Analysis of the chromosome variation within some natural populations of subterranean clover (Trifolium subterraneum L., Fabaceae) in Algeria

  • Zahira Bouziane Université Abbès Laghrour, Khenchela, Algérie
  • Rachida Issolah INRAA, CRP Mehdi Boualem, Division de Recherche sur les ressources phytogénétiques, BP 37, Baraki, Alger, Algérie
  • Ali Tahar Université Badji Mokhtar, Laboratoire de Biologie végétale et de l’environnement, Annaba, Algérie
Keywords: Chromosomes, Intraspecific variability, Karyotype, Subterranean clover, Trifolium subterraneum L.


Nine natural populations of subterranean clover (Trifolium subterraneum L.) coming from different eco-geographical sites of the North-East Algeria, have been studied for their chromosome number and karyotype features. The study is part of the evaluation and valorization of plant genetic resources of fodder and pastoral interest in Algeria. The results of mitosis detect two groups of populations, and reveal diversity in the number among and within populations. The Algerian populations of T. subterraneum are characterized by two chromosomic formulas. The first formula (2n=2x=16m) (median), more common in most of the studied populations, is in conformity with previous reports in this species. The karyotype of these populations is symmetrical for size and form. The second (2n=2x= 18m), is detected for the first time and described as a new chromosomal formula in T. subterraneum. The latter is relatively more frequent than the first one and characterizes the populations coming from high altitude areas. The karyotype (2n=2x=18m) is relatively symmetrical. At the level of the two established Karyotypes, satellites are highlighted at the first pair. A variation in the size and frequency of these satellites is observed. The species exhibits regular meiotic behaviour, confirming the presence of two basic chromosome numbers (x=8 and 9). The study also highlights the role of ecological factors (Altitude and Rainfall) of the originating environment of Algerian populations in the variation and evolution of chromosome numbers in T. subterraneum. The new cytogenetic data can be exploited in the taxonomy of the species in Algeria in order to select and develop this plant genetic resource in the agricultural field.


Allen O N, Allen E.K.1981. The leguminoseae, Macmillon.C.O, London.
Abdeddaim-Boughanmi k and Kaid-Harche M. 2009. Structure, ultrastructure of the Anther pollen microsporogenesis and morphology of pollen grains of two populations of Lygeum spartum L. in Algeria. Americ J. Agric. and Biol Sci. 4 (3) 201-205.
Abdelguerfi A, AbdelguerfI–Laouar M, M’hammedi Bouzina M, Guittonneau GG, Huguet T, Abbas K, Mebarkia A, Aouani M E and Madani T. 2006. Distribution et écologie de quelques Fabaceae spontanées d’intérêt pastoral et / ou fourrager en Algérie. Workshop international sur la Diversité des fabacées fourragères et de leurs symbiotes : Applications biotechnologiques, agronomiques et environnementales. Alger, 19-22 Février 2006: 27-36.
Alimardani F, Torabi S, Naghavi R, Ebrahimi A. 2014. Study of cytological among some Trifolium species of Iran. Interciencia. 39 (4) 147– 151.
Allard RW. 1965. Genetic systems associated with colonizing ability in predominantly self-pollinated species. In: Baker H.G. and Stebbins G.L. (Eds), Proc. First International Union of Biological Sciences Symp. Academic Press, New York, pp. 49– 75.
Allard RW, Adams J. 1969. Population studies in predominantly self-pollinating species. XII. Intergenotypic competition and population structure in barley and wheat. Am. Nat. 103: 621– 645.
Angulo MD, Sanchez de Rivera A.M. 1975. Studies on Trifolium subterraneum ecotypes. Cytologia, 40: 415– 423.
Angulo MD, Sanchez de Rivera.1977. Comparative chromosomal study of Spanich ecotyes and Autralien cultivars of Trifolium Subterraneum L. Cytologia 42: 473-482.
Angulo MD, Sanchez de Rivera.1983. Karyological studies on spanich taxa of Trifolium subterraneum L. Cytologia 48: 305-312.
Arano H, Saito H. 1980. Cytological studies in family Umbelliferae.V. Karyotypes of seven species in subtribe Seselinae. Chromosoma. 2 (17): 471–480.
Arslan E, Ertuğrul K, Tugay O, Dural H. 2012. Karyological studies of the genus Onobrychis Mill. and the related genera Hedysarum L. and Sartoria Boiss. & Heldr. (Fabaceae, Hedysareae) from Turkey. Caryologia. 65 (1): 11–17.
AxioVision 1999- 2009. By Carl Zeiss. Release 4.8.1.
Azizi N, Amirouche R, Amirouche N. 2016. Karyological investigations and new chromosome number reports in Bellevalia Lapeyrouse, 1808 and Muscari Miller 1758 (Asparagaceae) from Algeria. Comp Cytogen 10: 171-187. doi : 10.3897/CompCytogen.v10i1.6445.
Bacchetta G, Brullo S, Velari TC, Chiapella LF, Kosovel V. 2012. Analysis of the Genista ephedroides group (Fabaceae) based on karyological, molecular and morphological data. Caryologia. 65(1):47–61.
Baaziz K, Benamara-Bellagha M, Pustahija F, Brown CS, Siljak-Yakovlev S, Khalfallah N. 2014. First karyotype analysis, physical rDNA mapping and genome size assessment in four North African Astragalus taxa (Fabaceae). Turkish J. Bot., 38: 1248–1258. doi: 10.3906/bot-1405-40.
Baik N, Maamri F, Bandou H. 2017. Karylogical stydy and meiotic analysis of four species of Aegilops (Poaceae) in Algeria. Carylologia.70 (4) : 324-337.
Benhizia H, Benhizia Y, Ghernoub L, Siljak-Yakovlev S, Khalfallah N. 2013. Meiotic behaviour and karyotype features of endangered endemic fodder species Hedysarum perrauderianum (Fabaceae) in some populations from Algeria.Caryologia _Firenze. doi: 10.1080/00087114.2013.821838.
Benhizia H, Rached-Mosbah O, Benhizia Y, Kouachi A, KhalfallahN. 2003. Etude cytogenetique d’Hedysarum pallidum Desf. Espece endemique Nord-africaine, tolerante a l’antimoine. Universite de Constantine. Sciences et Technologie C. 20: 7–13.
Boubetra K, Amirouche N, Amirouche R. 2017. Comparative morphological and cytogenetic study of five Asparagus (Asparagaceae) species from Algeria including the endemic A. altissimus Munby. Turk J Bot. 41: 588-599. doi: 10.3906/bot-1612-63
Bleier, H. 1925 b. Chromosomenzahlen und Kern volumina in der Gattung Trifolium. Ber. Deutsch Bot. Ges. 43(5): 236-238.
Britten E. 1963. Chromosome number in the genus Trifolium. Cytologia, 28: 428-449.
Brock RD.1953. Species formation in Trifolium subterraneum. Nature, 171:939. doi 10.1038/171939a0. PMID: 13054785.
Cocks, P.S. 1992b. Evolution in sown populations of subterranean clover (Trifolium subterraneum L.)
in South Australia. Aust. J. Agric. Res. 43:1583–1595.
Contandriopoulos J. 1978. Contribution à l'étude cytotaxinomique des Sideritis section Empedoclea (Labiatae). Plant Syst. Evol. 129(4): 277-289.
Darlington CD, Janaki Ammal, E.K. 1945. Chromosome Atlas of cultivated plants. Allen and Unwin, London.
Darlington CD, Wylie AP. 1945. Chromosome Atlas for flowring plants, George Allen and Unwin Ltd., London.
Dyer AF. 1963. The use of lactopropionic orcein in rapid squash methods for chromosome preparations. Stain. Technol. 38: 85–90.
Ellison NW, Liston A, Steiner JJ, Williams WM, Taylor NL. 2006. Molecular phylogenetics of the clover genus (Trifolium– Leguminosae), Mol. Phylogenet. Evol. 39 (2): 688-705.
Eroğlu HE. 2015. Which chromosomes are subtelocentric or acrocentric? A new karyotype symmetry/asymmetry index. Caryologia. 68: 239- 245.
Eroğlu HE, Per S. 2016. Karyotype analysis of Zygoribatula cognata (Oudemans) (Acari: Oribatida: Oribatulidae). Turk Entomol. Derg. 40: 33-38.
Excel 2007. Windows 8. Microsoft office.
Falistocco E, Piccirilli M, Falcinelli M. 1987. Cytotaxonomy of Trifolium subterraneum L. Caryologia. 40: 123–130.
Falistocco E, Marconi G, Falcinelli M. 2013. Comparative cytogenetic study on Trifolium subterraneum (2n = 16) and Trifolium israeliticum (2n = 12). Genome 56: 307–313.
Gillett J M, Taylor NL. 2001. The World of Clovers. Iowa State University Press, Ames, Iowa, USA.
Gladstones JS, Collins WJ.1983. Subterranean clover as a naturalized plant in Australia. J. Aust. Inst. Agric. Sci. 49: 191–202.
Hayward MD, Breese EL. 1993. Population structure and variability. In: Hayward M.D., Bosemark N.O. and Romagosa I. (Eds), Plant breeding. Principles and Prospects. Chapman & Hall, London, pp. 16–29
Hezamzadeh Hijazi S M, Ziaeinasab M. 2006. Karyological study on some of species of Trifolium Genus in Iran. Iran. J. of Biol. 19 (3): 299 - 313.
Hejazi H, Mohsen S, Nasab MZ. 2010. Cytotaxonomy of some Onobrychis (Fabaceae) species and populations in Iran. Caryologia. 63(1):18–31.
Hutton EM, Peak JW. 1954. The Effect of autotetraploidy in five varieties of subterranean clover (Trifolium subterraneum L.) J.agrc .Res. 5: 356-364.
Issolah R, Abdelguerfi A. 1999 a. Variability within 31 spontaneous populations of Trifolium scabrum L., nature of relations with factors of the site of origin. Cahiers options méditerranéennes, 39: 123-127.
Issolah R, Abdelguerfi A. 1999b. Chromosome numbers within some spontaneous populations of Trifolium species in Algeria. Caryologia, 52: 151-154.
Issolah R. 2006. Synthese de travaux réalises sur des populations algeriennes de plusieurs espèces du genre Trifolium L. Workshop international « Diversité des Fabacees fourrageres et de leurs symbiotes : Applications Biotechnologiques, Agronomiques et Environnementales ». Alger, Algérie. 19-22 Février 2006: 81-83.
Issolah R, Benhizia H, Khalfallah N. 2006. Karyotype variation within some natural populations of Sulla (Hedysarum coronarium L., Fabaceae) in Algeria. Genet Resourc Crop. Evol. 53(8):1653– 1664.
Issolah R, Khalfallah N. 2007. Analysis of the morpho-physiological variation within some Algerian populations of Sulla (Hedysarum coronarium L.; Fabaceae). J. Biol. Sci., 7: 1082-1091.
Issolah R, Tahar A, Derbal N, Zidoun F, Ait Meziane MZ, Oussadi A, Dehiles I, Bradai R, Ailane M, Terki N, Aziez F, Zouahra, A, Djellal L. 2012. Caracterisation ecologique de l’habitat naturel du Sulla (Fabaceae) dans le Nord-Est de l’Algérie. Rev. Ecol. (Terre et Vie), 67: 295-304.
Issolah R, Bouazza L, Tahar A, Terki N, Dehiles I, Mansour B, Nagoudi T. 2015. Caracterisation ecologique de l'habitat naturel du trefle souterrain (Trifolium subterraneum L., Fabaceae) dans le Nord- Est de l'Algerie. Rev. Ecol., 70: 182-193.
Issolah R, Tahar A, Sadi S, Adjebi M, Alane F, Chelling- Siziani, Lebied M. 2016. Analysis of the behaviour and the chemical composition within populations of Trifolium subterraneum L. J.Biol.Sci., 16: 148-154. doi:10.3923/jb.2016.148.154.
Jahier J, Chevre AM, Delourme R, Eber F, Tangay AM. 1992. Techniques de cytogénétique végétale. INRA. Paris, pp. 1-184.
Kiran Y, Sahin A, Turkoglu I, Kursat M, Emre I. 2010. Karyology of seven Trifolium L. taxa growing in Turkey. Acta Biologica Cracoviensia Series Botanica, 52: 81-85.
Kliphuis E. 1962. Chromosome numbers of some annual Trifolium species, occuring in the Netherlands. Acta Bot. Neerland 11: 90-92.
Katznelson J. 1974. Biological flora of Israel. 5. The subterranean clovers of Trifolium subsect. Calycomorphum Katzn. Trifolium subterraneum L. (sensu lato). Isr J Bot 23: 69-108.
Katznelson J, Morley F. 1965 a. Speciation processus in Trifolium suberraneum L. Israel J. Bot. 14:15-35.
Levan A, Freda K, Sandberg AA 1964. Nomenclature for centromeric position on chromosomes. Hereditas. 52: 201- 220.
Marshall DR, Broue´ P. 1973. Outcrossing rates in Australian populations of subterranean clover. Aust. J. Agric. Res. 24: 863–867.
Masson P. 1997. Des prairies de très longue durée avec des espèces annuelles à ressemis spontané : les pâtures à trèfle souterrain. Fourrages, 153: 139-146.
Medoukali I, Bellil I, Khelifi D. 2015. Evaluation of Genetic Variability in Algerian Clover (Trifolium L.) Based on Morphological and Isozyme markers. Czech J. Genet. Plant Breed. 51(2): 50–61.
Muñoz-Rodríguez AF. 1995. Trifolium sect. Paramesus and Sect. Trifoliastrum in the Iberian Peninsula II. Karyological study. Stud Bot. 14:103–128.
Pecetti L, Piano E. 1998. Leaf size variation in subterranean clover (Trifolium subterraneum L. sensu lato). Gene.Res.crop. Evol. 45: 161–165
Pecetti L, Piano E. 2002. Variation of morphological and adaptive traits in subterranean clover populations from Sardinia (Italy). Gene.Res.crop. Evol. 00:0-1.
Piano E. 1984. Preliminary observations on the structure and variability of Sardinia populations of subterranean clover. Genet. Agr. 38: 75–90.
Piano E, Spanu F, Pecetti L. 1993. Structure and variation of subterranean clover populations from Sicily, Italy. Euphytica 68: 43–51.
Piano E, Pecetti L. 1996. Selecting subterranean clover varieties for Mediterranean environments in Italy. In: Parente G., Frame J. and Orsi S. (Eds), Grassland and land use system. Proc 16th EGF Meet. ERSA, Gorizia, pp. 283–286.
Piluzza G, Pecetti L, Bullitta S, Piano E. 2005. Discrimination among subterranean clover (Trifolium subterraneum L. complex) genotypes using RAPD markers. Genetic Resources and Crop Evolution 52, 193–199.
Pritchard A J. 1969. Chromosome numbers in some species of Trifolium. Austral. J. Agric. Res. 20: 883- 887.
Quezel P, Santa S. 1962. Nouvelle flore de l’Algerie et des régions desertiques meridionales. Tome I. Ed. CNRS, France.
Rossiter RC, Collins WJ. 1988 a. Genetic diversity in old subterranean clover (Trifolium subterraneum L.) populations in Western Australia. 1. Pastures sown initially to the Dwalganup strain. Aust. J. Agric. Res. 39: 1051–1062.
Rossiter RC, Collins WJ. 1988 b. Genetic diversity in old subterranean clover (Trifolium subterraneum L.) populations in Western Australia. 2. Pastures sown initially to the Mount Barker strain. Aust. J. Agric. Res. 39: 1063–1074.
Romero Zarco C. 1986. A new method for estimating karyotype asymmetry. Taxon 35:526-530.
Rutland J.P. 1941. The menton catalogue. A list of chromosome numbers of British plants. Suppl. 1. New Phytol. 40: 210.
Senn HA. 1938. Chromosome number relationship in the Leguminosae. Biblioth. Genet. 7: 175- 336. Siljak-Yakovlev S 1996. La dysploïdie et l’évolution du caryotype. Bocconea. 5: 211–220.
Siljak-Yakovlev S, Godelle B, Zoldos V, Vallès J, Garnatje T , Hidalgo O.2017. Evolutionary implications of heterochromatin and rDNA in chromosome number and genome size changes during dysploidy: A case stydy in Reichardia genus. Plosone 12(8): e0182318. 10.1371/ journal. Pone .0182318.
Singh R J. 2018. Practical manual on plant cytogenetics. CRC Press, Boca Raton, Boca Raton, FL 33487-2742 © 2018 by Taylor & Francis Group, LLC, pp 1-347.
Taylor NL. (Editor). 1985. Clovers around the world. In Clover science and technology. American Society of Agronomy, Madison, Wisc. pp. 1–6.
Uslu E. 2012. Karyology of nine Trifolium L. taxa from Turkey. Caryologia. 65(4): 304–310.
Vizintin L, Javornik B, and Bohanec B. 2006. Genetic characterization of selected 15 Trifolium species as revealed by nuclear DNA content and ITS rDNA region analysis, 16 Plant. Sci. 170: 859-866.
Yates JJ, Brittan NH. 1952. Cytological studies of subterranean clover (Trifolium subterraneum L.). Aust. J. Agric. Res. 3: 300–304. doi: 10.1071/ AR9520300.
Weselxen H. 1928. Chromosome number and morphology in Trifolium. Univ .of Calif. Publ. Agric. Sci.2: 255-376.
Zohary D, Katznelson J. 1958. Two species of subterranean clover in Israel. Aust. J. Bot. 6: 177–182. doi: 10.1071/BT9580177.
Zohary M, Heller D. 1984. The genus Trifolium. Israel Academy of Sciences and Humanities. Jerusalem. pp. 1-606.
How to Cite
Bouziane, Z., Issolah, R., & Tahar, A. (2019). Analysis of the chromosome variation within some natural populations of subterranean clover (Trifolium subterraneum L., Fabaceae) in Algeria. Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics, 72(4), 93-104.