Chromosome numbers and karyotype of five species from Seriphidium (Asteraceae)

Chromosome data are important for elucidating intergeneric relationships and delimiting infrafamilial tribes in plants. This paper reports karyological data for 12 Seriphidium (Bess.) Poljak. species from China, of these, five count as new reports and the others have been reported elsewhere. We also report the tetraploid level in S. schrenkianum for the first time. The karyotype formulae and types for the five new reports are: S. schrenkianum (Ledeb.) Poljak. 2n = 4x = 36 = 22m + 12sm + 2st, S. sublessingianum (Kell.) Poljak. 2n = 2x = 18 = 14m + 4sm and S. transiliense (Poljak.) Poljak. 2n = 2x = 18 = 4M + 6m + 2m(SAT) + 4sm + 2sm(SAT), belong to 2A type; S. cinum (Berg. ex Poljak.) Poljak. 2n = 2x = 18 = 8m + 10sm and S. sawanense Y. R. Ling et C. J. Humphries 2n = 2x = 18 = 10m + 2m(SAT) + 6sm, belong to 2B type. Furthermore, we estimated karyotype asymmetry in the 12 species by using an intrachromosomal asymmetry index and an interchromosomal asymmetry index. The data increase information about the karyosystematics of Seriphidium.     

Cytogenetics, geographical distribution, pollen stainability and fecundity of five diploid taxa of Santolina rosmarinifolia L. aggregate (Asteraceae: Anthemideae)

The chromosome analysis of Santolina rosmarinifolia subsp. rosmarinifolia, S. oblongifolia, S. semidentata subsp. semidentata, S. semidentata subsp. melidensis, S. canescens and the hybrid complex (S. rosmarinifolia subsp. rosmarinifolia, S. oblongifolia and their putative hybrids) shows that all the taxa are diploids (2n = 2x = 18; 18 + 1 or more B chromosomes, with 2n = 19, 20 only in the hybrid complex). The results show a conserved general structure of the karyotype (14m + 2sm + 2st), but in S. semidentata subsp. melidensis it is variable, with 14m + 2sm + 2st in ten individuals, 14m + (1m ? 1sm) + (1 m ? 1st) in nine individuals and 12m + (1m ? 1sm) + (1m ? 1st) + 2st + 1B in five individuals. Tetraploid individuals occurred in the diploid populations of S. rosmarinifolia subsp. rosmarinifolia and S. canescens, and their autopolyploid origin is discussed. Multivalent configurations at diakinesis, simple and double chromosome bridges and delayed disjunction of homologous and non-homologous chromosomes at anaphase I have negative effects on pollen stainability. The mean fructification percentage is moderate. The results suggest that the complex is a mosaic of introgressive hybrids.     

A cyto-evolutional study of Campanumoea Blume (Campanulaceae) and a possible pathway for secondary karyotype formation

Campanumoea is a small genus in the family Campanulaceae, with species divided into sections Campanumoea and Cyclocodon. Sixteen accessions from Campanumoea and related genera native to China were used to study their karyotype. The results showed that chromosome characteristics were different between the two sections. For Campanumoea, the karyotypic formula was 2n = 2X = 2m + 12sm + 2st = 16,3A and for Cyclocodon it was 2n = 2X = 6m + 12sm = 18,3B. These data, combined with chromosomal length characteristics, support the restoration of section Cyclocodon as a genus. However, the incorporation of section Campanumoea into Codonopsis requires more evidence. Comparison of chromosomal length and haploid set length revealed that chromosomal segment rearrangements occurred within sections of Campanumoea and between genera, with the difference within sections being greater than that between genera. Therefore, chromosomal segment rearrangements are present in Campanulaceae, implying that chromosomal segment rearrangement plays an important role in the evolution of diversity in Campanulaceae. By comparing the chromosomal characteristic in section Campanumoea and the genus Adenophora, we concluded that the secondary chromosome type such as n = 17, 18 would be derived by autopolyploidization of n = 9, and by chromosome fusion.     

Cytogenetics, Geographical Distribution, Pollen Stainability and Fecundity of Santolina impressa (Asteraceae: Anthemideae)

Santolina impressa (2n = 2x = 18) is an endemic species of Portugal, with restricted geographical distribution. The present study aimed to explore its chromosomal variation in respect to chromosome morphology, meiotic behaviour, and effects on pollen stainability and fecundity. Its karyotype formula was found to be either 12m + 2m^sat + 2sm^sat + 2st (75% of the individuals) or 12m + 2m^sat + 3sm + 1sm^sat (25% of the individuals). Univalents were observed in 29.21% of the meiocytes. Chromosome fragments due to breakage in the chromosome arm were observed in 10 meiocytes only (11.23% of the meiocytes). Chains and rings of trivalents were observed in 14.60% of the meiocytes (one trivalent per meiocyte was observed). Chains and rings of quadrivalents were observed in 21.34% and 11.23% of the meiocytes, respectively, with a range of 0–1 per cell. Nine plants (40.90% of the total) with 2n = 2x = 18 + 2B showed a quadrivalent configuration in diakinesis. Twenty abnormal anaphases with delayed disjunction of the four non-homologous and the two homologous chromosomes were observed. Simple chromosome bridges without fragments and interchromosomal adhesions were observed in 35.95% of the anaphases analyzed. B chromosomes showed bivalent association in diakinesis and their segregation at anaphase I was normal. Pollen was found to be fertile (mean ± s.d. = 89.57 ± 47.14%); the effect of univalent frequency and frequency of abnormal anaphase I on pollen stainability was strong and statistically significant.     

Cytogenetics of Alismatales s.s.: chromosomal evolution and C-banding

Ten species of Alismatales, five of Alismataceae, four of Limnocharitaceae and one of Hydrocharitaceae were studied with regard to chromosome number, chromosome morphology, and pattern of Giemsa C-bands. The genus Echinodorus had a diploid chromosome number of 22 for all species that were analyzed and a karyotypic formula of 2m + 20a. For the family Limnocharitaceae, Hydrocleys nymphoides had a diploid chromosome number of 16, Hydrocleys martii (4m + 2sm + 10a) had a diploid chromosome number of 16, Limnocharis flava had a diploid chromosome number of 20 and L. laforestii (4m + 16a) had a diploid chromosome number of 20. The only species of Hydrocharitaceae that was studied exhibited a karyotype that consisted of a diploid chromosome number of 28 and a karyotypic formula of 4m + 6sm + 4a. The distribution pattern of the C-banded karyotype in Echinodorus showed four blocks of constitutive heterochromatin in two smaller acrocentric pairs that corresponded to the heterochromatic NORs. In E. lanceolatus, 14 bands in the termini of the arms beyond the heterochromatic NORs of seven acrocentric pairs were also observed. Idiograms are presented and the karyotypic evolution patterns for the studied groups are discussed.     

A new species of Lysimachia (Myrsinaceae) from Dabieshan Mountain,China

A new species of Myrsinaceae, Lysimachia jinzhaiensis S.B. Zhou & K. Liu, is described and illustrated from Dabieshan mountain, Anhui, China. It is similar to L. christiniae in the prostrate stem, opposite leaves, yellow flowers born singly in leaf axils, but differs by being glabrous throughout or glandular on young parts, and having quadrangular stem, corolla with densely transparent glandular, orange-red corolla base with lobes being significantly overlapping and contorting to left in bud. Moreover, L. jinzhaiensis have a different karyotype, formulated as 2n = 2m + 2sm + 10st (2SAT) + 10t.     

Ploidy variation in Trigonobalanus verticillata (Fagaceae)

Cytological studies were carried out on two populations of Trigonobalanus verticillata in Yinggeling, Hainan Province, China (YGL), and Fraser’s Hill, Malaysia (FH). In the two populations, the pattern of interphase nuclei was of the simple chromocentre type, the mitotic prophases were of the proximal interstitial type and chromosome numbers were 2n = 2x = 14 (YGL) and 2n = 6x = 42 (FH), representing diploid and hexaploid respectively. The basic chromosome number of the species was x = 7; this is a unique number in the Fagaceae and in the Fagales as well. The characteristic karyotype of the diploid population in China was 2n = 14 = 8m + 4sm + 2st. Based on the new finding of the chromosome number in this study, we clarify the previous dispute about the basic chromosome number and propose that the two Asian Trigonobalanus species have closer affinity.     

XX/XO, a rare sex chromosome system in Potamotrygon freshwater stingray from the Amazon Basin, Brazil

Potamotrygonidae is a representative family of South American freshwater elasmobranchs. Cytogenetic studies were performed in a Potamotrygon species from the middle Negro River, Amazonas, Brazil, here named as Potamotrygon sp. C. Mitotic and meiotic chromosomes were analyzed using conventional staining techniques, C-banding, and detection of the nucleolus organizing regions (NOR) with Silver nitrate (Ag-NOR). The diploid number was distinct between sexes, with males having 2n = 67 chromosomes, karyotype formula 19m + 8sm + 10st + 30a, and fundamental number (FN) = 104, and females having 2n = 68 chromosomes, karyotype formula 20m + 8sm + 10st + 30a, and FN = 106. A large chromosome, corresponding to pair number two in the female karyotype, was missing in the male complement. Male meiotic cells had 33 bivalents plus a large univalent chromosome in metaphase I, and n = 33 and n = 34 chromosomes in metaphase II. These characteristics are consistent with a sex chromosome system of the XX/XO type. Several Ag-NOR sites were identified in both male and female karyotypes. Positive C-banding was located only in the centromeric regions of the chromosomes. This sex chromosome system, which rarely occurs in fish, is now being described for the first time among the freshwater rays of the Amazon basin.     

Considerations on karyotype evolution in the genera Imparfinis Eigenmann and Norris 1900 and Pimelodella Eigenmann and Eigenmann 1888 (Siluriformes: Heptapteridae)

Heptapteridae is one of the fish families of the order Siluriformes with a wide distribution throughout the basins of the Neotropical region. The genera Imparfinis and Pimelodella comprise few species and/or populations with some chromosome information. Specimens of Imparfinis schubarti, Imparfinis mirini, and Pimelodella meeki from different sites located in the Paranapanema River Basin/PR/Brazil were cytogenetically analyzed. The two species of the genus Imparfinis exhibited 2n = 58 and FN = 116: I. schubarti, with a karyotypic formula of 30m + 28sm and I. mirini with a karyotype formula of 36m + 22sm. P. meeki presented a karyotype of 2n = 46 characterized by 26m + 14sm + 6st and FN = 92, confirming a variability in the 2n of the Heptapteridae family. Both Imparfinis species exhibited interstitial NORs in pair 1, coincident with a secondary constriction; P. meeki presented NORs located in the terminal position on the short arm of pair 17. All AgNORs were coincident with 18S rDNA probe and CMA₃ positive. P. meeki showed a small amount of heterochromatin rich in AT and GC bases. The heterochromatin in I. schubarti was CMA₃ positive. In I. mirini the heterochromatin was DAPI-positive. Furthermore, the long arm of one of the chromosomes of pair 19 revealed the presence of heterochromatic heteromorphism only in male individuals. After meiotic analyses, this heteromorphism could be easily identified in the pachytene and metaphase I stages, and was heteropyknotic and DAPI positive. This feature may be an indication of initial differentiation of sex chromosomes in I. mirini, increasing the great karyotypic variability within this family of fish.     

New cytological data in Ophrys sect. Pseudophrys Godfery and comparative karyomorphological studies in Ophrys L. (Orchidaceae)

Eight species of Ophrys sect. Pseudophrys (Orchidaceae) were cytogenetically studied. The analysed species possess the most symmetrical karyotypes of the genus (M_CA ranged from 10.21 to 15.87 and CV_CL from 19.61 to 23.93) with 2n = 2x = 36, being composed of mainly metacentric chromosomes. The karyotype formulae were: 36m for Ophrys archimedea, 32m + 4sm for O. flammeola, 32m + 4sm for O. funerea, 36m for O. laurensis, 36m for O. lojaconoi, 34m + 2sm for O. lucifera, 34m + 2sm for O. obaesa and 36m for O. pallida. Karyotype morphometric characters were evaluated by calculating M_CA and CV_CL for the assessment of karyotype asymmetry, and CV_CI for the evaluation of heterogeneity in the position of the centromeres. The relationships between species were thus finally elucidated. The species characterised by wide distribution show greater karyomorphological distance than those with restricted distribution. The possible evolutionary role of chromosomal rearrangements as well as gene mutations in the speciation of Ophrys is discussed.     

Karyological studies on eight species of <Emphasis Type="Italic">Onobrychis</Emphasis> genus in Turkey

This study used karyological techniques to determine the chromosome numbers and morphology of eight species of Onobrychis L. (O. caput-galli (L.) Lam, O. aequidentata (Sibth. & Sm.) d’ Urv, O. fallax Freyn & Sint. var. fallax, O. lasiostachya Boiss, O. viciifolia Scop., O. oxyodonta Boiss. subsp. armena (Bois. & Huet) Aktoklu, O. hypargyrea Boiss. and O. cappadocica Boiss.). The results of this study determined the chromosome numbers of O. cappadocica as 2n = 16; O. viciifolia as 2n = 28 and the other species as 2n = 14 The karyotypes of species consisted of median-centromeric (m) or submedian-centromeric (sm) chromosomes. However, O. oxyodonta Boiss. subsp. armena (Bois. & Huet) Aktoklu was found to have only the median-centromeric (m) chromosomes. According to the results of the present study, of the eight Onobrychis taxa, only O. hypargyrea has a pair of satellite chromosomes (sat-chromosome). Furthermore, this study detected karyotype asymmetry.     

Pollen morphology of some taxa of Vicia L. subgenus Vicia (Fabaceae) from Turkey

In this study, the pollen morphology of 11 taxa belonging to Atossa (Alef.) Asch. & Graebner, Hypechusa (Alef.) Asch. & Graebner, Peregrinae Kupicha, Wiggersia (Alef.) Maxted, Vicia L. and Narbonensis (Radzhi) Maxted sections of the genus Vicia L. subgenus Vicia (Fabeae, Fabaceae) naturally growing in Turkey has been studied using Light Microscopy (LM) and Scanning Electron Microscopy (SEM) to evaluate the taxonomic relevance of pollen characters. Twelve morphometric characters are analysed with one-way analysis of variance and Tukey’s honestly significant difference test for multiple comparisons. Of the taxa studied V. narbonensis var. narbonensis (sect. Narbonensis) has the largest pollen grains (P = 51.98 μm × E = 30.52 μm) and V. lathyroides (sect. Wiggersia) has the smallest pollen grains (P = 27.71 μm × E = 20.14 μm). The pollen grains are subprolate to perprolate (P/E = 1.16–2.11), but the prolate shape occurs in the majority of the taxa. The regular pollen grains of all taxa are trizonocolporate, isopolar, and released in monads. Ornamentation of the mesocolpium is psilate-perforate in V. lathyroides (sect. Wiggersia), reticulate-rugulate in V. narbonensis var. narbonensis (sect. Narbonensis), (micro)reticulate in V. sericocarpa var. sericocarpa (sect. Hypechusa), V. sativa subsp. sativa (sect. Vicia) and V. grandiflora var. grandiflora (sect. Vicia), and reticulate-perforate in the remaining taxa. The apocolpium and colpus area are psilate or perforate in all taxa except V. sericocarpa var. sericocarpa (sect. Hypechusa) and V. sativa subsp. sativa (sect. Vicia), which exhibit the obscurely reticulate-perforate pattern. Several palynological features have taxonomic importance in sectional level: polar axis, equatorial diameter, pollen shape (P/E ratio), colpus length, colpus width, size of pori, porus length/width ratio, lumina diameter, muri thickness and ornamentation. The results also indicate that pollen characters can be useful in distinguishing the examined taxa.     

Karyotypic, palynological, and RAPD study on 12 taxa from two subsections of section Idaeobatus in Rubus L. and taxonomic treatment of R. ellipticus, R. pinfaensis, and R. ellipticus var. obcordatus

Twelve taxa belonging to two subsections of section Idaeobatus in Rubus L. from southwestern China were characterized by karyotypic, palynological, and random amplified polymorphic DNA (RAPD) data as follows: (1) The 12 taxa were all diploid species (2n = 2x = 14), among which the chromosome counts for R. mesogaeus var. oxycomus, R. subtibetanus, R. ellipticus var. obcordatus, R. inopertus var. echinocalyx, and R. stans were reported for the first time; (2) All taxa except for R. ellipticus and R. pinfaensis could be distinguished from each other by karyotype, pollen morphology, and RAPD markers. Karyotypes were mainly characterized by the difference in numbers and positions of submetacentric chromosomes and chromosomes with satellited pair, the index of the karyotypic asymmetry, and the ratio of the longest to the shortest chromosome. Pollen morphology were mainly characterized by the discrepancy in specific pollen size, P/E ratio, colpi width, distance between the apices of two ectocolpi, and exine ornamentation characters. The cluster results based on RAPD markers were consistent with morphology classification except for R. pinfaensis; (3) Based on the general data of karyotypic, palynological, and RAPD, R. ellipticus var. obcordatus should be treated as a species R. obcordatus, R. ellipticus and R. pinfaensis should be combined as R. ellipticus, and it was more reasonable to place the combinants and R. obcordatus into subsection Stimulantes rather than into subsection Pungentes.     

Karyological features and banding patterns in Arachis species belonging to the Heteranthae section

The section Heteranthae of Arachis is endemic to Brazil, occurring mainly in the semi-arid northeastern region. The section is considered derived within the genus and includes only annual herbs. Most previous cytological evaluations were restricted to chromosome numbers and morphology. The present approach comprised karyomorphological evaluation in 10 accessions from five species of this section, including standard staining and fluorochrome banding [chromomycin A3 (CMA)/4′,6-diamidino-2-phenylindole (DAPI)]. All accessions presented diploid chromosome numbers (2n = 20) with a prevalence of metacentric to submetacentric chromosome morphology. Arachis dardani, Arachis pusilla, and Arachis interrupta presented karyotypic formula 18m + 4sm and satellite type 2, while Arachis sylvestris and Arachis giacomettii presented 16m + 4sm and satellite type 10. Despite the conserved morphological features, higher diversity was detected in terms of size and number of GC-rich (CMA⁺) heterochromatic blocks among the species; however, all of them were located in the pericentromeric regions. The species A. pusilla presented the highest number of GC-rich blocks, present in all chromosomes of the complement. Based on the data obtained and considering literature data, we suggest that A. dardani and A. interrupta occupy a basal position in the group due to their moderate asymmetry and satellite type. At least in A. pusilla, the constitutive heterochromatin seems to have suffered recent modifications of its constitution, in contrast to other species that present pericentromeric CMA⁺ blocks in all chromosomes. A. giacomettii and A. sylvestris are closely related to each other and also similar to the previously studied Arachis seridoensis, revealing two clear-cut subgroups within the section from the karyological point of view.     

Cytogenetic study in Corydoras britskii (Siluriformes: Callichthyidae), using different chromosomal banding and fluorescence hybridization in situ with rDNA probes

Cytogenetic analyses were performed on Corydoras britskii from the Miranda River basin, an important river located in the Pantanal, Mato Grosso do Sul State, Brazil. The karyotype of this species comprises 90 chromosomes and a karyotype formula of 4m + 10 sm + 22 st + 54a. The nucleolus organizer regions were detected by impregnation with silver nitrate and FISH with an 18S rDNA probe on the short arm of three acrocentric chromosomes. The constitutive heterochromatin is distributed in pericentromeric and interstitial positions, and also associated with the NORs. The HinfI restriction endonuclease was used and showed homology with practically all types of heterochromatin observed in C. britskii, except for two interstitial heterochromatic blocks present in a subtelocentric pair. The fluorochrome staining evidenced six chromosomes with chromomycin-positive signals indicating that both the heterochromatin interspersed with NORs and some heterochromatic blocks were rich in GC base pairs. FISH using a 5S rDNA probe revealed the presence of these regions in only one subtelocentric pair in the interstitial position. The obtained data substantiate the karyotype diversity of the genus Corydoras and provide novel information about the composition of heterochromatin and location of 5S and 18S rDNA sites.     

Cytotaxonomy of autumnal flowering species of Hyacinthaceae from Algeria

Cytotaxonomic investigations of the autumn-flowering squills, Prospero autumnale (L.) Speta ≡ Scilla autumnalis L., Prospero obtusifolium (Poir.) Speta ≡ Scilla obtusifolia Poir., Barnardia numidica (Poir.) Speta ≡ Scilla numidica Poir., and Hyacinthoides lingulata (Poir.) Rothm. ≡ Scilla lingulata Poir. were performed in 20 populations from northern Algeria located between Tipasa and La Vieille Calle. Various chromosome numbers were found, including a new cytotype, 2n = 8, for the flora of Algeria, concerning plants identified as Prospero obtusifolium (Poir.) Speta [including P. fallax (Steinh.) Speta = S. autumnalis L. ssp. fallax (Steinh.) Batt.]. The numbers 2n = 14, 28, and 42 correspond, respectively, to diploid, tetraploid, and hexaploid levels of P. autumnale s.l. [including P. pulchellum (Munby) Speta ≡ Scilla pulchella Munby = S. autumnalis var. pulchella (Munby) Batt.], with x = 7. The cytotypes of Barnardia numidica (Poir.) Speta with 2n = 18 and Hyacinthoides lingulata (Poir.) Rothm. with 2n = 16 chromosomes were confirmed.     

Cytogenetic study on some Fritillaria species of Iran

Five species (14 ecotypes) belonging to three subgenera of ornamental-medicinal Iranian Fritillaria plant were chromosomally and karyotypically assessed, using squash technique and 1 % (w/v) aceto-orcein stain. All species were diploid (2n = 2x = 24) having mean chromosome length of 16.8 μm (14.2–18.6 μm). The satellites varied in number (1–4 pairs) and in size (1.27–3.01 μm): mostly locating on long arms. Four chromosome types (“m”, “sm”, “st”, “T”) formed 11 different karyotypic formulas: the latter is being reported for the first time in some ecotypes in either S1 or S4. Nine chromosomal parameters were calculated. ANOVA verified intra- and inter-specific chromosomal variation in examined Iranian Fritillaria species. Twelve different methods were used to assess the degree of karyotype asymmetry. Among those, one qualitative (Stebbins classification) and seven quantitative (TF %, CV_TL, DI, AsK %, A2, Rec, CG %) parameters verified that S2 and S5 species were recognized as having the most asymmetrical and symmetrical karyotypes, respectively.     

Cytomorphological diversity in some selected members of Poaceae from Parvati Valley in Kullu district of Himachal Pradesh,India

The present work includes detailed male meiotic studies on 46 species of grasses falling into 59 accessions from different localities of Parvati Valley in Kullu district of Himachal Pradesh in the altitudinal range of 1,100 to 2,750 m. All the species have been studied cytologically for the first time from the study area. The meiotic chromosome count of n = 14 for Calamagrostis emodensis is the first ever chromosome report. Three species, namely Agrostis alba (n = 21), Avena byzantina (n = 21) and Bromus inermis (n = 14) have been studied cytologically for the first time from India. New intraspecific diploid/polyploid cytotypes have been reported for Arthraxon serrulatus (2n = 4x = 32), Iseilema laxum (2n = 12x = 60), Digitaria albudens (2n = 8x = 72), Festuca kashmiriana (2n = 2x = 14) and Stipa orientalis (2n = 2x = 20). The existence of variable number of B-chromosomes (2n = 60 + 0-5B) has been reported for the first time in the 12x cytotype of Iseilema laxum. Secondary associations of chromosomes in the tetraploid cytotype of Cymbopogon martini (n = 20) indicated its secondary polyploid nature. As many as 18 species showed various meiotic anomalies such as the phenomenon of cytomixis involving inter PMC migration of chromatin material, chromatin stickiness, interbivalent connections, abnormal spindle activity, presence of bridges and laggards during anaphases and telophases and abnormal sporads. These meiotic abnormalities consequently yielded sterile and heterogeneous-sized fertile pollen grains. The polyploidy and aneuploidy have played an active role in the evolution of grasses.