Patterns, sources and ecological implications of clonal diversity in apomictic Ranunculus carpaticola (Ranunculus auricomus complex, Ranunculaceae)

Sources and implications of genetic diversity in agamic complexes are still under debate. Population studies (amplified fragment length polymorphisms, microsatellites) and karyological methods (Feulgen DNA image densitometry and flow cytometry) were employed for characterization of genetic diversity and ploidy levels of 10 populations of Ranunculus carpaticola in central Slovakia. Whereas two diploid populations showed high levels of genetic diversity, as expected for sexual reproduction, eight populations are hexaploid and harbour lower degrees of genotypic variation, but maintain high levels of heterozygosity at many loci, as is typical for apomicts. Polyploid populations consist either of a single AFLP genotype or of one dominant and a few deviating genotypes. genotype/genodive and character incompatibility analyses suggest that genotypic variation within apomictic populations is caused by mutations, but in one population probably also by recombination. This local facultative sexuality may have a great impact on regional genotypic diversity. Two microsatellite loci discriminated genotypes separated by the accumulation of few mutations ('clone mates') within each AFLP clone. Genetic diversity is partitioned mainly among apomictic populations and is not geographically structured, which may be due to facultative sexuality and/or multiple colonizations of sites by different clones. Habitat differentiation and a tendency to inhabit artificial meadows is more pronounced in apomictic than in sexual populations. We hypothesize that maintenance of genetic diversity and superior colonizing abilities of apomicts in temporally and spatially heterogeneous environments are important for their distributional success.     

Morphological differentiation within the Ranunculus cassubicus group compared to variation of isozymes, ploidy levels, and reproductive systems: implications for taxonomy

 In the partly apomictic Ranunculus cassubicus group, a subgroup of the R. auricomus complex, two species were studied by morphometric analyses: R. cassubicifolius W. Koch (with three diploid and two autotetraploid sexual populations), and R. carpaticola Soó (with three diploid sexual populations and a hexaploid apomictic one). Multidimensional scaling analyses (MDS) of individuals, boxplots and cluster analyses of populations revealed a differentiation of R. cassubicifolius and R. carpaticola, whereby in MDS the hexaploid apomictic individuals are partly intermediate between R. cassubicifolius and R. carpaticola. The cytodemes of R. cassubicifolius showed no morphological and only a weak genetic differentiation. A comparison of morphology, isozymes, reproductive system and ploidy levels showed only partly congruence of data sets in respect of grouping populations, thus illustrating the problem to find criteria for a taxonomic concept. A treatment of the apomictic population as a separate group is indicated by all data sets, afterwards R. cassubicifolius and diploid R. carpaticola represent two other well-defined groups. Canonical variate analysis including all characters confirmed the three suggested groups as significantly different and showed that a total of 89.3% of individuals are correctly classified; number of teeth of stem leaf segments and number of petals are the most discriminating characters. Herbarium studies confirm the morphological differentiation yielded from population samples. The three population groups are even better separated in a canonical variate analysis of isozyme data (presence/absence of 25 alleles) of the same material, here 92.6% of individuals are correctly classified. Morphology and isozyme data suggest that the hexaploid apomict originated from hybrids of R. cassubicifolius and diploid R. carpaticola and must be excluded from the sexual taxa; the final classification and naming of the apomicts must be left for further studies on a larger material. The sexual taxa should be classified as separate species. Herbarium studies indicate that R. carpaticola s.str. is widespread over the Carpathians and might include other populations hitherto ascribed to other microspecies as well. Received November 20, 2001; accepted May 10, 2002 Published online: September 13, 2002     

Diploid and autotetraploid sexuals and their relationships to apomicts in the Ranunculus cassubicus group: insights from DNA content and isozyme variation

 From the predominantly aposporous Ranunculus cassubicus group, a subgroup of the R. auricomus complex, two species with both diploid and tetraploid cytodemes (R. cassubicifolius W. Koch and R. carpaticola Soó) were known. Nine population samples of both species have been analyzed for variation of ploidy levels and isozymes. DNA image analysis showed that three populations of R. cassubicifolius from Bavaria and Salzburg are diploid, two from Lower Austria tetraploid. In R. carpaticola, two populations from Central Slovakia and one from Romania are diploid, one from northern Slovakia is hexaploid. The polyploid populations had somewhat smaller C-values than expected from diploids. Ploidy levels are consistent within populations, contradicting previous hypotheses that cycles of diploid-tetra-ploid-dihaploid apomictic individuals occur in natural populations of goldilocks. Isozyme-allozyme analysis of nine polymorphic loci showed that individual variation and genetic measures of di-ploid and tetraploid populations are equivalent to those of sexual taxa. Multiple allelism and unbalanced gene dosages in tetraploid R. cassubicifolius give evidence for autopolyploidy that is most probably of multiple origin. The observed excess of homozygotes and the high diversity between populations in the diploid populations of R. cassubicifolius are most probably due to geographical isolation of population groups, that might have happened during the Würm glaciation and might have promoted the separation of autotetraploids in the easternmost part of the distribution range. Genetic distance values analyzed by UPGMA of all sexual populations separated the two taxa and the cytodemes within R. cassubicifolius. Multidimensional scaling of individuals (including the apomicts) based on a presence/absence matrix of alleles confirmed this differentiation with an overlapping zone between R. cassubicifolius and R. carpaticola. The hexaploid R. carpaticola population showed reduced genotypic diversity, an increased number of heterozygotes and fixed heterozygosity as typical for apomictic mode of reproduction. This population shared alleles of both species without any specific ones, multidimensional scaling placed genotypes among R. cassubicifolius. Thus, the hexa-ploid apomicts might have originated from hybrids of R. cassubicifolius and R. carpaticola. In general, evolution of agamic lineages in the R. cassubicus complex might have been facilitated by autopoly-ploids, because they can provide a bridge between the reproductive systems that are otherwise isolated by ploidy levels, and also a starting point for spontaneous origin of apomixis. Received October 8, 2001; accepted May 10, 2002 Published online: November 7, 2002 Addresses of the authors: Elvia H?randl (E-mail: elvira.hoerandl@univie.ac.at), Department of Systematics and Evolution of Higher Plants; Johann Greilhuber (E-mail: Johann.Greilhuber@univie. ac.at), Department of Systematic Karyology and Embryology of Plants; both: Institute of Botany, University of Vienna, Rennweg 14, A-1030 Vienna, Austria.     

The role of hybridization, polyploidization and glaciation in the origin and evolution of the apomictic Ranunculus cassubicus complex

The Ranunculus cassubicus complex, comprising diploids and polyploids, is a good model for studying the role of hybridization and polyploidy in the origin of apomixis. Results from amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) analyses performed on 448 individuals were combined with evidence from morphology, isozymes, karyology and distribution. Our results indicated a unique hybrid origin for the apomictic hexaploid R. carpaticola from north-western Slovakia, involving two sexual parents: autotetraploid R. cassubicifolius from the northern pre-Alps, and diploid R. carpaticola from central Slovakia. The hybrids were intermediate to the parents, but unique alleles have resulted from genomic reorganisation in the allopolyploids, which might also have triggered apomixis. Their distribution patterns and estimated ages suggest that hybridization may be correlated with the last glacial period. Hybridization seems to be the major origination for apomicts in the R. cassubicus complex. Polyploidy creates novel sexual genotypes and acts as a springboard for the production of new hybrids, but it only results in a combination with hybridization in apomixis. In turn, asexuality has permitted the perpetuation and establishment of ecologically divergent hybrid genotypes.     

Cytogeography and reproduction of the Paspalum simplex polyploid complex

 Paspalum simplex is a grass distributed throughout the phytogeographic Chaco region in South America from which sexual diploid and apomictic tetraploid races have been reported. We analysed native populations to determine their homogeneity of ploidy level, and the relationship between geographic distribution, ploidy levels, and reproductive systems. The ploidy level was established for 379 plants from 32 wild populations. Tetraploidy and apomixis constitute the most common combination for this species all over the Chaco region. Apomictic hexaploid plants were found associated with 4x populations. Diploids were confined to a small sector of the region. One sexual triploid plant arose from seed harvested in a pure 2x population, and one apomictic 3x plant was found in a mixed 2x-4x population. The results suggest that P. simplex is a core agamic complex characteristic of the Chaco region from which other apomictic polyploid species of the subgenus Anachyris could have evolved. Received July 24, 2002; accepted September 12, 2002 Published online: December 11, 2002     

Morphometric analysis of the Bersama abyssinica Fresen. complex (Melianthaceae) in East Africa

 The morphological variation of the Bersama abyssinica complex in East Africa is analysed. A wide range of characters was examined both on specimens collected for this study and on a selection of type material. The character distribution of each character was examined individually. The ratios of some of the characters were calculated, and selected characters were plotted in pairwise combinations. Non-metric multidimensional scaling and principal component analysis were employed in the multivariate analyses. On the basis of the patterns observed in the numerical analyses as well as those in the field, a new combination, Bersama abyssinica Fresen. ssp. rosea (Hoyle) Mikkelsen is proposed. The presence of the two flower types described by White: (a) male flowers, with long stamens and short gynoecium and (b) female flowers, with short, abortive stamens and long gynoecium is confirmed, but found to be taxonomically insignificant. Received May 12, 2000 Accepted February 13, 2001     

Isozyme evidence for the allotriploid origin of Lycoris flavescens (Amaryllidaceae)

 Variation in isozyme patterns from ten populations of the Korean endemic Lycoris species was used to test the hypothesis that L. flavescens originated from natural hybridization between diploid L. chinensis and L. sanguinea var. koreana. Lycoris sanguinea var. koreana shows fixed heterozygosity at four of nine loci assayed, suggesting that this species is an allotetraploid instead of a diploid. Electrophoretic data suggest that Lycoris flavescens is an allotriploid species derived from the hybridization between diploid L. chinensis and tetraploid L. sanguinea var. koreana. The patterns of allelic distribution in populations of L. flavescens suggest multiple origins of the allotriploid. Within the L. flavescens complex, our isozyme data support the recognition of two taxa, L. flavescens and a recently recognized species, L. uydoensis. Received August 28, 2000 Accepted December 27, 2000     

Evolutionary history of the polyploid complex of Cardamine amara (Brassicaceae): isozyme evidence

 In samples from 56 populations of Cardamine amara, representing four diploid subspecies (subspp. amara, opicii, balcanica, and pyrenaea) and two tetraploid subspecies (subspp. austriaca and olotensis) from different parts of the European distribution area, four enzyme systems with 23 alleles were studied. These data, together with previous morphological and karyological data, suggest that the distribution and variation pattern within the species was strongly influenced by the last glacial period and postglacial migrations. Cardamine amara subsp. pyrenaea is monomorphic for a unique allele, and subsp. balcanica has a unique allele, too which, however, is not fixed in all populations of the taxon. Both taxa seem to be relic ones, although otherwise subsp. balcanica in respect of allelic spectrum much resembles subsp. amara. The other two diploid subspecies, subsp. amara and subsp. opicii, are not characterised by presence of unique alleles but differ in allele frequencies. The two tetraploid subspecies have different evolutionary histories. C. amara subsp. austriaca seems to be an autopolyploid derivative of subsp. amara which colonised open space offered by retreating glaciers in the Eastern Alps. C. amara subsp. olotensis from the Iberian Peninsula represents most probably a polyploid of preglacial time. Received June 22, 2001; accepted May 17, 2002 Published online: September 13, 2002     

Evolution, phylogeography, and taxonomy within the Viola alba complex (Violaceae)

 Taxa of the Viola alba complex were investigated using allozymes and morphometry. A taxonomic revision is presented. A wide delimitation of V. alba with only three morphological and geographical subspecies is suggested: (1) ssp. dehnhardtii distributed in the Mediterranean eastwards to Turkey; (2) ssp. alba flanking ssp. dehnhardtii in the north and east; and (3) ssp. cretica endemic to Crete. Ssp. cretica, up to now treated as a separate species, is particularly close to ssp. dehnhardtii. Viola cadevallii (NW Mediterranean) is included in the synonymy of ssp. dehnhardtii. Ssp. scotophylla (S Europe), ssp. thessala (Balkan), V. armena (Turkey), and V. besseri (Caucasus) are reduced to synonyms of V. alba ssp. alba. Viola pentelica (Greece) might represent transitional forms between ssp. alba and ssp. dehnhardtii. Glacial refugia for ssp. alba are suggested from the eastern Mediterranean via Turkey to the Caucasus, for ssp. dehnhardtii in the Mediterranean area in general, and for ssp. cretica in Crete. A key to the subspecies is provided. Taxonomic recombination: Viola alba Bess. ssp. cretica (Boiss. & Heldr.) Marcussen, comb. nov. Received June 17, 2002; accepted November 27, 2002 Published online: March 20, 2003     

Introgression of apomixis into sexual species is inhibited by mentor effects and ploidy barriers in the Ranunculus auricomus complex

Background and Aims

Apomictic plants maintain functional pollen, and via pollination the genetic factors controlling apomixis can be potentially transferred to congeneric sexual populations. In contrast, the sexual individuals do not fertilize apomictic plants which produce seeds without fertilization of the egg cells. This unidirectional introgressive hybridization is expected finally to replace sexuality by apomixis and is thought to be a causal factor for the wide geographical distribution of apomictic complexes. Nevertheless, this process may be inhibited by induced selfing (mentor effects) of otherwise self-incompatible sexual individuals. Here whether mentor effects or actual cross-fertilization takes place between diploid sexual and polyploid apomictic cytotypes in the Ranunculus auricomus complex was tested via experimental crosses.

Methods

Diploid sexual mother plants were pollinated with tetra- and hexaploid apomictic pollen donators by hand, and the amount of well-developed seed compared with aborted seed was evaluated. The reproductive pathways were assessed in the well-developed seed via flow cytometric seed screen (FCSS).

Key Results

The majority of seed was aborted; the well-developed seeds have resulted from both mentor effects and cross-fertilization at very low frequencies (1·3 and 1·6 % of achenes, respectively). Pollination by 4x apomictic pollen plants results more frequently in cross-fertilization, whereas pollen from 6x plants more frequently induced mentor effects.

Conclusions

It is concluded that introgression of apomixis into sexual populations is limited by ploidy barriers in the R. auricomus complex, and to a minor extent by mentor effects. In mixed populations, sexuality cannot be replaced by apomixis because the higher fertility of sexual populations still compensates the low frequencies of potential introgression of apomixis.Key words: Apomixis, Ranunculus auricomus, evolution, geographical parthenogenesis, crossing experiments, flow cytometry     

Isozyme evidence of the parental origin and possible fertility of the hybrid Potamogeton×fluitans Roth

 The hybrid Potamogeton×fluitans Roth is thought to have resulted from hybridization between P. lucens L. and P. natans L. This hybrid has only been recorded at a few locations in the British Isles. At most of these sites the number of individuals found have been quite low. The one exception to this being a population in the Moors River, Dorset and South Hampshire, England, where a substantially larger population exists. Using seven different enzyme systems, this study provided support for the putative parental origin of this hybrid. In addition, the population in the Moors River was shown to contain numerous multi-locus phenotypes suggesting that these individuals may be partially fertile and may backcross to one of its parental species, or alternatively undergo selfing or crossing to produce an F₂ generation. Received December 1, 2000 Accepted June 20, 2001     

Linkage mapping in apomictic and sexual Kentucky bluegrass ( Poa pratensis L.) genotypes using a two way pseudo-testcross strategy based on AFLP and SAMPL markers

The high versatility of the mode of reproduction and the retention of a pollen recognition system are the factors responsible for the extreme complexity of the genome in Poa pratensis L. Two genetic maps, one of an apomictic and one of a sexual genotype, were constructed using a two-way pseudo-testcross strategy and multiplex PCR-based molecular markers (AFLP and SAMPL). Due to the high ploidy level and the uncertainty of chromosome pairing-behavior at meiosis, only parent-specific single-dose markers (SDMs) that segregated 1:1 in an F₁ mapping population (161 out of 299 SAMPLs, and 70 out of 275 AFLPs) were used for linkage analysis. A total of 41 paternal (33 SAMPLs and 8 AFLPs) and 47 maternal (33 SAMPLs and 14 AFLPs) SDMs, tested to be linked in coupling phase, were mapped to 7+7 linkage groups covering 367 and 338.4 cM, respectively. The comparison between the two marker systems revealed that SAMPL markers were statistically more efficient than AFLP ones in detecting parent-specific SDMs (75% vs 32.4%). There were no significant differences in the percentages of distorted marker alleles detected by the two marker systems (27.8% of SAMPLs vs 21.3% of AFLPs). The pairwise comparison of co-segregational groups for linkage detection between marker loci suggested that at least some of the P. pratensis chromosomes pair preferentially at meiosis-I. Received: 31 August 2000 / Accepted: 12 January 2001     

Genetic diversity in sexual diploid and apomictic tetraploid populations of Paspalum notatum situated in sympatry or allopatry

Paspalum notatum is a subtropical grass widely distributed in the temperate areas of America. Diploids are sexual while polyploids give rise to clonal seeds through aposporous apomixis. RAPD markers were used to analyze the genetic structure of three natural populations: i) diploids reproducing sexually (R2X); ii) sympatric apomictic tetraploids collected in the vicinity of the diploids (R4X); iii) allopatric apomictic tetraploids growing in isolation (C4X). The apomictic reproduction rate was evaluated by the use of molecular markers in progeny tests, while chromosome-counting allowed the verification of ploidy levels. Data revealed that the R4X group presented a variation considerably higher than that observed for C4X. Jaccards coefficients were used to produce a cluster diagram using the UPGMA method. All but one tetraploid genotypes grouped together and were associated to diploid genotype A21. The possibility of occasional generation of novel tetraploid clones from the interaction between tetraploid and diploid individuals is discussed.     

Iridoid glucosides of the genus Veronica s.l. and their systematic significance

 The mode of reproduction, pollen production, chromosome numbers, genetic variation (RAPD, allozymes) and overall similarity were studied in 6 species of Hieracium sect. Alpina in the Tatry Mts. (the Western Carpathians, Slovakia). All species were confirmed to be agamospermous and, except of H. krivanense and H. slovacum, lacking pollen grains. For the first time, a chromosome number is reported for H. krivanense (2n=4x=36). Considerable genetic variation was revealed in H. alpinum and a correlation between geographic and genetic distances was found in this species. Between-population variation in RAPD and allozyme phenotypes was found in H. pinetophilum and H. crassipedipilum. In all other species, allozyme and RAPD variation was low or absent. With few exceptions, the species differ in their allozyme as well as RAPD patterns. The relatedness of one population of endemic H. slovacum and H. halleri was confirmed. It is shown, that Carpathian species of the H. fritzei group are derived from at least two ancestors. Received July 3, 2000; accepted June 24, 2002 Published online: November 20, 2002 Addresses of the authors: H. Štorchová, (e-mail: storchova@ueb.cas.cz) Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 135, CZ-165 00 Praha 6, Czech Republic. I. V. Bartish, J. Chrtek Jr., J. Kirschner, M. Tetera, J. Štěpánek, Institute of Botany, Academy of Sciences of the Czech Republic, CZ-252 43 Pruhonice, Czech Republic.     

Levels of genetic diversity at different stages of the domestication cycle of interior spruce in British Columbia

Concerns over the reductionist nature of the domestication of forest-tree species focus on the possibility of potential genetic erosion during this process. To address these concerns, genetic diversity assessments in a breeding zone the Province of British Columbia “interior” spruce (Picea glauca×engelmanni) program was conducted using allozyme markers. Genetic-variation comparisons were made between natural and production (seed orchard) populations as well as seed and seedling crops produced from the same breeding zone’s seed orchard. The natural population sample consisted of a total of 360 trees representing three stands within each of three watersheds present in the Shuswap-Adams low-elevation zone of interior British Columbia. Small amounts of genetic differentiation were observed among the nine natural populations (4%) and this was attributable to extensive gene flow Consequently, the sum of these nine populations was considered as a baseline for the genetic variation present in the breeding zone. The comparisons between the seed orchard and the breeding zone produced a similar percentage of polymorphic loci while the expected hetrozygosity (0.207 vs 0.210) and the average number of alleles per locus (2.7 vs 2.4) were slightly lower in the seed orchard. A total of seven natural populations’ rare alleles were not present in the orchard population, while one allele was unique to the orchard. The %P increased to 70.6% in the seedlot, but dropped to the natural populations level (64.7%) in the plantation. The observed increase in %P was a result of pollen contamination in the orchard. It is suspected that the reduction in the plantation was caused by an unintentional selection in the nursery. Simulated roguing in the orchard did not drastically reduce even if up to 50% of the orchard’s clones were rogued. However, roguing was associated with a reduction in the average number of alleles per locus (i.e., sampling effect). Received: 2 January 1996 / Accepted: 24 May 1996     

Genetic diversity within and among maize populations: a comparison between isozyme and nuclear RFLP loci

 In order to compare the potential of enzyme and DNA markers to investigate genetic diversity within and among populations, ten maize populations were characterized for (1) 20 isozyme loci and (2) restriction fragment length polymorphism (RFLP) for 35 probe-enzyme combinations. Each population was represented by a sample of at least 30 individuals. The average number of alleles detected per locus was clearly higher for RFLPs (6.3) than for isozymes (2.4). Similarly, total diversity was higher for RFLPs (0.60) than for isozymes (0.23). This difference is consistent with observations on inbred-line collections and can be related to the fact that many variations at the DNA level do not change the amino-acid composition or the global charge of proteins. By contrast, the magnitude of population differentiation, relative to the total diversity, was similar for isozymes (23%) and RFLPs (22%). This suggests that the isozyme and RFLP loci considered in this study are subject to similar evolutionary forces, and that both are mostly neutral. However, RFLPs proved clearly superior to isozymes both to (1) identify the origin of a given individual and (2) reveal a relevant genetic structure among populations. The higher polymorphism observed for RFLP loci and the greater number of these loci contributed to the superior discriminative ability of the RFLP data. Received: 1 June 1997 / Accepted: 3 November 1997     

Floral organogenesis of Chloranthus sessilifolius, with special emphasis on the morphological nature of the androecium of Chloranthus (Chloranthaceae)

 Floral organogenesis of Chloranthus sessilifolius K. F. Wu is described. The inflorescence primordium is dome-like in the beginning and then elongates, and bract primordia initiate almost decussately. Each floral primordium, arising from the axil of a bract, soon becomes a scale-like structure, with three primordia of androecial lobes originating from its abaxial part, and the gynoecial primordium in adaxial position. As the androecial lobes become more distinct, four thecae are already in differentiation, and the gynoecial primordium appears as a shallow disc. The androecial lobes do not extend their length until the thecae approach maturity and the stigma is differentiated. The androecial lobes are united at all the stages of development, and the entire androecium falls off as a unit at the end of anthesis. Based on these results, combined with published evidence from neobotany, palaeobotany and phylogenetic studies, the morphological nature of the androecium of Chloranthus is further discussed. Our studies support the viewpoint that the androecial structure of Chloranthus may have arisen by splitting of a single stamen with 2 marginal thecae. Received May 2, 2001 Accepted December 18, 2001     

Chloroplast DNA diversity within and among populations of the allotetraploid Prunus spinosa L.

High chloroplast DNA (cpDNA) diversity was found within and among populations of Prunus spinosa sampled from seven European deciduous forests. A study of 12% of the total chloroplast genome detected 44 mutations, which were distributed over 24 haplotypes; four were common to two or more populations and the rest were unique haplotypes. The most-abundant and widely distributed haplotype was H2 (frequency = 41% approximately). Six of the seven populations were polymorphic. All of the six polymorphic populations had ”private” haplotypes (frequency < 5%) in addition to common haplotypes. The UPGMA dendrogram demonstrated a correlation between populations and their geographical locations. The total diversity was high (h_T= 0.824) and a major portion of it was within populations (h_s= 0.663). The level of population subdivision for unordered alleles was low (G_ST= 19.5%) and for ordered alleles was lower (N_ST= 13.6%). No phylogeographic structure could be demonstrated in the present geographical scale. High polymorphism in the cpDNA of P. spinosa has to be considered carefully when planning phylogenetic studies involving this species. Received: 20 September 1999 / Accepted: 10 November 1999     

The arctic-alpine polyploids Cerastium alpinum and C. nigrescens (Caryophyllaceae) in a sympatric situation: breakdown of species integrity?

 The high polyploids Cerastium alpinum (8x) and C. nigrescens (12x) were investigated in a mixed population in central Norway to evaluate whether hybridisation has resulted in continuous variation in morphology and genetic markers, a hypothesis previously proposed to account for the extensive taxonomic confusion in this species group. Isozyme, fertility, and morphological (37 characters) variation were examined among 347, 265, and 237 plants, respectively. A PCO analysis based on 23 quantitative morphological characters identified two main groups, corresponding to C. alpinum and C. nigrescens. The groups were also clearly separated in isozyme markers, several qualitative morphological characters, and chromosome numbers. Only 20 plants (8.4%) were more or less intermediate in the PCO analysis. These plants had a parental or hybrid isozyme multilocus phenotype and typically few well-developed anthers, low pollen stainability, and no seed set. Several plants within the C. nigrescens group also had reduced pollen stainability. These results indicate that later-generation hybrids and/or backcrosses towards C. nigrescens are formed. Thus, interspecific, interploidal gene flow probably occurs, but at rates that are insufficient to break down species integrity. Received July 5, 2000 Accepted August 2, 2001     

Dosage effects in the endosperm of diplosporous apomictic Tripsacum (Poaceae)

 Imprinting in the endosperm of angiosperms, a phenomena by which expression of alleles differs depending on whether they originate from the male or female parent, has been shown to explain most failure of interploidy or interspecific crosses in plants. Because of imprinting, seeds develop normally only if a specific dosage is represented in the endosperm, with the relative contributions of genomes in the ratio of two maternal doses to one paternal dose (2m:1p). In Tripsacum, a wild relative of maize, all polyploids reproduce through the diplosporous type of apomixis. Diplospory results from meiotic failure in megasporocytes that develop into eight-nucleate unreduced female gametophytes. The male gametophytes remain unaffected. Flow cytometry was used to determine ploidy levels in the endosperm of both apomictic and sexual Tripsacum accessions. In both cases, fertilization appeared to involve only one sperm nucleus. Therefore, endosperm of apomictic Tripsacum develops normally even though the ratio of genomic contributions deviates from the normal 2m:1p ratio. Ratios of 2:1, 4:1, 4:2, 8:1 and 8:2 were observed, depending on both the ploidy level of the parents and the mode of reproduction. Thus, specific dosage effects are seemingly not required for endosperm development in Tripsacum. These findings suggest that evolution of diplosporous apomixis might have been restricted to species with few or no imprinting requirements, and the findings have strong implications regarding the transfer of apomixis to sexually reproducing crops. Received: 17 February 1997 / Revision accepted: 7 July 1997