Giemsa C-banded karyotypes of some diploidArtemisia species

Detailed C-banded karyotypes of eight diploidArtemisia species from three different sections are reported together with preliminary observations on three additional related diploid species. In the majority, the overall amount of banding is relatively low. Bands are mostly confined to distal chromosome regions; intercalary banding is virtually absent and centromeric heterochromatin is also scarce. With the exception ofA. judaica there is in general great uniformity in karyotype structure but considerable interspecific variation in total karyotype length (and hence DNA content) ranging from 44 µm inA. capillaris (2n = 18) to 99 µm inA. atrata (2n = 18).A. judaica (2n = 16; total karyotype length 97 µm) was distinguished by its karyomorphology, with one large non-banded metacentric chromosome pair and 7 pairs of smaller terminally banded meta- or submetacentric chromosomes.     

Cytological characterization of heterochromatin and rDNA inPinus radiata andP. taeda

Fluorochrome C-banding ofPinus radiata andP. taeda metaphase chromosomes showed many pericentromeric DAPI bands and interstitial CMA bands inP. radiata, and centromeric and interstitial CMA bands inP. taeda. Giemsa C-band patterns differed between the species with centromeric bands inP. radiata but no consistent bands inP. taeda. A karyotype ofP. radiata was developed based on banding patterns that distinguished all but two of the 12 pairs of chromosomes. In situ hybridization (ISH) using probes for high-copy ribosomal DNA (rDNA) showed 10 pairs of 18S–25S sites and two pairs of 5S sites in both species. Most of the sites were interstitial or centromeric.     

Karyotypic evolution,morphological variability and phylogeny inMedicago sect.Intertextae

Karyotype and external morphological analyses were conducted onMedicago ciliaris, M. intertexta, M. muricoleptis andM. granadensis which comprise theIntertextae section of the genusMedicago. All species were found to have 2n = 16 chromosomes (= 2 ×), including one pair of satellite chromosomes in each respective complement. Karyotypic evolution in theIntertextae involves changes in absolute chromosome size and in centromeric and relative size symmetry. Numerical taxonomic analyses were conducted independently on 17 karyotypic features and on 16 features of external morphology. The results of the two sets of analyses proved comparable, withM. ciliaris andM. intertexta forming a fairly close pair, and the remaining two species appearing to have more distant relationships to each other and to the first pair. These observations are consistent with the infertility relationships and chorologies of the species. It is suggested thatM. muricoleptis andM. granadensis are derived from theM. ciliaris/intertexta species complex withM. granadensis arising fromM. muricoleptis, or these two species independently evolving from a common species complex. Chromosomal and numerical analyses suggest thatM. ciliaris is the most primitive andM. granadensis the most derived species of theIntertextae.     

Allozyme variation in relation to morphology and taxonomy inMedicago sect.Spirocarpos subsect.Intertextae (Fabaceae)

Medicago intertexta andM. ciliaris have been controversially recognized as separate species. The only reliable diagnostic character, gland-tipped trichomes on the fruits inM. ciliaris, is controlled by presence of a single dominant allele, and such one-character taxonomies are debatable. Contributing to the difficulty,M. muricoleptis andM. granadensis, the other two species ofMedicago sectionSpirocarpos subsectionIntertextae, are sometimes confusingly similar toM. intertexta or to each other. Allozyme differences provided 95% verification of the suitability of the gland-tipped trichome character for separatingM. intertexta andM. ciliaris, thus corroborating their recognition as separate taxa. Several measures of allozyme variation indicated thatM. intertexta is more polymorphic than its sister species. Heterozygosity was also highest inM. intertexta, suggestive of a higher outcrossing rate, which is also consistent with larger floral size. Heterozygosity ofM. intertexta was concentrated in Sicily and nearby countries. Taxonomic difficulties in identifying SicilianM. intertexta are well known, and may be the result of interspecific hybridization and introgression.Medicago muricoleptis differed from the above two species in the frequency of several alleles, whileM. granadensis possessed numerous unique alleles consistent with its complete absence of genetic exchange with the other three substantially interfertile species.     

Taxonomic studies ofAsarum sensu lato

The karyotype and the C-banding pattern in two species ofHexastylis andAsarum epigynum were analysed in detail, and the results obtained were compared with those of the other species ofAsarum, Asiasarum andHeterotropa previously reported. The present results were partially different from the previous reports related to the karyotypes of these species. The karyotype observed in two species ofHexastylis (2n=26) was represented by ten pairs of metacentric chromosomes and three pairs of small subtelocentric chromosomes, which is very similar to that ofAsiasarum in eastern Asia. The C-banding patterns ofHexastylis andAsiasarum, however, were clearly different from each other. A striking difference was found in one of the three pairs of small subtelocentric chromosomes. A Formosan speciesAsarum epigynum had the somatic chromosome number 2n=12 and a highly asymmetrical karyotype composed of mainly subtelocentric chromosomes. These karyological features were remarkably different from those of the other groups inAsarum s.l.     

Karyotype evolution by centromeric fission inZamia (Cycadales)

The chromosome numbers of several species ofZamia from Mexico are reported.Z. paucijuga, distributed from central Oaxaca to Nayarit, has been found to have 2n = 23, 25, 26, 27 and 28. 2n = 28 is the highest chromosome number yet found in the cycads. Karyotypes of this species differ principally in the number of telocentric and metacentric chromosomes present in each; 2n = 23, 25, 26, 27 and 28 were found to have 5, 3, 2, 1 and 0 metacentric and 8, 12, 14, 16 and 18 telocentric chromosomes, respectively.Z. fischeri has been found to be 2n = 16,Z. furfuracea andZ. loddigesii 2n = 18.Zamia paucijuga on the basis of morphological and ecological characteristics, is considered to be an advanced member of this genus. Chromosome and karyotype evolution inZ. paucijuga may have occurred by centromeric fission of metacentric chromosomes; the karyotypes ofZ. paucijuga are strongly asymmetrical, suggesting that they evolved recently.     

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.     

In vitro propagation ofMedicago andLotus species by node culture

Summary Nodes ofMedicago sativa, Lotus corniculatus, Lotus tenuis, andLotus pedunculatus were cultured on MS basal media with different growth regulators. InM. sativa each node produced one shoot and the apical dominance was unaffected by high levels of cytokinins, and subsequent cycles of culture. Shoot development was stimulated by the presence ofN ⁶-isopentenyl-adenine in the culture medium and was dependent on the genotype of the explant. Shoot development was not affected by the original position of the node on the plant nor by the plant age. Shoots rooted in MS medium gelled with starch and containing 2 mg·liter⁻¹ indol-3-acetic acid. In the threeLotus species, node culture was a more effective technique than inM. sativa. The number of shoots per node increased with the time of culture and with the presence of 0.05 mg·liter⁻¹ of 6-benzylaminopurine. The highest number of shoots derived from one node was achieved inL. pedunculatus and inL. tenuis by culturing single nodes, whereas inL. corniculatus stem segments had to be totally covered by the medium for success. Rooting was easily achieved in MS medium with or without auxins.     

Karyosystematic studies on threeCrepis species (Asteraceae) endemic to Greece

This work examines the cytogeographical distribution, the morphological characters, and the karyotypes of threeCrepis species endemic to Greece (C. sibthorpiana, C. incana, andC. heldreichiana). C. sibthorpiana is diploid (2n = 2x = 8),C. incana is diploid (2n = 2x = 8) and tetraploid (2n = 4x = 16, 17), andC. heldreichiana is always dekaploid (2n = 10x = 40). The Giemsa positive bands, usually pairs of dots, are mainly centromeric inC. incana, while they are terminal inC. sibthorpiana (on the short arm of all chromosomes) and inC. heldreichiana (on both arms of all chromosomes). Intercalary C-bands are scarce and usually variable within karyotypes, individuals, and species. The most variable karyotype both in Feulgen and Giemsa preparations is that ofC. incana, in which also supernumerary chromosomes were observed, which are polysomic to standard set members. On the basis of morphological and karyological data the evolutionary relationships among the threeCrepis taxa are discussed.     

Tail length variation inMacaca fascicularis andM. mulatta

Based on collectors' measurements of 1,040 specimens, variation of relative tail length [RTL = (tail length/head and body length) × 100] has been studied inMacaca fascicularis andM. mulatta, two closely related species that replace one another in tropical Asia and subtropical Asia, respectively. RTL usually is greater than 90 inM. fascicularis and usually is less than 60 inM. mulatta; intermediate values occur in only 3.5% of specimens studied. Within each species, RTL is approximately equal in females and males. From infancy to adulthood, RTL tends to decline in both species. InM. fascicularis, RTL generally decreases with increasing latitude; inM. mulatta, RTL is approximately constant latitudinally. Where the geographic ranges of these two species meet at ca. 15°N in the Indochinese Peninsula, a few specimens have been collected in which RTL and dorsal pelage color are intermediate between those inM. fascicularis andM. mulatta. The observed pattern of variation suggests that the ranges ofM. fascicularis andM. mulatta formerly were separated by a zoogeographic barrier — perhaps during a Pleistocene glacial interval. After disappearance of the postulated barrier, the ranges of these two species apparently became contiguous and limited hybridization has occurred.     

Effects of trait plasticity on aboveground biomass production depend on species identity in experimental grasslands

Experimental studies with grassland species found a positive relationship between species richness and community biomass production, however the response of individual species was highly variable. The mechanisms behind these patterns are poorly understood. Here we studied aboveground biomass production and plasticity of growth characteristics of four legumes with similar morphology (Lotus corniculatus, Medicago lupulina, Onobrychis viciifolia, Trifolium hybridum) in experimental grasslands varying in species richness (1, 2, 4, 8, 16 and 60) and composition. We identified O. viciifolia and T. hybridum as species that reached higher biomass production in mixtures than expected from monoculture yields, while L. corniculatus and M. lupulina mostly had lower yields than expected. Variation of morphological traits across the species-richness gradient was lowest in the highly competitive O. viciifolia, but increased in the smaller species. The tall-growing O. viciifolia achieved higher biomass production by both, a higher number of plant individuals and an increase in mean mass per individual. Mean shoot number per individual remained constant, but individuals produced heavier shoots. The medium-sized T. hybridum also increased the number of plant individuals, but mean mass per individual did not respond to community species richness. The average mass per shoot was increased in mixtures, but the species developed less shoots per individual. Shoot length and stem weight ratio of T. hybridum increased with community species richness. Morphological changes in the less successful L. corniculatus and M. lupulina with a smaller growth stature were similarly directed as those of T. hybridum. The observed morphological changes are known as typical shade-avoidance mechanisms in dense vegetation. Our study shows that stress responses to changes in resource availability may be a mechanism to enforce higher aboveground biomass production of individual species in mixtures, but it depends on species identity whether trait plasticity is large enough to exceed stress-induced growth limitations.     

RFLP analysis for species separation in the generaBipolaris andCurvularia

Restriction fragment length polymorphism (RFLP) of the total DNA ofBipolaris andCurvularia species was analysed using arbitrarily chosen genomic clones of DNA fromCurvularia lunata andBipolaris maydis as probes. Clear differences among species in both genera, resulting in different banding positions, were obtained with some probe-enzyme combinations. Intraspecific polymorphism in banding positions with these probe-enzyme combinations was slight. These analyses allow discrimination between the species. DNA fingerprinting with intrageneric probes is a potentially useful tool for species separation and identification inBipolaris andCurvularia when coupled with another characteristic such as conidial morphology.Curvularia aeria comb. nov. was proposed forCurvularia lunata var.aeria on the basis of differences in RFLP banding patterns and differences in conidial morphology.     

Karyotaxonomical analysis in the genusAngelica (Umbelliferae)

Morphometric karyotype characters were studied in 25Angelica spp. (Umbelliferae, Apioideae) and in one species of the related genusTommasinia. For three species the chromosome numbers are new. In our study the majority of the species investigated are diploids with 2n = 22, some are tetraploids with 2n = 44 (for these tetraploids also diploid cytotypes are reported in the literature). Among the diploid species,A. miqueliana has a distinct karyotype consisting of submetacentric and acrocentric chromosomes only, the remaining diploids with 2n = 22 as well as tetraploids with 2n = 44 have rather symmetrical karyotypes, consisting of metacentric and submetacentric chromosomes. The very different chromosome number 2n = 28 has been found inA. gmelinii. Its karyotype includes two distinct groups of chromosomes: 8 pairs of rather large metacentrics and submetacentrics and 6 pairs of very short and asymmetrical chromosomes. Chromosome numbers and structures appear to be useful in the taxonomy of some intrageneric taxa inAngelica.     

Giemsa C-banded karyotypes of diploid and tetraploidHordeum bulbosum (Poaceae)

The similar-looking basic genomes ofHordeum bulbosum (2x and 4x) have five rather similar metacentric, one submetacentric, and one satellited choromosome. C-banding patterns are characterized by one or two centromeric, or juxtacentromeric, small to larger bands in most chromosomes, by bands at the nucleolar organizers, by small or very small telomeric bands, and by the nearly complete lack of intercalary bands. Banding pattern polymorphism is widespread. Banding patterns supported by chromosome morphology enable identification of homologues, and discrimination between non-homologues inH. bulbosum (2x). The C-banded karyotype ofH. bulbosum (4x) supports an autopolyploid origin, but it was possible to identify only homologues of submetacentrics and SAT-chromosomes.     

Karyotype and C-banding inTrigonella sectionFoenumgraecum (Fabaceae)

The six species of the sectionFoenum-graecum ofTrigonella have the same chromosome number, 2n = 16.T. gladiata andT. cariensis have fairly symmetrical karyotypes, while those ofT. foenum-graecum, T. berythea, T. macrorrhyncha andT. cassia are asymmetrical. C-bands are present in all six species but the number of bands and their positive vary considerably among the species. The karyotype evidence suggests that none of the available species of theFoenum-graecum section can be considered as the wild progenitor of fenugreek.     

Karyotypic analysis of Cajanus,Atylosia, and Rhynchosia species

Summary Somatic chromosomes of two cultivais of Cajanus cajan, eight species of Atylosia (A. albicans, A. cajanifolia, A. lineata, A. platycarpa, A. scarabaeoides, A. serica, A. trinervia and A. volubilis), and of Rhynchosia rothii were analysed. All species had 2n=22. Eight of the 10 species studied had two pairs of satchromosomes while A. scarabaeoides and A. sericea had only one sat-chromosome pair. Based on relative chromosome length (L%), arm ratio (pa-value) and presence or absence of secondary constriction, a karyotype formula for each species was formulated. Based on these parameters the chromosome pairs could also be assigned to groups ranging from 8 to 10 in different species. Except for the asymmetrical karyotype of A. albicans, the other species had rather moderately symmetrical karyotypes.     

Chromosome banding and genome size differentiation inProspero (Hyacinthaceae): Diploids

Prospero is a Mediterranean autumn-flowering genus ofHyacinthaceae commonly classified inScilla asS. autumnalis andS. obtusifolia. Extensive dysploid and polyploid variation has been reported. In the present study 77 diploid accessions from the western to the eastern part of the area of distribution, the major part being from continental Greece and Crete, have been analysed for karyotype structure and, in part, for genome size. Methods employed were acetocarmine staining, Giemsa C-banding, fluorochrome staining mainly with chromomycin A₃/DAPI, silver impregnation, and Feulgen densitometry. Banded idiograms were established with a computer assisted karyotype analysis procedure. Chromosome numbers were 2n = 8 inP. obtusifolium, and 2n = 12 and 14 inP. autumnale s. l. Dispensable euchromatic chromosome segments and different types of B chromosomes occurred. Among the cytotypes with 2n = 14 two karyotypes from Turkey differed from each other and from the rest in form, position of the nucleolar constriction, and in genome size. The remaining accessions were similar in karyotype shape but three levels of genome size could be discerned, the highest (1C = 7.50 pg) being found on the Iberian Peninsula, an intermediate one on Corsica and Malta, and the lowest (4.27 pg) in the Aegean. The karyotype with 2n = 12 had an intermediate genome size, and that ofP. obtusifolium a relatively low one. Heterochromatin amount was generally low, but some karyotypes showed characteristic banding patterns. The relationship between the chromosome complements with 2n = 14, 12 and 8 is discussed on the basis of idiograms and DNA amounts.The authors respectfully dedicate this papers to emer. o. Prof. Dr.Elisabeth Tschermak-Woess on the occasion of her 80th birthday.     

Isozyme variation and alleged progenitor-derivative relationships in theMedicago murex complex (Fabaceae)

Chromosomal studies ofMedicago lesinsii (n = 8) and its close relativeM. murex (n = 7) have led to the competing hypotheses that the latter is derived directly from the former, or that both originated from a common ancestor. In contrast to the relatively variableM. murex, M. lesinsii proved to be almost uniform isozymically, except that most populations of Greece differed by one allele from plants of the remainder of the range. This Greek variant ofM. lesinsii was indistinguishable from one of the isozyme variants ofM. murex. The greater level of allozyme variation inM. murex was consistent with its greater ecological amplitude and competitive ability. Also, this suggests thatM. murex is unlikely to have originated directly from the less variableM. lesinsii. The data suggest that either both species originated from a common ancestor, or that the n = 8 species evolved from the n = 7 species, a mode of chromosome evolution not previously hypothesized for the genus.     

Variation within and among the chloroplast genomes ofMelaleuca alternifolia andM. linariifolia (Myrtaceae)

Melaleuca alternifolia andM. linariifolia are commercially important Australian species harvested for their essential oils. Both species have relatively narrow and disjunct distributions on the central coast of eastern Australia. Variation in the chloroplast genome was assessed for eight individuals from each of twelve populations, representing the species' geographic range. Low nucleotide diversity withinM. alternifolia contrasted with high nucleotide diversity inM. linariifolia. CpDNA data are consistent with the southern population ofM. alternifolia being a hybrid population withM. linariifolia. The two species are sympatric in this region. Variation inM. linariifolia was geographically structured, with northern populations differing from southern populations by seven restriction site mutations, five length mutations and an inversion. There was no evidence of hybridisation of the cp genome of northernM. linariifolia with the partially sympatric speciesM. trichostachya. Intra- and interspecific variation in the chloroplast genomes ofM. alternifolia, M. linariifolia, andM. trichostachya indicate considerable potential for the use of intraspecific cpDNA studies in examining phylogenetic relationships in melaleucas.     

DNA evidence for multiple origins of intergeneric allopolyploids in annualMicroseris (Asteraceae)

Seventy populations of North American annualMicroseris, Stebbinsoseris, andUropappus species were examined for chloroplast and nuclear ribosomal DNA restriction site variability to determine the origin of the allotetraploid speciesS. heterocarpa andS. decipiens. Previously identified chloroplast DNA restriction site variants were used in concert with restriction site variation forNco I in the nuclear-encoded ribosomal DNA repeat. The presence of two, mutually exclusive restriction site gains were observed in diploid populations ofM. douglasii; these same variants were also found in populations of allotetraploidS. heterocarpa, indicating mutiple origins of this species from different maternal diploid populations ofM. douglasii. Variation in the rDNA repeat between the diploid annual species and the putative paternal genome ofU. lindleyi was found to be additive inS. heterocarpa. A similar relationship was observed for the origin ofS. decipiens; cpDNA restriction site variants found inM. bigelovii andM. douglasii were present inS. decipiens. The rDNANco I variants also were additive in this purported allotetraploid. These results confirm the reticulate evolutionary pattern inStebbinsoseris and provide another example of multiple origins of intergeneric allopolyploids.