Karyotypic studies in Solanum section Lasiocarpa (Solanaceae)

Mitotic chromosomes of 13 species included in Solanum sect. Lasiocarpa were studied. All species have 2n = 24. The chromosome numbers of S. stagnale, S. felinum, and S. repandum are reported for the first time. Statistical analyses of chromosomes, genome length, and centromere position yielded estimates of karyotype composition and asymmetry. A generalized karyotype of the section shows that most of the chromosomes are metacentric (73%) or submetacentric (25.6%). Two pairs of subtelocentric chromosomes are present only in S. sessiliflorum. Satellites are quite common, characterizing chromosomes for ten species and are attached to the short arms of either m or sm chromosomes. They could be seen in 10% to 70% of the cells. Only S. pseudolulo bears two satellited chromosomes, one of them attached to a long chromosome arm. Although the section is chromosomally quite homogeneous, species can be distinguished by karyotype formula, presence of satellites in a particular chromosome pair, and total chromosome length. There are no indications of major chromosomal rearrangements within the section. Using cytological data exclusively, cluster analysis indicates S. sessiliflorum is isolated in the section. Solanum candidum and S. vestissimum are somewhat isolated as well and have unique karyological traits. Solanum pectinatum has a peculiar karyotype, but in the phenogram it is not particularly separated. Karyotype features suggest that morphological differentiation was not always followed by chromosomal divergence. The origin of the domesticated S. quitoense is no further elucidated by our data.     

Chromosome diversity and evolution in Allium (Allioideae,Amaryllidaceae)

Despite extensive literature on the diversity of karyotypes in Allium is available, no attempt to analyse these data together, within a robust phylogenetic framework, has been carried out so far. Thus, we examined patterns and trends in chromosome evolution across the genus. Based on literature survey, karyo-morphometric features for 207 species belonging to 12 subgenera of Allium were obtained. Included in the data-set were basic chromosome number (x), somatic chromosome number (2n), total haploid (monoploid) chromosome length (THL) and three different measures defining karyotype structure: CV_CI, measuring how heterogeneous are centromeres positions in a karyotype, CV_CL and M_CA, quantifying interchromosomal and intrachromosomal karyotype asymmetry, respectively. Trends in karyotype evolution were analysed by phylogenetic regressions and independent contrasts. Mean karyotypes highlighted differences and similarities in karyotype structure between the 12 subgenera. Further differences were noted when the two parameters for analysing karyotype asymmetry were assessed. In addition, by examining the effects of increasing karyotype dimensions (a proxy for genome size) on karyotype structure and asymmetry, it was shown that in Allium species, the DNA was added proportionally to their arm lengths. Overall, p = 8 and somehow intermediate karyotype asymmetry levels seem to represent plesiomorphic character-states in Allium.     

Karyotype analysis in Stylidium crossocephalum (Angiospermae:Stylidiaceae)

Qualitative studies of karyotypes were carried out on 79 plants of Stylidium crossocephalum from 28 collection sites. The sample included 7 distinct karyotypic classes which could be resolved into 5 clearly distinguishable haploid karyotypes or genomes. Three genomes, J, K and L, were localised in an area covering a southern portion of the species range while the other two genomes, A and B, were found in a region 90km to the north. Two different genome heterozygotes were found, one characterised by a J/K karyotype, the other by an A/B karyotype. —Karyotypic studies of a synthetic hybrid and its two parents showed that the chromosomes of the constituent genomes of the hybrid could be readily and confidently discerned in the karyotypes of the parents. These studies demonstrated that the identification of genomes by the qualitative attributes of their constituent chromosomes constitutes a reliable analytical technique. — Statistical analyses using arm ratio and percentage length measurements of chromosomes from plants allocated to the two groups of genomes confirmed the differences between marker chromosomes and demonstrated additional statistically significant differences among the non marker chromosomes. Some of these statistical differences may be attributable to technical problems inherent in the karyotyping procedures. In two instances, however, they could be convincingly argued as real, demonstrating a further component of chromosome variation in S. crossocephalum.     

Constraints upon the organisation and evolution of chromosomes in Allium

Summary In terms of chromosome morphology, karyotype organisation, taxonomy and genetic relationship as judged from chromosome pairing in the Fl hybrid, A. cepa and A.fistulosum are two closely related species. But large variation in nuclear DNA amounts has occurred during the evolution of the two species. A comparison of the molecular composition of DNA in the two species has confirmed that the excess DNA acquired during evolution was predominantly repetitive sequences (sequences which do not encode genetic information). However, its distribution within the chromosome complements was equal in all chromosomes irrespective of the differences in chromosome size. The even distribution of the excess DNA within complements suggests strong constraints underlying evolutionary changes in genome organisation. The nature of the constraints is discussed, and it is shown that such constraints can influence the direction of karyotype evolution during speciation.     

Extreme degree of chromosome number variability in species of the spider genus Scytodes (Araneae, Haplogynae, Scytodidae)

Genus Scytodes includes most species of the spider family Scytodidae. Until now, 187 species of the genus have been described. In spite of this great diversity, only three Scytodes species were karyotyped so far. The present paper provides for the first time karyotype analysis of two synanthropic species, Scytodes fusca and Scytodes itapevi. Furthermore, new data on karyotype of Scytodes globula are also provided using conventional and differential cytogenetical procedures. The diploid number in the genus Scytodes varied considerably, namely from 2n = 13 to 2n = 31. The diploid number found in S. globula (2n♂ = 13) is the lowest in haplogyne spiders with monocentric chromosomes. Except S. globula, this number has been found only in one haplogyne spider with monocentric chromosomes, namely Ochyrocera sp. (Ochyroceratidae). On the contrary, the diploid number of S. fusca (2n♂ = 31) is one of the highest diploid numbers recorded in haplogyne spiders. The degree of intrageneric variation found in the genus Scytodes is the highest recorded in araneomorph spiders with monocentric chromosomes so far. Some karyotype characteristics (diploid number, chromosome morphology, total chromosome length, and distribution of constitutive heterochromatin) allowed us to postulate a close relationship between S. globula and S. itapevi. According to the karyotype data, S. fusca is not closely related to these two species. This conclusion corroborates a recent taxonomic work that grouped S. globula, S. itapevi, and other four Scytodes species in the ‘globula group’.     

A simple parameter of dispersion index that serves as an adjunct to karyotype asymmetry

In order to refine the measure of karyotype asymmetry a new chromosomal parameter of dispersion index is proposed that has the potential to decipher even the minor karyotypic variations, thus permitting further evolutionary gradations to the karyotype asymmetry classes of Stebbins. The higher the dispersion index, the more specialized would be the karyotype. The dispersion index takes into account the variance lor gradual change in chromosome size within a complementvis-a-vis variance for the position of centromere in a karyotypic totality. The dispersion index is calculated as the proportionate measure of centromeric gradient to the coefficient of variation for chromosome length; wherin centromeric gradient = length of median short arm — length of median chromosome. Thus, the three most important karyotypic criteriaviz., differences in: absolute chromosome size, position of centromere and relative chromosome size, are all covered in the proposed parameter. The effectiveness of dispersion index has been tested on a plant taxa,Papaver L., where karyomorphological details, nuclear DNA content, and morphotaxonomic parameters have been amply elucidated from an evolutionary stand point. It is hoped that dispersion index would find immense utility in delimiting species interrelationships particularly in the closely related taxa, when applied in conjunction with other systematic parameters.     

The karyotype and 5S rRNA genes from Spanish individuals of the bat species <Emphasis Type="Italic">Rhinolophus</Emphasis><Emphasis Type="Italic">hipposideros</Emphasis> (Rhinolophidae; Chiroptera)

The karyotype of individuals of the species Rhinolophus hipposideros from Spain present a chromosome number of 2n = 54 (NFa = 62). The described karyotype for these specimens is very similar to another previously described in individual from Bulgaria. However, the presence of one additional pair of autosomal acrocentric chromosomes in the Bulgarian karyotype and the differences in X chromosome morphology indicated that we have described a new karyotype variant in this species. In addition, we have analyzed several clones of 1.4 and 1 kb of a PstI repeated DNA sequence from the genome of R. hipposideros. The repeated sequence included a region with high identity with the 5S rDNA genes and flanking regions, with no homology with GenBank sequences. Search for polymerase III regulatory elements demonstrated the presence of type I promoter elements (A-box, Intermediate Element and C-box) in the 5S rDNA region. In addition, upstream regulatory elements, as a D-box and Sp1 binding sequences, were present in flanking regions. All data indicated that the cloned repeated sequences are the functional rDNA genes from this species. Finally, FISH demonstrated the presence of rDNA in nine chromosome pairs, which is surprising as most mammals have only one carrier chromosome pair.     

Rapid,localized amplification of a unique satellite DNA family in the rodent Microtus chrotorrhinus

A novel satellite DNA family (called MSAT-2570) was isolated and characterized from the rodent Microtus chrotorrhinus. With a length of 2,570 bp the repeat unit is among the largest yet reported in mammals and comprises a series of short direct and inverted repeats. These repeat motifs may prevent nucleosome formation or represent an endless source of genetic variation. Restriction enzyme digestion using the two pairs of isoschizomers HpaII/MspI and MboI/Sau3AI demonstrated tissue specific differences in satellite DNA methylation that may reflect variable chromatin conformation or differences in patterns of gene expression. The sex chromosomes of M. chrotorrhinus are unusually large in size among mammals, comprising 15%–20% of the karyotype and containing large blocks of heterochromatin. In situ hybridization of the satellite DNa revealed chromosomal localization predominantly to sex chromosome heterochromatin. A survey of related rodents including three congeneric species also with giant sized sex chromosomes demonstrated that MSAT-2570 is present only in the genome of M. chrotorrhinus. However, another previously reported satellite DNA also isolated from M. chrotorrhinus has been shown to reside on sex chromosome heterochromatin in one of the other three species, indicating that these giant blocks of heterochromatin are complex in structure and comprise multiple, unrelatined satellite DNA families.     

Karyotype and genome size variation in Ajuga L. (Ajugoideae–Lamiaceae)

Chromosome number changes and karyotype evolution play an important role in plant genome diversification and eventually in speciation. The genus Ajuga L. (Lamiaceae) has approximately 50 species distributed in temperate to subtropical regions. Four of these species are currently recognized in Korea (A. decumbens Thunb., A. multiflora Bunge, A. nipponensis Makino and A. spectabilis Nakai). Understanding the karyotype evolution in Ajuga has been hampered by the small size of their chromosomes and symmetrical karyotypes. Here we used classic Feulgen staining to establish chromosome numbers and construct karyotypes of the four species of Ajuga recognized in Korea and flow cytometry was used to study their variation in genome. The chromosome number of all investigated plants was 2n = 32. Still, the 2C DNA content ranged from 2.18 pg (A. decumbens) to 4.53 pg (A. multiflora). While the chromosome numbers were the same for all investigated species, the genome size variation could potentially be used as a taxonomic marker.     

Comparative cytogenetics between the species Passiflora edulis and Passiflora cacaoensis

Passiflora edulis Sims is the most economically important species of the genus Passiflora. A new species was described recently, Passiflora cacaoensis Bernacci & Souza, which displayed morphologic characteristics very similar to P. edulis. Due to the need for delimitation of the two species, karyomorphological and banding analyses were carried out. Both species have 2n = 18, with the same karyotype formula 16 m + 2sm. There was variation between the species regarding the location of satellites and the width of chromosome pairs 2, 4 and 8. C banding revealed the presence of constitutive heterochromatin in the centromeric and telomeric regions of all chromosomes in both species. However, only in P. cacaoensis did chromosomes 3 and 9 have a large quantity of heterochromatin. Fluorochrome banding revealed CMA⁺ bands only in the satellites, but no DAPI⁺ bands. Fluorescence in situ hybridisation (FISH) showed that in P. cacaoensis the rDNA 5S probe is located in a single site in the subterminal position of the long arm of chromosome 5. However, for the rDNA 45S probe, two sites were detected in terminal positions of the long arms of chromosome 7, with a bigger and stronger signal, and of chromosome 9. According to the asymmetry index and the quantity of heterochromatin, P. cacaoensis is a more basal species than P. edulis. The cytogenetic data indicate that P. cacaoensis is closely related to P. edulis, but is a different species.     

小麦属核型分析和BG染色体组及4A染色体的起源

应用植物有丝分裂染色体标本制备新方法和N—带技术对小麦属(Triticum)9个六倍体种(AABBDD),8个四倍体种(AABB,AAGG),3个二倍体种(AA,A~uA~u)及B组的可能供体沙融山羊草(Ae. shronensis)体细胞核型和N—带进行了分析。结果表明,小麦属全部为具中部或次中部着丝点染色体,核型属于“2A”类型,不对称性随倍性提高而有所增加。种问核型有一定差异。所有小麦B染色体组、G染色体组和4A染色体均显N—带,其它染色体则不显带或只显很浅的着丝点带。六倍体种B染色体组带型基本相同,四倍体小麦B组N—带种间有一定差异。提莫菲维小麦(T.Timopheevi)G组带纹数目和分布与B梁色体组有显著差别,作者认为两者非同源。沙融山羊草核型和带型都与小麦B组相近,是B组的可能供体。一粒系小麦A染色体组基本不显N—带,其中无与4A带型相同的染色体,4A起源尚待研究。     

Chromosomes tell half of the story: the correlation between karyotype rearrangements and genetic diversity in sedges,a group with holocentric chromosomes

Chromosome rearrangements may affect the rate and patterns of gene flow within species, through reduced fitness of structural heterozygotes or by reducing recombination rates in rearranged areas of the genome. While the effects of chromosome rearrangements on gene flow have been studied in a wide range of organisms with monocentric chromosomes, the effects of rearrangements in holocentric chromosomes—chromosomes in which centromeric activity is distributed along the length of the chromosome—have not. We collected chromosome number and molecular genetic data in Carex scoparia, an eastern North American plant species with holocentric chromosomes and highly variable karyotype (2n = 56–70). There are no deep genetic breaks within C. scoparia that would suggest cryptic species differentiation. However, genetic distance between individuals is positively correlated with chromosome number difference and geographic distance. A positive correlation is also found between chromosome number and genetic distance in the western North American C. pachystachya (2n = 74–81). These findings suggest that geographic distance and the number of karyotype rearrangements separating populations affect the rate of gene flow between those populations. This is the first study to quantify the effects of holocentric chromosome rearrangements on the partitioning of intraspecific genetic variance.     

Karyotype analysis and heterochromatin differentiation with Giemsa C-banding and fluorescent counterstaining inCephalanthera (Orchidaceae)

Detailed studies of the chromosomes of the three Austrian species of the genusCephalanthera showed them all to have basically similar karyotypes. BothC. damasonium (2n = 36) andC. longifolia (2n = 32) have three large and several classes of smaller chromosome pairs. The karyotype ofC. rubra (2n = 44) is composed of four large and several groups of smaller pairs. The heterochromatin in these species amounts to about 10% of total karyotype length. All the chromosomes have Giemsa-positive centromeres, but only a few have intercalary or terminal bands. Using differential fluorescent staining with DAPI/actinomycin D, quinacrine/actinomycin D (both A-T specific), and chromomycin A₃/distamycin A (G-C specific) three different types of major heterochromatic bands can be characterized in respect of their satellite DNA composition: highly A-T rich, slightly A-T rich, and very G-C rich. The chromosomes ofC. longifolia contain more A-T rich C-bands than those ofC. damasonium, while the latter's have more G-C rich heterochromatin. In both species several C-bands appear as secondary constrictions or gaps in the Feulgen-stained chromosomes, but most likely, in each species there is only one pair of chromosomes where the secondary constrictions function as nucleolus organizing regions. No major intraspecific variation could be observed except on one small chromosome pair ofC. longifolia which had a heteromorphic C-band in most individuals. Possible pathways of karyotype evolution involving polyploidy and Robertsonian events are discussed.     

Satellite DNA probes of Alstroemeria longistaminea (Alstroemeriaceae) paint the heterochromatin and the B chromosome,reveal a G-like banding pattern,and point to a strong structural karyotype conservation

Alstroemeria species present a well-conserved and asymmetric karyotype. The genus is divided into a Chilean clade, rich in heterochromatin, and a Brazilian clade, poor in heterochromatin. We investigated the distribution of the main repetitive sequences in the chromosomes of the Brazilian species A. longistaminea (2n = 16 + 0-6B) aiming to evaluate the role played by these sequences on the structural organization of the karyotype. In situ hybridization of the three most abundant retrotransposons, corresponding to ~ 45% of the genome, was uniformly distributed. Three satellite DNA sequences, representing near half of the whole satellite fraction (1.93% of the genome), were mainly concentrated on the heterochromatin and one of them painted the whole B chromosome. Noteworthy, some satellites were located on euchromatin, either dispersed or concentrated in clusters along the chromosomes, revealing a G-band-like pattern. The two satellites that presented more C-band- and G-band-like labeling were also hybridized in situ in two other Alstroemeria species. They revealed astonishing similar patterns of distribution, indicating an unusually structural karyotype conservation among Brazilian species.

     

Karyotype variation in 11 species of the Vernonieae Cass. tribe (Asteraceae Bercht. & J. Presl)

The Vernonieae tribe presents strong taxonomic delimitation problems as it is considered one of the most complex groups of the Asteraceae family, comprising approximately 1100 species distributed across 129 genera. In this study, a comparative analysis of the Vernonieae species was performed to understand the events involved in the chromosome evolution of these species and to further deduce their taxonomy. The representatives were cytogenetically characterized via analyses of morphology, karyotype asymmetry and differential staining with fluorochromes CMA and DAPI as well as FISH. According to morphometric data, all species showed symmetrical karyotypes with prevailing metacentric chromosomes, even in species belonging to different genera. Variability in diploid chromosome number was detected (2n = 18 to 2n = 60), and chromosome sizes were observed to be between 1.00 and 4.09 μm. Additionally, variation in the pattern of heterochromatin was observed mainly in relation to CMA⁺ bands, in which the number varied from 4 to 16 heterochromatic regions. Only one species, Vernonia scorpioides, presented positive DAPI bands, which were located in the terminal position in most of the chromosomes. The differences in the sizes and quantities of heterochromatic bands may be related to small structural rearrangements during karyotype evolution of the Vernonieae tribe.     

Karyomorphology of Maianthemum sensu lato (Polygonatae, Ruscaceae)

We report results of karyotype analyses using nine species of Maianthemum from China. The species, except M. atropurpureum (with 2n=72), had 2n=36, and the results support the earlier suggestion that Maianthemum has x=18 with 2n=36 in most species. The species examined, however, showed marked differences in karyotype, particularly in the numbers of metacentric, submetacentric, and acrocentric chromosomes as well as in the number of satellites. In addition, we distinguished three different modes based on the number of clear gaps in chromosome length variation: unimodal, bimodal, and trimodal. The unimodal variation (with no gap) was found in M. dahuricum and M. atropurpureum, the bimodal variation (with one gap) in M. tatsienense, and the trimodal variation (with two gaps) in M. bifolium, M. forrestii, M. japonicum, M. henryi, M. purpureum, and M. lichiangense. In the trimodal variation, the positions of the two gaps may differ from species to species. In addition, the frequency of acrocentric chromosomes per complement was generally higher in the trimodal variation than in the unimodal and bimodal variations. Results of our analyses, which had not been clearly presented prior to this, may provide a better understanding of species evolution in the tribe Polygonatae.     

Intra- and interspecific variations in nuclear parameters of two closely related species of Narcissus L.

Nuclear DNA amount, nuclear area, genome volume and karyotype length were analysed in different populations of two closely related species of Narcissus. There are intra- and interspecific variations in these parameters. 4C DNA amount and karyotype length, on one hand, and 2C DNA amount and telophase nuclear area, on the other, are not correlated. It seems that DNA content and chromosome length are independent parameters. However, 4C DNA content and karyotype volume are correlated, and are also correlated to different density estimations (4C DNA to Kar.length & 2C DNA to telophase area). These facts suggest that the relative length of the chromosomes is genetically controlled and that it is independent of the DNA that they contain. It seems that the interpopulational differences in DNA content are correlated with length changes of small segments in almost all chromosomes.     

Comparative cytogenetic studies in Sus verrucosus,Sus celebensis and Sus scrofa vittatus (Suidae,Mammalia)

Chromosome studies on the Javan warty pig (Sus verrucosus), the Sulawesi warty pig (S. celebensis) and a subspecies of the wild boar, S. scrofa vittatus, have revealed diploid chromosome numbers of 38. The morphology and C-band size of chromosome 10 are different in S. verrucosus and the two other species. Both S. verrucosus and S. celebensis have a Y chromosome that is larger than the Y chromosome of domestic and wild S. scrofa, and is submetacentric rather than metacentric. There are differences between all three species in the G-banding pattern of the long arm of the Y chromosome. The presence of 2n=38 chromosomes in the Javan warty pig and the Sulawesi warty pig provides new strong evidence that the basic chromosome number in the genus Sus is 38. The differences in karyotype between these pigs (chromosome 10 and the Y chromosome) confirm that they are separate species.     

Chromosomal location of 45S rDNA and dfr gene in Citrus sinensis

Dihydroflavonol 4-reductase (DFR) is a key enzyme in the anthocyanin biosynthesis. In this study, the localization of 45S rDNA and dfr gene (named as CsDFR-bo) of the blood orange (Citrus sinensis Osbeck cv. Ruby) on chromosomes was investigated by fluorescence in situ hybridization (FISH). A karyotype of C. sinensis was reconstructed based on the length of mitotic metaphase chromosomes. The 45S rDNAs were localized on chromosomes 2p and 7q. The detection ratios of 45S rDNA in the two chromosomes were 69.5 and 77.3 %, respectively. The CsDFR-bo was proved to be a single-copy gene and localized on chromosome 3p. The detection ratio of CsDFR-bo was 8.2 %.