Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics 76(3): 9-18, 2023 Firenze University Press www.fupress.com/caryologia ISSN 0008-7114 (print) | ISSN 2165-5391 (online) | DOI: 10.36253/caryologia-2238 Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Citation: Nassar, M., Sakhraoui, N., & Domina, G. (2023). Karyotype asymmetry in some Scilloideae (Hyacinthaceae) members from Algeria. Caryologia 76(3): 9-18. doi: 10.36253/caryologia-2238 Received: July 7, 2023 Accepted: January 18, 2024 Published: February 29, 2024 Copyright: © 2023 Nassar, M., Sakhraoui, N., & Domina, G. This is an open access, peer-reviewed article pub- lished by Firenze University Press (http://www.fupress.com/caryologia) 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 Karyotype asymmetry in some Scilloideae (Hyacinthaceae) members from Algeria Meryem Nassar1,4,*, Nora Sakhraoui2,4, Gianniantonio Domina3 1 Department of Natural Sciences and Life BP 26, University of 20 August 1955, El Hadaiek Road-Skikda 21000, Algeria 2 Department of Ecology and Environment BP 26, University of 20 August 1955, El Hadaiek Road-Skikda 21000, Algeria 3 Department of Agricultural, Food and Forest Sciences University of Palermo, Viale delle Scienze, Bldg 4, 90128, Palermo, Italy 4 Laboratory of Research in Biodiversity Interaction, Ecosystem and Biotechnology ‘LRIBEB’, University of 20 August 1955 BP 26, El Hadaiek Road-Skikda 21000, Algeria *Corresponding author E-mail: meryem4321@yahoo.fr, m.nassar@univ-skikda.dz Abstract. This cytogenetic study attempts to shed light on the karyomorphologi- cal and asymmetry data of species in the subfamily Scilloideae previously included in Scilla, namely Hyacinthoides lingulata (Poir.) Rothm., Prospero autumnale (L.) Speta, Prospero obtusifolium (Poir.) Speta, Barnardia numidica (Poir.) Speta, and Oncostema elongata (Parl.) Speta. These taxa are predominantly from the Skikda region (north- eastern Algeria). H. lingulata had a somatic chromosome number 2n=2x=16, P. autum- nale 2n=2x=14, P. obtusifolium 2n=2x= 8, B. numidica 2n=2x=18, and O. elongata 2n=2x=16. The indices of intrachromosomal (MCA, A1, and AsI) and interchromosomal (CVCL, CVCI, and A2) asymmetry revealed that H. lingulata has the most asymmetri- cal karyotype (1C), while P. autumnale has the most symmetrical karyotype (3A). P. obtusifolium has a relatively symmetrical karyotype (4A), while B. numidica and O. elongata have both an asymmetrical karyotype (1B). These findings differ from those previously reported for the same taxa in Algeria, hence indicating the substantial genetic variation that exists within the country. Keywords: chromosomes, bulbous, plants, genetic diversity, Scilla. INTRODUCTION The Hyacinthaceae family comprises exclusively herbaceous species, most of which are bulbous. In phylogenetic classification, this family is now included in the Asparagaceae (APG IV 2016), and its genera fall under the subfamily Scilloideae. However, the nomenclature of its genera is still wide- ly controversial. The species that belong to the former genus Scilla have also been split among several genera (Pfosser and Speta 1999). Three of them are related to the autumn squills of the North African flora, namely Prospero, Hyacinthoides, and Barnardia. The Hyacinthaceae family has a wide distribu- tion in Algeria, where it occurs with a certain concentration in the Tell Atlas http://www.fupress.com/caryologia https://doi.org/10.36253/caryologia-2238 https://doi.org/10.36253/caryologia-2238 http://www.fupress.com/caryologia https://orcid.org/0000-0002-1161-6957 https://orcid.org/0000-0002-9853-5702 https://orcid.org/0000-0002-9125-764X mailto:meryem4321@yahoo.fr mailto:m.nassar@univ-skikda.dz 10 Meryem Nassar, Nora Sakhraoui, Gianniantonio Domina region (Maire 1958; Quézel and Santa 1962). However, this family also includes endemic species shared between Algeria and neighboring countries, such as Albuca amoena (Batt.) J.C.Manning & Goldblatt, Ornithogalum sessiliflorum Desf. (both endemic to Morocco and Alge- ria), Hyacinthoides aristidis (Coss.) Rothm. (endemic to Algeria and Tunisia), and H. lingulata (Poir.) Rothm. (endemic to Morocco, Algeria, and Tunisia). The cytogenetic research carried out by Hamouche et al.(2010), Véla and de Bélair (2016), and Azizi et al. (2016a), has shed light on the karyotypes of various species that are proliferating in Algeria. However, tak- ing into account the variety of Algeria’s ecosystems, it’s possible that the genetic diversity found within Alge- ria’s populations is much greater than what was earlier thought. Previous research has shown that the genus Prospero has a significant amount of polymorphism, both in terms of the ploidy level and the fundamen- tal chromosomal number (2n=14, 2n=28, and 2n=42) (Hamouche et al. 2006; Hamouche et al. 2010; Jang et al. 2013; Jang et al. 2018). Two cytotypes, 2n=8 and 2n=16, have been found for H. lingulata, which primarily pro- liferates in diverse biotopes in the northwestern coastal region of Algeria (Hamouche et al. 2006; Hamouche et al. 2010). This underscores the necessity to extend and vary the sample regions in order to cover the widest pos- sible range of ecosystems. In light of this, we carried out cytogenetic research on some species of the Hyacinthaceae that are thriving in the Skikda area (northeastern Algeria), which have not been previously investigated. The following species were considered for the cytogenetic analyses: H. lingu- lata (Poir.) Rothm., P. autumnale (L.) Speta, P. obtusifo- lium (Poir.) Speta, B. numidica (Poir.) Speta, and O. elon- gata (Parl.) Speta. This study will allow us, on the one hand, to learn about the chromosomal composition of the studied taxa, and on the other hand, to shed light on the genetic diversity and karyological variation that may exist within Algeria’s populations. MATERIAL AND METHODS The plants selected for cytogenetic analysis were col- lected from their natural habitat (Figure 1) and identi- fied using the flora of North Africa (Maire, 1958), the GDB herbarium (https://gdebelair.com/), and confirmed Algerian occurrences documented on iNaturalist (htt- ps://www.inaturalist.org/observations?place_id=7300). The root tips were obtained from plants collected in nature (Table 1). Seed germination was not successful. After receiving a pretreatment with 0.05% colchicine, the roots were subsequently fixed in a combination of ethanol and acetic acid (3:1). Roots were then placed in a solution of hydrochloric acid (HCl 1N) and heated to 60 °C. This step lasted for 10 min. Following that, they were stained for at least 1h with Schiff’s reagent, and lastly, they were crushed in a drop of 3% acetic carmine. At least 10 plates per each population were studied. A series of photographs were taken of the best plates, and then different measures were carried out to determine the karyotype of each species. The chromosomal type was determined based on the terminology used by Levan et al. (1964). The karyotypes were accurately categorized using the Stebbins (1971) method. Different approaches were used to estimate additional parameters of karyotype asymmetry. HCL: Haploid total length, TF: Total form percentage (Huzi- wara 1962), AsI: Karyotype asymmetry index (Arano and Saito 1980), Syi: Karyotype asymmetry index, Rec: Chromosome size similarity index (Greihuber and Speta 1976), CVCL: Coefficient of variation in chromo- some length, CVCI: Coefficient of variation in centro- meric index, AI: Asymmetry index (Paszko 2006), A1: Intrachromosomal asymmetry, A2: Interchromosomal asymmetry (Romero Zarco 1986), MCA: Mean centro- meric asymmetry (Peruzzi and Eroğlu 2013). The Pear- son correlation between several asymmetry indices was calculated using IBM SPSS Statistics 24 software. Addi- tionally, the ideogram for each species was determined depending on their chromosomal size. RESULTS The karyotypes of five species from the subfamily Scilloideae have been thoroughly examined. Originally classified as Scilla by Maire (1958), these species have recently been reassigned to four distinct genera. Table 2 provides details on chromosomal numbers, total chro- mosome length, the ratio of long to short arms, and the total size of the haploid complement. H. lingulata, P. autumnale, P. obtusifolium, B. numidica, and O. elongata are all diploids with respective chromosome counts of 16, 14, 8, 18, and 16. The specific karyotypic formulas for each species are further documented in Table 2. The karyotypes of these species consist mostly of metacentric, submetacentric, subtelocentric, and even telocentric chromosomes, par- ticularly in the case of H. lingulata and B. numidica. The size of the chromosomes in B. numidica and H. lingu- lata varied from 1.27 μm to 6.49 μm depending on the species. H. lingulata stands out for having the largest haploid complement, which has an average size of 29.80 https://gdebelair.com/ https://www.inaturalist.org/observations?place_id=7300 https://www.inaturalist.org/observations?place_id=7300 11Karyotype asymmetry in some Scilloideae (Hyacinthaceae) members from Algeria μm. This size exceeds that of the other species. Follow- ing closely is O. elongata, which possesses a total hap- loid complement size of 28.28 μm. B. numidica possess- es chromosomes that are very tiny in size. However, its haploid complement measures 21.64 μm, which is larger than that of P. obtusifolium (16.60 μm) and P. autumnale (14.85 μm), as shown in Table 2. In order to assess the level of asymmetry in the karyotypes of the studied species, we used Stebbins’ cat- egorization together with the CVCL and MCA values. The specific asymmetry indexes used are outlined in Table 3. H. lingulata has much more prominent asymmetry char- acteristics compared to other species. The karyotype of this species is categorized as 1C according to Stebbins’ Figure 1. Illustration of the studied taxa showing differents parts of each plant: bulbs, flowers, and capsules,A: Hyacinthoides lingulata, B: Prospero autumnale, C: P. obtusifolium, D: Barnardia numidica, E: Oncostema elongata. Table 1. Localities of the studied taxa. Taxa Locality Habitat Latitudes Longitudes Altitude Month of collection Hyacinthoides lingulata Larbi Ben M’Hidi Coastal dune 36°53’08”N 7°00’55”E 40 m September Prospero autumnale Larbi Ben M’Hidi Coastal dune 36°53’08”N 7°08’01”E 40 m September Prospero obtusifolium Ramdan Djamel Olive grove 36°45’18”N 6°54’40”E 90 m December Barnardia numidica Filfilla Rocky cliffs 36°53’01”N 7°00’55”E 340 m November Oncostema elongata Ramdan Djamel Olive grove 36°45’18”N 6°54’40”E 90 m January 12 Meryem Nassar, Nora Sakhraoui, Gianniantonio Domina classification, with intrachromosomal asymmetry index MCA values of 59.4 and interchromosomal asymmetry index CVCL values of 48.4. In contrast, P. autumnale and P. obtusifolium, have significantly reduced asymmetry values. The intrachro- mosomal asymmetry indices (MCA) for P. autumnale and P. obtusifolium are 44.54 and 47.10, respectively. Simi- larly, their interchromosomal asymmetry indices (CVCL) are 18.53 and 17.14, respectively. The karyotypes of these two species are categorized as 3A and 4A, respectively. Although belonging to different genera, B. numidica and O. elongata exhibit very comparable asymmetry indices, which are characterized by rather high values. The intra- chromosomal asymmetry indices MCA for B. numidica and O. elongata are recorded as 52.57 and 51.45, respec- tively. The interchromosomal asymmetry index CVCL for B. numidica is determined as 42.01, whereas for O. elon- gata it is calculated as 45.69. Both karyotypes are classi- fied as 1B according to Stebbins’ classification (1971). DISCUSSION Hyacinthoides lingulata The earlier research carried out by Hamouche et al. (2006) and (2010) on populations from western Algeria assigned two different chromosomal numbers to this endemic taxon of North Africa. They were 2n=8 and 2n=16. This holds true when compared to the findings of the current investigation, which indicated that 2n=16. Furthermore, certain similarities in caryotype descrip- tions have been observed between the two populations. The main difference is that the Skikda population has two telocentric chromosomes but no satellite chromo- somes. According to Weiss Schneeweiss and Schneeweiss (2013), variation in caryotypical configuration between populations is indicative of chromosomal rearrange- ments, particularly those involving subtelocentric or tel- ocentric chromosomes that appear to result from chro- mosomal deletion or translocation. Previous research (Sato 1936) showed that members of the Scilloideae subfamily had various caryotypes even within the same species. It is worth noting that the presence of such vari- ability in karyotype organization does not always imply a significant morphological difference (Thompson 2005; Thompson 2020). Prospero autumnale The research conducted by Hamouche et al. (2010) on multiple populations from various locations in cen- tral Algeria revealed a high level of diversity within the species, which is closely associated with the altitude and environment in which they grow. Diploid (2n=2x=14), Table 2. Karyotype features of the studied taxa. Taxa 2n SC-LC (μm) LC/SC p (μm) q (μm) CL (μm) HCL CI (min- max) KF Hyacinthoides lingulata 16 1.54-6.49 4.21 0.75±0.1 2.97±0.17 3.72±0.26 29.80 0.05-0.44 1m+2sm+3st+2t Prospero autumnale 14 1.52-2.66 1.75 0.61±0.1 1.5±0.10 2.11±0.12 14.85 0.16-0.41 1m+msat+3sm+2st Prospero obtusifolium 8 3.17-4.82 1.52 1.08±0.2 3.04±0.1 4.14±0.22 16.16 0.21-0.32 2sm+2st Barnardia numidica 18 1.27-3.97 3.12 0.56±0.1 1.83±0.11 2.39±0.17 21.64 0.05-0.46 3m+2sm+3st+1t Oncostema elongata 16 1.79-5.38 3.00 0.85±0.06 2.66±0.12 3.51±0.17 28.28 0.13-0.40 2m+1sm+5st SC: shortest chromosome, LC: longest chromosome, p: mean short arm, q: mean long arm, CL: mean total chromosome length, HCL: total haploid chromosome length, CI: centromeric index, KF: karyotype formula, m: metacentric, sm: submetacentric, st: subtelocentric, t: telo- centric. Table 3. Asymmetry indices of the studies taxa. Taxa TF AsI Syi Rec CVCL CVCI AI A1 A2 MCA S Hyacinthoides lingulata 20.27 79.72 25.33 4.58 48.40 59.23 28.65 0.66 0.48 59.40 1C Prospero autumnale 28.98 71.01 37.33 5.57 18.53 29.23 5.40 0.56 0.18 44.54 3A Prospero obtusifolium 26.50 73.49 35.85 3.44 17.14 20.12 3.43 0.64 0.17 47.10 4A Barnardia numidica 23.77 76.22 30.76 5.43 42.01 51.89 21.79 0.57 0.42 52.57 1B Oncostema elongata 24.44 75.55 31.95 5.24 45.69 42.20 19.28 0.60 0.42 51.45 1B 13Karyotype asymmetry in some Scilloideae (Hyacinthaceae) members from Algeria Figure 2. Mitotic metaphase and idiograms of the studied taxa. A: Hyacinthoides lingulata 2n=16, B: Prospero autumnale 2n=14, C: P. obtusi- folium 2n=8, D: Barnardia numidica 2n=18, E: O. elongata 2n=16, scale bars=10 µm 14 Meryem Nassar, Nora Sakhraoui, Gianniantonio Domina tetraploid (2n=4x=24), and hexaploid (2n=6x=42) cyto- types were identified. The population of Skikda, which is located in northeastern Algeria, differs from the populations of Algiers, especially in terms of karyologi- cal description. It is distinguished by the presence of a metacentric chromosome (7) in addition to submetacen- tric and telocentric chromosomes, as well as a second- ary constriction on the long arm of chromosome 3. In contrast, the diploid population (2n=2x=14) from Algiers exhibits only submetacentric and subtelocentric chro- mosomes, in addition to a satellite on the short arm of chromosome 5. This implies that there is considerable variation even among diploid cytotypes (2n=2x=14) in Algeria, which might be due to chromosomal rearrangements such as inversions and translocations. Jang et al. (2013) and Jang et al. (2018) investigations both validated the existence of chromosomal rearrangements in the P. autumnale complex. In the P. autumnale complex, supernumer- ary forms such as B chromosomes and forms of ane- uploidy, particularly in diploid cytotypes (2n=2x=14+1) and hexaploid cytotypes (2n=6x=42+1), have been documented (Rejoin et al. 1980; Hamouche et al. 2010; Jang et al. 2018). In addition, Jang et al. (2013) showed the existence of four diploid cytotypes with three dis- tinct fundamental numbers, x=5, x=6, and x=7, based on their study of seventeen individuals from various countries bordering the Mediterranean Sea. Individuals with the cytotype 2n=2x=14 were assigned two chromo- somal descriptions, including one with submetacentric chromosomes (secondary constriction on chromosome 3), a subtelocentric chromosomes and a small metacen- tric chromosome. This is an exact match to the chromo- somal description discovered in this investigation. The possibility cannot be ruled out for the presence of other P. autumnale complex cytotypes in Algeria, given the country’s vast size and the vast diversity of its habitats and climates. Prospero obtusifolium Our findings regarding chromosomal number and formula are consistent with the studies conducted by Ebert et al. (1996), Hamouche et al. (2010), and Jang et al. (2013). These studies all identified a single cytotype (2n=8) in populations throughout the Mediterranean basin. Overall, this indicates that the species has suc- cessfully maintained significant stability in its karyotype composition, even when dispersed over different habitats and a wide geographic area. Barnardia numidica Our population exhibits a chromosomal formula distinct from that of the central Algerian population (2n=2x=18=4m+2sm-sat+3st) by possessing a telocen- tric pair but lacking a satellite. Both populations share metacentric and subtelo-centric chromosomes. The kar- yotype consists of two distinct sets of chromosomes, one containing small metacentric or submetacentric chro- mosomes and the other containing big subtelo-centric chromosomes (bimodal karyotype). The variation in size that was found might be attributed to hybridization, as shown by previous research (Speta 1979; Ebert et al. 1996; Hamouche et al. 2010). Other studies have indicat- ed that this heterogeneity is linked to higher amplifica- tion of distinct forms of heterochromatin within specific chromosomal sets (De la Herrán et al. 2001). Indeed, the existence of a bimodal karyotype has already been observed in Hyacinthaceae species such as Ornithogalum (Stedje 1989) and Galtonia (Forrest and Jong 2004). Oncostema elongata There is still no taxonomic consensus on the spe- cific delimitation within O. peruviana (Scilla peruvi- ana). Some authors attribute the variability observed in the CW Mediterranean to a single species (Almeida da Silva and Crespí 2013; Almeida da Silva et al. 2014), others prefer to consider different species (Dobignard and Chatelain 2010; APD 2023). Previous research has pointed to the presence of different chromosomal counts of 2n=16, 2n=15, and 2n=14 have been seen in different individuals of O. peruviana (Sato 1936). The same is true for O. hughii (Tineo ex Guss.) Speta, which has four distinct numbers (2n=17, 2n=19, 2n=20, and 2n=22). It is worth noting that these chro- mosomal count differences were discovered in plants that shared the same habitat (Sato 1936). In addition, the same author has shown that O. peruviana (L.) Speta has a stable karyotype with 2n=16=1M+4stSat+2m+1st. Although 2n=16 is thought to be the fundamental chro- mosome number in O. peruviana (Sato 1936; Battaglia 1950, Barone et al. 2021). Several asymmetry indices have been established since the work of Stebbins to evaluate karyotype asym- metry in its evolutionary state (Romero Zarco 1986). These are the first data on karyotype asymmetry for the several species included in this study. According to Peruzzi and Eroglu (2013), the intrachromosomal asym- metry index MCA and the interchromosomal asymmetry index CVCL are extensively utilized due to their abil- 15Karyotype asymmetry in some Scilloideae (Hyacinthaceae) members from Algeria ity to detect even the most subtle chromosomal varia- tions. Various asymmetry parameters have disclosed karyotype diversity among the studied species. Spe- cies in the genus Prospero, such as P. autumnale and P. obtusifolium, classified as types 3A and 4A, respectively, appear to have the most symmetrical karyotypes, indi- cating relatively stable karyotypes, according to Steb- bins’ classification. The karyotypes of B. numidica, O. elongata and H. lingulata, on the other hand, belong to categories 1B, 1B and 1C, respectively, indicating asym- metrical karyotypes. High CVCL and MCA values for all three species, especially H. lingulata, support this result (Table 3). The intrachromosomal asymmetry index MCA had a significant negative connection with Syi (-0.997**) and TF (-0.996**) and a positive correlation with AsI (0.996**) and HCL (0.917**) according to Pearson cor- relation. It also correlated positively with CVCL (0.093) and CVCI (0.516), although not significantly. The inter- chromosomal asymmetry index CVCL, on the other hand, had a strong and positive relationship with CVCI (0.83*), AI (0.99**), A2 (0.99**), and HCL (0.91**). There- fore, differences in asymmetry between chromosomes in the same set are associated with differences in size, which may result from changes in structure as well as differences in shape, particularly the location of the chromosomal centromere. The karyotypes of P. autumnale and P. obtusifolium are notable for their symmetrical structure, but those of B. numidica, O. elongata, and H. lingulata have sig- nificant asymmetry. According to Stebbins (1971), asym- metric karyotypes are seen as derived traits that have emerged more recently in evolutionary processes. They are often linked to plants that display specific morpho- logical features or are descendants of a more recent ori- gin. In the Mediterranean flora, variations in chromo- some size, base number, and symmetry between closely related species are not new. Many processes, such as polyploidy, aneuploidy, or dysploidy, can explain these changes (Stebbins 1971; Levin 2002; Guerra 2008; Choi et al. 2008) and morphological differences could result from the expression of these genetic variations (Thomp- son 2005; Thompson 2020). Species belonging to genera such as Reichardia, Brachyscome, Crepis and Geranium provide clear examples of this karyotypic variety (Sil- jak-Yakovlev 1996; Watanabe et al. 1999; Dimitrova and Greilhuber 2000; Martin et al. 2022). Interestingly, these variations are not limited to interspecific differences but extend to intraspecific levels, giving rise to the emer- gence of new chromosomal races (Levin 2002). Prospero autumnale (Parker et al. 1991, Ebert et al. 1996, Vaughan et al. 1997, Hamouche et al .2010), Ornithogalum tenuifo- lium (Stedje 1989), O. nutans (Cullen and Rattert, 1967), Albuca abyssinica (Stedje 1996), Bellevalia mauritanica, and Muscari neglectum (Azizi et al. 2016b) are notable examples in the Hyacinthaceae family. CONCLUSION In terms of karyotypic constitution and asymme- try indices, the cytogenetic data related to the examined species belonging to four genera of the subfamily Scil- loideae have revealed, on the one hand, that the popula- tions of the Skikda region are different from other popu- lations previously studied by other researchers, indicat- ing the important genetic diversity that exists in Algeria and on the other hand, significant differences between the species. This suggests that P. autumnale is the most stable species and H. lingulata is the most evolved spe- cies. However, molecular phylogenetic investigations are required to fully comprehend the relationship between the species under issue. ACKNOWLEDGMENT The authors would like to thank Dr Errol Véla from the University of Montpellier for the confirmation of the identification of Oncostema elongata. REFERENCES Almeida da Silva R, Rocha J, Silva A, García-Cabral I, Amich F, Crespí AL. 2014. 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N L (μm) S (μm) LT (μm) AR R value RL% F% CI CT 1 5.28±0.26 1.20±0.07 6.49±0.33 4.37±0.49 0.22±0.02 21.7±0.63 4.05±0.25 0.18±0.01 st 2 4.30±0.37 1.11±0.20 5.42±0.55 3.93±0.40 0.25±0.03 18.1±1.20 3.73±0.06 0.20±0.01 st 3 4.49±0.20 0.49±0.13 4.98±0.33 9.15±0.69 0.10±0.01 16.7±1.14 1.64±0.06 0.09±0.01 t 4 3.85±0.15 0.22±0.06 4.08±0.20 17.0±0.66 0.05±0.01 13.6±0.50 0.76±0.01 0.05±0.01 t 5 1.54±0.15 1.24±0.15 2.79±0.30 1.24±0.03 0.80±0.02 9.37±1.01 4.18±0.50 0.44±0.01 m 6 1.73±0.01 0.98±0.04 2.71±0.05 1.76±0.07 0.56±0.02 9.12±0.12 3.29±0.12 0.36±0.01 sm 7 1.49±0.16 0.26±0.11 1.75±0.26 5.57±0.30 3.74±0.17 5.89±0.19 0.88±0.38 0.15±0.07 st 8 1.03±0.09 0.50±0.01 1.54±0.11 2.07±0.10 0.48±0.02 5.19±0.38 1.71±0.06 0.33±0.01 sm Length of chromosome (L: long arm. S: short arm. LT: total length). F%: Form percentage of chromosome. CI: Centromeric index. AR: Arm ratio. R-value: S/L. RL: Relative length. CT: Chromosome type. Table 5. Karyomorphological analysis of Prospero autumnale. N L (μm) S (μm) LT (μm) AR R value RL% F% CI CT 1 2.03±0.19 0.63±0.04 2.66±0.22 3.18±0.23 0.31±0.02 17.95±0.13 4.25±0.26 0.23±0.01 st 2 1.97±0.03 0.38 ±0.08 2.35±0.12 5.55±0.27 0.18±0.01 15.82±0.13 2.52±0.13 0.16±0.01 st 3 1.36±0.05 0.94±0.08 2.31±0.13 1.41±0.06 0.70±0.03 15.55±0.40 6.38±0.01 0.41±0.01 m-sc 4 1.61±0.10 0.67±0.08 2.29±0.18 2.41±0.12 0.41±0.02 15.42±0.53 4.52±0.01 0.29±0.01 Sm 5 1.42±0.07 0.51±0.01 1.93±0.08 2.76±0.15 0.36±0.02 13.03±0.53 3.45±0.39 0.26±0.02 sm 6 1.20±0.05 0.57±0.05 1.77±0.11 2.14±0.12 0.46±0.02 11.96±0.79 3.85±0.39 0.32±0.01 sm 7 0.92±0. 15 0.59±0.03 1.52±0.1 8 1.53±0.20 0.65±0.08 10.23±0.13 3.98±0.26 0.39±0.03 m Length of chromosome (L: long arm. S: short arm. LT: total length). F%: Form percentage of chromosome. CI: Centromeric index. AR: Arm ratio. R-value: S/L. RL: Relative length. CT: Chromosome type. 18 Meryem Nassar, Nora Sakhraoui, Gianniantonio Domina Table 6. Karyomorphological analysis of Prospero obtusifolium. N L (μm) S (μm) LT (μm) AR R value RL% F% CI CT 1 3.49±0.11 1.32±0.28 4.82± 0.17 2.67±0.69 0.37±0.10 29.0± 1.04 7.98±1.73 0.27±0.04 Sm-Sat 2 2.99±0.11 1.48±0.32 4.47± 0.44 2.05±0.38 0.48±0.09 26.9± 2.66 8.91±1.96 0.32±0.04 sm 3 3.22±0.04 0.90±0.17 4.13± 0.13 3.65±0.75 0.27±0.05 24.8± 0.81 5.43±1.04 0.21±0.03 st 4 2.47±0.13 0.69±0.03 3.17± 0.17 3.55±0.01 0.28±0.01 19.0± 1.04 4.16±0.23 0.21±0.01 st Length of chromosome (L: long arm. S: short arm. LT: total length). F%: Form percentage of chromosome. CI: Centromeric index. AR: Arm ratio. R-value: S/L. RL: Relative length. CT: Chromosome type. Table 7. Karyomorphological analysis of Barnardia numidica. N L (μm) S (μm) LT (μm) AR R value RL% F% CI CT 1 3.20±0.10 0.77±0.04 3.97±0.02 4.27±0.11 0.23±0.01 18.3±0.09 3.56±0.17 0.19±0.01 st 2 3.20±0.37 0.18±0.10 3.39±0.27 19.2±2.00 0.05±0.01 15.6±1.25 0.86±0.48 0.05±0.01 t 3 2.79±0.12 0.41±0.11 3.20±0.31 6.70±0.62 0.14±0.02 14.8±0.76 1.92±0.19 0.12±0.01 st 4 2.33±0.23 0.79±0.12 3.12±0.19 2.94±0.69 0.33±0.07 14.4±0.19 3.65±0.57 0.25±0.04 sm 5 0.97±0.10 0.83±0.04 1.81±0.11 1.15±0.13 0.86±0.10 8.37±0.10 3.84±0.19 0.46±0.02 m 6 1.14±0.06 0.58±0.16 1.72±0.10 1.92±0.72 0.51±0.17 7.98±0.48 2.69±0.76 0.33±0.07 sm 7 0.99±0.04 0.68±0.06 1.68±0.10 1.50±0.07 0.66±0.08 7.79±0.48 3.17±0.28 0.40±0.01 m 8 0.81±0.06 0.62±0.04 1.43±0.02 1.26±0.18 0.78±0.11 6.64±0.10 2.88±0.19 0.43±0.03 m 9 1.02±0.04 0.24±0.04 1.27±0.07 0.95±0.25 0.25±0.05 5.87±0.09 1.15±0.19 0.19±0.03 st Length of chromosome (L: long arm. S: short arm. LT: total length). F%: Form percentage of chromosome. CI: Centromeric index. AR: Arm ratio. R-value: S/L. RL: Relative length. CT: Chromosome type. Table 8. Karyomorphological analysis of Oncostema elongata. N L (μm) S (μm) LT (μm) AR R value RL% F% CI CT 1 4.11±0.12 1.20±0.10 5.38±0.20 3.29±0.38 0.30±0.03 19.0±0.21 4.49±0.35 0.23±0.02 st 2 3.91±0.60 0.88±0.02 4,79±0.62 4,40±0,68 0.22±0,03 16,9±2,13 3,13±0.60 0,18±0.02 st 3 3,64±0.06 1,02±0,02 4,67±0,08 3,60±0,01 0.27±0.02 16.5±0.28 3.63±0.07 0.21±0.02 st 4 3,97±0.06 0,62±0.02 4.59±0.04 6.50±0.30 0.15±0.07 16.2±0.14 2.20±0.07 0.13±0.02 st 5 1,95±0.02 1.14±0.10 3.10±0.08 1.71±0.16 0.58±0.05 10.9±0.28 4.06±0.35 0.36±0.02 sm 6 1.18±0.06 0.80±0.02 1.99±0.06 1.45±0.07 0.68±0.03 7.05±0.21 2.85±0.02 0.40±0.02 m 7 1.47±0.10 0.46±0.02 1.93±0.12 3.27±0.08 0.30±0.11 6.84±0.42 1.63±0.07 0.32±0.04 st 8 1.10±0.10 0.68±0.08 1.79±0.18 1.58±0.04 0.62±0.01 6.34±0.64 2.42±0.28 0.38±0.02 m Length of chromosome (L: long arm. S: short arm. LT: total length). F%: Form percentage of chromosome. CI: Centromeric index. AR: Arm ratio. R-value: S/L. RL: Relative length. 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