Chromosome observations of three ranunculaceous genera in relation to their systematic positions

1. The present paper describes the observations of chromosome numbers and karyomorphology of 2 species of 2 endemic genera and I endemic species of Chinese Ranunculaceae: Asteropyrum peltatum (Franch.) Drumm et Hutch. 2n=16, x=8; Kingdonia unifolia Balf. f. et W. W. Sm. 2n=18, x=9 and Calathodes oxycarpa Sprague 2n=16, x=8. The chromosome counts of three ranunculaceous genera are reported for the first time. 2. The morphylogical, palynological and cytological date in relation to the systematic postition of Asteropyrum, Kingdonia and Calathodes within the family Ranunculaceae are diseussed and resulted in following conclusions: (1). On the basis of the basic number x=8 in Asteropyrum, it is further confirmed that this genus is distinct from the r elated genera such as Isopyrum, Dichocarpum and other allied taxa. The comparison of Asteropyrum with Coptis shows that they are identical in short chromosomes, with magnoflorina and benzylisaquinodine type of alkaloides, but different from coptis in the chromosome numbers (T-type), pantocolpate pollens, united carpels and the dorsi-ventral type of petioles. In view of these fundamental morphological and cytological differences, Asterop yrum is better raised to the level of Tribe. However Asteropyrum and Coptis may represent two divaricate evolutional lines of Thalictroideae. (2). The systematic position of the genus Kingdonia has been much disputed in the past. We support the view of Sinnote (1914), namely, the trilacunar in leaf traces “the ancient type”, appeared in the angiosperm line very early, while the unilacunar of Kingdonia may be derived from the trilacunar. On the basis of the chromosome numbers and morphylogical observation, the present writer accept Tamura’s and Wang’s treatment by keeping Kingdonia in Ranunculaceae instead of raising it to a family rank as has been been done by Forster (1961). Kingdonia and Coptis are similar in having short chromosome with x=9, but with one-seeded fruits; therefore it is suggested that placed into Thalictroideae as an independent tribe, indicating its close relationship with Coptideae. (3). Comparing with its allies, Calathodes being with out petals, seems to be more primitive than Trollius. But Calathodes differs from Trollius with R-type chromosomes in having T-type chromosome with x=8 and subterminal centromere. Those characteristics show that it is very similar to the related genera of Thalictroideae. But as Kurita already pointed out that most speci es of Ranunculus have usually large long chromosomes but some species have compar ativelly short chromosomes, therefore we regard T-type and R-type chromosomes appear independently in different subfamilies of Ranunculaceae. According to Tamura, G alathodes seems to be closely related to Megaleranthis, because of the resemblance in follicles. But due to lack of cytological data of the latter genus, the relationship between the two genera still is not clear pending further studies. From the fact that the morphology and chromosomes of the Calathodes differs from that of all other genera of the Helleboroideae, we consider Calathodes may form an independent tribe of its own with a closer relationship withTrollieae.9841     

Tribal relationships of Beesia, Eranthis and seven other genera of Ranunculaceae: evidence from cytological characters

The karyotypes of 16 species and one variety representing nine genera in the Ranunculaceae were analysed in order to obtain information on the placement of the genera Beesia and Eranthis at tribal rank in the family. Those of Beesia , Anemonopsis , Souliea , Cimicifuga , Actaea and Eranthis were found to be very similar to each other, but remarkably different from those of Caltha , Calathodes , Megaleranthis and Trollius with respect to chromosome size and morphology. From cytological data it is clearly evident that Beesia and Eranthis have a much closer affinity to Anemonopsis , Souliea , Cimicifuga and Actaea in the tribe Cimicifugeae than to Caltha , Calathodes , Megaleranthis and Trollius , which generally have been placed together in a single tribe, albeit with different tribal names such as Caltheae, Trollieae and Helleboreae. Therefore, Beesia and Eranthis may find better placement in the tribe Cimicifugeae. The long-presumed, mainly morphology-based close affinity between Eranthis and Helleborus is not supported by both cytological and palynological data. Cytologically, both Actaea and Eranthis appear to be the specialized genera in the Cimicifugeae, through having the most asymmetrical karyotypes in this tribe. Our results strongly support the recent re-definition of the tribe Actaeeae (as Cimicifugeae) to include Beesia and Eranthis , based on maximum parsimony analysis performed separately on molecular data.  © 2006 The Linnean Society of London. Botanical Journal of the Linnean Society , 2006, 150 , 267–289.     

Polymorphism of the Sphaerium corneum (Bivalvia, Veneroida, Sphaeriidae) revealed by cytogenetic and sequence comparison

Two populations of Sphaerium corneum were sampled from River Vilnelė and small pond in Vilnius, Lithuania. The chromosomes were studied using conventional Giemsa staining and karyometric analysis. Inter- and intra-individual variation in the diploid chromosome numbers was revealed and two different sources of chromosome variability were identified: B chromosomes and the structural changes of chromosomes of the basic (A) set. The chromosome set of the more common karyotypic form, 2 n  = 30, found in both populations, consists of all biarmed metacentric and meta-submetacentric chromosomes of gradually decreasing size. Small, biarmed, mitotically unstable B chromosomes were found in the cells of this karyotypic form. Specimens with 2 n  = 36 were found only in pond. No B chromosomes were detected in their cells. The karyotype is characterized by presence of two pairs of medium telocentrics and four pairs of small subtelocentrics. The remaining chromosomes are biarmed. Robertsonian fusions appear to be involved in formation of two karyotypic forms of S. corneum . DNA sequence analyses showed that ITS1 is identical in both karyotypic forms. On the other hand, differences in 16S sequence were revealed and two haplotypes, corresponding to two karyotypic forms, were identified. The present study opens new perspectives in establishing species-specific characters for confident identification of Sphaerium species and provides insights to the genetic intraspecific variability and possible mechanisms of speciation.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 53–64.     

Ribosomal, telomeric and heterochromatin sequences localization in the karyotype of Anemone hortensis

The karyotype of the Mediterranean species Anemone hortensis L. (Ranunculaceae) was characterized with emphasis on heterochromatin distribution and localization of ribosomal (18S−5.8S−26S and 5S rDNA) and telomeric repeats (TTTAGGG). Diploid chromosome complement, 2 n  = 2 x  = 16, common to all investigated populations, consisted of three acrocentric, one meta-submetacentric and four metacentric chromosomes ranging in size from 6.34 to 10.47 µm. Fluorescence in situ hybridization (FISH) with 18S and 5S rDNA probes revealed two 18S−5.8S−26S rDNA loci on a satellite and secondary constriction of acrocentric chromosome pair 2 and terminally on acrocentric chromosome pair 3, and two 5S rDNA loci in the pericentromeric region of meta-submetacentric chromosome pair 4 and in the proximity of the 18S−5.8S−26S rDNA locus on chromosome pair 2. The only GC-rich heterochromatin, as revealed by fluorochrome Chromomycin A3 staining, was that associated with nucleolar organizer regions, whereas AT-rich heterochromatin, stained with 4,6-diamino-2-phenylindole (DAPI), was distributed intercalarly and terminally on the long arm of all three acrocentric chromosomes, and terminally on chromosomes 4 and 5. FISH with Arabidopsis -type telomeric repeats (TTTAGGG) as a probe revealed two classes of signals, small dot-like and large bands, at chromosome termini exclusively, where they corresponded to terminal DAPI-stained heterochromatin. Heteromorphism of chromosome pair 4, which refers to terminal DAPI bands and FISH signals, was observed in populations of Anemone hortensis . Chromosome pairing during meiosis was regular with formation of localized chiasmata proximal to the centromere.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 150 , 177–186.     

Chromosomes of four species in three genera of Commelinaceae from China and their systematic implications

Chromosomes of four species in three genera of Commelinaceae from China were examined. Spatholirion longifolium had 2 n  = 20. This small genus is considered to be of a possible polyploid origin based on x  = 5, a basic number shared by its closest ally, the monospecific genus Streptolirion , with its only species having 2 n  = 10. The two genera were found to be very different from each other in their karyotypic constitution. The differences in both chromosome number (2 n  = 20 vs. 10) and karyotypic constitution support their separation as two independent genera. The remaining three species, Amischotolype hispida , Porandra ramosa and P. scandens , all had 2 n  = 36. In view of the occasional occurrence of 2 n  = 18 in the African Coleotrype , a genus very close to Amischotolype and Porandra in gross morphology, these three species are all very probably polyploid based on x  = 9. The high degree of karyotypic similarity of the three genera also strongly indicates their close affinity and supports their placement in the subtribe Coleotrypinae. © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 152 , 399–403.     

New or rarely reported chromosome numbers in taxa of subtribe Artemisiinae (Anthemideae, Asteraceae) from Mongolia

This study encompasses 25 chromosome counts of 18 species in the subtribe Artemisiinae (tribe Anthemideae) of the family Asteraceae, from Mongolia. Most (15 species) belong to Artemisia , the largest genus of the subtribe, whereas the others come from two genera very closely related to it: Ajania (two species) and Neopallasia (one species). Eleven counts are new reports, three are not consistent with previous reports and the remainder confirm scanty earlier information. The majority of the species have x  = 9 as their basic chromosome number, but there are some taxa with x  = 8. Ploidy levels range from 2 x to 6 x . The presence of B-chromosomes was detected in Ajania fruticulosa .  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 150 , 203–210.     

New chromosome counts in the Centaurea Jacea group (Asteraceae, Cardueae) and some related taxa

Twenty-seven chromosome counts are reported in 23 species of the genus Centaurea , mostly eastern endemic species of the Jacea group, which has become the core of the re-defined genus Centaurea . Twenty reports are new, one is a correction of a previous count, one is a confirmation of limited previous data and one represents a new basic number in the Centaureinae. The prevalence of the basic chromosome number x  = 9 among the Eastern sections of the Jacea group is confirmed, together with the close correlation between karyological data and classification of the genus. Two alternative hypotheses on the aberrant chromosome number (for the Centaureinae) found in C. behen are proposed. One of them, if verified, would confirm that a cycle of polyploidy and descending dysploidy is a key mechanism in the speciation of the group.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 145 , 345–352.     

Chromosomal variability in the genus Primula (Primulaceae)

Chromosome numbers and measurements were recorded in 47 individuals of ten taxa of Primula , representing eight species and two subspecies from ten populations. The basic numbers of chromosomes were x  = 8, 9 or 11, and they were mostly metacentric, medium-long to medium-small, ranging in length from c . 3.6 µm to 1.6 µm. Diploid chromosome numbers of 2 n  = 2 x  = 24 and 22 were scored for the first time in P. loeseneri Kitag. and P. prenantha Balf. & Sm., respectively. A new ploidy level of 2 n  = 4 x  = 44 was found in P. burmanica Balf. & King. The recorded diploid numbers of the remaining species confirmed earlier reports. Interspecific variability in chromosome numbers was correlated with heterogeneity in their mean length. Comparison of the data with those in the literature revealed that the observed variability of chromosomal characters was compatible with the other taxonomic criteria and supports the current taxonomic delimitation. Chromosomal variation at the diploid level is the predominant feature in Primula evolution, and allopatric speciation has played a major role in its specific diversity. Subgen. Aleuritia could represent the main clade from which the other subgenera have evolved. The origin of the chromosome numbers, geographical distribution and evolution of the species were assessed, and the parallel polyphyletic mode of evolution in the genus was confirmed.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 150 , 211–219.     

Cytotaxonomy of Begonia rubropunctata and B. purpureofolia (Begoniaceae)

The chromosome numbers of four Chinese Begonia species were counted for the first time as follows: 2 n  = 22 in B. rubropunctata , 2 n  = 18 in B. purpureofolia , 2 n  = 44 in B. pedatifida , and 2 n  = 22 in B. villifolia . The first two species had relatively large chromosomes with early condensed chromatin in the distal and interstitial or proximal regions, whereas the last two had small chromosomes with early condensed chromatin in the proximal regions. Although they have been reduced in the literature to synonyms of B. pedatifida and B. villifolia , respectively, B. rubropunctata and B. purpureofolia should be treated as distinct species because of the differences in chromosome numbers and other chromosome features.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 513–517.     

Chromosome number, karyotype, and taxonomic considerations on the enigmatic Sellocharis paradoxa Taubert (Leguminosae, Papilionoideae, Genisteae)

Until recently, Sellocharis paradoxa Taubert, the only species of this Genisteae genus, was known solely by the isotypes. Recent new collections in Rio Grande do Sul, southern Brazil, have enabled data on chromosome number and karyotype morphology to be obtained for the first time. S. paradoxa has 2 n  = 20 chromosomes, and a bimodal asymmetrical karyotype, composed of one pair of long ( c . 6.3 µm) metacentric, five pairs of shorter acrocentric, and four pairs of shorter telocentric chromosomes ranging from c . 3.7 to 2.7 µm. The chromosome number and karyotype morphology of S. paradoxa do not fit into the Genisteae pattern. Existing information is so far insufficient to answer evolutionary questions about its origin and phylogenetic relationships, but the uniqueness of this taxon, first indicated by its peculiar leaf arrangement, and now supported by its uncommon karyological constitution, strongly suggests that the suprageneric taxonomic position of S. paradoxa should be re-evaluated.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 223–226.     

Reduction of chromosome number in Eleocharis subarticulata (Cyperaceae) by multiple translocations

Eleocharis subarticulata is recorded as the third species of Cyperaceae with a reduced chromosome number ( n  = 3), following reports on Rhynchospora tenuis ( n  = 2) and Fimbristylis umbellaris ( n  = 3). For Eleocharis, the numbers recorded to date vary from 2 n  = 10 to 2 n  = c. 196, with x  = 5 as the possible basic number. The karyotype of E. subarticulata was studied using conventional staining (mitosis and meiosis), C-CMA₃/DAPI banding, and FISH with 45S rDNA and telomere probes. The chromosomes showed no primary constrictions, as expected in the holocentric chromosomes of Cyperaceae. The meiotic behaviour was abnormal, with a single multivalent ring of six chromosomes at metaphase I, resulting from multiple translocations. At anaphase I six chromatids migrated to each pole, evidencing the inverted meiosis, and these groups were also visible at metaphase II. The C-CMA₃/DAPI banding technique showed only four terminal GC-rich blocks. FISH with 45S rDNA probes revealed four terminal signals, probably associated with GC-rich blocks. The telomeric probe located terminal signals in all the chromosomes, besides a hybridization site in the middle of the large pair. The occurrence of ectopic telomeric sites has not been described previously for plants with holokinetic karyotypes and with reduced chromosome numbers. These data reinforce the hypothesis of the reduction in chromosome number by multiple translocations.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 149 , 457–464.     

Chromosome numbers in wild and semidomesticated Brazilian Capsicum L. (Solanaceae) species: do x = 12 and x = 13 represent two evolutionary lines?

Chromosome numbers were determined in 29 accessions of wild and semidomesticated Capsicum species from the EMBRAPA Hortaliças (Brazil) germplasm collection. 2 n  = 24 was found in C. baccatum var. praetermissum , C. chinense , C. flexuosum and C. parvifolium , while C. buforum , C. campylopodium , C. cornutum , C. schottianum , C. villosum var. villosum and five other native south-eastern Brazilian species not yet taxonomically named all had 2 n  = 26 chromosomes. These are the first chromosome number determinations for C. cornutum , C. schottianum , C. villosum var. villosum and the five other Brazilian taxa. Our data confirm published data for C. baccatum var. praetermissum , C. campylopodium , C. chinense , C. flexuosum and C. parvifolium , but not for C. buforum . The prevalence of 2 n  = 26 chromosomes among Brazilian species, along with the morphological and ecological characteristics presented by the wild and semidomesticated species occurring in Brazil, form a pattern different from that found in Andean wild and semidomesticated species. This supports the hypothesis that there are two different evolutionary lines in the genus and that the native south-eastern Brazilian species belong to the ancestral Capsicum gene pool. © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 259–269.     

New chromosome counts in the genus Cousinia (Asteraceae) from Iran

Chromosome counts are reported of 38 populations representing 24 species of the genus Cousinia from Iran, one of the main centres of speciation of the genus. Sixteen of the counts are new to science. Our results confirm that Cousinia exhibits a complex dysploid series ranging from x  = 13 to x  = 9. One of the basic chromosome numbers, x  = 9, is new for the genus and must be confirmed. Some considerations on the correlation between chromosome numbers and section classification are made.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 411–419.     

Ferulago trojana (Umbelliferae), a new species from western Turkey

Somatic chromosomes of four species of Ceratozamia , C. hildae , C. kuesteriana , C. mexicana and C. norstogii , and Stangeria eriopus , were observed and compared by the fluorescence in situ hybridization method using 5S ribosomal (rDNA) probes. The four Ceratozamia species and S. eriopus showed the same chromosome number of 2 n  = 16, and had similar karyotypes, comprising 12 metacentric (m), two submetacentric (sm) chromosomes and two telocentric (t) chromosomes. The four Ceratozamia species exhibited a proximal 5S rDNA site in the interstitial region of two m chromosomes. Stangeria eriopus exhibited a distal 5S rDNA site in the interstitial region of two m chromosomes, which probably indicates that the two genera differ in chromosome structure by at least one paracentric inversion. © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 145 , 499–504.     

A new divergent lineage of Daphnia (Cladocera: Anomopoda) and its morphological and genetical differentiation from Daphnia curvirostris Eylmann, 1887

The systematic biology of the subgenus Daphnia s . s . remains confused. Prior attempts at resolution used chiefly postabdominal claw morphology, chromosome numbers and rRNA gene sequences as characters for higher-level relations. Still, several taxa, such as Daphnia curvirostris Eylmann, 1878, have unclear affiliations. We addressed the position of D. curvirostris in this genus by estimating phylogenetic trees from a rapidly evolving protein coding gene (ND2), conducting broad geographical comparisons and carrying out detailed morphological comparisons. The Japanese ' curvirostris' was found to be a new divergent lineage in the subgenus Daphnia , and to possess distinctive morphological characteristics from D. curvirostris . We described this new species as Daphnia tanakai sp. nov. , and redescribed D. curvirostris . The polymorphic postabdominal claw morphology and the distinctive chromosome number of D. tanakai sp. nov. provided evidence for rapid evolution of these traits. Our new morphological, chromosomal and genetic assessment of Daphnia weakened the argument for division of the subgenus Daphnia ( Daphnia ) O. F. Müller, 1785 sensu Johnson, 1952, into two further subgenera.  © 2006 The Linnean Society of London, Zoological Journal of the Linnean Society , 2006, 146 , 385–405.     

Chromosomal polymorphism in the Atlantic dog-whelk, Nucella lapillus (Gastropoda: Muricidae): nomenclature, variation and biogeography

Nucella lapillus appears to be unique within the Muricidae, and indeed the Mollusca, in exhibiting a Robertsonian chromosomal polymorphism. The recorded diploid values range from 2 n  = 26 to 2 n  = 36, putatively brought about by fusion of smaller chromosomes to form five pairs of large metacentrics in the 2 n  = 26 form. In this study, the karyotypes of the numerically extreme forms (2 n  = 26 and 2 n  = 36) have been illustrated and described in detail, and a new scheme of nomenclature is proposed for the chromosomal rearrangements observed in N. lapillus . Chromosome numbers and karyotypes were recorded and analysed from 70 sites throughout its wide geographical range, mainly from around the UK coasts but also from sites in Norway, France, Spain, Portugal and the USA, showing remarkable variation in both diploid number and karyotypic configuration. The results provide new evidence that seven chromosomes in the 2 n  = 26 form can be involved in the Robertsonian rearrangements, but the maximum recorded diploid number remains 2 n  = 36. Inversions were confirmed in three chromosomes and one population was found to exhibit aneuploidy. Possible explanations for the geographical trends in karyotypic variation are discussed, but despite the advances in the karyology of N. lapillus , a simple solution to this enigmatic phenomenon remains elusive.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 87 , 195–210.     

Chromosomal criteria and their phylogenetic implications in the genus Onobrychis Mill. sect. Lophobrychis (Leguminosae), with special reference to Egyptian species

Chromosome numbers and measurements were recorded in 22 individuals from six populations of six species of Onobrychis , including the Egyptian species and most representatives of section Lophobrychis . The basic number of chromosomes was either x = 7 or x = 8 and the chromosomes were medium to medium-small, ranging in length, from c . 1.6 μm to 2.6 μm. Two new ploidy levels were found, 2 n = 4 x = 28 in O. bobrovii Grossh. and 2 n = 4 x = 32 in O. pulchella Schrenk. The origin of the chromosome numbers, geographical distribution and evolution of the species were assessed. Comparison of the data with those in the literature revealed that the observed interspecific variability among section Lophobrychis can be useful in taxonomic delimitation and demonstrates a complexity of evolution between the diploid and polyploid species. Section Lophobrychis has a comparatively highly derived organization and can be considered as a heterogeneous unit in the genus Onobrychis .  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 139 , 409–414.     

The cytology of Actinidia, Saurauia and Clematoclethra (Actinidiaceae)

Meiosis and mitosis of six Chinese Actinidiaceae were studied: Saurauia tristyla DC., S. miniata C. F. Liang & Y. S. Wang, Actinidia chinensis Plach., A. deliciosa (Cheval.) C. F. Liang & A. R. Ferguson, A. indochinensis Merr. and Clematoclethra lasioclada Maxim. The chromosome numbers of Saurauia tristyla and S. miniata were 2 n  = 6 x  = 78, establishing a base chromosome number of x  = 13 in the genus, differing from the previous report of x  = 15. The chromosome number of Clematoclethra was first reported to be 2 n  = 4 x  = 48 ( x  = 12), while that of Actinidia was x  = 29, consistent with previous reports. The base chromosome number of Clematoclethra ( x  = 12) was derived from an aneuploid decrease from Saurauia ( x  = 13). Actinidia (x  = 29) was derived from the palaeotetraploid ( x  = 14), which was formed through the increase of the basic chromosome number x  = 13 to x  = 14 by aneuploidy and through the breakage of a centromere to add one more new chromosome. The chromosome data in Actinidia were consistent with the geographical and morphological evidence for the evolution of the three genera. The tropical American and Asian disjunct distribution pattern and the diversity of base chromosome numbers of Saurauia further support the probability that the genus was an early divergent from a common ancestor of Actinidia and Clematoclethra .  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 147 , 369–374.     

Karyological and taxonomic notes on Ophrys (Orchidoideae, Orchidaceae) from the Iberian Peninsula

Karyological information on Iberian Ophrys species is very limited. This paper provides the haploid and diploid chromosome numbers of 11 taxa of sect. Pseudophrys and sect. Ophrys , both of which are well represented in the Iberian Peninsula, and two taxa from Tunisia. The first data on chromosome numbers for O. vasconica (2 n  = 72, 74), O. ficalhoana (2 n  = 36), O. picta (2 n  = 36), O. sphegifera ( n  = 18, 2 n  = 36, 38) and O. passionis (2 n  = 36) are also presented, confirming the stability of the chromosome number in Ophrys . In addition, populations of the group O. omegaifera ( O. dyris and O. vasconica ), together with tetraploidy, pentaploidy and the existence of aneuploid phenomena, are reported for the first time in Iberia. The basic diploid number is always 2 n  = 36. The karyotypes of several species were analysed. Evolutionary trends in Ophrys chromosomes are discussed. Taxonomic and phytogeographical details are provided on several species or groups of species from Iberia.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society, 2003 , 142 , 395−406.