Comparative karyotype analysis in Haplopappus Cass. and Grindelia Willd. (Asteraceae) by double FISH with rRNA specific genes

The genera Grindelia Willd. and Haplopappus Cass. belong to the family Asteraceae - Astereae and are distributed in America and South America, respectively. Previous cytotaxonomic studies showed for South American species of Grindelia 2n=12 and for Haplopappus 2n=10 and 2n=12. Both Grindelia species (G. anethifolia, G. prunelloides), newly analyzed with molecular-cytological methods, exhibited symmetric karyotypes (AsI %=55.46 and 55.95) with metacentric chromosome sets (5m + 1m-sat) and 2n=12 chromosomes. The NOR was detected after fluorescence in situ hybridization (FISH) with 18/25S rDNA in the satellite chromosome 2. In contrast H. Happlopappus glutinosus, H. grindeloides and H. stolpii showed exclusively a higher asymmetric index (66.83%, 67.01% and 68.87%, respectively) with submetacentric chromosome sets (4sm + 1sm–sat). The sat-chromosomes 3 of H. glutinosus and H. grindelioides were both significantly different in their length from chromosomes 2 and 4. Furthermore in Grindelia the FISH with 5S rDNA could estimate signals in the short arms of chromosomes 3 or 4, that were not significantly differentiated in their length. Contrary to these findings in Grindelia, the position of 5S rDNA in Haplopappus was detected in the long arms of chromosome 1 (H. grindelioides and H. stolpii) and chromosome 2 (with two different loci) and chromosome 4 of H. glutinosus. The lengths of all measured chromosome arms with 5S rDNA were significantly different to those of the neighbours in the karyotypes. The two-color FISH of 5S and 18/25S rDNA had provided clear karyotypic markers for three (Haplopappus glutinosus) and two (H. grindelioides and H. stolpii) chromosomes. The number and position of rDNA signals were relatively highly conserved in the investigated five species without the double marked chromosome 2 of H. glutinosus, which shows an evolutionary dynamic of this 5S rRNA specific gene cluster. This investigation supports the assumption that the evolution of New World members of Grindelia and Haplopappus has not been accompanied by large karyotypic changes, but small chromosomal rearrangements have undoubtedly occurred (e.g. 5S rDNA localizations).     

Cytogenetic studies in South American species of Serjania (Sapindaceae: Paullinieae)

Abstract

Serjania Mill. (Paullinieae) is considered the most important neotropical genus of Sapindaceae due to species number and its widespread distribution. In this study, 14 species belonging to three sections were analyzed using conventional staining, C/CMA/DAPI banding, and fluorescence in situ hybridization (FISH) with a 18S-5.8S-26S rDNA probe. New chromosome counts are reported for Serjania crassifolia, Serjania platycarpa, and Serjania regnellii, all with 2n = 24, which is remarkably constant for Serjania. The karyotypes are moderately asymmetric, and variations observed in A1 and A2 indices show resemblances between S. platycarpa, Serjania hebecarpa, and S. crassifolia, and between Serjania communis, Serjania gracilis, and S. regnellii. The banding pattern was homogeneous in Serjania. C/DAPI bands (AT-rich sites) were not clearly evidenced, but changes in the number and position of GC-rich sites (CMA bands) were observed. These segments were associated with 18S-5.8S-26S rDNA sites. The significance of the results is discussed in relation to chromosomal data available for the genus and in regard to the infrageneric treatment of Serjania.     

Molecular cytogenetic studies of the “Xanthocephalum group” (Asteraceae)

Fourteen North American members of the “Xanthocephalum group” were studied by classical and molecular cytogenetics. Location and number of rDNA sites were determined by FISH. For the 5S rDNA, a probe was obtained from Prionopsis ciliata. Most species were diploid (2n?=?12), although Isocoma menziesii, Grindelia hirsutula, G. robusta, both varieties of G. stricta, and one population of G. camporum were tetraploid (2n?=?24). Diploid Grindelia and Prionopsis ciliata were 5m?+?1sm, tetraploids 10m?+?2sm, except G. hirsutula (8m?+?4sm), and Isocoma and Olivaea 6m?+?2sm and 3m?+?3sm, respectively. Most species had satellites on the short arms of m pairs: two in tetraploids and P. ciliata and one in diploids. Satellites were associated with two CMA⁺/DAPI^? bands in diploid species and four bands in tetraploids and in P. ciliata. rDNA loci (two in diploids to four in tetraploids) may be indicative of ploidy level. Grindelia tetraploids could have originated recently by autopolyploidy. Chromosome duplication was followed by modifications in the genome structure, resulting in higher heterochromatin amounts not associated with NORs. There is only one 5S site per basic genome in para or pericentromeric regions. Although not always large, chromosome variation has accompanied the evolutionary divergence of the taxa studied.     

Do polyploids require proportionally less rDNA loci than their corresponding diploids? Examples from Artemisia subgenera Absinthium and Artemisia (Asteraceae,Anthemideae)

Abstract

Fluorescent in situ hybridisation (FISH) of 5S and 18S-5.8S-26S ribosomal DNA was carried out in two species of the genus Artemisia, belonging to the subgenera Artemisia (A. medioxima) and Absinthium (A. lagocephala), each one showing both low and high ploidy levels (2x, 4x and 16x, and 2x and 6x, respectively). Both species have a base chromosome number of x = 9. Linkage of both rDNA genes has been observed confirming previous results. Diploid A. lagocephala (2n = 18) shows three rDNA loci, and the hexaploid six. Also in A. medioxima, the number of rDNA loci does not increase in the proportion given by the ploidy level, and a relative loss is found. In this species, the diploid population shows two rDNA loci, the tetraploid four, and the hexaidecaploid has around 20. The results evidence a relative loss of rDNA loci and heterochromatin, a phenomenon that is more pronounced at higher ploidy levels. Nevertheless, the DAPI banding pattern of A. lagocephala does not follow this trend, as it shows a spectacular increase of heterochromatic bands at the hexaploid level. These results are discussed in the light of possible chromosome restructuring and gene silencing mechanisms that take place during polyploidy, and more especially allopolyploid formation.     

Cytogenetic characterization and nuclear DNA content of three North African endemic Centaurea species

Three endemic Centaurea species from North Africa are investigated for the first time by chromomycin fluorochrome banding for GC-rich DNA distribution, fluorescence in situ hybridization for physical mapping of rRNA genes, and flow cytometry for genome-size assessment. Investigated species belong to three different sections and possess three basic chromosome numbers: C. tougourensis subsp. tougourensis 2n = 4x = 36 (x = 9), C. musimonum 2n = 2x = 20 (x = 10), and C. maroccana 2n = 2x = 24 (x = 12). The number and distribution of chromomycin positive bands (CMA⁺) and 18S-5.8S-26S (35S) rDNA loci were different among investigated species and ranged from 6 to 80 chromomycin bands and from 2 to 6 35S rDNA loci. The three species have just one 5S rDNA locus at intercalary position on a separate chromosome pairs, except in the case of C. musimonum in which both rDNA loci were localized on the same chromosome. All rDNA loci were co-localized with CMA⁺ bands, except three 35S in C. musimonum. Genome size ranged from 2C = 1.66 to 2C = 2.86 pg in diploid species (C. musimonum and C. maroccana, respectively) and to 2C = 4.51 pg in tetraploid C. tougourensis subsp. tougourensis.     

华中植物区系新资料

该文报道了产鄂西和湘西北的双子叶植物省级分布新记录1个属和8个种。湖北分布新记录有龙珠(Tubocapsicum anomalum)、直梗高山唐松草(Thalictrum alpinum var. elatum)、虫莲(Sanguisorba filiformis)、腺地榆(S. officinalis var. glandulosa)、平叶酸藤子(Embelia undulata)、广西地海椒(Physaliastrum chamaesarachoides)、东北薄荷(Mentha sachalinensis)、黄鼠狼花(Salvia tricuspis),其中龙珠对应的龙珠属(Tubocapsicum)是湖北新记录属;湖南分布新记录有宝兴藨寄生(Gleadovia mupinense)。根据IUCN标准,宝兴藨寄生属濒危(EN)物种。这些新记录的发现丰富了华中植物区系资料,在植物保护方面也有一定意义。     

Physical mapping of 18S-5.8S-26S and 5S ribosomal RNA gene families in three important vetches (Vicia species) and their allied taxa constituting three species complexes

The most-important vetch species, Vicia narbonensis (narbon vetch, section Faba), Vicia villosa (hairy vetch, section Cracca) and Vicia sativa (common vetch, section Vicia) and their close relatives (often difficult to circumscribe into distinct taxa) constitute respectively, Narbonensis, Villosa and Sativa species complexes in the genus Vicia. The distribution of the 18S-5.8S-26S (18S-26S) and 5S ribosomal RNA (rRNA) gene families on the chromosomes of 19 (2n=2x=10,12,14) of the 24 species and subspecies belonging to the three species complexes, and Vicia bithynica (2n=12, section Faba) and Vicia hybrida (2n=12, section Hypechusa) was studied by fluorescence in situ hybridization (FISH) with pTa 71 (18S-26S rDNA) and pTa 794 (5S rDNA) DNA clones. Computer – aided chromosome analysis was performed on the basis of chromosome length, the arm-length ratio and the position of the hybridization signals. The positions of the four (2+2) signals of the two rRNA gene families were similar between each of the three, as well as two subspecies of V. narbonensis and Vicia johannis, respectively. Two major 18S-26S rDNA loci were found in the nucleolus organiser regions (NORs) of each of the species except V. hybrida, where it was present in two out of four SAT chromosomes. In addition to major NORs, two minor loci have been physically mapped at the centromeric regions of chromosomes of group 1 in Vicia amphicarpa, Vicia macrocarpa and V. sativa, and two NORs of group 5 in V. hybrida, and on the long arms of group 4 in V. bithynica. Two or four 5S rDNA loci, observed in the short arms of groups 2–4 and 5, and 18S-26S rDNA loci were located in different chromosomes of all the species within the Narbonensis and Villosa species complexes, and Vicia angustifolia of the Sativa species complex. In the remaining six species of the Sativa species complex, and V. bithynica and V. hybrida, the two or four 5S rDNA sites were present in chromosomes which harbor 18S-26S rRNA genes. The tandemly repeated 5S rDNA sites, located at the proximal part of the long arm of groups 3–5, were diagnostic for V. angustifolia, Vicia cordata, Vicia incisa, V. macrocarpa, Vicia nigra and V. sativa of the Sativa species complex. In V. amphicarpa of the same complex, the tandem repeats were located at the distal part of the long arms of group 3. Variability in the number, size and location of two ribosomal DNA probes could generally distinguish species within the Narbonensis and Sativa species complex, V. bithynica and V. hybrida. With respect to the four species of the Villosa species complex the karyotypes could not be identified individually on the basis of the distribution of two ribosomal gene families in three out of seven pairs of chromosomes. Received: 18 October 2000 / Accepted: 20 March 2001     

Molecular cytogenetic analyses and phylogenetic studies in the Nicotiana section Tomentosae

Phylogenetic schemes based on changing DNA sequence have made a major impact on our understanding of evolutionary relationships and significantly built on knowledge gained by morphological and anatomical studies. Here we present another approach to phylogeny, using fluorescent in situ hybridisation. The phylogenetic scheme presented is likely to be robust since it is derived from the chromosomal distribution of ten repetitive sequences with different functions and evolutionary constraints [GRS, HRS60, NTRS, the Arabidopsis-type telomere repeat (TTTAGGG)_n, 18S-5.8S-26S ribosomal DNA (rDNA), 5S rDNA, and four classes of geminiviral-related DNA (GRD)]. The basic karyotypes of all the plant species investigated Nicotiana tomentosiformis, N. kawakamii, N. tomentosa, N. otophora, N. setchellii, N. glutinosa (all section Tomentosae), and N. tabacum (tobacco, section Genuinae) are similar (x=12) but the distribution of genic and non-genic repeats is quite variable, making the karyotypes distinct. We found sequence dispersal, and locus gain, amplification and loss, all within the regular framework of the basic genomic structure. We predict that the GRD classes of sequence integrated into an ancestral genome only once in the evolution of section Tomentosae and thereafter spread by vertical transmission and speciation into four species. Since GRD is similar to a transgenic construct that was inserted into the N. tabacum genome, its fate over evolutionary time is interesting in the context of the debate on genetically modified organisms and the escape of genes into the wild. Nicotiana tabacum is thought to be an allotetraploid between presumed progenitors of N. sylvestris (maternal, S-genome donor) and a member of section Tomentosae (T-genome donor). Of section Tomentosae, N. tomentosiformis has the most similar genome to the T genome of tobacco and is therefore the most likely paternal genome donor. It is known for N. tabacum that gene conversion has converted most 18S-5.8S-26S rDNA units of N. sylvestris origin into units of an N. tomentosiformis type. Clearly if such a phenomenon were widespread across the genome, genomic in situ hybridisation (GISH) to distinguish the S and T genomes would probably not work since conversion would tend to homogenise the genomes. The fact that GISH does work suggests a limited role for gene conversion in the evolution of N. tabacum. Received: 8 November 1999; in revised form: 23 February 2000 / Accepted: 1 March 2000     

CHROMOSOME NUMBERS IN PERITYLE AND RELATED GENERA (PERITYLANAE-COMPOSITAE)

Chromosome numbers are presented for 28 species of the genus Perityle, one putative inter-sectional hybrid, two species of Amauria, one species of Eutetras, and one species of Pericome. For Perityle, initial counts are recorded for 12 species of sect. Laphamia (n = 16, 17, 18, 36, ca. 102) and 11 species of sect. Perityle (n = 11, 12, 13, 16, 17, 18, 19, 34, 51). Chromosome numbers for the two species of Amauria (n = 18) are first reports for the genus. Including the current information, chromosome numbers have been recorded for 37 of the approximately 50 species recognized for Perityle. At least 24 taxa have numbers of n = 17, suggesting a base chromosome number of x = 17 for Perityle.     

Heterochromatin and rDNA patterns in Solanum species of the Morelloid and Dulcamaroid clades (Solanaceae)

The heterochromatin distribution and the position of 18-5.8-26S, and 5S rDNA loci were determined in 13 species of Solanum of the Morelloid and Dulcamaroid clades. The CMA/DAPI staining and FISH were employed. Two types of constitutive heterochromatin were determined: CMA⁺/DAPI^? associated to NOR and CMA⁺/DAPI^? distributed as terminal bands. In the Morelloid clade, CMA⁺/DAPI^? bands were found in five species while in the Dulcamaroid clade, only S. angustifidum presented this feature. In the Morelloid clade, two to four 18-5.8-26S rDNA loci occupied terminal positions and two rDNA 5S loci were found with variable positions (terminal, intercalary, and centromeric). In the Dulcamaroid clade, two terminal 18-5.8-26S rDNA loci were detected with the exception of S. salicifolium which possessed four such loci and two to four 5S rDNA loci. Solanum crispum is the only species possessing the 5S in synteny with 18-5.8-26S rDNA loci. Karyotype features chromosome banding pattern as well as the location of ribosomal genes which varied among the species, reflecting the chromosome differentiation and evolutionary divergence. The findings obtained contributed to the development of tools that can be used for establishing chromosomic homeologies among species and hence to clarify their taxonomic relationships.     

Rhodotorula lamellibrachii sp. nov., a new yeast species from a tubeworm collected at the deep-sea floor in Sagami Bay and its phylogenetic analysis

A new species of the genus Rhodotorula was isolated from a tubeworm (Lamellibrachia sp.) collected at a depth of 1156 m in Sagami Bay, Japan. Strain SY-89 had physiological properties quite similar to R. aurantiaca. Two phylogenetic trees, one based on internal transcribed spacer (ITS) regions and 5.8S rDNA sequences and the other based on the D1/D2 region of the large subunit (26S) rDNA sequences, united strain SY-89 to the type strain of Sakaguchia dacryoides through a considerable evolutionary distance. Strain SY-89 was differentiated from S. dacryoides by the G+C content of the nuclear DNA and differences in the ability to utilize specific carbon and nitrogen compounds. The low complementarity of strain SY-89 DNA to that of the type strain of S. dacryoides confirmed that this strain was genetically unrelated to previously known species. The tubeworm isolates are described as R. lamellibrachii sp. nov. The type strain of R. lamellibrachii is strain SY-89 (= JCM 10907). R. lamellibrachii formed a cluster with Erythrobasidium hasegawianum, R. lactosa, S. dacryoides and Sporobolomyces elongatus on the ITS and 5.8S rDNA phylogenetic tree. These five species shared a signature sequence in 26S rDNA, although this relationship was not supported by phylogeny based on the D1/D2 region of 26S rDNA.     

Chromosome investigations in annual Medicago species (Fabaceae) with emphasis on the origin of the polyploid Medicago rugosa and M. scutellata

Chromosome number variations play an important role in the genus Medicago. In addition to polyploidy there are cases of dysploidy as evidenced by two basic numbers, x = 8 and x = 7, the latter limited to five annual species having 2n = 14. Annuals are diploid with the exception of Medicago scutellata and Medicago rugosa which have 2n = 30 and are considered the result of crosses between the 2n = 16 and 2n = 14 species. However, this hypothesis has never been tested. This study was carried out to investigate the 2n = 14 and 2n = 30 karyotypes and verify the allopolyploid origin of M. scutellata and M. rugosa. Fluorescence in situ hybridization (FISH) of rDNA probes and genomic in situ hybridization (GISH) were performed. FISH showed that all five diploids with 2n = 14 have one pair of 45S and one pair of 5S rDNA sites. M. scutellata displayed four sites of 45S and four sites of 5S rDNA, while in M. rugosa only one pair of each of these sites was found. GISH did not produce signals useful to identify the presumed progenitors with 14 chromosomes. This result suggests alternative evolutionary pathways, such as the formation of tetraploids (2n = 32) and subsequent dysploidy events leading to the chromosome number reduction.     

Molecular-cytogenetic studies of ribosomal genes and heterochromatin reveal conserved genome organization among 11 Quercus species

Genomes of 11 Quercus species were characterized using cytogenetic (Giemsa C-banding, fluorochrome banding), molecular-cytogenetic (fluorescence in situ hybridization, FISH, to ribosomal genes) and molecular (dot-blot for ribosomal gene-copy number assessment) techniques. Ribosomal genes are the first DNA sequences to be physically mapped in oaks, and the copy number of the 18S-5.8S-26 S rRNA genes is estimated for the first time. Oak karyotypes were analysed on the basis of DAPI banding and FISH patterns; five marker chromosomes were found. In addition, chromosomal organization of ribosomal genes with respect to AT- and GC-differentiated heterochromatin was studied. Fluorochrome staining produced very similar CMA/DAPI banding patterns, and the position and number of ribosomal loci were identical for all the species studied. The 18S-5.8S-26 S rRNA genes in oak complements were represented by a major locus at the subterminal secondary constriction (SC) of the only subtelocentric chromosome pair and a minor locus at paracentromeric SC of one metacentric pair. The only 5 S rDNA locus was revealed at the paracentromeric region of the second largest metacentric pair. A striking karyotypic similarity, shown by both fluorochrome banding and FISH patterns, implies close genome relationships among oak species no matter their geographic origin (European or American) or their ecophysiology (deciduous or evergreens). Dot-blot analysis gave preliminary evidence for different copy numbers of 18S-5.8S-26 S rRNA genes in diploid genomes of Q. cerris, Q. ilex, Q. petraea, Q. pubescens and Q. robur (2700, 1300, 2200, 4000 and 2200 copies, respectively) that was correlated with the size polymorphism of the major locus. Received: 26 February 1999 / Accepted: 16 March 1999     

Nuclear ribosomal DNA phylogeny and its implications for evolutionary trends in Mexican Bursera (Burseraceae)

The genus Bursera (Burseraceae) is one of the most diversified and abundant groups of plants of the tropical dry forests of Mexico. In order to provide a basis for better understanding of its evolutionary biology, we reconstructed a phylogeny of 57 species and varieties using the nucleotide sequences of the internal transcribed spacer regions (ITS1 and ITS2) of 18S–26S and the 5.8S coding region of nuclear ribosomal DNA. We used four species of the allied genera Commiphora and Boswellia and one species of Spondias (Anacardiaceae) as outgroups. Our results support the views that Bursera is monophyletic and more closely related to Commiphora than to Boswellia. The division of Bursera into sections Bullockia and Bursera is also strongly supported by our phylogeny. Several other subclades also had high bootstrap values, especially within section Bursera. We use the phylogeny as a basis for discussing evolutionary tendencies in bark, leaves, breeding systems, and fruits.     

A STUDY OF TRIPLOID AND 3 X – 1 ANEUPLOID PLANTS IN THE GENUS GOSSYPIUM

Cytological comparisons were made of triploid and 3x – 1 plants of Gossypium hirsutum (haplo 17 and 18) X G. aridum, G. armourianum, G. harknessii, and G. raimondii. Tests and observations led to these conclusions: (1) Chromosome conjugation varied significantly from plant to plant and date to date within plants. (2) The D genome chromosomes of G. hirsutum are closer in homology to G. raimondii than to the other species tested. (3) The chromosomes of G. aridum, have closer homology to the A genome of G. hirsutum than do the chromosomes of the other D species tested. (4) The D genome of G. hirsutum has a small translocation as compared to the genomes of the four D species studied.     

Data reassessment in a phylogenetic context gives insight into chromosome evolution in the giant genus Solanum (Solanaceae)

Chromosome data are fundamental in evolution. However, there has been no attempt to synthesize and evaluate the significance of such information from a phylogenetic perspective in the giant genus Solanum, which was the aim of this work. New and published information of the main cytotaxonomic features (chromosome number, polyploidy, total length of the haploid complement, mean chromosome length, mean arm ratio, karyotype formula, nuclear DNA amount, number/position of rDNA sites) was compiled and mapped onto an embracing Solanaceae phylogeny, performing Ancestral States Reconstruction. There were 506 Solanum species with chromosome counts (49.7% from an estimated total of 1,018 spp.), with x?=?12 being the most frequent number (97%). Species with karyotypes represent 18.8%, while 8% have been studied with any molecular cytogenetic technique. Chromosome characters showed transitions associated with supported nodes, some of which have undergone fewer transitions than others. The common ancestor of all Solanum was a diploid with 2n?=?24, a karyotype with st and/or t chromosomes, 2C DNA content of 1–1.2 pg, one locus of 18–5.8–26S rDNA and one of 5S, both loci being asyntenic. The chromosomal variables behave as homoplastic, with reversions in all branches. The analysed characters were sorted from more to less conserved: asynteny of rDNA loci; number of sites of 18–5.8–26S; chromosome number; karyotype formula; number of 5S loci. This pattern of chromosomal evolution distinguishes Solanum from closely related genera and from genera from other families with a similar number of species.     

THE FEMALE GAMETOPHYTE OF THREE SPECIES OF GRINDELIA AND OF PRIONOPSIS CILIATA (COMPOSITAE)

In the four species studied, Grindelia stricta ssp. blakei, G. arizonica, G. lanceolata, and Prionopsis ciliata (Haplopappus ciliatus), the female gametophyte develops according to the Polygonum (normal) type from the chalazal megaspore of a row of four. In most cases only two antipodal cells are formed, the micropylar one originally containing two nuclei. The number of nuclei increases in both cells; one or both antipodal cells typically grow laterally into the integument, often extending to near the surface of the ovule. This resembles the condition previously reported in Grindelia squarrosa. Since the four species of Grindelia have similar antipodal outgrowths, such outgrowths may be considered typical for this genus. The fact that Prionopsis resembles Grindelia in regard to outgrowths from the antipodal cells and differs in this respect from typical Haplopappus helps to justify its separation from Haplopappus, and perhaps its suggested merger with Grindelia.     

Genetic diversity of four new species related to southwestern Sichuan buckwheats as revealed by karyotype,ISSR and allozyme characterization

The genetic diversity of four new species related to southwestern Sichuan buckwheats was examined using karyotypes, allozymes and intersimple sequence repeats (ISSR). Karyotype analysis showed that Fagopyrum wenchuanense, F. qiangcai and F. pugense are diploid (2n = 2x = 16) while F. crispatofolium is tetraploid (2n = 4x = 32). The karyotype evolutionary trend of buckwheat indicated that F. crispatofolium, F. gracilipes var. odontopterum and F. gracilipes seemed to be more advanced than F. wenchuanense, F. qiangcai, F. pugense, F. esculentum, F. cymosum and F. tataricumare. Three polymorphic enzyme systems were used for the examination of variation, including peroxidase, esterase and superoxide dismutase. Consistent banding patterns were obtained for esterase and superoxide dismutase, while peroxidase produced a large number of sharp bands. A total of 18 ISSR primers were selected for the analysis and showed high variations among the species. Allozymes and ISSR markers were utilized to estimate the genetic distance among accessions and to draw phylogenetic trees. Our data provide evidence of a high degree of genetic diversity among southwestern Sichuan buckwheats. In addition, both cultivars and wild types showed a high degree of divergence suggesting a complex domestication process in this crop. This study provides a better understanding of evolutionary mechanisms and genetic relationships in four new species of buckwheat.     

CHROMOSOME NUMBERS IN THE LEGUMINOSAE. III. SPECIES OF THE SOUTHWESTERN UNITED STATES AND MEXICO

Turner, B. L., and Olin S. Fearing. (U. Texas, Austin.) Chromosome numbers in the Leguminosae. III. Species of the Southwestern United States and Mexico. Amer. Jour. Bot. 47(7) : 603–608. Illus. 1960.—Chromosome counts for 43 species of the Leguminosae from the southwestern United States and Mexico have been reported. These include first reports for 42 taxa of which 16 are for the subfamily Mimosoideae. Olneya tesota (2n = 18) is the only new generic count listed. Chromosome reports of particular significance include a single polyploid count for a North American species of Acacia, as well as diploid and tetraploid counts for closely related taxa in this genus. Four species of the genus Schrankia were found to be diploid with In = 26, indicating a base of x = 13 instead of the x = 8 reported by some previous workers. Leucaena pulverulenta was found to have a diploid count of 2n = 56 indicating a base of x = 14.     

Genome differentiation in Aegilops. 3. Evolution of the D-genome cluster

 Six polyploid Aegilops species containing the D genome were studied by C-banding and fluorescence in situ hybridization (FISH) using clones pTa71 (18S-5.8S-26S rDNA), pTa794 (5S rDNA), and pAs1 (non-coding repetitive DNA sequence) as probes. The C-banding and pAs1-FISH patterns of Ae. cylindrica chromosomes were identical to those of the parental species. However, inactivation of the NOR on chromosome 5D with a simultaneous decrease in the size of the pTa71-FISH site was observed. The N^v and D^v genomes of Ae. ventricosa were somewhat modified as compared with the N genome of Ae. uniaristata and the D genome of Ae. tauschii. Modifications included minor changes in the C-banding and pAs1-FISH patterns, complete deletion of the NOR on chromosome 5D^v, and the loss of several minor 18S-5.8S-26S rDNA loci on N^v genome chromosomes. According to C-banding and FISH analyses, the D^cr1 genome of Ae. crassa is more similar to the D^v genome of Ae. ventricosa than to the D genome of Ae. tauschii. Mapping of the 18S-5.8S-26S rDNA and 5S rDNA loci by multicolor FISH suggests that the second (X^cr) genome of tetraploid Ae. crassa is a derivative of the S genome (section Emarginata of the Sitopsis group). Both genomes of Ae. crassa were significantly modified as the result of chromosomal rearrangements and redistribution of highly repetitive DNA sequences. Hexaploid Ae. crassa and Ae. vavilovii arose from the hybridization of chromosomal type N of tetraploid Ae. crassa with Ae. tauschii and Ae. searsii, respectively. Chromosomal type T1 of tetraploid Ae. crassa and Ae. umbellulata were the ancestral forms of Ae. juvenalis. The high level of genome modification in Ae. juvenalis indicates that it is the oldest hexaploid species in this group. The occurrence of hexaploid Ae. crassa was accompanied by a species-specific translocation between chromosomes 4D^cr1 and 7X^cr. No chromosome changes relative to the parental species were detected in Ae. vavilovii, however, its intraspecific diversity was accompanied by a translocation between chromosomes 3X^cr and 3D^cr1. Received July 24, 2001 Accepted October 1, 2001