Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics 76(4): 23-37, 2023 Firenze University Press www.fupress.com/caryologia ISSN 0008-7114 (print) | ISSN 2165-5391 (online) | DOI: 10.36253/caryologia-1924 Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Citation: Sebkhi, Z., Issolah, R., Mel- zi, N., Benmouhoub, H., & Mefti, M. (2023). Karyological analyses in several Algerian populations of six species of the genus Vicia L. (Fabaceae). Caryo- logia 76(4): 23-37. doi: 10.36253/caryo- logia-1924 Received: January 09, 2023 Accepted: December 29, 2023 Published: March 14, 2024 Copyright: © 2023 Sebkhi, Z., Issolah, R., Melzi, N., Benmouhoub, H., & Mefti, M. 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. Karyological analyses in several Algerian populations of six species of the genus Vicia L. (Fabaceae) Zahia Sebkhi1,4, Rachida Issolah1,*, Nabila Melzi2, Hassina Ben- mouhoub3, Mohamed Mefti4 1 INRAA, CRP Mehdi Boualem, Division de Recherche sur les Ressources phytogéné- tiques, BP 37, Baraki, 16200, Alger, Algérie 2 Université Saad Dahleb, Blida 1, Algérie 3 Laboratoire d’Ecologie et environnement, Faculté des sciences de la nature et de la vie, Université de Béjaïa, 06000 Béjaïa, Algérie 4 ENSA, LRGB, Alger, Algérie *Corresponding author. E-mail: rachida.issolah@yahoo.com Abstract. As part of the evaluation and valorization of plant genetic resources of fod- der and pastoral interest in Algeria, seventeen (17) natural populations belonging to six (06) species of the genus Vicia (Vicia sativa, Vicia disperma, Vicia monardii, Vicia ohchroleuca, Vicia onobrychioides and Vicia lutea), originated from different ecologi- cal regions in the North-Eastern of the country, were considered. The populations have been the subject of mitotic and meiotic studies. Haploid and diploid numbers and chromosome measurements were determined. Original results were observed for the first time in some species. In fact, chromosome counts have revealed some new chro- mosome numbers. The first number (2n=14, n=7) was observed in the endemic sub- species of Algeria, V. ochroleuca subsp. atlantica and in the species Vicia onobrychioides. The second number (n=6) was observed in the species Vicia disperma. Within Vicia monardii, the three new chromosome numbers, previously observed only in mitosis by our research team, were confirmed for the first time through the present meiosis study (n=6, 7 and 8), indicating that they are A type chromosomes. The base num- ber x=7 is the most frequently observed number in the six Vicia species. The observed chromosome numbers would be related to some ecological factors (altitude, rainfall) of the origin environment of the populations. Chromosome measurements and estab- lished karyotypes were determined for the first time in V. monardii, V. onobrychioides and V. ochroleuca subsp. atlantica. Chromosome size and karyotype formula are vari- able among the studied species and subspecies. Karyotypes vary from symmetrical to asymmetrical and the intrachromosomal asymmetry is higher than interchromosomal one. The new cytogenetic data would contribute to a better understanding of the evolu- tion mechanism of the species in the genus Vicia L. Keywords: chromosomes, endemic, karyotype, plant genetic resources, Vicia L. http://www.fupress.com/caryologia https://doi.org/10.36253/caryologia-1924 https://doi.org/10.36253/caryologia-1924 https://doi.org/10.36253/caryologia-1924 http://www.fupress.com/caryologia http://www.fupress.com/caryologia mailto:rachida.issolah@yahoo.com 24 Zahia Sebkhi et al. INTRODUCTION The Leguminosae (Fabaceae) is the third larg- est angiosperm family in terms of species number after Asteraceae and Orchidaceae, comprising over 770 genera and 19500 species (Lewis et al. 2005, 2013). The family is morphologically, physiologically and ecologically diverse, representing one of the most spectacular examples of evolutionary diversification in plants (LPWG 2017). The genus Vicia L. is a member of the legume tribe Vicieae of the subfamily Papilionoideae (Kupicha1976). The exact number of species in the genus Vicia L. is quite difficult to estimate, due to cytological and mor- phological differences (Kartal et al. 2020). It is including approximately 210 species that are widely distributed in temperate regions of Europe, Asia and America (Hanelt and Mettin 1989). Archeological evidence suggests that the main center of diversification of genus Vicia is the Mediterranean region (Raveendar et al. 2015). Most of species of the genus Vicia are annual but a few of them belonging to the section Cracca are perennial (Yamanoto 1973). Recently, the genus Vicia has been recognized for its vital role in sustainable agriculture (Han et al. 2021). The most common classification is that of Kupicha (1976) revised by Maxted (1993). Kupicha (1976) divided the genus Vicia into two subgenera, Vicilla and Vicia with 17 and 5 sections, respectively. Subgenus Vicia is smaller than Vicilla but it is more coherent and includes the more agriculturally important species (Ruffini Cas- tiglione et al. 2011). Maxted (1993) subdivided the subge- nus Vicia into 9 series, 38 species, 14 subspecies and 22 varieties. The subgenus Vicilla is considered more primi- tive and diverse than subgenus Vicia (Kupicha 1976 and Maxed 1993). According to Quezel and Santa (1962), the Algerian flora is represented by 26 species and 18 subspecies cor- responding to the genus Vicia L. Two subspecies, Vicia ochroleuca subsp. atlantica and Vicia ochroleuca subsp. baborensis are respectively quite rare and very rare in Algeria. More recently, Dobignard and Chatelain (2012) report 39 taxa within the genus Vicia in Algeria, includ- ing two endemic subspecies, V. ochroleuca subsp. atlan- tica and V. ochroleuca subsp. baborensis. The prospecting and collection mission carried out, in 2016, in north- eastern Algeria, showed the frequency of Vicia sativa, followed by Vicia disperma and the rarity recorded in some encountred species such as Vicia monardii, Vicia narbonensis and Vicia ochroleuca (subsp. atlantica) (Issolah et al. 2022). Vicia sativa L. is a variable genus comprising of sev- eral subspecies; it is most commonly called Vicia sativa complex (Cvs) or sativa aggregate (Jauzein 1995; Kartal et al. 2020; Benlioglu 2021). Members of this complex are morphologically, karyologically and ecologically variables and were considered to be in active evolution (Potokina et al. 2000; Shiran and Raina 2001; El-Bok et al. 2015), making identification difficult and confusing. Several karyological studies have been carried out in Vicia. They had an important role in improvement and solving of several taxonomic problems between the relat- ed species (Lavia et al. 2009; Murti et al. 2012). There is a considerable variability in haploid nuclear DNA content (1.8-13.3 pg) and basic chromosome number (2n=10, 12 or 14) between Vicia species (Raina and Narayan 1984; Maxed 1995). This makes the genus an interesting model for the study of plant genome and Karyotype evolution (Navratilova et al. 2003). Most of them are diploids with a basic number x =5, 6 or 7 (Maxted 1991), while only six of them are polyploids (Cremonini et al. 1992). In Algeria, karyological studies on spontaneous populations of the genus Vicia L. are very rare. Several species are totally unknown. The main objectives of this study are to determine the number and size of chromo- somes within some species of the genus Vicia in Algeria, in order to contribute to a better understanding of the mechanism of evolution of the different species in this genus. This work follows previous studies conducted on the evaluation and valorization of diversity in spontaneous and local fodder legumes in Algeria (Issolah and Abdel- guerfi 1999; Bouziane et al. 2019; Chabouni et al. 2019; Issolah et al. 2006, 2012, 2015, 2018, 2022). MATERIAL AND METHODS Plant material Following a prospecting mission carried out by INRAA through the North-East of Algeria (Issolah et al. 2022), several species belonging to the genus Vicia were collected. Seventeen (17) populations belonging to six (06) species of the genus Vicia L. (V. sativa subsp. sativa (04), V. sativa subsp. macrocarpa (03), Vicia disperma (03), Vicia monardii (04), V. ohchroleuca subsp. atlantica (01), Vicia onobrychioides (01) and Vicia lutea (01) were the subject of the present study (Table 1). Mitotic study Diploid chromosome numbers were counted from the seedling root tips which were germinated in Petri dishes on filter paper at room temperature. The root tips meristems (about 1 cm long; zone of active division) 25Karyological analyses in several Algerian populations of six species of the genus Vicia L. (Fabaceae) were excised in the morning between 7.30 am – 8.30 am periods of active cell division. Firstly, root tips were pre- treated for 02 h with α- bromonaphtalene (1%) at room temperature. Then, they were fixed in ethanol chloro- form acetic acid (6:3:1) during 24 h at 4 °C. Root tips were hydolysed with 1 N HCL and were stained using lactopropionic orcein solution (Dyer 1963). Chromosome counts in mitosis metaphase and kar- yotype analyses were obtained usually based on five best plates (metaphase cells) of chromosomes, for each popu- lation. Each chromosome was identified on the basis of its total chromosome length. Chromosomal nomenclature was carried out according to Levan et al. (1964). Idi- ograms were constructed by arranging the chromosomes in homologous pairs by order of their length. For the numerical characterization of the karyo- types, the following parameters were calculated: The length of long arm (L), short arm(S), total chromosome length (LT= L+S) and the relative length (RL ‰) = 1000x TL/∑ TL (Levan et al. 1964). To determine chromosome type and centromere position, two parameters were calculed: arm ratio (r = L/S) and centromeric index (CI%=S/LT x 100) according to the nomenclature of Levan et al. (1964). To determine the asymmetry of the karyotype, the interchromosomal asymmetry was determined by calcu- lating the coefficient of variation of chromosome length CVCL (Paszko 2006), Rec index (Venora et al. 2002) and the interchromosomal asymmetry index A2 (Zarco 1986). The intrachromosomal asymmetry was deter- mined by calculating, the mean centromeric asymmetry MCA (Peruzzi and Eroglu 2013), the percentage of karyo- type asymmetry index AsK% (Arano 1963), total form percentage TF% (Huziwara 1962), the Syi index (Greil- huber and Speta 1976), the intrachromosomal asymme- try index A1 (Zarco1986) and the degree of asymmetry of karyotype A (Watanabe et al. 1999). Meiotic study A meiotic analysis was conducted in order to con- firm the results obtained during mitosis and determine the chromosome type of the newly observed chromo- some in the considered Algerian populations (Vicia L.). On this purpose, a trial was set-up at the INRAA exper- imental station. The protocol adopted is a total rand- omization. The sowing was carried out on 15 November 2018 at the rate of twenty seeds (individuals), for each population. In April 2018, flower buds of different sizes were col- lected for each population and immediately fixed in a solution of Carnoy acetic ethanol (3:1 v/v). The hydroly- sis was performed using 1N HCL for 1 to 3 min at 60°C. The staining was done with lactopropionic orcein at room temperature. The meiotic behavior was analyzed. The observations of the mitotic plates were made using a Primo Zeiss Star microscope and photographed with a digital camera attached to this microscope. The Table 1. Ecological characteristics of natural habitats of several spontaneous populations within some Vicia L. species in Algeria. Populations Species Subspecies Origin Altitude (m) Rainfall (mm) 11/14 65/15 75/15 63/16 55/14 73/15 77/15 49/14 80/14 79/15 40/16 48/16 51/16 66/16 52/16 82/14 83/15 V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. disperma DC. V. disperma DC. V. disperma DC. V. monardiii Boiss V. monardiii Boiss V. monardiii Boiss V. monardiii Boiss V. ochroleuca Spreng. V. onobrychioides L. V. lutea L. subsp. sativa subsp. sativa subsp.sativa subsp.sativa subsp.macrocarpa subsp. macrocarpa subsp. macrocarpa - - - - - - - subsp. atlantica - subsp..vestita Boumerdes Bejaia Bejaia Bejaia Bejaia Bejaia Bejaia Bejaia Bejaia Tizi-Ouzou Bouira Bejaia Bejaia Bouira Bejaia Tizi-Ouzou Blida 25 450 550 160 1250 860 410 550 90 1030 800 560 595 730 1140 750 230 850 950 950 950 1100 700 800 700 750 1100 500 700 700 500 1100 950 650 Source: (Issolah et al. 2022, completed). 26 Zahia Sebkhi et al. analysis of cytogenetic data was made with the Axio- vision software (1999-2009). The different karyotype cal- culations were made with Excel (2007). RESULTS Mitotic analyses In this study, the chromosome numbers and detailed chromosome measurements were determined for the natural populations of Vicia species in Algeria. Mitotic studies revealed that all studied populations were diploid (figure 1). Vicia sativa subsp. sativa At the metaphase stage, chromosomal counts revealed the existence of a chromosomal number (2n=2x=12) for all analysed populations of Vicia sativa subsp. sativa. However, a second chromosome number (2n=2x=10) was found in population 11/14 with a high frequency (80%) in the cells of the same individual and in different individuals of this population (figure1). This indicates a chromosomic variation within and between the populations of Vicia sativa L. The shortest chromosome length is 1.9 µm (popu- lation n° 11/14); the longest one is 4.84 µm (population n° 63/16). The mean value of the total length TLG of all the studied populations is 3.65 µm. The centromeric index varies between 23.4 to 38.53 and relative lengths vary from 87.51 to 222.97. The chromosomes observed in Vicia sativa subsp. sativa populations were mainly sub- metacentric or subtelocentric types (Table 3). For the karyotype asymmetry, the lowest value of intrachromosomal asymmetry MCA is 30.63, the high- est one is 47.63. The lowest value of interchromosomal asymmetry CVCL is 9 and the highest value is 30 (Table 7). The idiograms were drawn based on centromeric index and arranged in the decreasing size order (Figure 2). Vicia sativa subsp. macrocarpa The present study showed that 2n=2x=12 is the chromosome number of all populations of Vicia sativa subsp. macrocarpa (Table 2). The shortest chromosome length is 2.25 µm (population n° 77/15), the longest one is 4.85µm (population n° 55/14). The mean value of the total length TLG of all studied population is 3.46 µm, the centromeric index varies between 21, 2 and 37.92 and the relative lengths vary from 111 to 206.7 (Table 4). For karyotype asymmetry, the lowest value for intrachromosomal asymmetry MCA is 36.68, the highest one is 49.0. The lowest and highest values of interchro- mosomal asymmetry CVCL are 13 and 20, respectively (Table 7). The karyotype formula is very variable between pop- ulations of the same subspecies in Vicia sativa (Table 3, 4). The idiograms of Vicia sativa subsp. sativa and Vicia sativa subsp. macrocarpa were illustrated on the basis of their centromeric index and arranged in descending order of the chromosomal size (Figure 2). Vicia monardii Within this rare species (Quezel and Santa 1962), our observations showed that populations n°51/16 and n°40/16 have a stable number of chromosomes (2n=14), while populations n°48/16 and n°66/16 have two chro- mosomes numbers (2n=12 and 2n=14) and (2n=14 and 2n=16), respectively (Table 2). In Vicia monardii, karyological measurements were determined for the first time in the present study. The shortest chromosome length is 2.76 µm (population n°48/16), the longest chromosome is 6.37 µm (population n° 66/16). The mean value of the total length TLG of all studied population is 4.40. The centromeric index var- ies between 23.07 and 37.9 and the relative lengths vary from 87.45 to 196.29 (Table 5). For the karyotype asymmetry, the lowest value of MCA intrachromosomal asymmetry is 36, the highest one is 44. The lowest value for interchromosomal asymmetry CVCL is 15.4 and the highest value is 24.22 (Table 7). The karyotype formula is variable between the pop- ulations of Vicia monardii. The observed chromosomes are mainly submetacentrics (Table 5). The idiograms were illustrated on the basis of the centromeric index and arranged in descending order of chromosome size (Figure 2). Vicia ochroleuca subsp. atlantica The somatic chromosome number of Vicia ochro- leuca subsp. atlantica is 2n =2x=14 (Figure1). It is a new number observed for the first time in this endemic spe- cies in Algeria. Karyological measurements were made for the first time within this subspecies. It is characterized by chro- mosomes with lengths of 3.19 - 4.86 µm and an average value of the total length of 4.14 µm. The centromeric 27Karyological analyses in several Algerian populations of six species of the genus Vicia L. (Fabaceae) index varies from 25.92 to 38.91 and the relative length varies from 109.96 to 167.52 µm. The karyotype formula is 2n=14=2m+12sm (Table 6). For the intrachmosomal asymmetry MCA and inter- chromosomal asymmetry CVCL, the values are 39.61 and 14.5, respectively (Table 7). The idiogram was illustrated on the basis of the centromeric index and arranged in descending order of chromosome size (Figure 2). Vicia onorychioides For the polymorphic species Vicia onobrychioides, the somatic chromosome number is 2n =2x=14. It is a new number observed in this species (figure 1). The shortest chromosome length, the longest chro- mosome length, the mean chromosome length and the total haploid length are 2.91, 4.55, 3.67and 25.69 µm, respectively. The centromeric index varies from 22.63 to 38.14 µm, respectively. The relative length varies from113.27 to 177.11. The karyotype formula is 2n =14= 10sm+2m+2st (Table 6). The values of intrachromosomal asymmetry MCA and interchromosomal asymmetry CvCL are 39 and 16, respectively. The idiogram was illustrated on the basis of the centromeric index and arranged in descending order of chromosome size (Figure 2). Table 2. Chromosome number within 17 Algerian populations of the genus Vicia L. Population Species Subspecies Chromosomes numbers 2n n 11/14 V. sativa subsp. sativa 10 and 12 5 and 6 65/15 V. sativa subsp. sativa 12 6 75/15 V. sativa subsp. sativa 12 6 63/16 V. sativa subsp. sativa 12 6 55/14 V. sativa subsp. macrocarpa 12 6 73/15 V. sativa subsp. macrocarpa 12 6 77/15 V. sativa subsp. macrocarpa 12 6 49/14 V. disperma - 14 7 80/14 V. disperma - 14 7 79/15 V. disperma - - 6 and 7 40/16 V. monardii - 14 - 48/16 V. monardii - 12 and 14 6 and7 51/16 V. monardii - 14 - 66/16 V. monardii - 14 and 16 7 and 8 52/16 V. ochroleuca subsp. atlantica 14 - 82/14 V. onobrychioides - 14 7 83/15 V. lutea subsp. vestita - 7 Figure 1. Mitosis observed in natural populations of the genus Vicia in Algeria: A - Vicia sativa subsp. sativa (population 11/14, 2n = 12) ; B- Vicia sativa subsp. sativa (population 11/14, 2n = 10); C- Vicia sativa subsp. macrocarpa (population 55/14, 2n = 12); D- Vicia ochroleuca ( population 52/16, 2n = 14); E- Vicia onobrychioides ( population 82/14, 2n = 14); F- Vicia disperma  ( population 49/15 , 2n= 14); G  - Vicia monardii (population 48/16, 2n = 12); H- Vicia monardii (population 66/16, 2n = 16) . Scale bar 5 µm. 28 Zahia Sebkhi et al. Vicia disperma The somatic chromosome number of Vicia disperma is 2n =2x=14 for the populations 49/15 and 80/16 (Table 2). DISCUSSION The present study showed a high variation in the number, the asymmetry of chromosomes and the karyo- type formula within Algerian populations of different species in the genus Vicia (Vicia sativa, Vicia monardii, Vicia disperma, Vicia ochroleuca, Vicia onobrychioides and Vicia lutea). The chromosome number of Vicia sativa. subsp. macrocapa is 2n=12 for all Algerians populations. These results were in agreement with those of Raina and Rees (1983), Meriç and Dane (1999), Bisht et al. (1998), Raina et al. (2001), Basbag et al. (2013), Osman et al. (2020), Kartal et al. (2020) and Benlioglu (2021). However, Kar- yotype formula and quantitative analysis are variable among the populations, considered in the present study. In Vicia sativa subsp. sativa, we recorded two chro- mosome numbers (2n=10 and 12). Similar results were reported by El-Bok et al. (2014) in Tunisian acces- sions of the same subspecies. According to Raina and Rees (1983), the multi basic chromosome number is a common phenomenon in the genus Vicia which was assigned as Rebertsonia translocation. The number (2n=12) has been reported by several authors in differ- ent ecotypes of V. sativa subsp. sativa (Meriç and Dane 1999; Navratilova et al. 2003; Gaffazardeh-Namazi et al. 2008; El-Bok et al. 2015; Martin et al. 2018; Osman et al. 2020). Ladizinsky and Shefer (1982) revealed that 2n=10 cytotypes were found in secondary and artificial habitats while 2n=12 cytotypes were found in natural vegetation among dwarf shrubs or in the maquis. Within Vicia sativa aggregate, different experi- ments from several areas revealed that three chromo- Vicia sativa subsp. sativa Vicia sativa subsp. macrocarpa Vicia monardii Vicia ochroleuca Vicia onobrychioides 5µm 75/15, n=6 2 2 2 (population 11/14, 2n = 10); 11/14 11/14, n=5 11/14, n=6 63/16, n=6 65/15, n=6 55/15, n=6 11/14 73/15, n=6 77/15, n=6 66/16, n=7 11/14 51/16, n=7 40/16, n=7 48/16, n=7 n n=7 48/16, n=6 52/15, n=7 11/14 82/14, n=7 Figure 2. Idiograms of Algerian populations corresponding to four species in the genus Vicia L. B M ei o si s o b s e r v e d i n n a t u r a l p o p u l a ti o n s o f t h e g A M ei o si s o b s e r v e d i n n a t u r a l p o p u l a ti o n s o f t h e g M e i o s i s o b s e r v e d i n n a t u r a l p o p u l a t i o n s o f t h E s e r v e d i n n a t u r a l p o p u l a ti o n s o f t h e g D M ei o si s o b s e r v e d i n n a t u r a l p o p u l a ti o n C H s e r v e d i n n a t u r a l p o p u l a ti o G s e r v e d i n n a t u r a l p o p u l a ti o F e r v e d i n n a t u r a l p o p u l a ti o n s o f t h e g e K r v e d i n n a t u r a l p o p J s e r v e d i n n a t u r a l p I s e r v e d i n n a t u r a l p o p u l a ti o Figure 3. Meiosis observed in natural populations of the genus Vicia in Algeria. A- Vicia sativa subsp. sativa (population 11/14, n = 5); B- Vicia sativa subsp. sativa (population 11/14, n = 6); C- 55/14 Vicia sativa subsp. macrocarpa (population 55/14, n = 6); D- 48/16 Vicia monardii (population 48/16, n =7 ); E- Vicia monardii (population 48/16, n = 6); F- Vicia monardii (population 66/16, n = 7); G- Vicia monardii (population 66/16, n = 8); H- Vicia dis- perma (79/15 , n = 6); I- Vicia disperma ( population 80/14, n = 7); J- Vicia lutea (population 83/15, n = 7); K- Vicia onobrychioides (population 82/14 , n = 7) . Scale bar 5 µ. 29Karyological analyses in several Algerian populations of six species of the genus Vicia L. (Fabaceae) some number (2n=10,12 and 14) were reported by several authors (Ladiznsky 1978; Yamanoto and Plitman 1980; Frediani et al. 2004; Arslan 2012; El -Bok et al. 2015; Martin et al. 2018; Kartal et al. 2020). However, the most reported chromosome number is 2n=12 (Ladizensky 1978; Ladizensky and Temkin 1978). Very little work has been done on the species Vicia monardii. The present study follows and completes pre- vious preliminary work (mitosis) carried out by our research team (Melzi 2018) on the same Algerian popu- lations belonging to Vicia monardii. The preliminary results of mitosis (2n=12, 14 and 16) were confirmed through the present study and completed by the study of meiosis, indicating the effective presence of three chro- mosomes numbers (n=6, 7 and 8) in Vicia monardii, confirming that the observed chromosomes are indeed of type A. Furthermore, in Vicia sativa and Vicia monar- dii, the chromosome numbers 2n=10 and 2n=12 were found only in populations originating from regions of low altitude and relatively high rainfall, whereas the number 2n=16 was found in populations originating from regions of high altitude and relatively low rainfall. Therefore, the variability of the chromosome number within and between the populations observed in Vicia sativa and Vicia monardii, would be linked to the eco- logical factors (altitude, rainfall) of the environment of origin of the considered populations. In Vicia onobrychioides and the endemic subspecies V. ochroleuca subsp. Atlantica, the chromosome counts Table 3. The measurement data of chromosome pairs in Algerian populations of V. sativa subsp. sativa. Pop Pair L(µm) (±SD) S (µm) (±SD) TL (µm) ∑TL TLG RL0/00 r Ci Ct Karyotype formula   1 3.19(0.24) 1.08(0.26) 4.27 19.15 3.83 222.97 2.95 25.29 sm 11/14 2 2.98(0.35) 1.1(0.26) 4.08 213.05 2.7 26.96 sm n=5 3 2.7(0.27) 1.07(0.16) 3.77 196.86 2.52 28.38 sm 4sm+1st   4 2.74(0.29) 0.86(0.17) 3.6 188 3.18 23.88 st   5 2.29(0.24) 1.14(0.20) 3.43 179.11 2 33.32 sm 11/14 1 3.5(0.37) 1.27(0.33) 4.77 21.71 3.62 219.71 2.75 26.62 sm n=6 2 3.24(0.11) 1.2(0.30) 4.44 204.51 2.7 27.02 sm   3 3.13(0.24) 1.05(0.23) 4.18 192.53 2.98 25.11 sm   4 2.8(0.61) 0.92(0.21) 3.72 171.34 3.04 24.73 st 5sm+1st   5 1.8(0.66) 0.9(0.31) 2.7 124.36 2 33.33 sm   6 1.3(0.59) 0.6(0.06) 1.9 87.51 2.16 31.57 sm 63/16 1 3.63(0.53) 1.21(0.11) 4.84 22.36 3.73 216.45 3 25 sm n=6 2 3.25(0.7) 1.18(0.22) 4.43 198.12 2.75 26.63 sm 3 3.08(0.32) 1.04(0.27) 4.12 184.25 2.96 25.24 sm 6sm   4 2.65(0.50) 0.96(0.21) 3.61 161.44 2.76 26.59 sm   5 2.11(0.26) 0.86(0.17) 2.9 129.69 2.45 29.65 sm   6 1.68(0.30) 0.78(0.17) 2.46 110.01 2.15 31.7 sm 65/15 1 3.49(0.62) 1.28(0.29) 4.77 21.63 3.61 220.53 2.72 26.83 sm n=6 2 3.39(0.69) 1.04(0.19) 4.43 204..8 3.25 23.47 st 3 2.83(0.65) 0.9(0.28) 3.73 172.44 3.14 24.12 st 4sm+2st   4 2.37(0.62) 0.91(0.26) 3.28 151.64 2.6 27.74 sm   5 2.18(0.67) 0.77(0.12) 2.95 136.38 2.83 26.1 sm   6 1.69(0.54) 0.78(0.16) 2.47 114.19 2.16 31.57 sm 75/15 1 2.92(0.51) 1.61(0.42) 4.53 20.81 3.47 217.68 1.81 35.54 sm n=6 2 2.43(0.21) 1.46(0.42) 3.89 186.92 1.66 37.53 m   3 2.53(0.36) 1.11(0.35) 3.64 174.91 2.27 30.49 sm   4 2.36(0.24) 1.16(0.26) 3.52 169.14 2.03 32.95 sm 4sm+2m   5 1.85(0.11) 1.16(0.39) 3.01 144.64 1.59 38.53 m   6 1.47(0.20) 0.75(0.11) 2.22 106.68 1.96 33.78 sm Abreviations: long arm length (L), short arm length (S), total chromosome length (TL), mean value of total length (TLG), relative length (RL), arm ratio (r), centromeric index (Ci), chromosome type (Ct), median(m), submedian (sm), subterminal(st), standard deviation (SD). 30 Zahia Sebkhi et al. (mitoses) showed, for the first time, 2n=14. Bolkhoskik et al. (1974) reported a chromosome number of 2n =12 for Vicia ochroleuca and Vicia onobrychioides. In the Algerian populations of Vicia disperma and Vicia lutea, the haploid chromosome number recorded is n= 7. This result is in agreement with the reports of Choii (1971) and El Allaoui- Faris (2011), concerning these same species. However, our study highlighted also a new number (n=6), reported for the first time in Vicia disperma. The diploid number (2n =14) was recorded by sever- al authors in Vicia disperma and Vicia lutea (Raina and Ress 1983; Jauzein1995; Venora et al. 2008; Bas Bag et al. 2013). Through this study, we found that the number 2n=14 is more frequent in Algerian populations of the genus Vicia L. According to Holling and Satce (1974) and Tabour et al. (2002), 2n=14 is the most common chromosome number in the genus Vicia L. Thus, Raina and Ress (1983) indicated that the chromosome num- ber 2n=14 is the most primitive in the genus Vicia and reported that the numbers 2n=10 and 2n=12 appeared later, by chromosomal rearrangement. Concerning the chromosome size of the Vicia sativa subspecies, our results (1.9 - 4.84 µm) are very similar to those recorded by Benlioglu (2021) in Turkey (1.68-4.88 µm). However, they are relatively lower than the results reported by El-Bok et al. (2014) and Gaffazardeh-Namazi et al. (2008) on Tunisian (1.71 - 6 µm) and Iranian acces- sions (2.89 - 5. 69 µm) within the same subspecies. Concerning the chromosome size observed in V. sativa subsp. macrocarpa (2.25 - 4.85 µm), it appears to be lower than that reported by Osman et al. (2020) (7.9 - 15.71 µm) and similar to that found by Benlioglu (2021), in some wild populations of the same species, in Turkey (2.54 - 4.98 µm). Within the Algerian populations of the genus Vicia L., the chromosome size of the populations is relatively different from one species to another. According to their size, we can classify the chromosomes as follows: Vicia sativa, Vicia onobrychioides, Vicia ochroleuca and Vicia monardii. Vicia sativa presents the smallest chromosome size and Vicia monardiii is characterised by the largest one. Akpinar and Bilaloglu (1997) signalized that sub- species of Vicia sativa have smaller chromosomes and a lower DNA content than other species of the genus Vicia. Exception made for Vicia sativa, detailed chromo- some measurements and degrees of karyotype asymme- try, indicated through the present study, would be deter- mined, for the first time, in some species (Vicia monar- dii, Vicia ochroleuca and Vicia onobrychioides). Table 4. The measurement data of chromosome pairs in Algerian populations of V. sativa subsp. macrocapa. Pop Pair L(µm) (±SD) S(µm) (±SD) TL (µm) ∑TL TLG RL0/00 r Ci Ct Karyotype formula 73/15 1 2.74(1.94) 0.78(0.13) 3.52 18.49 3.08 190.37 3.51 22.15 st n=6 2 2.6(0.62) 0.84(0.07) 3.44 186.04 3.09 24.41 st 3 2.44(0.52) 0.75(0.09) 3.19 172.52 3.25 23.51 st 2sm+4st   4 2.26(0.31) 0.73(0.13) 2.99 161.7 3.09 24.41 st   5 2.1(0.57) 0.81(0.15) 2.91 157.38 2.59 27.83 sm   6 1.57(0.39) 0.87(0.23) 2.44 131.96 1.8 35.65 sm 55/14 1 3.82(0.53) 1.03(0.33) 4.85 23.6 3.93 205.5 3.7 21.23 st n=6 2 3.38(0.15) 1.02(0.28) 4.4 186.44 3.31 23.18 st   3 2.8(0.42) 1.16(0.30) 3.96 167.79 2.41 29.29 sm   4 2.9(0.31) 0.94(0.15) 3.84 162.71 3.08 24.47 st 1m+2sm+3st   5 2.45(0.38) 0.99(0.21) 3.44 145.76 2.47 28.77 sm   6 1.95(0.43) 1.16(0.38) 3.11 131.77 1.68 37.92 m 77/15 1 2.94(0.50) 1.25(0.42) 4.19 20.27 3.38 206.7 2.35 29.83 sm n=6 2 2.82(0.25) 1.03(0.25) 3.85 189.93 2.73 26.75 sm   3 2.32(0.41) 1.34(0.36) 3.66 180.56 1.73 36.61 sm 6sm   4 2.43(0.40) 0.94(0.16) 3.3 162.8 2.58 28.48 sm   5 1.96(0.41) 1.06(0.23) 3.02 148.99 1.84 35.1 sm 6 1.44(0.45) 0.81(0.11) 2.25 111 1.77 36 sm Abreviations: long arm length (L), short arm length (S), total chromosome length (TL), mean value of total length (TLG), relative length (RL), arm ratio (r), centromeric index (Ci), chromosome type (Ct), median(m), submedian (sm), subterminal(st), standard deviation (SD). 31Karyological analyses in several Algerian populations of six species of the genus Vicia L. (Fabaceae) The presence of different karyotypes formulas within Algerians populations of the genus Vicia may be due to the ecological differences characterizing their geographic origins. According to Benlioglu (2021), the changes in the structure of chromosome morphology can be explained as a gradual alteration which occurred through the evolution of the karyotype during natural or manual selection. In Vicia sativa subsp. sativa, the karyotype formulas reported in the present study, are different from those reported by other authors: 1m +5 st (Namazi et al. 2008; El- Bok et al. 2014), 1m+1sm+3st (El -Bok et al. 2014), 3 m +3st (Osman et al. 2020), 3m+3sm, 2m+3sm and 2sm+4st (Benlioglu 2021), but similar (5sm+1st) to that reported by Martin et al. (2018). The chromosomes observed in Algerian populations of genus Vicia were mainly submetacentric. According to Zuo and Yuan (2011), the predominance of submeta- centric chromosomes indicated that these populations might have retained some of their primitive wild traits. Table 5. The measurement data of chromosome pairs in Algerian populations of Vicia monardii. Pop Pair L(µm) (±SD) S(µm) (±SD) TL (µm) ∑TL TLG RL0/00 r Ci Ct Karyotype formula 66/16 1 4.60(1.00) 1.77(0.5) 6.37     179.43 2.59 27.78 sm   n=7 2 3.86(0.89) 1.76(0.40) 5.62   185.3 2.19 31.31 sm   3 3.82(0.62) 1.42(0.26) 5.24 123.18 2.69 27.1 sm   4 3.54(0.81) 1.44(0.44) 4.98 35.5 5.07 144.68 2.45 28.91 sm 6sm+1m   5 3.04(0.54) 1.72(0.30) 4.76     138.29 1.76 36.13 sm     6 2.82(0.70) 1.72(0.74) 4.54     131.9 1.63 37.88 m     7 2.56(0.53) 1.42(0.34) 3.98     115.63 1.8 35.67 sm   51/16 1 4.26(0.86) 1.38(0.36) 5.64 30.6 4.37 184.07 3.08 24.46 st   n=7 2 3.69(0.41) 1.48(0.32) 5.17 68.73 2.49 28.62 sm     3 3.56(0.47) 1.2 (0.25) 4.76 155.35 2.96 25.21 sm     4 3.44(0.46) 1.27(0.54) 4.71 153.72 2.70 26.96 sm 6sm+1st   5 2.98(0.17) 1.23(0.22) 4.21 137.40. 2.42 29.21 sm     6 2.41(0.54) 0.98(0.27) 3.39 110.63 2.45 28.9 sm     7 1.76(0.31) 1.00(0.09) 2.76 90.08 1.76 36.23 sm   40/16 1 3.66(0.54) 1.35(0.49) 5.01 28.2 4.02 177.91 2.71 26.94 sm n=7 2 3.34(0.78) 1.08(0.20) 4.42 156.69 3.09 24.43 st   3 3.20(0.56) 1.11(0.15) 4.31 153.05 2.88 25.75 sm     4 2.62(0.6) 1.38(0.60) 4.00 142.04 1.89 34.5 sm 5sm+1m+1st   5 2.63(0.32) 1.16(0.34) 3.79 134.58 2.26 30.6 sm     6 2.25(0.68) 1.24(0.13) 3.49 123.93 1.81 35.53 sm     7 1.95(0.28) 1.19(0.14) 3.14 111.5 1.63 37.9 m   48/16 1 4.80(0.61) 1.44(0.04) 6.24 31.8 4.54 196.29 3.33 23.07 st   n=7 2 4.01(0.56) 1.23(0.03) 5.24 164.83 3.26 23.47 st     3 3.53(0.47) 1.33(0.28) 4.86 152.88 2.65 27.36 sm   4 3.13(0.06) 1.46(0.12) 4.59 144.38 2.14 31.8 sm 5sm+2st   5 2.99(0.34) 1.29(0.23) 4.28 134.63 2.31 30.14 sm     6 2.51(0.33) 1.27(0.13) 3.80 119.53 1.97 33.42 sm     7 1.88(0.87) 0.90(0.28) 2.78 87.45 2.08 32.37 sm   48/16 1 2.92(0.25) 1.33(0.25) 4.23 24.3 4.04 174.36 2.18 31.44 sm 5sm+1st n=6 2 3.35(0.37) 1.41(0.21) 4.76 196.20 2.37 29.62 sm   3 3.34(0.27) 1.03(0,13) 4.37 180.13 3.24 23.56 st   4 3.03(0.15) 1.17(0.05) 4.2 173.12 2.58 27.85 sm   5 2.82(0.08) 1.02(0.18) 3.84 158.28 2.76 26.56 sm   6 2.00(0.50) 0.86(0.07) 2.86 117.88 2.32 30.07 sm Abreviations: long arm length (L), short arm length (S), total chromosome length (TL), mean value of total length (TLG), relative length (RL), arm ratio (r), centromeric index (Ci), chromosome type (Ct), median(m), submedian (sm), subterminal (st), standard deviation (SD). 32 Zahia Sebkhi et al. The predominance of subtelocentric chromosomes in V. sativa subsp. macrocarpa populations (73/15 and 55/14) indicated asymmetrical Karyotypes. According to Hanelt and Mettin (1989), the subtelocentric chromo- somes are predominant in the subgenus Vicia and it may have an evolutionary significance. All the chromosomes of the population 77/15 core- sponding to V.sativa subsp. macrocarpa are sub median. According to Paszko (2006), the karyotype of the last population V. sativa subsp. macrocarpa is considered as symmetrical. Several authors (Maxed et al. 1991; Kamel 1999; Weber and Shifino-Wittman 1999; Navratilova et al. 2003) demonstrated that Vicia sativa subsp. sativa is the only subspecies of Vicia sativa that has a metacen- tric pair in its Karyotype. However, our study showed Table 6. The measurement data of chromosomes pairs in Algerian populations of Vicia onobrychioides and Vicia ochroleuca subsp. atlantica. Pop Species BL(µm) (±SD) BC( µm) (±SD) LT ∑LT TLG RL0/00 R Ci Ct Karyotype formula 52/16 V. ochroleuca subsp. atlantica 1 3.60(0.58) 1.26(0.19) 4.86 29.01 4.14 167.52 2.85 25.92 sm   n=7 2 3.48(0.63) 1.26(0.05) 4.74 163.39 2.76 26.58 sm   3 3.17(0.69) 1.32(0.27) 4.49 154.77 2.4 29.39 sm   4 2.85(0.38) 1.27(0.24) 4.12 142.01 2.24 30.82 sm 6sm+1m   5 2.86(0.51) 1.05(0.20) 3.91 134.78 2.72 26.85 sm     6 2.26(0.43) 1.44(0.17) 3.70 127.54 1.56 38.91 m     7 2.03(0.27) 1.6(0.09) 3.19 109.96 1.75 36.36 sm   82/14 V. onobrychioides 1 3.52(0.82) 1.03(0.28) 4.55 25.69 3.67 177.11 3.41 22.63 st   n=7 2 2.81(0.42) 1.3(0.30) 4.11 159.98 2.16 31.63 sm     3 2.66(0.34) 1.28(0.17) 3.94 153.36 2.07 32.48 sm     4 2.66(0.38) 1.04(0.21) 3.70 144.02 2.55 28.10 sm 5sm+1m+1st   5 2.33(0.21) 1.04(0.20) 3.30 128.45 2.24 31.51 sm     6 2.10(0.21) 1.08(0.11) 3.18 123.97 1.94 33.96 sm     7 1.8(0.18) 1.11(0.15) 2.91 113.27 1.62 38.14 m   Abreviations: long arm length (L), short arm length (S), total chromosome length (TL), mean value of total length (TLG), relative length (RL), arm ratio (r), centromeric index (Ci), chromosome type (Ct), median(m), submedian (sm), subterminal (st), standard deviation (SD). Table 7. The asymmetry index values in natural populations of Vicia species in Algeria. Pop Species 2N Ask TF Syi Rec A1 A2 A CVcl MCA 11/14 V. sativa subsp. sativa 10 72.58 27.41 37.77 89.69 0.47 0.09 0.45 9 45.16 11/14 V. sativa subsp. sativa 12 72.63 27,36 37.8 75.85 0.15 0.3 0.45 30 45.02 63/16 V. sativa subsp. sativa 12 73.34 26.97 36.81 76.99 0.14 0.24 0.46 24 46.38 65/15 V. sativa subsp. sativa 12 73.74 26.25 35.47 75.57 0.53 0.24 0.47 24 47.63 75/15 V. sativa subsp. sativa 12 65.16 34.83 53.09 76.56 0.4 0.22 0.3 22 30.63 55/14 V. sativa subsp. macrocarpa 12 73.3 26.69 36.45 81.09 0.47 0.16 0.46 16 45.56 73/15 V. sativa subsp. macrocarpa 12 74.14 21.14 34.21 87.55 0.61 0.13 0.49 13 49.01 77/15 V. sativa subsp. macrocarpa 12 68.62 31.72 46.32 80.62 0.46 0.2 0.36 20 36.68 40/16 V. monardii 14 69.77 30.22 43.21 78.88 0.39 0.15 0.39 15.40 39 48/16 V. monardii 12 71.88 28.11 38.96 84.94 0.43 0.16 0.44 16.00 44 48/16 V. monardii 14 71.87 28.05 38.95 72.77 0.86 0.24 0.44 24.22 44 51/16 V. monardii 14 72.12 27.87 38.73 77.60 0.39 0.23 0.44 23.00 44 66/16 V. monardii 14 68.44 31.7 46.1 79.61 0.20 0.16 0.36 16.40 36 82/14 V. onobrychioides 14 69.59 30.67 43.92 80.65 0.44 0.16 0.39 16 39 52/16 V. ochroleuca subsp. atlantica 14 69.8 30.19 43.25 85.27 0.37 0.14 0.39 14.5 39.61 Abreviations: karyotype asymmtry index(Ask), total form percentage (TF), the index of karyotype symmetry (Syi), the symmetric index (Rec), the intrachromosomal asymmetry index (A1), interchromosomal asymmetry index(A2), degree of asymmetry of karyotype (A), coef- ficient of variation of the centromeric index (CVCL), mean centromeric asymmetry (MCA). 33Karyological analyses in several Algerian populations of six species of the genus Vicia L. (Fabaceae) the presence of one pair of metacentric chromosomes in Vicia sativa subsp. macrocarpa (population n° 55/14). This type of chromosome is also indicated by El Bok et al (2014), Osman et al. (2020) with 3m+2sm+1st and Ben- lioghlu (2021) with 4sm+2st and 3 m +3sm. In Vicia monardii, the symmetrical karyotype is rep- resented by the population 66/16, while asymmetrical kar- yotypes characterize the populations 40/16, 48/16 and 51/16 with a predominance of subtelocentric chromosomes. The study of karyotype asymmetry is one of the most important parameters in the karyomorphology (Astuti et al. 2017; Shamsolshoara et al. 2020; Martin et al. 2018). The intrachromosomal asymmetry gradu- ally increases with centromere shift from median point to terminal point, while the interchromosomal asymme- try increases with more chromosome size heterogeneity (Martin et al. 2018). In the present study, karyotype asymmetry was assessed on the basis of quantitative indices. The val- ues of some indices (MCA, CVcl, Ask %, A1, A2 and A) increase with increasing asymmetry while the values of some indices (TF%, Syi and Rec) decrease with increas- ing asymmetry ( Zuo and Yuan 2011; Eroglu et al. 2013; Atlay et al. 2017). Based on the following asymmetry indices: MCA, AsK, TF, Syi and A, the population 75/15 (V. sativa ssp. sativa) presents the most symmetrical karyotype, while the population 73/15 (V. sativa subsp. macrocarpa) is characterized by the most asymmetrical one. However, the asymmetric karyotypes are different in interchro- mosomal asymmetries. The population 11/14 (2n=10) (V. sativa subsp. sativa) is the most symmetrical karyotype with respect to three indices (CVCL, Rec and A2), where- as the population 48/16 (2n=14) corresponding to Vicia monardii species, is the most asymmetrical karyotype, with respect to only two indices (Rec and A2). The results of the karyotype asymmetry index anal- ysis showed that the chromosomes of the studied species in the genus Vicia vary from median to subterminal. The karyotypes vary from symmetrical to asymmetrical and the intrachromosomal asymmetry was higher than interchromosomal one. These results were in agreement with the reports of Martin et al. (2018) and Benlioglu (2021), within the genus Vicia. According to Kamel et al. (1999), the evolution of Kar- yotype might be inferred from symmetry to asymmetry as a result of pericentric inversion or unequal translocation. The differences in the asymmetry of karyotype were great in the genus Vicia, for which it may be assumed that diversity of the genus has been accompanied by very small changes in the structure of the chromosome (El- Bok et al. 2014). According to Altay et al. (2007), the dif- ference in chromosome morphologies may contribute to the variation of the genera, sections and species. In addition, a number of studies have been carried out, using different methodological approaches to gain a better understanding of the complex phylogenetic rela- tionships between the different species of the Vicia genus. Thus, our results indicated that x = 7 is the most fre- quently observed number in the six Vicia species. Previ- ous work has confirmed that x = 7 is the ancestral num- ber of the genus Vicia (Shiran et al. 2014). For their part, Metin and Hanelt (1964) hypothesised that x = 7 is the most likely chromosome number in the genus Vicia and the numbers x= 6 and x= 5, observed in some species, are derived base numbers. Other authors think that x = 5 is the basic number and x = 6 and 7 are derived num- bers (Schubert et al. 1986). From a phylogenetic point of view, the results of Schaefer et al. (2012) showed that the phylogenetic rela- tionships between species in the genus Vicia are as fol- lows: In section Cracca, Vicia disperma is closely related to Vicia ochroleuca and Vicia monardii, but the degree of relationship between Vicia disperma and Vicia ochro- leuca is less than that found between Vicia disperma and Vicia monardii; these two species (Vicia disperma and Vicia monardi) are the most closely related species. In the Sativa section, Vicia sativa is distant from Vicia lutea, but it is closely related to Vicia angustifolia. On the other hand, Vicia onobrychioides in the Peduncu- latea section is distant from all the other studied species in the Cracca and Vicia sections (Schaefer et al. 2012). According to Shiran et al. (2014), V. sativa subsp. sativa is distant from V. sativa subsp. macrocarpa but it is closely related to V. sativa subsp. angustifolia and these two subspecies (subsp. sativa and subsp. macrocarpa) are distant from Vicia lutea and Vicia disperma. Shiran and Raina (2001) and Shiran et al. (2014) revealed that within the Vicia sativa complex two line- ages are evident in all phylograms. Lineage 1 consists of V. sativa subsp. macrocarpa and V. sativa subsp. angus- tifolia, while lineage 2 includes V. sativa subsp. sativa, V. sativa subsp. cordata, V. sativa subsp. amphicarpa, V. sativa subsp. incisa and V. sativa subp. nigra (Shiran and Raina 2001; Shiran et al. 2014). More recently, results have described a close rela- tionship between Vicia macrocarpa and Vicia narbonen- sis (Osman et al. 2020). CONCLUSION The present study highlighted the characteristics of the chromosomes in the natural populations belonging 34 Zahia Sebkhi et al. to six (06) species (Vicia sativa, Vicia disperma, Vicia monardii, Vicia ohchroleuca, Vicia onobrychioides and Vicia lutea) of the genus Vicia, coming from different eco-geographical zones of North-Eastern Algeria, and the relationships which would be exist with some eco- logical factors of the environment of origin (altitude and rainfall). The results carried out in mitosis and meiosis, showed the presence of some new chromosomal num- bers in Algerian populations of the genus Vicia L. The first number (2n=14, n=7) was observed for the first time in the endemic subspecies Vicia ochroleuca subsp. atlan- tica (Population n° 52/16) and the species Vicia onobry- chioides (Population n° 82/14). The second number (n=6) was observed in Vicia disperma (Population n° 79/15). In Vicia monardii, the three new numbers of chro- mosomes previously observed in mitosis (2n=12, 14 and 16), were confirmed through the study of meiosis (n=6, 7 and 8), indicating that they are indeed A-type chromo- somes. This study has shown that the base number x=7 is the most frequently observed number in the six Vicia species. The observations showed that chromosome num- bers 2n=10 and 2n=12 are more frequently encountered within populations located in regions of relatively low altitude and relatively high rainfall, respectively in the two species Vicia sativa and Vicia monardii. The number 2n =16 is only found in populations of Vicia monardii originating from high altitude and relatively low rainfall regions. Exeption made for Vicia sativa, detailed chromo- some measurements and degrees of karyotype asymme- try would be determined for the first time in the follow- ing species: Vicia monardii, Vicia ochroleuca and Vicia onobrychioides. The karyological variations observed in Algerian populations corresponding to some species of the genus Vicia are clearly detectable in the chromosomal mor- phologies. The chromosomes vary from median to sub- terminal and the karyotype varies from symmetrical to asymmetrical. Ecological conditions, in particular the altitude factor of the geographical origin of the popula- tions, would have an effect on the changes in chromo- some structure. This research is a contribution to the evaluation and valorization of plant genetic resources in Algeria, par- ticularly in the genus Vicia L. The analysis of chromo- somal diversity, based on new data, allowed to answer some questions related to the mechanism of evolution of the species belonging to the genus Vicia L. The charac- terization carried out could play an important role in the conservation and use of these genetic resources through a plant breeding programme. REFERENCES Akpinar N, Bilaloğlu R. 1997. Cytological investigations of certain species of Vicia L. Turk.J. Biol. 21(2): 197- 207. Altay D, Eroğlu HE, Hamzaoğlu E, Koc M. 2017. Karyo- type analysis of some taxa of Dianthus section Ver- ruculosi (Caryophyllaceae, Sileneae).  Turk.J.Bot. 41: 367-374. Arano H. 1963. Cytological studies in subfamily Cardu- oideae (Compositae) of Japan XIV. The karyotype analysis on genus Artemisia (3). Bot. Mag. Tokyo. 76: 459-465. Arslan E, Ertugrul K, Öztürk AB. 2012. Karyological studies of some species of the genus Vicia L. (Legu- minosae) in Turkey. Caryologia. 65(2):106-113. Astuti G, Roma-Marzio F, Peruzzi L. 2017. Traditional karyomorphological studies: can they still provide a solid basis in plant systematics.  Flora Mediterr.  27: 91-98. Axio Vision 1999-2009. By Carl Zeiss. Release 4.8.1. Başbağ M, Hoşgören H, Aydin A.2013. Vicia taxa in the flora of Turkey. Anadolu Tarim Bilim. Derg. 28 (1): 59-66. Benlioglu B. 2021. Cytogenetic diversity and characteriza- tion of Vicia sativa subspecies. Legum. Res. 44: 501- 507. Bisht MS, Kesavacharyulu K, Raina SN. 1998. Nucleolar chromosome variation and evolution in trie genus Vicia. Caryologia. 51(2): 133-147. Bolkhoskikh Z, Grif V, Matvejeva T, Zakharyeva O.1974. Chromosome numbers of flowering plants. Koenig- stein. 322-325. Bouziane Z, Issolah R, Tahar A. 2019. Analysis of the chromosome variation within some natural popula- tions of subterranean clover (Trifolium subterraneum L., Fabaceae) in Algeria. Caryologia. 72(4): 93-104. https://doi.org/1013128/cayologia-164. Chabouni A, Issolah R, Benabdelkader E, Tahar A. 2019. Comportement de quelques populations oasiennes de luzerne pérenne (Medicago sativa L.) dans une zone hyperaride du sud de l’Algérie. Fourrages. 238: 181- 187. Chooi WY. 1971. Variation in nuclear DNA content in the genus Vicia. Genetics. 68: 195-211. Cremonini R, Funari S, Mazzuca S. 1992. Cytology of Vicia species: nuclear structure, Karyological analysis and DNA content. Chromatin. 1: 135-146. https://doi.org/1013128/cayologia-164 35Karyological analyses in several Algerian populations of six species of the genus Vicia L. (Fabaceae) Dobignard A, Chatelain C. 2012. Index of synonymous flora North Africa.V4. Dicotyledonea: Fabaceae- Nymphaeceae. Dyer AF. 1963. The use of lactopropionitic orcein in rapid squash methods for chromosome preparations .Stain. Technol. 38(2): 85-90. El Allaoui-Faris FE, Tahiri H, Molina J.A, EL-Aissami A. 2011. Nombre chromosomique de quelques plantes à fleur. Lagascalia. 31: 69-76. El-Bok S, Zoghlami-Khelil A, Brahim TB, Ouji A, Has- sen H, Lamine O, Jabri C, Douggari R, El –Gazzah M. 2014. Chromosome number and karyotype analy- sis of some taxa of Vicia genus (Fabaceae): revision and description. Int. J. Agric. Biol. 16: 1067-1074. El-Bok S, Zoghlami-Khelil A, Dougari R, Jabri C, Lamine O, El –Gazzah M. 2015. Vicia sativa subsp. sativa. (Fabaceae): New taxonomic division in Tunisia based on karyological data. Pak. J. Agric. Sci. 52(2): 279-283. Excel, 2007. Windows . Microsoft office. Eroğlu HF, Simsek N, Noc M, Hamzaoğlu E. 2013. Kary- otype analysis of some Minuartia L. (Caryophyllace- ae) taxa. Plant Syst. Evol. 299: 67-73. Frediani M, Maggini F, Gelati MT, Cremonini R. 2004. Repetitive DNA sequences as for phylogenetic analy- sis in Vicia genus. Caryologia. 57(4): 379-386. Gaffazardeh- Namazi L, Badrzadeh M, Asghari-Zakaria R. 2008. Karyotype of several Vicia species of Iran. Asian J. Plant Sci. 7: 417-420. Greilhuber J, Septa F.1976. C –banded karyotypes in the Scilla hoheenackeri group, S.persica and Puschkinia (Liliaceae). Plant Syst.Evol. 126: 149-188. Han S, Sebastin R, Wang X, Lee KJ, Cho GT, Hyun DY, Chung JW. 2021. Identification of Vicia native to South Korea using molecular and morphological characterestics. Front. Plant Sci. 12: 608559. Hanelt P, Mettin D. 1989. Biosytematics of the genus Vicia L. (Leguminosae). Ecol. Syst. 20:199-223. Hollings E, Stace CA. 1974. Karyotype variation and evo- lution in the Vicia sativa aggregate. New Phytologist. 73(1): 195-208. Huziwara Y. 1962. Karyotype analysis in some genera of Compositae. VIII. Further studies on the chromo- some of Aster. Am. J. Bot. 49(2): 116-119. Issolah R, Abdelguerfi A. 1999. Chromosome numbers within some spontaneous population of Trifolium species in Algeria. Caryologia. 52: 151-154 Issolah R, Benhizia H, Khalfallah N. 2006. Karyotype variation within some natural populations of sul- la (Hedysarum coronarium L., Fabaceae) in Alge- ria. Genet. Resour. Crop Evol. 53(8): 1653-1664. Issolah R, Benhizia H, Khalfallah N. 2006. Karyotype variation within some natural population of Sul- la (Hedysarum coronarium L., Fabaceae) in Alge- ria. Genet. Resour. Crop Evol. 53(8): 1653-1664. Issolah R, Bouazza L, Tahar A, Terki N, Dehiles I, Man- sour B, Nagoudi T. 2015. Caracterisation écologique de l’habitat naturel du trèfle souterrain (Trifolium subterraneum L., Fabaceae) dans le Nord-Est de l’Algerie. Rev Ecol-Terre Vie. 70(2): 182-193. Issolah R, Sebkhi Z, Bouziane Z, 2022. Ecological Charac- terization of the natural habitats of some Vicia L. species (Fabaceae) in Northeastern Algeria. Pak. J. Bot. 54(6): 2253-2261. http://doi.org/10.30848/PJB2022-6(23). Issolah R, Tahar A, Derbal N, Zidoun F, Meziane MZA, Oussadi A, Djellal L. 2012 . Caractérisation écologique de l’habitat naturel du Sulla (Fabaceae) dans le nord-est de l’Algérie. Rev Ecol-Terre Vie. 67(3): 295-304. Issolah R. (2018). Diversité et valorisation des ressources fourragères en Algérie: cas des genres Trifolium L. et Hedysarum L. (Fabaceae).  Issolah R. et Abbas K., éd. INRAA (2018) Actes du Congrès national sur les res- sources phytogénétiques: évaluation, conservation et valorisation, 22-23.- Jauzein P.1995.Flore des champs cultivés. Ed INRA. Paris. Kamel EA. 1999. Karyological studies on some Taxa of the genus Vicia L. Cytologia. 64: 441-448. Kartal GK, Senbek G, Karaca M, Acikgoz E. 2020. Hybridization studies in Vicia sativa complex. Euphytica. 216: 29-39. Kupicha FK. 1976. The infrageneric structure of Vicia L. Notes Roy. Bot. Gard Edinb. 34: 287-326. Ladizensky G, Temkin R. 1978. The cytogenetic structure of Vicia sativa aggregate. J. Appl. Genet. 53: 33-42. Ladizesky G. 1978. Chromozomal Polymorphism in wild populations of Vicia sativa L. Caryologia. 31(2): 233- 241. Ladizinsky G and Shefer Y. 1982. Polyploidy in the Vicia sativa aggregate. New Phytol. 91(3): 541-547. Lavia GI, Ortiz A.M, Fermandez A. 2009. karyotype studies in wild germplasm of Arachis (Leguminosae). Genet. Resour. Crop Evol. 56(6): 755-764. Levan A, Fredga K, Sandberg AA.1964. Nomenclature for centromeric position on chromosomes. Hereditas. 52(2): 201-220. Lewis GP, Schrire B D, Mackinder BA, Rico L, Clark R. 2013. Linear sequence of legume genera set in a phy- logenetic context. A tool for collections management and taxon sampling. S. African J. Bot. 89: 76-84. Lewis GP, Schrire B, Mackinder B, Lock M. 2005. Leg- umes of the world. London: Kew Press. LPWG. 2017. A new subfamily classification of the legu- minosae based on a taxonomically comprehensive phylogeny. Taxo. 66(1): 44-77. http://doi.org/10.30848/PJB2022-6(23 36 Zahia Sebkhi et al. Martin E, Yildiz KH, Kahraman A, Binzat OK, Eroğlu HE. 2018. Detailed chromosome measurements and Karyotype asymmetry of some Vicia (Fabaceae) taxa from Turkey. Caryologia. 71(3): 224-232. Maxted N, Callimassia MA, Bennett MD. 1991. Cytotax- onomic studies of Eastern Mediterrannean Vicia spe- cies (Leguminosae). Plant Syst. Evol. 177(3): 221-234. Maxted N. 1993. A phonetic investigation of Vicia L. sub- genus Vicia (Leguminosae- Vicieae). Bot. J. Linn. Soc. 111(2): 155-182. Maxted N. 1995. An ecogeographical study of Vicia sub genus Vicia. Systematic and ecogeographic studies on cropgenepools. IPGRI. Rome. Italy. P8. Melzi N. 2018. Etude caryologique chez quelques popula- tions algériennes de l’espèce Vicia monardiiiBoiss. & Reut. Mémoire Master. Univ Blida (Algérie). 48 p. Meriç Ç, Dane F. 1999. Karyological studies on Vicia sati- va subsp.incisa (Beib.) Arc. var. incisa. Turk. J. Bot. 23(1): 63-68. Mettin D, Hanelt P. 1964. Cytosystematic investigations in the species groupe around Vicia sativa L. I.  Die Kulturpflanze. 12: 163-225. Murti RH, Kim HY, Yeoung YR. 2012. Morphologi- cal and anatomical characters of ploidy mutants of Strawberry. Int. J. Agric. Biol. 14(2): 204-210. Navrátilová A, Neumann P, Macas J. 2003. Karyotype analysis of four Vicia species using in situ hybridiza- tion with repetitive sequences. Ann. Bot. 91(7): 921- 962. Osman SA, Ali HB, El-Ashry ZM, El- Khodary SE. 2020. Karyotype variation and Biochimical analysis of Vicia species. Bull Natl Res Cent. 44(1): 1-8. Paszko B. 2006. A critical review and a new proposal of karyotype asymmetry indices. Plant Syst. Evol. 258 (1): 39-48. Peruzzi L, Eroğlu HE. 2013. Karyotype asymmetry: again, how to measure and what to measure? Comp. Cytogenet. 7: 1-9. Potokina E, Vaughan D.A, Eggi EE, Tomooka N. 2000. Population diversity of the Vicia sativa agg.(Fabaceae) in the flora of the former USSR deduced from RAPD and seed protein analyses. Genet. Resour. Crop Evol. 47(2): 171-183. Quezel P, Santa S. 1962. Nouvelle flore de l’Algérie et des régions désertiques méridionales. Tome I. Ed. CNRS, France. Raina S N, Narayan RKJ. 1984. Changes in DNA compo- sition in the evolution of Vicia species. Theor. Appl. Genet. 86(1): 187-192. Raina SN, Mukai Y, Kawaguchi K, Goel S, Jain A. 2001. Physical mapping of 18S-5.8 S and 5S ribosomal RNA gene families in three important vetches (Vicia species) and their allied taxa constituting three spe- cies complexes. Theor. Appl. Genet. 103(6): 839-845. Raina SN, Ress H. 1983. DNA variation between and within chromosome complements of Vicia species. Heredity. 51(1): 335-346. Raveendar S, Lee GA, Jeon YA, Lee JR, Lee YJ, Cho JH, Cho JT. Park JH. Ma KH, Chung JW. 2015. Cross – amplification of Vicia sativa subsp. sativa microsatel- lites across 22 other Vicia species. Molecules. 20(1): 1543-1550. Ruffini Castiglione MM, Frediane MT, Gelati MT, Veno- ra G, Giorgetti L, Caputo P, Cremonini R. 2012. Cytological and molecular characterization of Vicia barbazitae Ten and Guss. Protoplasma. 249(3): 779- 788. Schaefer H, Hechenleitner P, Santos-Guerra A, de Seque- ira M. M, Pennington R. T, Kenicer G, Carine M. A. 2012. Systematics, biogeography, and character evolu- tion of the legume tribe Fabeae with special focus on the middle-Atlantic island lineages. BMC Evolution- ary Biology. 12:1-19. Schubert I, Rieger R, Michaelis A. 1986. Structural and numerical manipulation of the Vicia faba karyotype  : Results and perspectives. Biol. Zbl. 105: 9-17. Shamsolshoara Y, Javadi H, Miri S.M. 2020. Karyomor- phological study of seven species of the genus Astra- galus (Fabaceae) from Iran. Iran J Bot. 26(2): 172-178. Shiran B, Kiani S, Sehgal D, Hafizi A, ul-Hassan T, Chaudhary M. Raina S. N. 2014. Internal transcribed spacer sequences of nuclear ribosomal DNA resolv- ing complex taxonomic history in the genus Vicia L. Genet. Resour. Crop Evol. 61: 909-925. Shiran B, Raina SN. 2001. Evidence of rapid evolution and incipient speciation in Vicia sativa species com- plex based on nuclear and organellar RFLPs and PCR analysis. Genet. Resour. Crop Evol. 48(5): 519-532. Tabur S, Civilek S, Bagci E. 2002. Cytotaxonomic stud- ies on some Vicia L. species growing in the estern mediterranean and southern Aegean regions II. Acta Bot. Hung. 44(1-2): 185-204. Venora G, Blangiforti S, Ruffini Castiglione M, Pignone D, Losavio F, Cremonini R.2002. Chromatin organi- zation and computer aided Karyotyping of Triticum durum Desf.cv Timilia. Caryologia. 55(1): 91-98. Venora G, Ravalli C, Cremonini R.2008. The karyotype as tool to identify plant species: Vicia species belonging to Vicia subgenus. Caryologia. 61(3): 300-319. Watanabe K, Yahara T, Denda T, Kosuge K. 1999. Chro- mosomal evolution in the genus Brachyscome (Aster- aceae, Astereae): Statistical tests regarding correlaion between changes in karyotype and habit using phylo- genetic information. J. Plant Res. 112(2): 145-161. http://Desf.cv 37Karyological analyses in several Algerian populations of six species of the genus Vicia L. (Fabaceae) Weber LH, Shifino –Wittman MT.1999.The Vicia sativa L. aggregate (Fabaceae) in southern Brazil. Genet. Resour. Crop Evol. 46(3): 207-211. Yamanoto K, Plitman AU. 1980. Isozyme Polymorphism in species of the genus Vicia (Leguminosae). Jap. J. Genet. 55(3): 151-164. Yamanoto K. 1973. Karyotaxonomical studies on Vicia L. on the karyotype and character of some annual spe- cies of Vicia. Jap. J. Genet. 48(5): 315-327. Zarco CR. 1986. A new method for estimating Karyotype asymmetry. Taxon. 35(3): 526-530. Zuo L, Yuan Q. 2011. The difference between the hetero- geneity of the centromeric index and intrachromo- somal asymmetry. Plant Syst. Evol. 297(1): 141-145. First cytogenetic study of the Somphong’s rasbora (Trigonostigma somphongsi) (Perciformes, Cyprinidae), a critically endangered species in Thailand Surachest Aiumsumang1, Chavalit Vidthayanon2, Sitthi Kulabtong3, Alongklod Tanomtong4, Sumalee Phimphan1,* Evaluation of the evolutionary process within Populus caspica species from Hyrcanian forests by karyotype analysis Fereshteh Asadi-Corom1,*, Farhad Asadi2, Hossein Mirzaie-Nodoushan1 Chromosomal and genome size variations in Opium poppy (Papaver somniferum L.) from Afghanistan Sayed Zia Rasekh, Ghasem Karimzadeh* Karyological analyses in several Algerian populations of six species of the genus Vicia L. (Fabaceae) Zahia Sebkhi1,4, Rachida Issolah1,*, Nabila Melzi2, Hassina Benmouhoub3, Mohamed Mefti4 Cytogenotoxic and antimicrobial effects of Nezara viridula (L.) (Hemiptera: Heteroptera: Pentatomidae) alcoholic extracts Nicoleta Anca Şuţan1, Mircea Bărbuceanu2, Daniela Bărbuceanu1,*, Ionica Deliu1 Apogamous Isoetes coromandelina L.f. (Isoetaceae) with asynaptic meiosis V. Irudayaraj1,2, A. Benniamin3,*, S. Arokia Raj1 Evaluation of the antigenotoxic potential of fresh bovine whey in onion meristematic roots exposed to Quizalofop-P-tefuryl Florica Colă1, Elena Bonciu1,*, Mugurel Colă1, Nicoleta Anca Șuțan2