to obtain the differ- ent measurements indicated in this work. The IdeoKar software was used for calculating karyotype parameters (Mirzaghaderia and Marzangib 2015). RESULTS AND DISCUSSION The karyomorphological results for A. maritima subsp. maritima and A. maritima subsp. bolosii are pre- sented in Tables 2 and 3, respectively. The studied popu- lation of A. maritima subsp. maritima was identified as tetraploid, with a chromosomal number of 2n=36. The subspecies has a total haploid length of 67.54 µm. The total length of chromosomes ranges from 2.50 µm to 5.14 µm, and the ratio between the long arm and short arm varies from 1.12 to 3.31. The karyotype consists of 18 pairs of chromosomes, of which eleven (1, 2, 3, 4, 5, 6, 8, 10, 12, 15, 16) are metacentric, three (9, 14, 17) are submetacentric, and four (7, 11, 13, 18) are subtelocen- tric. The chromosomal formula of A. maritima subsp. maritima is 2n=4x=36=11m+3sm+4st (Figure 1). A. maritima subsp. bolosii has nine pairs of chro- mosomes, four pairs (1, 3, 6, 9) are metacentric, three pairs (2, 4, 7) are submetacentric, and two pairs (5, 8) are subtelocentric. Based on these data, it was deter- mined that this subspecies is diploid, with x=9 basic Table 1. The location of the two A. maritima subspecies. Subspecies Locality Latitudes Longitudes Altitude Month of collection A. maritima subsp. maritima Larbi ben M’Hidi 36°53’33”N 7°00’25”E 10 m November 2022 A. maritima subsp. bolosii Stora 36°53’11”N 6°53’55”E 4 m August 2021 Table 2. Karyomorphological analysis of A. maritima subsp. mar- itima. N L (μm) S (μm) LT (μm) AR RL% CI CT 1 3.02±0.11 2.12±0.71 5.14±0.80 1.42 7.59 40.30 m 2 2.90±0.28 1.88±0.16 4.78±0.44 1.54 7.03 39.44 m 3 2.60±0.07 1.98±0.08 4.58±0.14 1.31 6.74 43.32 m 4 2.60±0.10 1.88±0.06 4.48±0.16 1.38 6.59 42.33 m 5 2.18±0.04 1.94±0.08 4.12±0.11 1.12 6.09 47.05 m 6 2.32±0.21 1.74±0.50 4.06±0.70 1.33 6.00 40.33 m 7 2.96±0.16 0.98±0.08 3.94±0.20 3.02 5.83 23.18 st 8 2.42±0.06 1.48±0.08 3.90±0.20 1.63 5.77 37.80 m 9 2.50±0.10 1.34±0.08 3.84±0.12 1.86 5.68 35.22 sm 10 2.28±0.06 1.56±0.10 3.84±0.16 1.46 5.48 40.50 m 11 2.84±0.08 0.88±0.08 3.72±0.15 3.22 5.47 23.86 st 12 2.06±0.13 1.50±0.08 3.56±0.15 1.37 5.24 40.20 m 13 2.50±0.24 0.82±0.08 3.32±0.34 3.04 4.91 24.22 st 14 2.00±0.17 1.18±0.40 3.18±0.57 1.69 4.71 37.30 sm 15 1.80±0.17 1.18±0.36 2.98±0.50 1.52 4.41 39.87 m 16 1.74±0.21 1.08±0.32 2.82±0.52 1.61 4.18 38.57 m 17 1.88±0.11 0.90±0.40 2.78±0.50 2.08 4.12 32.55 sm 18 1.92±0.11 0.58±0.20 2.50±0.30 3.31 3.69 23.36 st Length of chromosome (L: long arm, S: short arm, LT: total length), AR: Arm ratio, RL: Relative length, CI: Centromeric index, CT: Chromosome type. Table 3. Karyomorphological analysis of A. maritima subsp. bolosii. N L (μm) S (μm) LT (μm) AR RL% CI CT 1 2.79±0.06 2.39±0.11 5.07±0.17 1.16 14.1 46.07 m 2 2.96±0.16 1.56±0.08 4.52±0.24 1.89 13.0 34.61 sm 3 2.40±0.18 1.64±0.20 4.04±0.39 1.46 11.6 40.67 m 4 2.55±0.18 1.37±0.12 3.92±0.31 1.86 11.2 35.03 sm 5 2.98±0.25 0.82±0.07 3.80±0.33 3.63 10.9 21.65 st 6 2.14±0.21 1.58±0.11 3.72±0.32 1.35 10.7 42.49 m 7 2.28±0.27 1.36±0.08 3.64±0.35 1.67 10.4 37.38 sm 8 2.52±0.10 0.64±0.06 3.16±0.16 3.93 9.11 20.61 st 9 1.80±0.23 1.22±0.13 3.02±0.37 1.47 8.65 40.55 m Length of chromosome (L: long arm. S: short arm. LT: total length), AR: Arm ratio, RL: Relative length,CI: Centromeric index, CT: Chromosome type. 70 Meryem Nassar, Nora Sakhraoui, Gianniantonio Domina chromosomes number and a karyotype formula of 2n=2x=18=4m+3sm+2st (Figure 2). The chromosom- al lengths range from 3.02 µm to 5.07 µm, the haploid genome is 34.87 µm long, and the long arm to short arm ratio ranges from 1.16 to 3.93. These findings agree with prior research on the basic number x=9 and the degree of ploidy. Among the 28 species of Anthemis studied in North Africa (from Morocco and Tunisia), 23 were diploid, and the remaining species were tetraploid (2n=4x=36), including A. maritima subsp. maritima (Tunisian popu- lation) (Oberprieler 1998). All karyotypes were symmetrical, with metacentric, submetacentric, and subtelocentric chromosomes and at least two satellites. In our case, for both subspecies, all chromosomes were metacentric, submetacentric, and subtelocentric without satellites or secondary constric- tions. Generally, species in the genus Anthemis contain at least one chromosomal pair bearing a satellite (Ober- prieler 1998; Goula et al. 2022). The absence of satellites for the two studied taxa could be due to strong chromo- somal condensation that prevented their appearance. The asymmetry index (AsI) for A. maritima subsp. maritima and A. maritima subsp. bolosii is 63.40% and 64.02%, respectively. According to Stebbins’ classifica- tion (1971), both karyotypes are symmetrical 2A and 1A, types respectively. Other research on the karyology of the genus Anthemis has already confirmed the domi- nance of the base x=9 (Inceer and Hayirlioglu-Ayaz 2007; Chehregani and Mehanfar 2008; Javadi et al. 2013; Qari Figure 1. Karyotype of Anthemis maritima subsp. maritima. A: The plant, B: Metaphase plate, scale bars= 10 µm, C: Idiogram. 71Karyomorphology of two subspecies of Anthemis maritima (Asteraceae) from Algeria et al. 2016; Shariat et al. 2021; Goula et al. 2022). The base x=9 appears to be constant in the genus Anthemis, imply- ing that disploidy, which is often the origin of chromo- somal number changes, remains very low. Several asymmetry indices were calculated to assess the evolutionary linkages between the two subspecies, with values presented in Table 4 for intrachromosom- al asymmetry indices (A1, TF%, CVCI, and MCA) and interchromosomal indices (A2 and CVCL). These criteria allowed for a more accurate description of their distinct karyotypes. The intrachromosomal asymmetry indices A1, CVCI, and MCA are higher in the diploid subspecies (A. maritima subsp. bolosii) than in the tetraploid sub- species (A. maritima subsp. maritima). In contrast, the tetraploid subspecies (A. maritima subsp. maritima) pos- sess higher interchromosomal indices A2 and CVCL than the diploid subspecies (Table 4). The MCA and CVCL indices are considered reli- able quantitative parameters capable of estimating even minor intrachromosomal and interchromosomal variations in the chromosomal complement (Peruzzi et al. 2009; Peruzzi and Eroglu 2013; Baeza et al. 2015; Genc and Firat 2016; Phukan and Saha 2019; Eroglu and Budak 2020). CVCL and MCA values for A. mar- itima subsp. maritima and A. maritima subsp. bolosii are 19.65, 17.36, 24.3, and 28, respectively. When com- pared to its tetraploid relative, the diploid subspecies had higher CVCI and MCA values, indicating more vari- ability in centromere location. The tetraploid subspecies, on the other hand, exhibits higher CVCL and A2 values, Figure 2. Karyotype of Anthemis maritima subsp. bolosii. A: The plant, B: Metaphase plate, scale bars= 10 µm, C: Idiogram. 72 Meryem Nassar, Nora Sakhraoui, Gianniantonio Domina indicating more variation in chromosomal size. Fur- thermore, the IA index (Paszko 2006), another statistic parameter used to assess karyotype coherence, is high- er in the tetraploid subspecies (4.94) than in the dip- loid subspecies (4.61). A higher IA index value reflects increased chromosomal variability. As a result, it appears that the karyotype of A. maritima subsp. mar- itima has evolved more than that of A. maritima sub- sp. bolosii. Additionally, Stebbins’ (1971) classification classified the karyotypes of the subspecies A. maritima subsp. bolosii and A. maritima subsp. maritima in the symmetrical classes 1A and 2A, respectively, indicating some similarity between the two taxa but also a more primitive karyotype of the diploid subspecies compared to the tetraploid one. According to Oberprieler (1998), the karyotypes of Hiorthia section species are consist- ently of type A2, with less variation in intrachromo- somal symmetry but higher variation in interchromo- somal symmetry. As a consequence, our findings are consistent with previous research, particularly for the tetraploid subspecies. The differences in interchromo- somal indices between the two subspecies are quite considerable, as are the differences in intrachromo- somal indices, especially MCA, which can be induced by chromosomal mutations (translocations and inversions) resulting in changes between the two taxa. Despite the fact that 82.14% of North African species are diploid (Oberprieler, 1998), practically all species of the section Hiorthia, to which the A. maritima complex belongs, have been found to be tetraploid (Oberprieler, 1998; Presti et al. 2010). Mitsuoka and Ehrendorfer (1972) raised doubts about limiting the A. maritima complex to its tetraploid cytotype, suggesting that this plant may also possess a diploid cytotype, like its Moroccan coun- terpart, the Anthemis pedunculata complex. Thus, our data show unequivocally that the subspe- cies A. maritima subsp. bolosii represents the diploid cytotype for the A. maritima complex. Furthermore, A. pedunculata and A. maritima, are the only species that exhibit both diploid and tetraploid cytotypes. Phyloge- netic investigations have also shown that the two com- plexes are closely related (Oberprieler 2001; Presti et al. 2010; Oberprieler et al. 2022b). The similarity in terms of chromosome size and type between the karyotypes of both subspecies leads us to propose the following hypothesis: the tetraploid cytotype of A. maritima may have resulted from either duplication (autopolyploidy) of the diploid cytotype of A. maritima subsp. bolosii or potentially from an allopolyploidy event involving A. maritima subsp. bolosii and another related species. Knowing that hybridization between related Anthemis species is not new as evidenced by various studies (Uitz 1970; Mitsuoka and Ehrendorfer 1972; Nagl and Ehren- dorfer 1974; Oberprieler 1998; Oberprieler 2001; Tison and De Foucaul 2014; Oberprieler et al. 2022b). This is also the case for the tetraploid A. cupaniana, which is native to Sicily, and has been shown to be the result of allopolyploidization (during the pleistocene). Specifi- cally, this involved a maternal parent from the A. creti- ca group and a paternal parent from the North African complex A. pedunculata (Horthia section), namely the diploid Sicilian A. ismelia (Oberprieler et al. 2022b). The comparative palynological study conducted by Oberprieler (1998) suggested that the subspecies A. mar- itima subsp. bolosii is the only diploid subspecies (based on pollen size) among the three subspecies in the A. maritima complex, and A. maritima subsp. maritima and A. maritima subsp. pseudopunctata were identified as tetraploid. Furthermore, Algeria is the only territory where all three representatives of the A. maritima complex have been found. As mentioned above, A. maritima subsp. bolosii is interesting due to its rarity resulting from its limited distribution along coastal rocks and cliffs. All these information support the hypothesis that A. mar- itima subsp. bolosii is the ancestor of the A. maritima complex and that Algeria is the center of its diver- sification, especially since the Algerian population (2n=4x=36=11m+3sm+4st) and the Tunisian population (2n=4x=36=11m+3sm+4stsat) (Oberprieler 1998) of A. maritima subsp. maritima appear to be identical from a cytological point of view. According to Greuter (1979), Table 4. Asymmetry indices of A. maritima subsp. maritima and A. maritima subsp. bolosii. Subspecies HCL TF% AsI% CVCL CVCI AI A1 A2 MCA S Cl A. maritima subsp. maritima 67,54 36,5 63.4 19,65 25,18 4,94 0.41 0.19 24,3 2A A. maritima subsp. bolosii 34.87 35.97 64.02 17.36 26.56 4,61 0.42 0.17 28 1A HCL: Haploid total length, TF: Total form percentage (Huziwara 1962), AsI: Karyotype asymmtry index (Arano and Saito 1980), CVCL: Coefficient of variation of chromosome length, CVCI: Relative variation in centromeric index, AI: Asymmetry index (Paszko 2006), A1: Intrachromosomal asymmetry, A2: Interchromosomal asymmetry (Romero Zarco 1986), MCA: Mean centromeric asymmetry (Peruzzi and Eroğlu 2013), S Cl: Stebbins classification (Stebbins 1971). 73Karyomorphology of two subspecies of Anthemis maritima (Asteraceae) from Algeria populations of the same taxon that are geographi- cally separated but have karyotype stability are rem- nants of an ancient Mediterranean flora present during the Messinian period. Therefore, A. maritima complex might represent one of the groups that are ancient poly- ploids. In contrast to the diploid subspecies, the tetra- ploid A. maritima subsp. maritima is found in a number of coastal locations in Algeria (Quézel and Santa 1963; Boulemtafes et al. 2018) and across the Mediterranean area (Oberprieler 1998, 2001; Oberprieler et al. 2009). According to Winter et al. (1999), polyploid species spread more easily than their diploid ancestors, which grow rare and eventually survive only in isolated places shielded from hybridization. Stebbins (1971) asserted that polyploid plants are more adapted to changing environmental circumstances than their diploid ances- tors. This adaptation has evolved over millions of years as a result of genetic changes such as mutations and genetic recombination (Adams and Wendel 2005; Alix et al. 2017). Indeed, such a hypothesis requires strong molecular evidence to be confirmed, especially given that the data presented in this study is preliminary and insufficient to prove it. CONCLUSION The karyomorphological data of the Algerian popu- lation of A. maritima subsp. maritima and the subspe- cies A. maritima subsp. bolosii are reported for the first time and are the only data for the Algerian Anthemis taxa. Morphological differences between the two species previously documented by numerous authors are close- ly related to karyological differences, namely the ploidy level, proportions of each chromosomal type, and chro- mosomal formula. However, a few similarities between the two subspecies have been identified, including the degree of asymmetry and chromosomal size. As a result, A. maritima subsp. bolosii might be the origin of the A. maritima complex. 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Sofi², Aijaz H. Ganie², Manzoor A. Shah², Namrata Sharma¹ Comparative cytogenetics of four endemic Capoeta (Teleostei: Cyprinidae) species from Anatolia, Türkiye Sevgi Unal-Karakus1,*, Muhammet Gaffaroglu2, Muradiye Karasu-Ayata3 A karyomorphological comparison of seven species of Achillea L. from Kurdistan of Iran Fatemeh Nezhadi1, Farzad Fayaz2,*, Ezzat Karami2, Hooshmand Safari3, Abdol Rahman Rahimi2 Genotoxicity of a synthetic plant growth regulator, Forchlorfenuron (CPPU), on human lymphocytes using chromosome aberration assay Ayşe Yavuz Kocaman1,*, Berna Yakar2 Phenthoate toxicity evaluation in root meristem of Pisum sativum L. Sazada Siddiqui Karyomorphology of two subspecies of Anthemis maritima (Asteraceae) from Algeria Meryem Nassar1,4,*, Nora Sakhraoui2,4, Gianniantonio Domina3 Cytogenetic effects of Tribulus terrestris L. on meristematic cells of Allium cepa L. and Vicia faba L. Ali Bouzekri1,2,*, Meryem Nassar1,2, Souheila Slimani1,2, Zohra Chekroud1,2 New chromosomal data, karyotype asymmetry and polyploid variations of some Gundelia (Asteraceae) species from Turkey Esra Martin1, Metin Armağan2, Halil Erhan Eroğlu3*, Aslı Doğru-Koca4, Osman Tugay5, Golshan Zare6, Osman Kola7, Mahmut Miski8, Nur Tan9, Ernst Vitek10 Allelopathic and toxicological effects of Origanum vulgare L. essential oil Lejla Husić, Adisa Parić, Aner Mesic* Cytogenetic analysis in Tetragonopterus franciscoensis (Characiformes): another piece to the karyoevolutionary puzzle of tetra fishes Mauricio Barros Fernandes, Jamille de Araújo Bitencourt, Joandson Calixto dos Santos, José Henrique Galdino*, Paulo Roberto Antunes de Mello Affonso