Evolution of polyploidy in cultivatedNarcissus subgenusNarcissus

With the availability of vegetatively-propagated ancestral cultivars ofNarcissus together with their modern descendants, the majority of which have known dates of introduction, the evolution of polyploidy in cultivation during the past hundred years is traced using previously published chromosome counts. From an original range of largely diploid species triploid cultivars arose initially, followed later by tetraploids. After an early gradual increase which was similar to those of diploids and triploids the rate of introduction of new tetraploid cultivars accelerated rapidly from the 1920's onwards, with the result that tetraploids now comprise nearly 75% of the cultivars of which the chromosome numbers are known. Higher polyploid cultivars are very rare (5x) or unknown (6x, 8x etc.). The horticulturally optimal tetraploid level inNarcissus differs from the optimal levels of ploidy inHyacinthus (3x–4x) andTulipa (2x) which have growth habits that are similar to those ofNarcissus and which are also subject to similar breeding and propagation methods, but the optimal chromosome numbers of all three genera are comparable (2n=24−31). As their chromosomes are of an equivalent order of size there is thus an optimal DNA amount affecting growth rate and horticultural value in these genera which must be approximately the same in each one, but which is reached at different levels of ploidy depending on differences in basic chromosome number. A loss of vigour and/or growth rate rate with a consequent lack of favourable artificial selection probably occurs if the optimal DNA amount is greatly exceeded.     

DNA amounts of roses (<Emphasis Type="Italic">Rosa</Emphasis> L.) and their use in attributing ploidy levels

The aims of the investigation were to characterise variability among the DNA amounts of roses and assess the predictability of ploidy levels from DNA amounts. Chromosome numbers in the genus Rosa range from 2n = 2x = 14 to 2n = 8 x = 56 and aneuploidy is rare. Published 2C DNA amounts range from 0.78 pg in R. xanthina Lindl. and R. sericea Lindl. (2n = 2x = 14) to 2.91 pg in R. canina L. (2n = 5x = 35). In this investigation, DNA amounts were estimated by flow cytometry of leaf nuclei stained with propidium iodide, using Petroselinum crispum (2C DNA amount = 4.46 pg) as the internal calibration standard. Ploidy levels based on DNA amounts (DNA ploidy) were assigned by comparing their DNA amounts with published DNA amounts and identifying peaks and intervening discontinuities in frequency distributions of DNA amounts. 2C DNA amounts ranged from 0.83 pg in R. ecae (2x = 2x = 14) to 3.99 pg in R. acicularis (2n = 8 x = 56). Differences in the 1Cx-values (2C DNA amount/ploidy values) were found among the taxonomic sections of Rosa. Ploidy levels could be confidently assigned to most species and cultivars, but the ploidy of some specimens in the section Caninae was uncertain for reasons attributed to genomic diversity and aneuploidy. Cytochimerism was detected in three cultivars of R. x alba. DNA ploidy was determined in 384 specimens representing 74 species and 5 horticultural classes.     

The distribution and localization of an UDP-glucose: Flavonol 3-O-glucosyl transferase activity in pollen

Summary The occurrence of UDP-glucose: flavonol 3-O-glucosyltransferase activity in pollen extracts of various plant species was tested. In case ofAlnus, Quercus, Narcissus andTulipa pollen high enzyme activity could be detected. The high level of enzyme activity inTulipa pollen made short time extraction experiments possible, which showed that the O-glucosyltransferase activity might be located in the pollen wall, possibly in the exine.Abbreviations UDP-glucose uridine diphospho-D-glucose - UDP-rhamnose uridine diphospho-L-rhamnose - DTE dithioerythritol     

Chromosome numbers and DNA ploidy levels of selected species ofHieracium s.str. (Asteraceae)

Chromosome numbers and /or ploidy levels are reported for 44 species and subspecies ofHieracium s.str. from the following European countries: Andorra, Austria, Bulgaria, Czech Republic, France, Italy, Montenegro, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland and Ukraine. The chromosome numbers/DNA ploidy levels ofH. bocconei (2n ～ 4x),H. bupleuroides subsp.leviceps (2n = 27),H. caesioides subsp.caesioides (2n = 27),H. basifolium (H. caesium agg., 2n = 36),H. plumbeum (H. caesium agg., 2n = 36),H. glaucum subsp.nipholepium (2n= 27),H.gouanii (2n = 18),H. gymnocerinthe (2n = 27),H. ramondii (2n = 27),H. recoderi (2n = 18),H. stelligerum (2n = 18), andH. tomentosum (2n = 18, 2n ～ 2x, 2n ～ 3x) were determined for the first time. New ploidy levels are reported forH. cerinthoides s.str. (2n = 27),H. humile (2n = 36), andH. tommasinianum (2n = 27).     

Comparative karyomorphology and interrelationships ofSelaginella in Japan

Karyomorphological comparisons were made of 16 native and cultivated species ofSelaginella in Japan. The somatic chromosome numbers are 2n=16 inS. boninensis; 2n=18 inS. doederleinii, S. helvetica, S. limbata, S. lutchuensis, S. nipponica, S. selaginoides, S. tama-montana, andS. uncinata; 2n=20 inS. biformis, S. involvens, S. moellendorffii, S. remotifolia, andS. tamariscina; 2n=30 inS. rossii; and 2n=32 inS. heterostachys. The interphase nuclei of all species examined are uniformly assigned to the simple chromocenter type. The metaphase karyotype of 2n=16 (x=8) is 8 m (=median centromeric chromosomes)+8(st+t)(=subterminal and terminal). The group of the species having 2n=18 (x=9) is heterogeneous karyomorphologically: The karyotype ofS. nipponica is 2n=18=6 m+12(st+t),S. tama-montana 10 m+2 sm(=submedian)+6(st+t), andS. uncinata 6 m+7 sm+5(st+t). Although the remaining five species have the common karyotype 8 m+4 sm+6(st+t), the values of mean chromosome length are variable. Another group of the specles having 2n=20 (x=10) is homogeneous, since all species have the same karyotypes 8 m+4 sm+8(st+t) and have similar chromosome size. The karyotype of 2n=30 is 12 m+6 sm+12(st+t) and is suggested to be a triploid of x=10, and 2n=32=16m+16(st+t), a tetraploid of x=8. Thus, three kinds of basic chromosome numbers, x=8, 9, 10 are present in JapaneseSelaginella examined, and their karyomorphological relationships are discussed.     

Evolution alpiner Populationen vonEuphrasia (Scrophulariaceae): Chromosomenzählungen an diploiden und polyploiden Sippen aus den Ostalpen

Chromosome counts are reported for several E. Alpine taxa ofEuphrasia sect.Euphrasia. First records of diploidy for small-flowered taxa are 2n = 22 forE. inopinata andE. sinuata, related toE. minima (4 x). Aberrant E. AlpineE. hirtella is 2 x, just as the typical W. Alpine populations of this species. Tetraploidy, 2n = 44, has been found inE. pumila, close toE. stricta (also 4 x). The limitation of ploidy levels within sect.Euphrasia to 2 x and 4 x on the chromosome base number x = 11 is confirmed.

     

Polyploidy and aneuploidy inOphrys,Orchis, andAnacamptis (Orchidaceae)

Studies on chromosome numbers and karyotypes in Orchid taxa from Apulia (Italy) revealed triploid complements inOphrys tenthredinifera andOrchis italica. InO. tenthredinifera there is no significant difference between the diploid and the triploid karyotypes. The tetraploid cytotype ofAnacamptis pyramidalis forms 36 bivalents during metaphase I in embryo sac mother cells. Aneuploidy was noticed inOphrys bertolonii ×O. tarentina with chromosome numbers n = 19 and 2n = 38. There were diploid (2n = 2x = 36), tetraploid (2n = 4x = 72), hexaploid (2n = 6x = 108) and octoploid (2n = 8x = 144) cells in the ovary wall of the diploid hybridOphrys apulica ×O. bombyliflora. Evolutionary trends inOphrys andOrchis chromosomes are discussed.     

New chromosome numbers for plant taxa endemic to the Balearic Islands

Mitotic chromosome numbers are reported from 25 vascular plant taxa, endemic to the Balearic Islands that are poorly known cytogenetically. The chromosome numbers ofAnthyllis vulneraria subsp.balearica (2n=12),Cymbalaria fragilis (2n=56), andPolygonum romanum subsp.balearicum (2n=40) were determined for the first time. A new chromosome number was found in several populations ofAnthyllis hystrix (2n=70) suggesting that this species is decaploid, in contrast to an earlier work reporting a higher ploidy level (2n=12x=84). The new chromosome number 2n=32 was reported inHypericum hircinum subsp.cambessedesii. It is suggested that the previous count (2n=40) could be explained by the presence of anomalous pentaploid cells in some tissues, contrating with the presence of a regular tetraploid complement (2n=32). Cytogenetic observations suggest thatSibthorpia africana has a diploid chromosome complement of 2n=18, with 0–2 accessory chromosomes. Accessory chromosomes are also reported forPhlomis italica, being the first record of B chromosomes in this genus. Chromosomal instability was found inGalium crespianum andG. friedichii species, with three numbers 2n=44, 55 and 66. Two cytotypes differing in ploidy level were documented within single plants. It is suggested that both species share a regular complement of 2n=44 and that the past hybridization events and formation of regenerating roots from the typical rootstock ofG. crespianum andG. friedrichii could be involved in the genesis of chromosome variants through partial endopolyploidy and concomitant somatic segregation.     

Morphometric and karyological analysis of a population ofSesleria sadleriana Janka in the Biele Karpaty Mountains (Slovakia)

Morphometric and karyological analysis was employed to prove the occurrence of a population ofSesleria sadleriana Janka, at the locality “Vr?atec” in the Biele Karpaty Mts. (Slovakia). Morphological characters of this population were analysed statistically (ANOVA, PCA) and compared with those of a population ofS. sadleriana in Austria (Hainburg an der Donau) and a population ofS. albicans Kit. exSchult. in the Czech Republic (Javo?í?ko). The two populations ofS. sadleriana did not differ from each other in the uppermost leaf length, spike length, glume length, lemma length, awn length of lemma and palea length. On the other hand, they differed significantly in these morphological characters from the population ofS. albicans. Karyological analysis demonstrated that both populations ofS. sadleriana consisted of octoploid plants (2n=8x=56). In addition to chromosome counting, flow cytometry was employed to screen ploidy levels. The experiment demonstrated a positive correlation between the ploidy estimated by chromosome counting and DNA flow cytometry inS. sadleriana andS. albicans. Flow cytometry proved extremely useful for ploidy screening in large numbers of plants. The locality “Vr?atec” represents the northernmost locality ofS. sadleriana known so far.     

The allopolyploid origin ofSedum rupestre subsp.rupestre (Crassulaceae)

InSedum rupestre L. a polyploid series (x = 16) occurs in which aneuploid chromosome numbers and odd levels of ploidy prevail. The most common and widely distributed cytotype,S. rupestre subsp.rupestre, is 2n = 112. Plants resemblingS. rupestre subsp.rupestre can be obtained by hybridizing the tetraploid cytotypes ofS. forsterianum Sm. (2n = 48) andS. rupestre subsp.erectum 't Hart (2n = 64). Comparison of these artificial hybrids with their parents and a large number of plants ofS. rupestre subsp.rupestre (2n = 112) from nature showed thatS. rupestre subsp.rupestre and the artificial hybrids are morphologically indistinguishable, and intermediate betweenS. forsterianum andS. rupestre subsp.erectum. MorphologicallyS. rupestre subsp.rupestre is closer to subsp.erectum than toS. forsterianum. Chloroplast DNA restriction patterns ofS. rupestre subsp.rupestre, however, resembleS. forsterianum more closely. The combined results of the hybridization experiments, the analysis of the cpDNA restriction patterns, and the morphological variation indicate the allopolyploid origin ofS. rupestre subsp.rupestre. Natural hybrids inSedum (Crassulaceae) 4.     

Chromosome numbers, karyotypes and nuclear DNA contents from perennial polyploid groups ofCerastium (Caryophyllaceae)

Somatic chromosome numbers have been determined for the followingCerastium taxa:C. eriophorum (2n = 36),C. alpinum (2n = 72),C. transsylvanicum (2n = 108),C. arcticum (2n = 108),C. latifolium (2n = 36),C. carinthiacum (2n = 36),C. banaticum (2n = 36),C. arvense subsp.glandulosum (2n = 36),C. arvense subsp.arvense (2n = 72) andC. fontanum (2n = 144). Karyotypes of three diploid species (C. eriophorum, C. banaticum andC. latifolium), belonging to three different taxonomic groups, were analysed and found to be similar. The relative nuclear DNA contents of all taxa were determined by flow cytometry and, for five species, also by Feulgen cytophotometry. The values obtained by the two methods are similar. A comparison of nuclear DNA contents among diploids shows that values differ significantly between different taxonomic groups, and are correlated with average chromosome size. Within closely related polyploid groups nuclear DNA amounts increase from 2x- to 4x- and 6x taxa as 1 : 1.4 : 2.4 in theC. alpinum complex, whereas DNA amounts are doubled comparing 2x- and 4x-subspecies in theC. arvense complex.     

The neotropical angiosperm familiesBrunelliaceae andCaryocaraceae: First karyosystematical data and affinities

Chromosome numbers are polyploid, 2n = 28 inBrunellia comocladiifolia andB. mexicana, and 2n = 46 inCaryocar brasiliense, C. microcarpum andC. villosum. The chromosome are small in both genera, with a length of ca. 1,6-0,4µm. Interphase nuclei correspond to the prochromosomal and the chromocentric type, respectively. This is in conformance with the systematic placement ofBrunelliaceae intoCunoniales, and ofCaryocaraceae intoTheales. Brunellia exhibits affinities to various other orders ofRosidae (andHamamelididae), and is suggested to be primarily apetalous. On a comparative basis, the chromosome numbers found in both families are interpreted as paleopolyploid (4 x and 6 x). This apparently is in correspondence with their rather primitive features, systematic isolation, relatively depauperate status, and evidently great age.     

The karyology of some neotropicalStyracaceae

Chromosome numbers and karyomorphological characters have been investigated inPamphilia andStyrax (Styracaceae). Counted for the first time in the genus, two species ofPamphilia were found to have 2n = 16. The twoStyrax spp. investigated share withPamphilia the same chromosome number, a peculiar condensation behaviour of the chromosomes (Fig. 1a–c) and the same type of semi-reticulate interphase nucleus, results which indicate a close relationship of the two genera. The base number inStyracaceae is probably x = 8 (2n = 2x = 16) with stabilized triploids inHalesia andPterostyrax (2n = 3x = 24). A preliminary comparison withSapotaceae andEbenaceae does not allow a general karyological characterisation of the orderEbenales.     

The annual species ofAdonis (Ranunculaceae)—a polyploid complex

The five annual species ofAdonis L., sect.Adonis, growing in Israel, form a series of diploid, tetraploid and hexaploid species. Their somatic chromosome numbers are 2n = 16 inA. annua L.,A. dentata Del. andA. palaestina Boiss., 2n = 32 inA. microcarpa DC., 2n = 48 inA. aestivalis L.; counts forA. dentata, A. palaestina andA. microcarpa are new records. There are indications that alloploidization may have been involved in the process of speciation in sect.Adonis. A taxonomic survey of the 8 species of the section reveals that a higher ploidy level is usually combined with a larger distribution area.     

Ploidy-dependent genomic stability in the tissue cultures of ornamental Phlox drummondii Hook

Comparisons of the chromosome numbers, 2C nuclear DNA amounts and karyomorphology were made in explant cultures of diploid (2n = 2x = 14) and autotetraploid (2n = 4x = 28) Phlox drummondii. In 6–36 week old calli derived from diploid internodal segment explants, and in cells of root tips regenerated from such callus, marked differences were observed in chromosome number. The chromosome numbers ranged from 2n = 14 to 2n = 100 and DNA amounts from 8.20 to 63.20 pg in the diploid derived callus, while the extent of variation was much reduced in the regenerated roots. In contrast, the autotetraploid cultures were characterised by the maintenance of the same chromosome number and DNA amounts as the mother plant. Modified chromosome structures were not apparent in any of the cultures. The possible reasons for the chromosomal instability at the diploid level and stability at the tetraploid level are discussed.     

DNA ploidy levels and intraspecific DNA content variability in Romanian fescues (Festuca, Poaceae) measured in fresh and herbarium material

DNA ploidy level estimates are presented for 11 species and two natural hybrids ofFestuca sampled from 39 locations in Romania. Altogether 48 living samples (22 of them also as one-year-old herbarium specimens) and additional 65 one-year-old herbarium specimens were analyzed using flow cytometry with DAPI staining. The following DNA ploidy levels were assessed:F. callieri (4x),F. ovina (2x),F. pallens (2x),F. polesica (2x),F. pseudodalmatica (2x, 4x, 5x),F. pseudovaginata (2x),F. pseudovina (2x),F. pseudovina ×F. rupicola (4x),F. rupicola (6x),F. vaginata (2x),F. vaginata ×F. valesiaca (2x),F. valesiaca (2x) andF. xanthina (2x).Festuca pseudovaginata is reported for the flora of Romania for the first time. Measurements of one-year-old herbarium specimens produced significantly smaller sample/standard ratios (P<0.001) and higher coefficients of variance (P < 0.001) when compared with fresh plant samples. Several species exhibited marked intraspecific nuclear DNA content variability, documented by non-overlapping double-peaks or bimodal peaks in simultaneous measurements. A maximum difference of about 9.2% in relative DNA contents was observed inF. pallens, 5.5% inF. polesica, 4.2% inF. vaginata, and 3.8% inF. rupicola. DiploidF. xanhina (Festuca sect.Eskia) had 1.39–1.58 times higher monoploid relative DNA content in relation to the species of sectionFestuca.     

Chromosome analysis ofHaemanthus andScadoxus (Amaryllidaceae)

Chromosome analysis of nine species ofHaemanthus (2n = 16) and four species ofScadoxus (2n = 18), using conventional stains, Quinacrine fluorescence and C-banding, has shown that the two genera do not possess significant amounts of constitutive heterochromatin. The two genera are closely related and differ in respect of a translocation which has resulted in the dysploid reduction in chromosome number from 2n = 18 inScadoxus to 2n = 16 inHaemanthus.     

Karyology of theScorzonera (Compositae) species from the Iberian Peninsula

A karyological study of 15 taxa ofScorzonera L. from the Iberian Peninsula has been made. The chromosome numbers found inS. hispanica var.pinnatifida, S. baetica, S. reverchonii, S. angustifolia, S. laciniata var.calcitrapifolia and var.subulata (2n = 14) are new. Diploid cytotypes with 2n = 14 and 2n = 12 prevail, andS. hispanica var.crispatula is the only taxon which exhibits autopolyploidy (2n = 14, 28). x = 7 is considered to be the base chromosome number within the genus, with x = 6 being derived from it by translocation. This and detailed karyotype analyses allow to group the Iberian Peninsula species ofScorzonera into three groups.     

Cytotaxonomic study of genusHymenasplenium (Aspleniaceae) in Xishuangbanna, Southwestern China

The gametic chromosome numbers of sevenHymenasplenium (Aspleniaceae) species from Xishuangbanna, Yunnan Prov., China, were investigated. All the examined individuals ofH. obscurum, H. cheilosorum andH. latipinnum were sexual diploids with n=39 chromosomes. Intraspecific cytological variation was found inH. excisum, which has a sexual diploid (n=39) and a tetraploid (n=78). Only a triploid apogamous cytotype (n=ca.117) was found inH. laterepens. Hymenasplenium apogamum showed the most complicated intraspecific variation and included a sexual diploid (n=39), a sexual tetraploid (n=78) and an apogamous triploid (n=ca.117). This work reports for the first time the sexual diploids ofH. cheilosorum andH. apogamum, which are only apogamous elsewhere in east Asia, Himalayas and Indochina. These results may indicate that this area is one of the diversity centers ofHymenasplenium. Most of the above species have chromosome numbers based on x=39. In contrast,H. costarisorum contains a sexual diploid (n=36) and a sexual tetraploid (n=72), indicating that its basic number is x=36.     

Determination of the ploidy level in the genusHieracium subgenusPilosella (Hill) S.F. Gray by flow cytometric DNA analysis

Ploidy levels of 8 taxa of the genusHieracium subgenusPilosella collected from the same locality were determined by means of flow cytometric estimation of relative nuclear DNA content. The DNA anaysis of isolated nuclei stained with propidium iodide was performed with a flow cytometer equipped with an argon-ion laser. The diploid speciesH. lactucella (2n=18) was used as internal standard. As a rule 20 individuals per taxon were analysed. The flow cytometric analysis proved to be a rapid, accurate and sensitive method for screening of ploidy levels. Considering the known cytological data, the following ploidy levels were determined:H. aurantiacum 4x,H. pilosella 4x,H. piloselloides 4x,H. bauhini 5x,H. caespitosum 5x,H. stoloniflorum 4x,H. brachiatum 4x and 6x,H. leptophyton 7x.H. piloselloides, H. bauhini andH. caespitosum yield relative DNA contents which are the exact multiples of the haploid DNA content ofH. lactucella. In the case ofH. pilosella the haploid DNA content is lower than that predicted from the diploid, namely 0.43 instead of the expected 0.5. The haploid 1x DNA contents of bothH. brachiatum andH. leptophyton range between the value found forH. pilosella and those for the other investigated species, confirming their hybrid origin.