Biosystematic studies on the genusIsoetes (Isoetaceae) in Japan. III. Variability within qualitative and quantitative morphology of spores

Qualitative and quantitative characteristics of mega- and microspores from all the cytotypes of JapaneseIsoetes are described based on the voucher specimens whose chromosome numbers were known. InI. japonica, the hexaploid possessed reticulate megaspores and levigate microspores, while the octaploid and the heptaploid had echinate microspores. Mega- and microspores of the hexaploid and the octaploid were of normal appearance, while those of the heptaploid displayed polymorphism. The tetraploid and the hexaploid ofI. sinensis resembled each other, since they both possessed cristate megaspores and echinate microspores. Echinate megaspores and levigate microspores characterized the diploidI. asiatica. The spore size was largely variable within each cytotype, while the size of the megaspores varied more than that of the microspores. The microspore length was closely correlated with polyploid level. InI. sinensis, the mean microspore length of the tetraploid was 27.6 μm while that of the hexaploid was 31.9 μm, hence these two cytotypes were easily distinguishable. In the hexaploidI. japonica, variations in mega-and microspore size displayed geoclinal variation showing a positive correlation (r=0.43–0.55) with the longitude and the latitude of the populations. A palynological key for cytotypes is presented.     

Biosystematic study ofSedum L. subgenusAizoon (Crassulaceae)

Chromosome numbers of 114 individuals from twelve populations ofSedum aizoon L. var.aizoon (Crassulaceae) are reported. They include 37 different chromosome numbers ranging from 2n=71 to 124. Although the chromosome number variation has been found in all populations examined, no correlation with geographical distribution could not be found. Various kinds of meiotic irregularities, i.e., multivalents, univalents, chromosome lagging, and polysporous “tetrad” formation have been found. These irregularities lead to the formation of gametes with various chromosome numbers. All aneuploid plants set seeds and seem to reproduce sexually. The extensive aneuploidy in var.aizoon seems to be caused by the unequal chromosome segregation in meiosis and the subsequent fertilization of gametes with various chromosome numbers.     

Basic chromosome numbers of terrestrial orchids

The chromosome numbers of forty-one Brazilian species belonging to 11 genera of preferentially terrestrial orchids (subfamilies Cypripedioideae, Spiranthoideae, Orchidoideae, and Vanilloideae) were examined. Previous records for these subfamilies were reviewed in order to identify the ancestral chromosome numbers of terrestrial orchids. The variation observed within the subfamilies Spiranthoideae (2n=28, 36, 46, 48 and 92), and Orchidoideae (2n=42, 44, ca. 48, ca. 80, 84, and ca. 168) was similar to that previously reported in the literature. In the subfamily Spiranthoideae, some species of Prescottia (subtribe Prescottiinae) and some genera of Spiranthinae showed a bimodal karyotype with one distinctively large pair of chromosomes. The analysis of chromosome numbers of the genera in subfamilies revealed the predominance of the polyploid series 7, 14, 21, 28, 42 with a dysploid variation of ±1 in each ploidy level. These results suggest that the basic chromosome number of terrestrial orchids is x₁=7 for the subfamilies Spiranthoideae and Orchidoideae, as well as other Epidendroid orchids, and that the majority of the genera are composed of palaeopolyploids.     

First report of chromosome numbers of the Carlemanniaceae (Lamiales)

The Carlemanniaceae comprises two small genera that are restricted to East Asia: the Carlemannia and Silvianthus. These genera were previously placed in the Rubiaceae or Caprifoliaceae, but are now considered a distinct family that is probably related to the Oleaceae in the Lamiales. The family is still poorly understood with respect to its morphological characteristics. Here, we present the first report of the chromosome numbers of the family using species from both genera, i.e., Carlemannia tetragona, Silvianthus bracteatus ssp. bracteatus, and S. bracteatus ssp. clerodendroides. The species were compared with the chromosome numbers of Oleaceae and associated families using a Bayesian tree that was generated from rbcL and ndhF sequence data from Genbank. C. tetragona had 2n = 30 (x = 15), whereas the two subspecies of Silvianthus had 2n = 38 (x = 19). Comparisons of chromosome numbers support the distinctness of the Carlemanniaceae, not only from the Oleaceae (x = 11, 13, 23), but also from the Tetrachondraceae (x = 10, 11), a family that is possibly related to the Carlemanniaceae and/or Oleaceae in the Lamiales. The notable difference in chromosome number between Carlemannia and Silvianthus, as well as the differences in other characteristics (pollen, seed, and fruit morphology), suggests that the family split early in its evolution.     

Cytological studies on meiotic configurations of intraspecific aneuploids ofCarex blepharicarpa (Cyperaceae) in Japan

Chromosome configurations at meiotic metaphase I ofCarex blepharicarpa were determined for 245 individuals collected from 11 localities in the Chugoku District of Japan. Nine intraspecific aneuploids, 2n=26–33 and 41, were found. The most common diploid number was 29, and found in 73 individuals. No clear geographical pattern was suggested by a distribution of these aneuploids. A consecutive series of chromosome numbers from 2n=26 to 32 was found in two populations from Okayama Prefecture. At meiotic metaphase I, univalents and multivalents were found, and one to three trivalents were observed in each aneuploid. Heteromorphic chain trivalents comprising large, medium and small chromosomes were prevalent. Homomorphic trivalents, which are thought to be originated from chromosome duplications caused by unreduced gametes, were found in only one individual with 2n=41. The high frequency of heteromorphic trivalents in this species indicates that most aneuploidy probably results from fission and/or fusion of chromosomes.     

国产毛茛科驴蹄草属两种植物的细胞学研究

为探讨国产毛茛科(Ranunculaceae)驴蹄草属(Caltha L.)植物的细胞学特征,对驴蹄草(C.palustris L.)3个居群和花葶驴蹄草(C.scaposa Hook.f.&Thoms.)5个居群进行了细胞学研究。驴蹄草贵州纳雍居群的染色体数目为2n=32(四倍体),两个云南中甸居群的染色体数目均为2n=64(八倍体)。花葶驴蹄草四川红原、康定、石渠居群的染色体数目均为2n=32(四倍体),该数目为首次报道;西藏林芝和云南德钦居群的染色体数目均为2n=64(八倍体)。驴蹄草的染色体比花葶驴蹄草大。这两种植物的32或64条染色体分别以4条或8条为单位大致能够排列为8组同源染色体,但同一组内的染色体经常具有明显的异形性(heteromorphy),不同居群的核型组成多少具有差异。同时,还分析了驴蹄草和花葶驴蹄草的不同倍性细胞型在我国的地理分布式样。     

Biosystematic study ofSedum L. SubgenusAizoon (Crassulaceae)

Variation in chromosome number and morphological characters ofSedum aizoon L. var.floribundum Nakai were investigated to analyze, correlations between them. Geographical variation in chromosome number was also examined. Chromosome numbers of 189 individuals from 55 localities were counted as 2n=32, 33, 34, 48, 61, 64, 78, 80, 84, 85, 88, 93, 94, 95, 96, 97 and 102. InSedum subgenusAizoon, which has the basic number of X=8, var.floribundum of the speciesaizoon showed a polyploid series from tetraploid to dodecaploid. Tetraploids were found most frequently in this variety. More than two chromosome numbers were found in all the populations with 2n=61 or more. Tetraploids were mainly distributed in the eastern part of Japan, and higher polyploids (higher than hexaploid level except octaploids) were distributed in the western part of Japan. Thirteen morphological characters were examined in 119 individuals belonging to 30 populations. The principal component analysis and the cluster analysis of these characters indicated that populations with similar chromosome numbers were not always morphologically similar. Ten morphotypes can be recognized based on the combination of chromosome number and morphological characters. ThusS. aizoon var.floribundum can be regarded as a polyploid-aneuploid complex.     

Biosystematic studies inTulipa sect.Eriostemones (Liliaceae)

Evolutionary relationships and infra- and interspecific variability within the species ofTulipa sect.Eriostemones Boissier were studied. Measurements of 35 morphological characters were used for principal component and canonical variate analyses. Crossing experiments and chromosome counts were carried out. Two axes of morphological variation were encountered within the principal component plot. Variation of subsect.Biflores was traced along one axis, the species of subsect.Australes as well asSaxatiles were found along the other axis. The nomenclature of names of subsect.Biflores was considered on the basis of their geography, specific morphological characteristics and polyploidy. Conspecificity of names of all species was tested based on both morphology and crossability.     

Biosystematic studies in European species ofGladiolus (Iridaceae)

Representative specimens of the EuropeanGladiolus species (excl.G. palustris) were used to study variability and relationships. 25 characters were measured for cluster, principal component and Wells hybrid analyses (Figs. 1–3). Three (with PCA) respective five (cluster analysis) main groups are revealed. Bulb characteristics appear to be very useful for indentification. The subspecific status ofG. communis subsp.byzantinus is confirmed. Hybrids of four different parental combinations were detected in our collection. Most species include polyploid series. Partly 3x, 4x, 6x, 8x, 12x and aneuploid cytotypes but no diploids were found. Polyploids as pioneers and hybridization of sympatric species are apparently linked to migrations during the glacial periods.     

Studies on the alteration of chromosome copy number and cell division potential in a dnaA mutant of Escherichia coli

Summary The dnaA167 mutant of Escherichia coli, N167, maintains, on the average, two replicating chromosomes per cell at the perimissive growth temperature of 30°C and only one per cell at the higher permissive growth temperature of 38°C. When the growth temperature of this mutant is changed from 30° to 38°C the cells rapidly readjust their chromosome copy number from two to one. I have examined the kinetics of this transition with reference to DNA replication and cell division. My results indicate that this mutant uncouples cell division from chromosome duplication to achieve the appropriate copy number, suggesting that the dnaA gene product may be involved in the coordination between these two cellular events.     

Cytogeography of<Emphasis Type="Italic"> Ixeris nakazonei</Emphasis> (Asteraceae,Lactuceae) in the Ryukyu Archipelago of Japan and Taiwan

The cytogeographical structures of Ixeris nakazonei, a putative hybrid between I. debilis (6x) and I. repens (2x), were investigated in the Ryukyu Archipelago and Taiwan. In the Ryukyus, I. debilis occurs on Miyakojima Island of the southern Ryukyus and northward, while I. repens occurs on all islands except for Iriomotejima and Yonagunijima Islands. I. nakazonei, comprises six polyploid cytotypes, 3x, 4x, 5x, 6x, 7x and 8x, based on x=8. Four cytotypes from 3x to 6x occur in the central Ryukyus, while four cytotypes from 5x to 8x occur in the southern Ryukyus. The higher polyploids of I. nakazonei tend to be distributed in the more southerly area. Tetraploids of I. nakazonei always co-occur with I. debilis and I. repens, supporting the hybrid origin of this cytotype. Considering the chromosome number, octoploids, which predominate in the southern Ryukyus and Taiwan, may have derived directly from hybridization between I. debilis and I. repens. Odd-numbered polyploids of I. nakazonei, 3x, 5x and 7x, are relatively rare. Their chromosome numbers indicate that triploids and heptaploids are hybrids between the tetraploid of I. nakazonei and I. repens, and between the octoploid of I. nakazonei and I. debilis, respectively. Pentaploids of I. nakazonei in the central and southern Ryukyus are, respectively, hybrids between the tetraploid of I. nakazonei and I. debilis and between the octoploid of I. nakazonei and I. repens, indicating that pentaploids of I. nakazonei have at least two independent origins.     

Steric hindrance and its effect on chromosome movement inVicia faba somatic cells

Summary The arrangement of chromosome arms in metaphases and anaphases has been studied inVicia faba root meristem cells. During metaphase, the long chromosome arms are aligned parallel to the spindle axis. As a consequence, at the onset of anaphase, one chromatid can move straight ahead to the spindle pole whereas the other has to invert its orientation. Specially in narrow cells it has been observed frequently that some chromatids move in a reverse orientation to the pole, i.e., they move telomere-first instead of centromere-first. This behaviour results in a chromatid which protrudes beyond the main group of late anaphase or telophase chromatids. It is dicussed that the most likely explanation for the phenomenon is that in narrow cells chromatid behaviour is influenced by steric hindrance by the tightly packed surrounding chromatids and microtubules. When there is insufficient room, some chromatids are unable to make the required U-turn. Under such conditions the kinetochore of a non-inverted chromatid pulls the chromatid in a reverse orientation to the pole. An alternative explanation, i.e., protruding chromatids being the result of a neocentric activity at the telomere end of a reverse-directed chromatid or the lateral associations of spindle microtubules, failed to find support by electron microscopical studies.     

Analysis of metaphase I chromosome association in species of the genus Aegilops

Summary Metaphase-I chromosome associations in every diploid and polyploid species of the genus Aegilops were studied using C-banding in order to analyse the cytogenetic behaviour of the whole complement as well as of specific genomes in different polyploid species. Differences were observed in the frequency of associations per cell among different species of the same ploidic level and even between species sharing the same genomic constitution. Differences were also found between different genomes within the same polyploid species and between the same genome when present in several diploid and polyploid species. Several factors proposed as having an influence on the frequency of metaphase-I associations, such as chromosome morphology, C-heterochromatin content, genetic control and genome interactions, are discussed. Most of the polyploid Aegilops species showed a diploid-like behaviour at metaphase I although multivalents involving homoeologous associations were occasionally observed in Ae. biuncialis, Ae. juvenalis and Ae. crassa(6x); therefore, the Aegilops diploidising genetic system is not equally effective in all polyploid species.     

Karyomorphological studies onWoodwardia sensu lato of Japan

Karyomorphological comparisons were made of five species of JapaneseWoodwardia. There were no marked differences at interphase and prophase among the five species.Woodwardia japonica, W. prolifera, andW. unigemmata were diploid with 2n=68 and the formulas of their metaphase karyotypes uniformly 4m(median centromeric chromosomes)+12sm(submedian)+52(st+t)(subterminal and terminal).Woodwardia orientalis was tetraploid with 2n=136 and 8m+24sm+104(st+t), and the ratio of each chromosomal type to total complement was identical to that of three diploid species. These four species had several characteristics in common:x=34, the longest chromosome of sm, and a mean chromosome length over 3.0 μm. AlthoughWoodwardia orientalis showed some similarity toW. prolifera, it seems to be an allotetraploid which originated by chromosome doubling of a hybrid ofW. prolifera and a diploid species as yet karyomorphologically unknown.Woodwardia kempii was tetraploid with 2n=124 and 8m+24sm+92(st+t), and differed from the others in havingx=31, the longest chromosomes of t, and a mean chromosome length under 3.0 μm. This species has been classified as an independent genus,Chieniopteris, and our karyomorphological study supports this treatment.     

Karyotype of North American shortnose sturgeon Acipenser brevirostrum with the highest chromosome number in the Acipenseriformes

The shortnose sturgeon Acipenser brevirostrum was revealed to have a larger number of chromosomes than previously reported for other sturgeon species. Its chromosome number ranged from 362 to 372 (of ten specimens examined), showing intraindividual variation. The karyotype of metaphase with the highest chromosome number (372) consisted of 89 pairs of macrochromosomes and 97 pairs of microchromosomes (fundamental number; NF=550). Although the microchromosomes were relatively shorter than the macrochromosomes, most of them had discernible arms and centromeres. Silver-stained nucleolar organizer regions (Ag-NORs) were localized on the telomeric regions of 5 pairs of chromosomes (Ag-NORs=10): 4 were made up of small meta/submetacentrics and 1 of acrocentrics. Polyploidy of A. brevirostrum should be hexaploid based on the karyotype, numerous chromosomes, Ag-NORs, and previously reported large genome size (ca. 13pg DNA/cell).Supplementary material to this paper is available in electronic format at http://dx.doi.org/10.1007/s10228-004-0257-z     

Structural variability and chromosome numbers variation in natural populations ofScilla autumnalis (Liliaceae)

Chromosome variability within three populations ofScilla autumnalis was analysed. Among a total of 167 individuals we have detected one triploid structural heterozygote, 26 B-containing (with two different B-types), 11 structural variants (in the fifth chromosome pair), and one asynaptic. The possible relation between both types of chromosome variability (structural and chromosome numbers) is discussed.     

Cytological studies of triploids and their progeny from male-sterile (ms1) soybean

Summary Triploids (2n=3X=60) were obtained from genetic male-sterile (ms1 ms1) soybean [Glycine max (L.) Merr.] plants. Meiosis, pollen fertility, and chromosome number of their progeny were studied. Studies of meiosis in fertile and sterile triploids revealed no distinguishable differences in chromosome associations. Male-sterile plants formed coenocytic microspores characteristic of the ms1 mutant. Restitution of some dyad and tetrad nuclei were observed in male-sterile plants. Chromosomes of the triploids tended to occur in trivalents during diakinesis and metaphase I (MI), but multivalents, bivalents, and univalents also were observed. Average types and frequencies of chromosome associations per cell in diakinesis and MI from 542 pollen mother cells were 0.004 IX + 0.06 VI + 0.002 V + 0.005 IV + 16.99 III + 1.79 II + 5.03 I. Some secondary associations, nonhomologous pairing, and aberrant nucleolar distributions occasionally were observed. Such behavior support the hypothesis of duplicated genomes and the polyploid origin of soybean. Pollen fertility in male-fertile triploid plants (Ms1 ms1 ms1) varied from 57% to 82%, with an average of about 71%. Chromosome numbers of progenies obtained from these fertile triploids varied from 2n=40 to 2n=71, and exhibited a near-random distribution, with the majority (about 60%) being between 56 and 65. Progenies of the fertile triploids gave segregation ratios for the ms1 allele, which confirmed the Ms1 ms1 ms1 genotype.Joint contribution: Agricultural Research Service, U.S. Department of Agriculture, and Journal Paper No. J-11672 of the Iowa Agriculture and Home Economics Experiment Station, Ames, IA 50011, USA, Project 2471     

New chromosome number inRutaceae

Detailed chromosome counts have been made in 61 species, belonging to 33 genera ofRutaceae. 30 of these species are reported here for the first time. For 18 species at least one previous publication gives a chromosome number differing from that reported here. Such discrepancies are, in most cases, due to errors in counting or identification of the material. By critically reviewing the literature on each particular case, it appears possible to eliminate most of the false data. On the basis of the present results, the base number x = 10 is proposed for the genusRuta. Cytogenetics ofRutaceae, I.     

Taxonomic studies of nectrioid fungi in Japan. I: The genus Neonectria

Five species of the genus Neonectria (Nectriaceae, Hypocreales) collected from Japan are described with discussion and illustrated. Among them, one found on dead twigs of a broad-leaved tree is described as a new species, Neonectria tokyoensis (anamorph: Cylindrocarpon tokyoense). Neonectria cinnamomea and Neo. discophora are newly added to the Japanese mycobiota. Nectria azureo-ostiolata, recorded in 1977 from Japan, is reexamined as a synonym of Neo. jungneri. Neonectria radicicola, teleomorph of Cy. destructans, a well-known soil-borne plant pathogen in Japan, is newly recorded from Japan. Additional distribution records are provided for the Neonectria species hitherto recorded in Japan.     

In vitro culture ofBellevalia romana (L.) Rchb. I. Plant regeneration through adventitious shoots and somatic embryos

Summary Callus induction, adventitious shoot and root formation, and somatic embryogenesis were investigated in root, cotyledon and mesocotyl cultures ofBellevalia romana (L.) Rchb. grown on a synthetic nutrient medium containing different plant hormones. The combination of naphtaleneacetic acid plus benzylaminopurine was very effective in causing callus growth and plant regeneration from mesocotyl explants. On the contrary 2,4-dichlorophenoxyacetic acid caused suppression of shoot bud development in the same type of callus. Both cotyledon and root derived calli showed a low growth rate and did not regenerate shoots but only roots. Differentiation of somatic embryos which eventually developed into plantlets was promoted by 2,4-dichlorophenoxyacetic acid in suspension cultures. The results are discussed in relation to studies on nuclear behaviour during different morphogenetic pathways.