Nuclear DNA Content as an Index Character Discriminating Taxa in the Genus Petunia sensu Jussieu (Solanaceae)

Nuclear DNA contents over the total range of the genus Petuniasensu Jussieu comprising 20 taxa of Petunia sensu Wijsman (2n = 2 x = 14) and 32 taxa of Calibrachoa(2 n = 2 x = 18) wereestimated by flow cytometry after staining the nuclei with propidiumiodide (PI) or 4',6-diamidino-2-phenylindole (DAPI). With respectto nuclear DNA content, taxa of Petunia sensu Wijsman seemedto be homogeneous (2C = 2.60 to 3.41 pg), but Calibrachoa taxawere clearly separated into two groups: (1)C. parviflora andC. pygmaea(1.56 to 1.91 pg); and (2) remaining members of Calibrachoa(2.84to 3.26 pg). Taxa of Petunia sensu Wijsman exhibited largerPI/DAPI ratios (relative fluorescence intensity with PI stainingto that with DAPI staining) than Calibrachoa species exceptC.parviflora and C. pygmaea. This suggests that Petunia sensuWijsman has nuclear DNA with more adenine-thymine rich regionsthan Calibrachoa. Copyright 2000 Annals of Botany Company Calibrachoa, 4',6-diamidino-2-phenylindole (DAPI), flow cytometry, nuclear DNA content,Petunia , PI/DAPI ratio, propidium iodide (PI), Solanaceae     

Nuclear DNA Content in Twelve Species of Alstroemeria L. and Some of their Hybrids

Nuclear DNA content (2C-value), estimated through flow cytometryusing propidium iodide (PI), was shown to vary from 36.5 pgto 78.9 pg among 29 accessions of 12Alstroemeria species (2n=2x =16). The extremes were found inA. magnifica ssp.magnificaand inA. ligtu ssp.simsii , both belonging to the Chilean speciesgroup. The four Brazilian species exhibited less variation innuclear DNA content (49.8–56.4 pg), than the eight Chileanspecies (36.5–78.9 pg). Nuclear DNA content was positivelycorrelated (r =0.92,n =7,P <0.01) with the total chromosomelength. It was also positively correlated (r =0.85,n =5,P <0.01)with the length of C-bands, when only the Chilean species wereconsidered. When both karyotype parameters, length of non-C-bandedchromosome regions (x) and length of C-bands (y) were determined,it was possible to predict the nuclear DNA content (z) withthe formula z=0.65x +1.31y-0.45 (R ²=0.97,P =0.004). The DAPI fluorescence of most accessions was proportional tothe PI fluorescence (r =0.98,P <0.001), except for one accessionofA. ligtu , that had a relatively high PI/DAPI ratio (1.88).The PI/DAPI ratios of the Brazilian species were lower (1.59–1.67)than those of the Chilean species (1.68–1.88), which mightreflect a difference in base pair composition. Four groups ofspecies could be distinguished on the basis of fluorescencevalues. Diploid interspecific hybrids were shown to have a DNAcontent intermediate to the values of the parents involved.Both the PI and the DAPI fluorescence values of these hybridsapproximated the mid parent values. Tetraploids, derived fromselfing of diploids, had PI and DAPI fluorescence values thatwere twice that of the diploid hybrids. It was possible to distinguishaneuploids from euploids based on fluorescence values. Alstroemeria ; aneuploidy; C-banding; DAPI; evolution; flow cytometry; genome size; geophytes; karyotypes; Inca Lily; nuclear DNA; propidium iodide     

Genome size and environmental factors in the genus Pinus

Nuclear 1C DNA content in haploid megagametophyte tissue of 18 North American and one exotic Pinus species was determined using scanning microspectrophotometry. The nuclear DNA content in root meristematic cells of Zea mays L. ssp. mays, inbred line Va35 (4C = 10.31 pg) was used as a standard. DNA content measured by microspectrophotometry was verified using laser flow cytometry with two additional standards, Hordeum vulgare cv. Sultan (2C = 11.12 pg) and P. eldarica (2C = 47.30 pg). DNA values obtained by both methods were significantly correlated (r = 0.987). The 1C nuclear DNA content ranged from 21 pg to 31 pg. The ratio of DNA content in embryo tissue of P. eldarica to that in megagametophyte tissue was 1.72 by scanning microspectrophotometry and 1.74 by laser flow cytometry. To date, this is the most comprehensive data set available for North American Pinus species. Relationships between genome size of 18 North American Pinus species and climatic factors and indices of growth were investigated using regression and correlation analyses. Positive correlations were observed between nuclear DNA content and growth indices, minimum seed-bearing age, and seed dimensions. Strong negative correlations were observed between nuclear DNA content and two climatic factors, the lowest mean annual and monthly precipitation (excluding January) and the highest mean monthly spring air temperature. These correlations suggest that the large genome size and its variation in Pinus are adapted responses to the habitats of these species.     

Variable DNA content of <Emphasis Type="Italic">Cyclamen persicum</Emphasis> regenerated via somatic embryogenesis: rethinking the concept of long-term callus and suspension cultures

By flow cytometric experiments in Cyclamen persicum both with propidium iodide (PI) and DAPI (4′,6-Diamidino-2-phenylindol)-staining we were able to present (1) a new estimation for the absolute DNA content in the range of 3.17 pg DNA/2C and (2) ploidy abnormalities which were detected in the pathway of somatic embryogenesis. These aberrations might have arisen from ploidy mutations or abnormal polyploidization processes. Morphologically normal somatic embryos contained the smallest proportion of individuals with ploidy abnormalities, but also a regenerated plant in the green house displayed a significantly elevated DNA content. The results are discussed in the framework of future application of in vitro propagation techniques based on regeneration from fastly growing undifferentiated cells like long-term callus and suspension cultures.     

Substantial Genome Size Variation in Taraxacum stenocephalum (Asteraceae, Lactuceae)

There are only a few exceptions to the rule that polyploidy in Taraxacum is associated with agamospermy. One of them is the sexual, tetraploid species Taraxacum stenocephalum. Incidentally, remarkable variation in karyology was found in this species. The present study aims to confirm this variation by an extensive screen of nuclear DNA content. Individuals from two large populations in the Lesser and Greater Caucasus, Georgia were analyzed using flow cytometry to ascertain intraspecific nuclear DNA content variation. Across the whole data set comprising all 159 individuals, a 1.223-fold difference was detected based on propidium iodide (PI) analyses. To verify this finding, we compared flow-cytometric data obtained using DAPI (4′,6-diamidino-2-phenylindole) and PI staining using a representative subset of individuals. This comparison revealed a 1.194-fold difference in DNA content for DAPI and a 1.219-fold difference for PI. Mean nuclear genome size in absolute terms (2C value ± s.d.) was estimated at 4.38?±?0.21 pg, ranging from 4.01 pg to 4.89 pg, despite the invariable chromosome counts of 2n?=?32. A regression analysis comparing the datasets for DAPI and PI staining found a strong correlation between data obtained by the DAPI and PI dyes (R?=?0.976; P?=?0.0001). Simultaneous high-resolution flow-cytometric analyses also proved the accuracy of our findings. We discuss possible sources of these large differences in DNA content within Taraxacum stenocephalum. Further research is needed to identify the source of this remarkable variation.     

Nuclear DNA Amounts in Roses

Nuclei isolated from young leaves were stained with propidiumiodide (PI) and their fluorescence intensities were measuredby flow cytometry. The ratio of fluorescence intensities offour calibration standards and 34 roses to an internal standard,parsley (Petroselinum crispum), provided a basis for estimatingthe DNA amounts of P. crispum and rose. The 2C DNA amount ofP. crispum(2 n = 22) was estimated as 4.46 pg (s.d. ±0.08 pg). The 2C DNA amounts of diploid roses (2n = 14) variedbetween subgenera, sections and cultivars, and ranged from 0.78pg (s.d. ± 0.08 pg) in Rosa xanthina and R. sericea(sectionPimpinellifoliae) to 1.29 pg (s.d. ± 0.08 pg) in ‘Félicitéet Perpétue’ (Hybrid Sempervirens). Within eachsection, the DNA amounts of diploid species were similar. Inthe sections Carolinae and Cinnamomeae, DNA amounts were proportionalto ploidy numbers. In the Pimpinellifoliae, DNA amounts of tetraploidswere disproportionately larger than those of diploids whichsuggests that they originated as hybrids with species of sectionswith larger DNA amounts. Ratios of the fluorescence intensitiesof nuclei of roses to P. crispum(internal standard) were alsomeasured using 4',6-diamidino-2-phenylindole (DAPI) which bindspreferentially to AT base pairs. These DAPI ratios were lowerthan, but closely correlated (r² = 0.997) with PI ratios. Fluorescenceintensities of either PI or DAPI-stained nuclei of roses canbe used as rapid indicators of ploidy if variation in the DNAamounts between different taxonomic groups is taken into account.Copyright 2000 Annals of Botany Company Flow cytometry, nuclear DNA amounts, Petroselinum crispum, phenolics, Rosa, roses     

Karyology and nuclear DNA quantification of four species ofChaetomorpha (Cladophorales,Chlorophyta) from the western Atlantic

Chromosome numbers are given for four species ofChaetomorpha from the warm temperate and tropical western Atlantic. The basic chromosome number is six, with three median and three submedian chromosomes.Chaetomorpha species represent a polyploid series, with numbers of 12, 18 and 24 found in the present study. Microspectrophotometry data for each species were quantified by reference to standards with known DNA contents. Results indicate similar 2X =1C=12 genome sizes forC. aerea (0.20 pg) andC. brachygona (0.26 pg), and forC. antennina (0.53 pg) andC. melagonium (0.58 pg). These findings are compared with karyological features ofCladophora species to characterize the karyology of the cladophoralean genome.     

Nuclear DNA content of the Solanum spp. in the series Etuberosa as determined by laser flow cytometry

Nuclear DNA content was determined in three accessions of Solanum brevidens, three accessions of S. etuberosum, and one accession of S. fernandezianum, which are diploid (2n = 2×= 24), closely related, non tuber-bearing wild potato species belonging to the series Etuberosa (Solanaceae). The plants were grown in vitro at 18°C or at 25°/22°C (day/night). S. brevidens was also grown in soil in the glasshouse at 25°/19°C (day/night), and in growth chambers at 18°C or 32°C. Leaf nuclei were isolated using a chopping method and stained with propidium iodide. Chicken red blood cells (CRBC; 2.33 pg) were added to the samples of nuclei as internal standards. The fluorescence of plant nuclei relative to CRBC was measured with an EPICS PROFILE flow cytometer. The 2C values of in vitro-grown S. brevidens and S. etuberosum were similar (1.48–1.54 pg, depending on the accession), but they were smaller than the 2C value of S. fernandezianum (1.63 pg). The 2C values of S. brevidens and S. etuberosum were generally smaller than those of the diploid species S. berthaultii (1.60–1.61 pg) and the diploid clones of S. tuberosum (1.60–1.72 pg). A similar relative difference of nuclear DNA content was found also between tetraploid S. brevidens and tetraploid S. tuberosum (2C = 3.15–3.16 pg and 3.50–3.62 pg, respectively). High (32°C) and low (18°C) growth temperatures caused abnormal changes in morphology and reduced fertility in S. brevidens in the growth chamber. The 2C values of S. brevidens grown at 25°/19°C (day/night) or at 32°C were similar, whereas the 2C values were c. 10% lower at 18°C.     

Nuclear DNA content of Pinus sylvestris (L.) as determined by laser flow cytometry

Nuclear DNA content was determined in nuclei isolated from needles, stems and roots of in vitro grown seedlings and from megagametophytes and embryo of mature seeds in three accessions of Pinus sylvestris L. One accession was from Inari, northern Finland at timber line, and two accessions were from the Alpine region in Italy. Nuclei were mechanically isolated by a chopping method, stained with propidium iodide, and DNA content was determined using an EPICS PROFILE laser flow cytometer. Nuclei isolated from leaves of barley (Hordeum vulgare L. cv. Sultan; 2C=11.12 pg) were used as an internal standard for measurement of pine nuclei. Mean 1C nuclear DNA content of P. sylvestris was 27.88 pg as determined from megagametophyte tissue. Mean 2C value was 52.25 pg as determined from stem and root tissue, and 55.58 pg as determined from embryo tissue. The ratio of 2C to 1C value was 1.87 and 1.99, respectively. Extracts of nuclei from needles contained propidium iodide-absorbing debris which may have interfered with measurements and resulted in lower 2C values than those obtained from stem and root.     

DNA content of wheat monosomics at interphase estimated by flow cytometry

 Two complete, independently maintained sets of 21 monosomic wheat lines derived from cv. ‘Chinese Spring’ were analyzed for their DNA content at the G1 stage with flow cytometry. The DNA content of individual chromosomes was estimated by subtracting the value of a monosomic line from that of euploid wheat. Our data show that the estimated 2C DNA of individual wheat chromosomes in 21 monosomics at the G1 stage ranges from about 0.58 pg in chromosome 1D to approximately 1.12 pg in chromosome 3A. The A genome (2C=6.15 pg) seems to contain more DNA than the B (2C=6.09 pg) and D (2C=5.05 pg) genomes. Analysis of variance showed significant differences (α=0.01) in DNA content both among homoeologous groups and among genomes. Our estimates of interphase DNA content of wheat chromosomes from monosomic lines were poorly correlated to the chromosome sizes at metaphase (r=0.622, P≤0.01). This poor correlation might be due to differential coiling among chromosomes during cell division, possible bias of fluorochrome binding to heterochromatin, or heterogeneity among monosomic lines. Finally, flow cytometry may aid but cannot replace cytological checks in aneuploid maintenance. Received: 21 January 1997 / Accepted: 23 June 1997     

Genome size variation inCajanus cajan (Fabaceae): A reconsideration

A recent investigation of genome size in certain samples of the pigeonpea,Cajanus cajan, indicates values from 1.55 pg to 1.99 pg (1C level), which is 1.29-fold variation between accessions. In the present analysis those of these accessions which had particularly high or low DNA contents in that study were subjected to a reanalysis using propidium iodide and DAPI flow cytometry and Feulgen densitometry. Only minor differences in genome size, not more than 1.047-fold, were found with flow cytometry, and no significant differences were obtained with Feulgen densitometry. The previously reported genome size cannot be confirmed. It is about half as large and was determined in the present study as 0.825 pg (1C, propidium iodide flow cytometry,Glycine max as standard) and 0.853 pg (1C, Feulgen densitometry,Allium cepa andPisum sativum as standards), respectively.     

Nuclear DNA content of Vitis vinifera cultivars and ploidy level analyses of somatic embryo-derived plants obtained from anther culture

Flow cytometry was employed to determine the ploidy level of Vitis vinifera L. somatic embryo-derived plants obtained from anther culture. Only one among the 41 analysed plants (2.4%) presented somaclonal variation (tetraploidy); the other plants were diploid. No significant differences (P≤0.05) were detected between diploid and parental field plants. No haploid or aneuploid plants were observed. The nuclear DNA content of nine V. vinifera cultivars was also estimated using flow cytometry. A non-significant variation was found among the cultivars, with DNA content ranging from 1.17 pg/2C (cv. ‘Tinta Barroca’ and ‘Viosinho’) to 1.26 pg/2C (cv. ‘Cabernet Sauvignon’). These results and previous studies on other Vitis species suggest that Vitis genome is stable with regard to nuclear DNA content.     

Genome size and A-T rich DNA in selachians

The nuclear DNA content of 23 selachian species (10 Batoidea, 11 Galeomorphii, and 2 Squalomorphii) was histophotometrically studied. Their genome sizes range from 7.5 pg/N in Raja fillae (Batoidea) to 34.1 pg/N in Oxynotus centrina (Squalomorphii).Results show slight differences in the pattern of quantitative variations between the superorders Batoidea and Galeomorphii; Squalomorphii preserve their peculiar wide interspecific variability at the intrafamilial level, with values sited between 13.1 and 34.1 pg/N.In 21 species also the DNA base composition was determined by means of DAPI. The study shows that in the species examined the DAPI positive fraction varies from a minimum of 27.7% in Oxynotus centrina, which possesses the largest genome size among all the Selachians studied, to a maximum of 72.5% in Carcharhinus limbatus. As a whole the data show an inverse correlation between the DNA content and the DAPI positive fraction, a condition common to all cold-blooded vertebrates.The low percentage of DAPI positive DNA found in Oxynotus centrina could be attributable to a lower stainability by the fluorochrome caused by a higher chromatin condensation in the erythrocytes.The validity of the DAPI method was verified by comparison with the biochemical assay according to the thermal denaturation method in 6 selachian species.     

COMPARISON OF PLANT DNA CONTENTS DETERMINED BY FEULGEN MICROSPECTROPHOTOMETRY AND LASER FLOW CYTOMETRY

An improved procedure is reported for determining DNA amounts of plant nuclei. Nuclei stained with propidium iodide, isolated from chopped plant leaves, were passed through an Ortho Cytofluorograph with a Lexel model 95 argon laser (514 nm) and the fluorescence measured, integrated, and recorded using an Ortho 2140 Data Acquisition computer. All nuclear samples were mixed with nuclei of Sultan barley (2C DNA content = 11.12 pg [picogram]) as an internal standard. DNA contents of ten plant species, ranging from 2C = 1.7 pg to 36.1 pg measured by flow cytometry, correlated strongly (r = 0.99, slope = + 1.00) with DNA contents determined from Feulgen-stained nuclei of the same species using microspectrophotometry. The flow cytometric procedures were sufficiently sensitive to detect differences in DNA content between inbred lines of corn and their F1 hybrids. Our results obtained with improved procedures, specifically using propidium iodide as a fluorochrome and plant nuclei instead of chicken erythrocytes as an internal standard, demonstrate that laser flow cytometry can be a precise, rapid, and reliable method for determining nuclear DNA content of plants.     

Karyotypes, chromosome bands and genome size variation in New Zealand endemic gymnosperms

Karyotype studies on 20 taxa of gymnosperms endemic to New Zealand show a wide diversity of chromosome number and form. Fluorochrome banding with DAPI and CMA reveals a depauperate pattern of bands with CMA and no reliable banding with DAPI. Characteristically one pair of chromosomes shows a prominent CMA band, which may or may not be associated with a secondary constriction. A band size polymorphism was observed in all plants ofDacrycarpus dacrydioides, irrespective of the sex of the plant. Measurements of genome size by flow cytometry show a range of values from 12.3 pg to 40.0 pg DNA per 2C nucleus. Intraspecific variation in genome size was observed inManoao colensoi.     

Quantitation of Absolute Pneumocystis carinii Nuclear DNA Content. Trophic and Cystic Forms Isolated from Infected Rat Lungs are Haploid Organisms

The Pneumocystis carinii carinii DNA content in nuclei of trophic forms and cysts (spore cases) containing 2, 4, or 8 intracystic bodies, were compared using quantitative fluorescence image analysis. The nuclear DNA content was found to be lower than the theoretical limits of Feulgen cytophotometry. Several fluorescent DNA dyes provide brighter staining, but these techniques suffer from nonspecific binding to other cellular components, such as RNA. It was demonstrated that the thick glycocalyx surfaces of trophic forms and the cyst walls of P. carinii organisms, as well as the cell wall of S. cerevisiae, bound all fluorescent dyes tested to varying degrees. Hence in this study, measurements were performed on cells in which the outer surfaces of organisms were first removed with lyticase. Two stains that appeared most specific for DNA, DB181 and 4′,6-diamidino-2-phenylindole (DAPI), were used for quantitations; lower deviations of fluorescence intensities were observed with DB181. Haploid wild type Saccharomyces cerevisiae and cdc-28 temperature-sensitive mutant cells, accumulated at the restrictive temperature (37° C), were used as quantitative internal standards for estimating the absolute nuclear DNA content of P. carinii. Haploid wild type and mutant nuclei stained with DAPI had the same relative fluorescence intensities. The P. carinii nuclear DNA content of trophic forms and individual intracystic bodies (spores), regardless of life cycle stage, were not different. The mean values obtained were 6.9 and 6.7 fg DNA/nucleus with DB181 and DAPI, respectively (approximately 9.26 and 8.99 Mbp nucleotides, respectively). Since these would include 2C (G-2 phase) and S-phase nuclei, a 1C population of nuclei was selected by histogram distributions of DB181-stained nuclei. Almost all nuclei analyzed in all life cycle stages fell within this population. The 1C mean of 6.55 fg DNA/nucleus (median, 6.62 fg DNA/nucleus) was estimated as representing 8.79 Mbp nucleotides, assuming only A-T binding of the dye and taking into account the G+C content of S. cerevisiae and P. carinii. A 4C (G-2-phase diploid nuclei) population was not detected in histograms of DB181- or DAPI-stained nuclei. The P. carinii nuclear DNA content values obtained in this study were similar to those independently obtained by calculating the total DNA in the organism's chromosomes resolved by electrophoretic techniques. Together, the data on total chromosome numbers and the estimated DNA content of those chromosomes, with our quantitation of nuclear DNA content of different life-cycle stages demonstrate that P. carinii carinii isolated from infected rat lungs are haploid organisms.     

Chromosome CPD(PI/DAPI)- and CMA/DAPI-Banding Patterns in Allium cepa L.

Chromosome banding patterns of Allium cepa L. were obtained by using fluorescent dye combinations chromomycin A₃ (CMA) + 4",6-diamidino-2-phenylindole (DAPI), DAPI + actinomycin D (AMD) and propidium iodide (PI) + DAPI. In A. cepa,telomeric heterochromatin displayed dull fluorescence after staining with DAPI and DAPI/AMD. After joint staining with the GC-specific CMA and AT-specific DAPI, the CMA-positive fluorescence of the NOR region and the telomeric bands of C-heterochromatin was observed. In combination with DAPI, PI, a dye with low AT/GC specificity, produced almost uniform fluorescence of chromosomal arms and heterochromatin, whereas the NOR-adjoining regions displayed bright fluorescence. Denaturation of chromosomal DNA (2 × SSC, 95°C for 1–3 min) followed by renaturation (2 × SSC, 37°C, 12 h) altered the chromosome fluorescence patterns: specific PI-positive bands appeared and the contrast of CMA-banding increased. Bright fluorescence of NOR and adjoining regions was also observed in the case. Three-minute denaturation led also to a bright PI-positive fluorescence of telomeric heterochromatin. The denaturation of chromosomal DNA before staining results in changes of the DAPI fluorescence pattern and in the appearance of bright DAPI fluorescence in GC-rich NOP regions. The mechanisms underlying the effects of denaturation/renaturation procedures on chromosome banding patterns obtained with different fluorochromes are discussed.     

Flow cytometric analysis of nuclear genome of the Ethiopian cereal tef [Eragrostis tef (Zucc.) Trotter]

Flow cytometric analysis of nuclear DNA content was performed by using nuclei isolated from young leaf tissue of tef (Eragrostis tef). The method was very useful for rapid screening of ploidy levels in cultivars and lines of tef representing the phenotypic variability of this species in Ethiopia. The results of the analysis showed that all cultivars were tetraploid. Flow cytometry was also used to determine nuclear DNA content in absolute units (genome size) in four tef cultivars. Nuclei isolated from tomato (Lycopersicon esculentum, 2C=1.96 pg) were used as an internal reference standard. The 2C DNA content of individual tef cultivars ranged from 1.48 to 1.52 pg (1C genome size: 714 Mbp-733 Mbp), the differences among them being statistically nonsignificant. The fact that the nuclear genome of tef is only about 50% larger than that of rice should make it amenable for analysis and mapping at the molecular level.     

Genome size variation in Central European species of Cirsium (Compositae) and their natural hybrids

BACKGROUND AND AIMS: Nuclear DNA amounts of 12 diploid and one tetraploid taxa and 12 natural interspecific hybrids of Cirsium from 102 populations in the Czech Republic, Austria, Slovakia and Hungary were estimated. METHODS: DAPI and PI flow cytometry were used. KEY RESULTS: 2C-values of diploid (2n = 34) species varied from 2.14 pg in C. heterophyllum to 3.60 pg in C. eriophorum (1.68-fold difference); the 2C value for the tetraploid C. vulgare was estimated at 5.54 pg. The DNA contents of hybrids were located between the values of their putative parents, although usually closer to the species with the smaller genome. Biennial species of Cirsium possessed larger nuclear DNA amounts than their perennial relatives. Genome size was negatively correlated with Ellenberg's indicator values for continentality and moisture and with eastern limits of distribution. A negative relationship was also detected between the genome size and the tendency to form natural interspecific hybrids. On the contrary, C-values positively corresponded with the spinyness (degree of spinosity). AT frequency ranged from 48.38 % in C. eriophorum to 51.75 % in C. arvense. Significant intraspecific DNA content variation in DAPI sessions was detected in C. acaule (probably due to the presence of B-chromosomes), and in tetraploid C. vulgare. Only the diploid level was confirmed for the Pannonian C. brachycephalum, generally considered to be tetraploid. In addition, triploidy was discovered for the first time in C. rivulare. CONCLUSIONS: Considerable differences in nuclear DNA content exist among Central European species of Cirsium on the diploid level. Perennial soft spiny Cirsium species of wet habitats and continental distributions generally have smaller genomes. The hybrids of diploid species remain diploid, and their DNA content is smaller than the mean of the parents. Species with smaller genomes produce interspecific hybrids more frequently.     

DNA contents,giemsa banding,and systematics inScilla bifolia,S. drunensis,andS. vindobonensis (Liliaceae)

DNA contents have been determined cytophotometrically in the three Central European, relatedScilla speciesS. bifolia (2n = 18, 2 x, 1 C = 6.2 pg),S. drunensis (2n = 36, 4 x, 1 C = 12.8 pg), andS. vindobonensis (2n = 18, 2 x, 1 C = 9.4 pg). The tetraploid speciesS. drunensis contains twice as much DNA as the diploidS. bifolia. However, the diploid speciesS. vindobonensis differs in DNA content fromS. bifolia by a factor of about 1.5. This difference is largely due to euchromatic DNA, although the higher DNA content inS. vindobonensis is combined with higher heterochromatin content. The data indicate thatS. bifolia andS. drunensis on the one hand, andS. vindobonensis on the other hand are phyletically well separated. Previous taxonomic conclusions from morphology as well as C-banding are thus corroborated.Evolution ofScilla and Related Genera, V.