Research Article 
Corresponding author: Rachid Amirouche ( ramirouche@usthb.dz ) Academic editor: Alexander Belyayev
© 2022 Naila Chahinez Boukhebache, Nabila Amirouche, Rachid Amirouche.
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Citation:
Boukhebache NC, Amirouche N, Amirouche R (2022) Cytotaxonomic investigations on species of genus Narcissus (Amaryllidaceae) from Algeria. Comparative Cytogenetics 16(1): 5576. https://doi.org/10.3897/compcytogen.v16.i1.78852

This paper provides new cytotaxonomic data on the genus Narcissus Linnaeus, 1753, in Algeria. Populations of seven taxa, N. tazetta Linnaeus, 1753, N. pachybolbus Durieu, 1847, N. papyraceus Ker Gawler, 1806, N. elegans (Haworth) Spach, 1846, N. serotinus sensu lato Linnaeus, 1753, including N. obsoletus (Haworth) Steudel, 1841, and N. cantabricus De Candolle, 1815, were karyologically investigated through chromosome counting and karyotype parameters. N. tazetta and N. elegans have the same number of chromosomes 2n = 2x = 20 with different karyotype formulas. Karyological and morphological characteristics, confirm the specific status of N. pachybolbus and N. papyraceus, both are diploids with 2n = 22 but differing in asymmetry indices. The morphotypes corresponding to N. serotinus sensu lato show two ploidy levels 2n = 4x = 20 and 2n = 6x = 30 characterized by a yellow corona. Some hexaploid cytotypes have more asymmetric karyotype with predominance of subtelocentric chromosomes. They are distinguished by orange corona and may correspond to N. obsoletus. Other cytotype 2n = 28 of N. serotinus was observed in the North Western biogeographic sectors. N. cantabricus was found to be diploid with 2n = 2x = 14, which is a new diploid report in the southernmost geographic range of this polyploid complex.
Amaryllidaceae, chromosomes, karyotype, Narcissus, NorthAfrica, polyploidy
The extended family of the Amaryllidaceae J. S. Hilaire, 1805, is one of the largest families of Asparagales. Among the subfamily Amaryllidoideae Burnett, 1835, species of tribe Narcisseae H.C. Lam et De Candolle, 1806, distributed in about 11 sections (
In the section Tazetteae, four species were recognized in the Algerian flora (
Although belonging to two different sections, N. elegans and N. serotinus would be involved as parents in the origin of natural hybrids such as N. obsoletus (Haworth) Steudel, 1841, and N. miniatus DonnisonMorgan, Koopowitz, Zonneveld, 2005, this latter species was discovered in Southern Spain (
Similar ambiguity arose in Algeria for Narcissus cantabricus De Candolle, 1815, of the section Bulbocodii. This species has been considered first under N. bulbocodium subsp. monophyllus (Durieu) Maire, 1931, then later, as a distinct species (
Despite its central biogeographic position in the southwestern Mediterranean region, Algeria is characterized by an obvious lack of cytotaxonomic data leading to controversies about status and circumscription of many taxonomic units particularly within the Asparagales (
The aim of this study is to fill the gap in the karyological data that links between the floras of the western Mediterranean region. It focuses on the main taxa of genus Narcissus recognized in the flora of Algeria, namely N. tazetta, N. elegans, N. serotinus sensu lato, N. pachybolbus, N. papyraceus and N. cantabricus. Chromosomal counting, structural parameters of the karyotype and the geographical distribution of the polyploidy have been done for each species. Karyological data were linked to morphological and chorological criteria in order to improve taxonomic and nomenclatural knowledge on the genus Narcissus in Algeria.
Plant material used in this study consists of 32 natural populations of genus Narcissus sampled in contrasting ecological conditions along the eastwest biogeographic gradient of the northern Algeria (Table
Coordinates, altitude and bioclimate of the collecting sites in northern Algeria.
Locality  Altitude (m)  Geographic coordinates  Bioclimate^{†}  Collected species^{‡} 

Beni Bahdel  760  34°42'30.49"N, 01°31'08.33"W  Subhumid  N. cantabricus 
Ain Ftouh  831  34°43'23.00"N, 01°27'13.00"W  Subhumid  N. elegans / N. serotinus s.l.* 
Ahfir  1202  34°46'56.40"N, 01°24'54.70"W  Subhumid  N. serotinus s.l. 
Mansourah  1160  34°50'12.60"N, 01°02'20.90"W  Subhumid  N. cantabricus 
ElOurit  739  34°25'00.00"N, 01°16'00.00"W  Subhumid  N. pachybolbus 
Emir Abdelkader  460  35°13'34.50"N, 01°23'33.50"W  Subhumid  N. pachybolbus 
Tessala  801  35°16'09.90"N, 00°46'16.80"W  Subhumid  N. elegans 
Boutlélis  291  35°34'11.40"N, 00°54'00.00"W  Semi arid  N. elegans / N. serotinus s.l. 
Santa Cruz  319  35°42'36.40"N, 00°39'51.10"W  Semi arid  N. elegans 
Miliana  570  36°18'45.60"N, 02°16'22.06"E  Subhumid  N. tazetta 
Mouzaïa  110  36°28'00.00"N, 02°41'00.00"E  Subhumid  N. tazetta 
Chréa  1000  36°28'16.50"N, 02°55'37.40"E  Humid  N. tazetta 
Chenoua  15  36°36'23.00"N, 02°22'21.00"E  Subhumid  N. elegans 
Sainte Salsa  20  36°35'31.00"N, 02°26'58.00"E  Subhumid  N. elegans / N. serotinus s.l. 
Hammam Mélouane  142  36°29'51.70"N, 03°03'29.60"E  Humid  N. tazetta 
Ain Tagourait  219  36°35'00.00"N, 02°37'00.00"E  Subhumid  N. elegans / N. serotinus s.l. 
Béni Messous  50  36°46'44.00"N, 02°58'30.10"E  Subhumid  N. elegans 
Baraki  22  36°39'58.00"N, 03°05'30.00"E  Subhumid  N. tazetta 
Baïnem  248  36°48'00.00"N, 02°58'00.00"E  Subhumid  N. serotinus s.l. 
Bologhine  25  36°48'24.20"N, 03°02'24.50"E  Subhumid  N. papyraceus 
El Alia  30  36°43'12.00"N, 03°10'00.00"E  Subhumid  N. papyraceus 
Yakouren  700  36°43'49.90"N, 04°27'51.00"E  Humid  N. tazetta 
Tizi Tghidet  750  36°44'48.00"N, 04°26'55.00"E  Humid  N. tazetta 
Adekar  500  36°41'00.00"N, 04°40'00.00"E  Humid  N. elegans 
Mont Gouraya  540  36°46'07.20"N, 04°49'50.00"E  Subhumid  N. elegans 
El Aouana  74  36°46'00.00"N, 06°33'00.00"E  Humid  N. elegans 
Aït Ali (Ziama)  970  36°37'04.40"N, 05°28'44.10"E  Humid  N. serotinus s.l. 
Djebel Ouahch  983  36°24'24.50"N, 06°40'32.50"E  Subhumid  N. tazetta 
Sidi Khélifa  864  36°21'08.90"N, 06°17'01.40"E  Subhumid  N. tazetta 
Oued Djenane  302  36°49'17.10"N, 08°37'30.10"E  Humid  N. tazetta 
El Aïoun  282  36°49'04.80"N, 08°37'29.40"E  Humid  N. tazetta 
Tabarka (Tunisia)  80  36°52'21.70"N, 08°43'53.70"E  Humid  N. tazetta 
Habits and flowers of species of genus Narcissus from Algeria. N. tazetta: A, B Sidi Khélifa C, D Hammam Mélouane E Yakouren F–G Tizi Tghidet. N. pachybolbus: H–K ElOurit. N. papyraceus: L–M Bologhine. N. elegans: N–Q. N. serotinus: R–S Ain Ftouh. N. obsoletus: T–U Sainte Salsa. N. cantabricus: V–X Mansourah. Photos by Rachid Amirouche.
Chromosomal analysis was based on metaphase plates of roottip cells from cultivated bulbs. Young roots (6–10 mm long) were pretreated with 1% colchicine for 5–6 hours at room temperature, then fixed in ethanolacetic acid (3:1) for 48 hours and conserved at 4 °C in ethanol 70°.The protocol was adapted from the Feulgen method (
Comparison of the studied species of Narcissus based on the main diagnostic criteria.
Section  Tazetteae  Serotini  Bulbocodii  

Species  N. tazetta  N. pachybolbus  N. papyraceus  N. elegans  N. serotinus sensu lato  N. cantabricus 
Bulb length (mm)  28–58  39–77  37–62  15–38  13–22  19–21 
Bulb width (mm)  15–58  37–68  30–55  12–34  7–20  10–15 
Color of the tunic  black brown  black  black brown  black  black  black 
Leaf number at flowering  2–8  3–5  3–6  1  0  1–5 
Synanthous versus hysteranthous  synanthous  synanthous  synanthous  synanthous  hysteranthous  synanthous 
Length of scape (mm)  80–510  204–496  370–672  102–523  85–240  104–137 
Length of spathe (mm)  32–70  30–50  35–50  17–44  15–30  18–25 
Number of flowers per scape  3–12  9–15  6–13  1–5  1 rarely 2  1 
Hypanthial tube length (mm)  23–44  19–39  14–36  14–30  13–24  23–47 
Hypanthial tube shape  cylindric  cylindric  cylindric  subcylindric narrow  subcylindric  obconic–funnel 
Corona color  yellow–orange  white  white  olive yellow / greenish orange  variable yellow to orange  White rarely white–yellowish 
Corona size  medium  medium  medium  small  small  very large 
Color of tepals  white yellow  white  white  greenish white  greenish white  white 
Pedicel length (mm)  18–52  19–40  27–62  9–40  11–25  3–4 
Stamen position  emergent / not emergent  emergent  not emergent  not emergent  not emergent  emergent 
Karyomorphometric measurements and the homologous chromosome ordering were made using the KaryoType Software 2.0 (
In order to link karyological data to morphological relationships between the studied species, multivariate analyses were carried out using the diagnostic descriptors of vegetative and reproductive parts, some from personal observations (Table
List and abbreviations of the morphological characters used in the multivariate analysis.
Quantitative characters  Shape of the scape  
Bl  Bulb length (mm)  SScyl  cylindrical slightly ridged 
Bw  Bulb width (mm)  SSang  angular ribbed 
Ln  Leaf number  Section of the scape  
Scl  Scape length (mm)  SSfill  filled 
Spl  Spathe length (mm)  SSfist  fistilous 
Nf  Number of flowers by scape  Shape of pedicel  
Pl  Pedicel length (mm)  SPs  smooth 
Hl  Hypanthial tube length (mm)  SPa  angular 
Ns  Number of scape sheath/ scape  Color of tunic bulb  
Ow  Ovary width (mm)  TBcol1  black 
Ol  Ovary length (mm)  TBcol2  brown 
Tl  Outer Tepal length (mm)  Color of corona  
Tl/w  Ratio tepal length / width (mm)  Corcol1  orange bright 
Tel  Tunic extension wrapping the scape (mm)  Corcol2  yelloworange 
Ch  Corona height (mm)  Corcol3  Yellowlemon 
Corcol4  white  
Qualitative characters  Corcol5  orange / orange greenish  
Leaves at flowering  Corcol6  yellow / yellow greenish  
Syn  Synanthous  Shape of hypanthial tube  
Hyst  Hysteranthous  Hysh1  subcylindric large 
Color of the tepals  Hysh2  subcylindric narrow  
Tc1  White  Hysh3  cylindric 
Tc2  yellow  Hysh4  obconic funnel 
Chromosome numbers, ploidy level and characteristics of the karyotypes of the examined populations are summarized in Table
Chromosome number, ploidy level and karyotype characteristics of the examined populations of genus Narcissus in Algeria.
Species/ Populations  Ind/ cells  x  2n  Pl  Karyotype formula  THL  Asymmetry indices  

Stebbins  A1  A2  MCA  CV_{CL}  CV_{CI}  
Narcissus tazetta L.  
Tabarka (Tunisia)  4/16  10  20  2x  4m + 10sm (2sat) + 6st  114.75  3B  0.54  0.36  38.85  35.83  27.56 
Oued Djenane  5/20  10  20  2x  2m + 8sm + 10st  126.12  3B  0.62  0.40  46.68  39.93  29.23 
Sidi Khélifa  3/8  10  20  2x  10sm + 10st  126.17  3B  0.66  0.38  50.27  38.45  20.54 
El Aïoun  4/12  
Hammam Mélouane  7/31  
Yakouren  3/15  
Baraki  5/38  
Mouzaïa  5/15  
Narcissus pachybolbus Dur.  
Emir Abdelkader  5/21  11  22  2x  6m (2sat) + 6sm (2sat) + 8st + 2t  151.92  3B  0.53  0.43  40.18  43.06  40.73 
ElOurit  3/10  
Narcissus papyraceus Ker Gawl.  
Bologhine  6/36  11  22  2x  6m (2sat) + 12sm + 4st  115.50  3B  0.55  0.38  39.86  37.62  29.57 
El Alia  3/10  
Narcissus elegans (Haw.) Spach  
Ain Tagourait  4/8  10  20  2x  2m + 2sm + 14st + 2t (2sat)  145.23  3B  0.72  0.29  58.73  29.00  46.78 
Boutlélis  3/14  10  20  2x  2m + 4sm + 14st  125.32  2B  0.69  0.32  54.15  31.94  32.92 
Santa Cruz  4/9  
Sainte Salsa  7/28  
Béni Messous  3/9  
Chenoua  3/20  
Tessala  4/8  
Narcissus serotinus L.  
Aït Ali  2/30  5  20  4x  2m + 6sm + 12st  66.01  3B  0.69  0.33  55.29  34.40  39.40 
Ain Ftouh  6/10    28                   
Boutlélis  4/10  
Sainte Salsa  6/10  5  30  6x  6m + 6sm + 18st  78.89  3C  0.58  0.39  43.53  39.34  35.20 
Ain Tagourait  4/12  5  30  6x  1M + 11m + 6sm + 12st  101.89  3B  0.47  0.37  34.86  37.15  38.07 
N. obsoletus (Haw.) Steud  
Ain Ftouh  4/10    30                   
Boutlélis  5/15  
Sainte Salsa  3/10  
Narcissus cantabricus DC.  
Mansourah  5/15  7  14  2x  6m + 4sm + 4st  67.80  3A  0.45  0.27  31.33  26.91  29.16 
Somatic metaphases of some species of genus Narcissus from Algeria. A–D N. tazetta: A 2n = 20 El Aïoun B 2n = 20 + 1 Sidi Khélifa C 2n = 20 + 2 Sidi Khélifa D 2n = 20 + 1 Oued Djenane E 2n = 20 Tabarka F N. pachybolbus 2n = 22 Emir Abdelkader G N. papyraceus 2n = 22 Bologhine H–I N. elegans: H 2n = 20 Sainte Salsa I 2n = 20 Ain Tagourait J–M N. serotinus s.l. : J–K 2n = 30 Ain Tagourait, Sainte Salsa L 2n = 28 Ain Ftouh M 2n = 20 Aït Ali. N–O N. cantabricus: N 2n = 14 O 2n = 14 + 1 Mansourah. Black arrows indicate satellites. White arrows indicate supernumerary chromosomes. Scale bar: 10 μm.
Ideograms of the studied species of genus Narcissus in Algeria A N. tazetta 2x (El Aïoun, Yakouren, Hammam Mélouane, Baraki, Sidi Khélifa, Mouzaïa) B N. tazetta 2x Oued Djenane C N. tazetta 2x Tabarka D N. pachybolbus (Emir Abdelkader, El Ourit) 2x. E N. papyraceus (Bologhine, El Alia) F N. elegans 2x (Boutlélis, Tessala, Béni Messous, Chenoua, Sainte Salsa, Santa Cruz) G N. elegans 2x Ain Tagourait H N. serotinus s.l. 6x Ain Tagourait I N. serotinus s.l. 6x Sainte Salsa J N. serotinus s.l. 2n = 28 Ain Ftouh K N. serotinus s.l. 4x. Aït Ali L N. cantabricus 2x Mansourah. Arrows indicate satellites. Scale bar: 10 µm.
≡ Narcissus tazetta subsp. tazetta
This species has many heterotypic synonyms. It is widespread in the north of Algeria and shows a high polymorphism with regard to the color of the perianth and corona (Fig.
Chromosome numbers of the studied species of genus Narcissus from Algeria compared to reports from the literature.
Species  This study  Reports from the literature  

N. tazetta L.  2n = 20 2n = 20 + 1  2n = 14, 20, 22, 24, 28, 30, 32 

2n = 20 


2n = 10, 20, 21, 22, 30, 31, 32 


N. pachybolbus Dur.  2n = 22  2n = 22 

2n = 36 


N. papyraceus Ker Gawl.  2n = 22  2n = 22  Brandham (1942), 
N. elegans (Haw.) Spach  2n = 20  2n = 20 

2n = 30 


N. serotinus L.  2n = 20 2n = 28 2n = 30  2n = 10 

2n = 10 (15) 


2n = 20 


2n = 30 


N. obsoletus (Haw.) Steud.  2n = 30  2n = 30 (20, 29, 31, 45) 

2n = 30 


N. cantabricus DC.  2n = 14 2n = 14 + 1  2n = 14 

2n = 28 

≡ Narcissus tazetta subsp. pachybolbus (Durieu) Baker, 1888
≡ Narcissus papyraceus subsp. pachybolbus (Durieu) D.A. Webb, 1978
Narcissus pachybolbus is narrowly distributed in NW Algeria mainly in the region of Tlemcen. Two populations were sampled at Emir Abdelkader and El Ourit. Both are diploids with 2n = 2x = 22 and share the same karyotype formula 6m (2sat) + 6sm (2sat) + 8st + 2t (Table
≡ Narcissus tazetta subsp. papyraceus (Ker Gawler) Baker, 1888
This species has long been confused with the spontaneous N. pachybolbus due to strong similarities in the flower. N. papyraceus is an ancient cultivated species locally naturalized in Algeria. Two populations were found in the cemeteries of Algiers at Bologhine (ex. Saint Eugène) (Fig.
≡ Hermione elegans Haworth, 1831
N. elegans is encountered mainly in the Tell of the biogeographical sectors of Oranie, Algiers and the Kabylies. Seven representative populations were karyologically examined. The same diploid somatic chromosome number 2n = 20 are observed in all the samples with x = 10 (Table
= Narcissus serotinus var. emarginatus Chabert, 1889
Including N. obsoletus (Haworth) Steudel, 1841
≡ Hermione obsoleta Haworth, 1819
N. serotinus sensu lato is found in the same biogeographical areas than N. elegans, however with a much smaller occurrence. Sometimes, the two species grow in sympatry as in Ain Ftouh, Boutlélis, Ain Tagourait and Sainte Salsa. Five populations belonging to N. serotinus s.l. were examined and three chromosome numbers were observed, 2n = 20, 2n = 28 and 2n = 30 (Table
= Narcissus bulbocodium subsp. monophyllus (Durieu) Maire, 1931
For this baeticorifan species, two populations were sampled in NW Algeria, on clayeymarly slope in Mansourah forest near Tlemcen and on the edge of Lake Beni Bahdel towards the AlgerianMoroccan border. A diploid chromosome number was established 2n = 2x = 14 (Table
In order to link karyological and morphological data of the Algerian species, Principal Components Analysis (PCA) were performed on the basis of the main taxonomic criteria (see Table
Principal Component Analysis of the main taxa of genus Narcissus in Algeria A overall scatter plot of 186 individuals representative of all the studied species B loading of the 24 quantitative and qualitative morphological and floral traits on the circle of correlations (see Table
All of the ten Algerian populations belonging to N. tazetta share the same chromosome number 2n = 20 with sometimes one or two B chromosomes. This somatic number was previously reported by
Due to their morphological similarities,
Morphologically N. tazetta, N. pachybolbus and N. papyraceus constitute three distinct clusters (Fig.
Narcissus elegans and N. serotinus s.l. have been described in all ancient floras of Algeria (
Principal components analysis focused on populations of Narcissus elegans and N. serotinus sensu lato A scatter plot on the first two PC of individuals of each taxon B loading of the morphological variables on the circle of correlations (see Table
In our study, N. elegans has a constant somatic chromosome number 2n = 20 reported also in the literature but often without mention of the base number (
The Algerian populations belonging to N. serotinus sensu lato, display three somatic chromosome numbers 2n = 20, 2n = 28 and 2n = 30. The karyotype formula and the ideograms let suppose a base number x = 5 and consequently tetraploid and hexaploid levels. The tetraploids (2n = 20) were encountered in Sicily (
The presence of N. cantabricus in Algeria, was subject to controversy with N. bulbocodium. N. cantabricus was not mentioned previously in the floras of North Africa.
Overall, this work has contributed with new information supplementing our knowledge on chromosome numbers, karyotypes and ploidy levels of species of the genus Narcissus. The relationships between karyological and morphological characteristics made it possible to confirm and/or update the nomenclature and the taxonomy of species of genus Narcissus in Algeria. Therefore, seven main taxa have been recognized. Into the section Tazetteae, N. tazetta and N. elegans are diploids showing 2n = 2x = 20, while N. pachybolbus and N. papyraceus have 2n = 2x = 22 chromosomes. Section Serotini is represented by both tetraploid and hexaploid N. serotinus (2n = 20, 2n = 30) and also by the hexaploid N. obsoletus (2n = 30). These two species are very similar morphologically and have long been confused with each other in the field. Among N. serotinus type, tetraploids are rare comparatively to hexaploids. The distribution of N. obsoletus (6x) is widespread from west to east through various habitats. N. cantabricus show 2n = 2x = 14 and one recurrent B chromosome and constitute the southernmost diploids, providing new element for our understanding of the distribution of polyploidy within this species.
This work has received a financial assistance from the University of Sciences and Technology Houari Boumediene (USTHB, Algiers, Algeria). It was conducted in the framework of the Project on “Asparagales in Algeria” (PRFU No. D01N01UN160420180016) of the Team Biosystematics, Genetics and Evolution. We are grateful to S. Benhouhou, the manager of the Official Herbarium of ENSA (Algiers). The authors wish to thank also the two anonymous reviewers for their suggestions and comments that have improved our manuscript.
Nabila Amirouche https://orcid.org/0000000299511753
Rachid Amirouche https://orcid.org/0000000234088093