The origin of polyploid genomes of bluegrasses <Emphasis Type="Italic">Poa</Emphasis> L. and Gene flow between northern pacific and sub-Antarctic Islands

The involvement of present-day diploid bluegrass species in the formation of polyploid genomes was investigated using comparison of sequences of internal transcribed spacers ITS1 and ITS2, and the 5.8S rRNA sequence. It was demonstrated that highly polyploid New Zealand bluegrasses, P. cita (2n = 84; ca. 96 to 100), P. chathamica (2n = 112), and P. litorosa (2n 263–266) formed separate highly supported clade together with tetraploids (2n = 28) P. intrusa, P. anceps, and P. triodioides (Austrofestuca littoralis). Among the diploid species (2n = 14), the closest relatives of these species, as well as of the polyploid species of section Poa, are the genomes of Eurasian species P. remota, P. chaixii (sect. Homalopoa), P. densa (sect. Bolbophorum), and P. sibirica (sect. Macropoa). Nuclear genomes of polyploid Stenopoa, Tichopoa, Oreinos, and Secundae are definitely related to the genome of Arctic species P. pseudoabbreviata (sect. Abbreviatae). On the contrary, judging by the genes for nuclear 45S rRNA, genomes of diploid P. trivialis (sect. Pandemos), P. annua, and P. supina (sect. Ochlopoa both) are only remotely related to the genomes of highly polyploid species (p-distances between them and other bluegrass species from different sections of subgenus Poa constitute 6–10% and 11–15%, respectively). The conclusion on the relationships between highly polyploid and diploid bluegrass species was tested using analysis of synapomorphic mutations in the 5.8S rRNA gene. It was demonstrated that genomes of Poa eminens (2n = 42) and P. schischkinii (2n = 70) (sect. Arctopoa both) were noticeably different in ITS regions from the genomes of the members of the type subgenus Poa. A comparison of the Arctopoa ITS regions showed that the differences between them constituted only 0.2%. At the same time, p-distances between the Arctopoa ITS and those from the species belonging to other sections of the genus Poa varied from 5 to 14%. South American species P. chonotica (sect. Andinae) (= Nicoraepoa chonotica) (2n = 42) was found to be related to Arctagrostis, Festucella, and Hookerochloa, being at the same time quite distant from the other species of the genus Poa. Polymorphic in chromosome number highly polyploid species of Northern Hemisphere, P. arctica (2n = 42 to 106), P. turneri (2n = 42, 63 to 64), and P. smirnowii (2n = 42, 70) (sect. Malacanthae) are relative to a large group of tetraploid (2n = 28) endemic bluegrass species from New Zealand and sub-Antarctic islands (P. novaezelandiae and allied species).     

Genome relationships in polyploid Poa pratensis and other Poa species inferred from phylogenetic analysis of nuclear and chloroplast DNA sequences.

The genus Poa comprises approximately 500 species that occur throughout the world, including the widely grown Kentucky bluegrass (P. pratensis L.). Hybridization and polyploidization have played a prominent role in the evolution of this complex genus, but limited information is available regarding genome relationships in Poa. Thus, we amplified, cloned, and compared the DNA sequences of 2 nuclear genes (CDO504 and thioredoxin-like protein) and 2 chloroplast genome loci (ndhF and trnT-trnF) from 22 Poa species. Four distinct classes of sequences corresponding to 4 putative homoeologous loci from each nuclear gene were found within polyploid P. pratensis. Nuclear sequences from 15 other Poa species were found to group with at least 1 P. pratensis homoeolog, whereas 6 species displayed sequences not present in P. pratensis. The nuclear genome phylogenies presented here show the first evidence of diverse and related genomes in the genus Poa.     

Evolution of Internal Transcribed Spacer Region of Nuclear Ribosomal DNA in Allopolyploids of Aegilops

Hybridization with subsequent polyploidy is a prominent process in evolution of higher plants, but few data address the evolution of homeologous sequences after polyploidy. The internal transcribed spacer (ITS) of nuclear ribosomal DNA (nrDNA) from eleven allopolyploid species in Aegilops was investigated by PCR amplification and direct sequencing. The sequences obtained were used to study the evolution of ITS region in allopolyploid species. The length of ITS region varied from 599 to 606 bp and the number of variable sites was 93, i.e. 51 and 42 for ITS1 and ITS2 re spectively. Some polymorphic sites were observed in polyploid species, and this indicated that the ancestral sequences had not been homogenized completely by concerted evolution. Distance matrix analysis of diploid and polyploid species by neighbor-joining method, using Triticum monococcum as outgroup, resulted in well-resolved neighbor-joining tree indicating that the ITS regions of UUMM and UUSS genome ( sect. Vertebrata) were homogenizing toward those of UU ancestal genome. This result is in agreement with the results of ctyogenetics of Aegilops. On the other hand, the neighbor joining tree including the D-genome group species (sect. Cylindropyrum and sect. Polyeides ) com prised three clades (CC-DDCC, UU-DDMM-DDMMSS-DDMMUU and MM-DDMvMv), which sug gested that concerted evolution was homogenizing the ITS region of the polyploid derivatives to either of their ancestors.     

Phylogenetic relationships in Elymus (Poaceae: Triticeae) based on the nuclear ribosomal internal transcribed spacer and chloroplast trnL-F sequences

To estimate the phylogenetic relationship of polyploid Elymus in Triticeae, nuclear ribosomal internal transcribed spacer (ITS) and chloroplast trnL-F sequences of 45 Elymus accessions containing various genomes were analysed with those of five Pseudoroegneria (St), two Hordeum (H), three Agropyron (P) and two Australopyrum (W) accessions. The ITS sequences revealed a close phylogenetic relationship between the polyploid Elymus and species from the other genera. The ITS and trnL-F trees indicated considerable differentiation of the StY genome species. The trnL-F sequences revealed an especially close relationship of Pseudoroegneria to all Elymus species included. Both the ITS and trnL-F trees suggested multiple origins and recurrent hybridization of Elymus species. The results suggested that: the St, H, P, and W genomes in polyploid Elymus were donated by Pseudoroegneria, Hordeum, Agropyron and Australopyrum, respectively, and the St and Y genomes may have originated from the same ancestor; Pseudoroegneria was the maternal donor of the polyploid Elymus; and some Elymus species showed multiple origin and experienced recurrent hybridization.     

Chloroplast DNA variation in diploid and polyploid species of Bromus (Poaceae) subgenera Festucaria and Ceratochloa

Summary Chloroplast DNA (cpDNA) restriction endonuclease patterns are used to examine phylogenetic relationships between Bromus subgenera Festucaria and Ceratochloa. Festucaria is considered monophyletic based on the L genome, while Ceratochloa encompasses two species complexes: the B. catharticus complex, which evolved by combining three different genomes, and the B. carinatus complex, which is thought to have originated from hybridization between polyploid species of B. catharticus and diploid members of Festucaria. All species of subgenus Ceratochloa (hexaploids and octoploids) were identical in chloroplast DNA sequences. Similarly, polyploid species of subgenus Festucaria, except for B. auleticus, were identical in cpDNA sequences. In contrast, diploid species of subgenus Festucaria showed various degrees of nucleotide sequence divergence. Species of subgenus Ceratochloa appeared monophyletic and phylogenetically closely related to the diploid B. anomalus and B. auleticus of subgenus Festucaria. The remaining diploid and polyploid species of subgenus Festucaria appeared in a distinct grouping. The study suggests that the B. catharticus complex must have been the maternal parent in the proposed hybrid origin of B. carinatus complex. Although there is no direct evidence for the paternal parent of the latter complex, the cpDNA study shows the complex to be phylogenetically very related to the diploid B. anomalus of subgenus Festucaria.     

Genomes,multiple origins,and lineage recombination in the Glycine tomentella (Leguminosae) polyploid complex: histone H3-D gene sequences

Relationships among the various diploid and polyploid taxa that comprise Glycine tomentella have been hypothesized from crossing studies, isozyme data, and repeat length variation for the 5S nuclear ribosomal gene loci. However, several key questions have persisted, and detailed phylogenetic evidence from homoeologous nuclear genes has been lacking. The histone H3-D locus is single copy in diploid Glycine species and has been used to elucidate relationships among diploid races of G. tomentella, providing a framework for testing genome origins in the polyploid complex. For all six G. tomentella polyploid races (T1-T6), alleles at two homoeologous histone H3-D loci were isolated and analyzed phylogenetically with alleles from diploid Glycine species, permitting the identification of all of the homoeologous genomes of the complex. Allele networks were constructed to subdivide groups of homoeologous alleles further, and two-locus genotypes were constructed using these allele classes. Results suggest that some races have more than one origin and that interfertility within races has led to lineage recombination. Most alleles in polyploids are identical or closely related to alleles in diploids, suggesting recency of polyploid origins and spread beyond Australia. These features parallel the other component of the Glycine subgenus Glycine polyploid complex, G. tabacina, one of whose races shares a diploid genome with a G. tomentella polyploid race.     

Cytogenetic and phylogenetic studies of diploid and polyploid members of tribe Anemoninae (Ranunculaceae)

The ancestry, phylogenetic differentiation and systematic classification of the worldwide-distributed genus Anemone have been debated for many years. In this paper 11 Anemone, three Pulsatilla species and Hepatica nobilis were subjected to detailed karyotype analysis with the aim of obtaining new cytogenetic data that will contribute to karyotype evolutionary studies of the tribe Anemoninae. The results are interpreted in a phylogenetic context, established from the intergenic nontranscribed spacer (NTS) of 5S rDNA and internal transcribed spacer (ITS) of 35S rDNA. One to three 35S and one to three 5S rDNA loci are present in diploid and polyploid taxa. The 35S rDNA loci are located terminally on the short arm of acrocentric chromosomes, while for 5S rDNA there is no preferential chromosomal position as it exhibits terminal, subterminal, interstitial or pericentromeric positions, and is located either on acrocentric or metacentric chromosomes. The karyotype of hexaploid A. baldensis (2n = 6x = 48) is presented for the first time, and A. sylvestris is proposed as one of its putative parental species. Chromosome fusion/translocation is proposed as the key mechanism involved in reduction of the basic chromosome number from 8 in the Anemone subgenus to 7 in the Anemonidium subgenus. The cytogenetic data obtained are mainly supported by ITS and NTS phylogeny. Diversification of the genus Anemone was accompanied by a large reduction of heterochromatin, from the Mediterranean anemones that have large amounts of heterochromatin to the New World anemones without any detectable heterochromatic blocks.     

Genome analysis of Elytrigia pycnantha and Thinopyrum junceiforme and of their putative natural hybrid using the GISH technique.

Genomic in situ hybridization (GISH), using genomic DNA probes from Thinopyrum elongatum (Host) D.R. Dewey (E genome, 2n = 14), Th. bessarabicum (Savul. & Rayss) A. Love (J genome, 2n = 14), Pseudoroegneria stipifolia (Czern. ex Nevski) Love (S genome, 2n = 14), and Agropyron cristatum (L.) Gaertner (P genome, 2n = 14), was used to characterize the genome constitution of the polyploid species Elytrigia pycnantha (2n = 6x = 42) and Thinopyrum junceiforme (2n = 4x = 28) and of one hybrid population (2n = 5x = 35). GISH results indicated that E. pycnantha contains S, E, and P genomes; the first of these was closely related to the S genome of Ps. stipifolia, the second was closely related to to the E genome of Th. elongatum, and the third was specifically related to A. cristatum. The E and P genomes included 2 and 10 chromosomes, respectively, with S genome DNA sequences in the centromeric region. GISH analysis of Th. junceiforme showed the presence of two sets of the E genome, except for fewer than 10 chromosomes for which the telomeric regions were not identified. Based on these results, the genome formula SSPsPsEsEs is proposed for E. pycnantha and that of EEEE is proposed for Th. junceiforme. The genomic constitution of the pentaploid hybrid comprised one S genome (seven chromosomes), one P genome (seven chromosomes), and three E genomes (21 chromosomes). The E and P genomes both included mosaic chromosomes (chromosomes 1 and 5, respectively) with the centromere region closely related to S-genome DNA. On the basis of these data, the genome formula SPSESEE is suggested for this hybrid and it is also suggested that the two species E. pycnantha and Th. junceiforme are the parents of the pentaploid hybrid.     

Untersuchungen zur Karyologie und Mikrosporogenese vonPelargonium sect.Pelargonium (Geraniaceae)

The chromosome numbers of the 24 species of sect.Pelargonium were determined from field collected and cultivated plants of known localities in S. Africa. Twelve species are diploid (2n = 22), eight tetraploid (2n = 44), one hexaploid (2n = 66), and three octoploid (2n = 88). The chromosome numbers correlate well with the proposed subdivision of sect.Pelargonium. Its chromosomes are relatively small (1.0–1.5 µm) in comparison to most of the other sections, and its diploid karyotype is considered to be primitive. The occurrence of the basic number x = 11 in this section, in other sections of the genus, and in related genera (Monsonia, Sarcocaulon) leads to the conclusion that x = 11 probably is basic for the whole genus. — The pollen meiosis, microsporogenesis and pollen fertility of the diploid species is normal, with the exception of one, possibly young taxon from the Greyton Nature Reserve. The tetraploid species could be of autoploid origin, the higher polyploids exhibit a mixed auto-alloploid nature. — The 20 diploid and tetraploid species have a relatively small distribution range, most of them occur in the SW. Cape Province of South Africa. This area may therefore be considered as the centre of origin of the genus. Three of the four high polyploid species occupy rather large areas.

Untersuchungen zur Karyologie und Mikrosporogenese der GattungPelargonium, 1.     

Is genome downsizing associated with diversification in polyploid lineages of Veronica?

The study of genome size evolution in a phylogenetic context in related polyploid and diploid lineages can help us to understand the advantages and disadvantages of genome size changes and their effect on diversification. Here, we contribute 199 new DNA sequences and a nearly threefold increase in genome size estimates in polyploid and diploid Veronica (Plantaginaceae) (to 128 species, c. 30% of the genus) to provide a comprehensive baseline to explore the effect of genome size changes. We reconstructed internal transcribed spacer (ITS) and trnL‐trnL‐trnF phylogenetic trees and performed phylogenetic generalized least squares (PGLS), ancestral character state reconstruction, molecular dating and diversification analyses. Veronica 1C‐values range from 0.26 to 3.19 pg. Life history is significantly correlated with 1C‐value, whereas ploidy and chromosome number are strongly correlated with both 1C‐ and 1Cx‐values. The estimated ancestral Veronica 1Cx‐value is 0.65 pg, with significant genome downsizing in the polyploid Southern Hemisphere subgenus Pseudoveronica and two Northern Hemisphere subgenera, and significant genome upsizing in two diploid subgenera. These genomic downsizing events are accompanied by increased diversification rates, but a ‘core shift’ was only detected in the rate of subgenus Pseudoveronica. Polyploidy is important in the evolution of the genus, and a link between genome downsizing and polyploid diversification and species radiations is hypothesized. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 178 , 243–266.     

Molecular phylogeny of Larrea and its allies (Zygophyllaceae): reticulate evolution and the probable time of creosote bush arrival to North America.

Nucleotide sequences of Rubisco Large Subunit (rbcL) and the internal transcribed spacers (ITS) of nrDNA were obtained for the five species of Larrea and one species each of Bulnesia (ITS only) and Plectrocarpa (rbcL only). Parsimony analyses were conducted, including sequences from seven genera of Zygophyllaceae reported by other authors-Kallstroemia, Zygophyllum, Augea, Fagonia, Pintoa, Guaiacum, and Porlieria. The main conclusions of the present study are (1) the Argentine endemic Plectrocarpa tetracantha belongs to the subfamily Larreoideae (New World Clade); (2) all three phylogenies obtained from rbcL, ITS, and combined data sets show a close relationship between the tetraploid L. cuneifolia (sect. Bifolium) and the diploid multifoliolate pair L. nitida-L. ameghinoi (sect. Larrea), which could result from a possible intersectional hybrid origin of the tetraploid; (3) L. divaricata (sect. Bifolium) and L. tridentata (sect. Bifolium) form a highly supported monophyletic group, which agrees with previous cytogenetic and molecular evidence; and (4) the rate of nucleotide substitution of rbcL was estimated based on geological and fossil records. Under the molecular clock hypothesis, nucleotide sequence divergence between L. divaricata and L. tridentata suggests a Late Neogene (8.4 to 4.2 mybp) time of arrival of the diploid ancestors of L. tridentata to North American deserts.     

Studies on the origin and evolution of tetraploid wheats based on the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA

In this study, the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA in the tetraploid wheats, Triticum turgidum (AABB) and Triticum timopheevii (AAGG), their possible diploid donors, i.e., Triticum monococcum (AA), Triticum urartu (AA), and five species in Aegilops sect. Sitopsis (SS genome), and a related species Aegilops tauschii were cloned and sequenced. ITS1 and ITS2 regions of 24 clones from the above species were compared. Phylogenetic analysis demonstrated that Aegilops speltoides was distinct from other species in Aegilops sect. Sitopsis and was the most-likely donor of the B and G genomes to tetraploid wheats. Two types of ITS repeats were cloned from Triticum turgidum ssp. dicoccoides, one markedly similar to that from T. monococcum ssp. boeoticum (AA), and the other to that from Ae. speltoides (SS). The former might have resulted from a recent integression event. The results also indicated that T. turgidum and T. timopheevii might have simultaneously originated from a common ancestral tetraploid species or be derived from two hybridization events but within a very short interval time. ITS paralogues in tetraploid wheats have not been uniformly homogenized by concerted evolution, and high heterogeneity has been found among repeats within individuals of tetraploid wheats. In some tetraploid wheats, the observed heterogeneity originated from the same genome (B or G). Three kinds of ITS repeats from the G genome of an individual of T. timopheevii ssp. araraticum were more divergent than that from inter-specific taxa. This study also demonstrated that hybridization and polyploidization might accelerate the evolution rate of ITS repeats in tetraploid wheats.     

The genome composition of hexaploid Psammopyrum athericum and octoploid Psammopyrum pungens (Poaceae: Triticeae).

The genomic constitution of two species in the genus Psammopyrum, i.e., Ps. athericum (2n = 6x = 42) and Ps. pungens (2n = 8x = 56), was studied by genomic in situ hybridization (GISH). In Ps. athericum, one diploid chromosome set hybridized to a genomic probe from Pseudoroegneria ferganensis (St genome), one diploid set to a probe from Agropyron cristatum (P genome), and one diploid set to a probe from Thinopyrum junceiforme (EbEe genomes) or Th. bessarabicum (Eb genome). Substituting the St-genome probe with an L-genome probe from Festucopsis serpentinii resulted in exactly the same hybridization pattern, suggesting a genomic constitution of EStP or ELP for Ps. athericum. The same probes used on Ps. pungens showed two diploid sets of chromosomes hybridizing to the St-genome probe, one diploid set hybridizing to the P-genome probe, and one diploid set hybridizing to the EbEe-genome probe. The L-genome probe hybridized to approximately 14 of the chromosomes that were labeled by the St-genome probe. Hence the genomic constitution for Ps. pungens is proposed to be EStStP or EStLP.     

On polyphyly of the former section Ochlopoa and the hybridogenic section Acroleucae (Poa,Poaceae): insights from molecular phylogenetic analyses

In this study, sequence data from the inert nuclear region ITS1‐5.8S rDNA‐ITS2 and the chloroplast region trnL–F, as well as a few morphological characters, are analysed to the relationships among known annual Poa (bluegrasses). It is shown that all taxa from the Poa annua aggregate distinguished by lemma characters and growth form have identical ITS and trnL–trnF sequences, all ITS sequences of this aggregate are the same as thethose of P. supina, and all trnL–trnF sequences are homologous with those of P. infirma. Furthermore, no differences were found between unusual morphotypes of P. supina with short spinules on their panicle branches and typical plants, but Siberian samples were found to have a slightly differentiated trnL–trnF region. These results suggest a hybrid origin of the Asian annual bluegrasses. Their maternal genome is close to that of P. sect. Homalopoa, but their ITS sequences are different. Some ITS sequences from annual Asian bluegrasses are resolved among representatives of P. sect. Stenopoa while for other (morphologically closely similar) species they fall in a clade with representatives of P. sect. Malacanthae. The latter group is distant from P. sect. Ochlopoa and is better treated as a separate section, viz P. sect. Acroleucae. The American annual bluegrasses are heterogeneous and also rather distant from P. sect. Ochlopoa. Poa chapmaniana, a species with cleistogamic flowers, is nested among the basal Subantarctic sections, far away from the taxa with which it has previously been considered related. It is indeed closer to P. sect. Ochlopoa than to other annual American bluegrasses. Thus, the studied annual species in fact belong to four independent evolutionary lines (or six including the separate genus Eremopoa and the Turkish Poa jubata), one of which, Acroleucae, has gone through three reticulation events. As in previous studies, our analysis did not support the generic status of P. sect. Ochlopoa.     

Cytotaxonomic observations in tropical Vaccinieae (Ericaceae)

Chromosome numbers are reported for 36 accessions representing 31 species from nine genera of the tribe Vaccinieae, family Ericaceae. The plants are tropical and come from Southeast Asia and Central and South America. The taxonomy of the tribe is outlined in these regions. Genera are often poorly defined and taxa were chosen to reflect the range of variation of the Vaccinieae. Most Southeast Asian Vaccinium species were diploid (2 n = 24) as were those of Agapetes subgenus Agapetes (apart from the Himalayan A. flava ), Agapetes scortechinii and Costera endertii. All other accessions were found to be polyploid. The correlation between polyploidy, geographical distribution and the possession of an 'anatomical complex' of the leaf and stem in Vaccinieae of New Guinea and the neotropics is discussed.     

A cytological study ofPelargonium sect.Eumorpha (Geraniaceae)

The chromosome numbers of seven species ofPelargonium sect.Eumorpha have been determined from material of known wild origin, and karyotypic comparisons have been made. Within the section there is variation in basic chromosome number (x = 4, 8, 9, 11), variation in chromosome size, and two species have polyploid races. The three species with chromosome numbers based on x = 11 have the smallest chromosomes (1.0–1.5 µm); chromosomes are larger (1.0–3.0 µm) in the other species.P. elongatum has the lowest chromosome number in the genus (2n = 8).P. alchemilloides is exceptional in that it has four cytotypes, 2n = 16, 18, 34 and 36, and the form with 2n = 36 has large chromosomes (2.0–5.0 µm). Evidence from a synthesized hybrid suggests thatP. alchemilloides with 2n = 16 may be of polyploid origin. The three species based on x = 11 appear to be more closely related to species from other sections ofPelargonium that have the same basic chromosome number and small chromosome size, rather than to other species of sect.Eumorpha.     

Genome composition and origin of the polyploid Aegean grass Avenula agropyroides (Poaceae)

Aim The Aegean is a hotspot of plant biodiversity, with its island biota harbouring a large number of endemic taxa. To investigate the relationship between biogeography, polyploid speciation and genomics in the Aegean we used the biogeographically isolated highly polyploid eastern Mediterranean grass species Avenula agropyroides (2n = 70) as an example of complicated polyploid origin. Location Mediterranean, Aegean. Methods To clarify the origin of A. agropyroides, we conducted chromosome studies using repetitive DNAs as hybridization probes in fluorescent in situ hybridization experiments, chromosome banding methods and DNA sequence analyses of plasmid‐cloned nuclear ribosomal (nr) ITS1–5.8S–ITS2 DNA. Results Decaploid A. agropyroides had near‐autopolyploid karyotype structure and contained characteristic sequence motifs of nrDNA repeats not encountered in any of the diploids studied. Special repeat types found in one of its accessions (Crete) showed that A. agropyroides originated from a diploid species with a hybrid background. One of the genomes involved was close to both that of extant species (Avenula aetolica, Avenula compressa, Avenula hookeri, Avenula schelliana, Avenula versicolor) distributed mostly in the eastern Mediterranean to Asia and North America and also to the west Mediterranean (Avenula bromoides). The other resembled that of exclusively western Mediterranean species (Avenula albinervis, Avenula levis, Avenula marginata, Avenula sulcata). Main conclusions Avenula agropyroides represents a remarkable polyploid in the eastern Mediterranean, conserving the genome structure of a diploid species that no longer exists. This highlights how the Aegean has been less affected than other Eurasian regions by the repeated shifts of climatic zones and vegetation belts since the Late Tertiary.     

Ancestry of American polyploid Hordeum species with the I genome inferred from 5S and 18S-25S rDNA

* BACKGROUND AND AIMS: The genus Hordeum exists at three ploidy levels (2x, 4x and 6x) and presents excellent material for investigating the patterns of polyploid evolution in plants. Here the aim was to clarify the ancestry of American polyploid species with the I genome. * METHODS: Chromosomal locations of 5S and 18S-25S ribosomal RNA genes were determined by fluorescence in situ hybridization (FISH). In both polyploid and diploid species, variation in 18S-25S rDNA repeated sequences was analysed by the RFLP technique. * KEY RESULTS: Six American tetraploid species were divided into two types that differed in the number of rDNA sites and RFLP profiles. Four hexaploid species were similar in number and location of both types of rDNA sites, but the RFLP profiles of 18S-25S rDNA revealed one species, H. arizonicum, with a different ancestry. * CONCLUSIONS: Five American perennial tetraploid species appear to be alloploids having the genomes of an Asian diploid H. roshevitzii and an American diploid species. The North American annual tetraploid H. depressum is probably a segmental alloploid combining the two closely related genomes of American diploid species. A hexaploid species, H. arizonicum, involves a diploid species, H. pusillum, in its ancestry; both species share the annual growth habit and are distributed in North America. Polymorphisms of rDNA sites detected by FISH and RFLP analyses provide useful information to infer the phylogenetic relationships of I-genome Hordeum species because of their highly conserved nature during polyploid evolution.     

Chromosome numbers and their taxonomic implications in the genusPoa of Japan

As part of the work leading to a taxonomic revision ofPoa of Japan, chromosome numbers of nearly all indigenous species of JapanesePoa, together with those of some putative interspecific hybrids, were examined. The following chromosome numbers were found:Poa annua, 2n=28;P. crassinervis, 2m=28;P. acroleuca, 2n=28;P. hisauchii, 2n=28;P. nipponica, 2n=28;P. crassinervis, 2n=28;P. acroleuca, 2n=35;P. tuberifera, 2n=28;P. fauriei, 2n=28;P. radula, 2n=42;P. hakusanensis, 2n=70;P. hayachinensis, 2n=42;P. malacantha var.shinanoana, 2n=63≈98;P. yatsugatakensis, 2n=ca. 71≈74, 77;P. sachalinensis, 2n=63, ca. 64, ca. 74;P. matsumurae, 2n=28;P. ogamontana, 2n=42;P. sphondylodes, 2n=28;P. viridula, 2n=42, 49, 56;P. nemoralis, 2n=70;P. glauca, 2n=42, 49, 56;P. eminens, 2n=42. It became clear that information obtained from chromosome counts is quite helpful in clarifying species boundaries in several species aggregates, such as thePoa acroleuca-hisauchii-nipponica aggregate. Results of the examination of morphological features of the voucher specimens in various species aggregates with taxonomic difficulties were reported, and the needs of some amendments for the species delimitation appearing in current floristic manuals were pointed out. A summary of chromosome counts so far made for JapanesePoa was tabulated. ThePoa flora of Japan characteristically lacked diploid plants.