Interspecific hybridization among Hieracium species in New Zealand: evidence from flow cytometry

Hieracium pilosella: (Asteraceae) was accidentally introduced to New Zealand about 100 years ago. Since then it has become an aggressive weed, and an unexpected degree of genetic and genome size variation has been detected; features that might result from interspecies hybridization. We investigated the possibility that H. pilosella has hybridized with related taxa. Of the four other subgenus Pilosella species introduced to New Zealand, H. praealtum is the most abundant and, on morphological and distributional evidence, most likely to be the other parent. Flow cytometry was used to estimate relative genome size for 156 Hieracium plants collected from the wild. Plants assigned to either parental or hybrid morphotypes were found to comprise tetraploid and pentaploid individuals using genome size measurements, and this was confirmed with direct mitotic chromosome counts for a subset of plants. The haploid DNA content of H. praealtum was approximately 22% larger than that of H. pilosella. Putative hybrids that were tetraploid had mean genome sizes equivalent to two H. pilosella and two H. praealtum haploid chromosome sets, implying they were hybrids arising from the fertilization of two reduced gametes. Similar results were obtained from tetraploid hybrids produced by controlled pollination. However, the majority of field hybrids were pentaploid with a genome size equivalent to four H. pilosella and one H. praealtum haploid chromosome sets. We infer that these are not first-generation hybrids but represent successful backcrossing with H. pilosella and/or hybrid-hybrid crossing, and that sexual tetraploid hybrids have been the parents. We note that populations putatively of H. pilosella often comprise apomictic pentaploid hybrids. Significantly, our data indicate the emergence of sexual hybrids that provide further opportunity for gene flow among taxa in this complex.     

Incongruent plastid and nuclear DNA phylogenies reveal ancient intergeneric hybridization in Pilosella hawkweeds (Hieracium, Cichorieae, Asteraceae)

Phylogenetic relationships for Hieracium subgen. Pilosella were inferred from chloroplast (trnT-trnL, matK) and nuclear (ITS) sequence data. Chloroplast markers revealed the existence of two divergent haplotype groups within the subgenus that did not correspond to presumed relationships. Furthermore, chloroplast haplotypes of the genera Hispidella and Andryala nested each within one of these groups. In contrast, ITS data were generally in accord with morphology and other evidence and were therefore assumed to reflect the true phylogeny. They revealed a sister relationship between Pilosella and Hispidella and a joint clade of Hieracium subgenera Hieracium and Chionoracium (Stenotheca) while genus Andryala represented a third major lineage of the final ingroup cluster. Detailed analysis of trnT-trnL character state evolution along the ITS tree suggested two intergeneric hybridization events between ancestral lineages that resulted in cytoplasmic transfer (from Hieracium/Chionoracium to Pilosella, and from the introgressed Pilosella lineage to Andryala). These chloroplast capture events, the first of which involved a now extinct haplotype, are the most likely explanation for the observed incongruencies between plastid and nuclear DNA markers.     

Chromosomes carrying meiotic avoidance loci in three apomictic eudicot Hieracium subgenus Pilosella species share structural features with two monocot apomicts

The LOSS OF APOMEIOSIS (LOA) locus is one of two dominant loci known to control apomixis in the eudicot Hieracium praealtum. LOA stimulates the differentiation of somatic aposporous initial cells after the initiation of meiosis in ovules. Aposporous initial cells undergo nuclear proliferation close to sexual megaspores, forming unreduced aposporous embryo sacs, and the sexual program ceases. LOA-linked genetic markers were used to isolate 1.2 Mb of LOA-associated DNAs from H. praealtum. Physical mapping defined the genomic region essential for LOA function between two markers, flanking 400 kb of identified sequence and central unknown sequences. Cytogenetic and sequence analyses revealed that the LOA locus is located on a single chromosome near the tip of the long arm and surrounded by extensive, abundant complex repeat and transposon sequences. Chromosomal features and LOA-linked markers are conserved in aposporous Hieracium caespitosum and Hieracium piloselloides but absent in sexual Hieracium pilosella. Their absence in apomictic Hieracium aurantiacum suggests that meiotic avoidance may have evolved independently in aposporous subgenus Pilosella species. The structure of the hemizygous chromosomal region containing the LOA locus in the three Hieracium subgenus Pilosella species resembles that of the hemizygous apospory-specific genomic regions in monocot Pennisetum squamulatum and Cenchrus ciliaris. Analyses of partial DNA sequences at these loci show no obvious conservation, indicating that they are unlikely to share a common ancestral origin. This suggests convergent evolution of repeat-rich hemizygous chromosomal regions containing apospory loci in these monocot and eudicot species, which may be required for the function and maintenance of the trait.     

Genome size variation and species relationships in Hieracium sub-genus Pilosella (Asteraceae) as inferred by flow cytometry

BACKGROUND AND AIMS: Hieracium sub-genus Pilosella (hawkweeds) is a taxonomically complicated group of vascular plants, the structure of which is substantially influenced by frequent interspecific hybridization and polyploidization. Two kinds of species, 'basic' and 'intermediate' (i.e. hybridogenous), are usually recognized. In this study, genome size variation was investigated in a representative set of Central European hawkweeds in order to assess the value of such a data set for species delineation and inference of evolutionary relationships. METHODS: Holoploid and monoploid genome sizes (C- and Cx-values) were determined using propidium iodide flow cytometry for 376 homogeneously cultivated individuals of Hieracium sub-genus Pilosella, including 24 species (271 individuals), five recent natural hybrids (seven individuals) and experimental F(1) hybrids from four parental combinations (98 individuals). Chromosome counts were available for more than half of the plant accessions. Base composition (proportion of AT/GC bases) was cytometrically estimated in 73 individuals. KEY RESULTS: Seven different ploidy levels (2x-8x) were detected, with intraspecific ploidy polymorphism (up to four different cytotypes) occurring in 11 wild species. Mean 2C-values varied approx. 4.3-fold from 3.53 pg in diploid H. hoppeanum to 15.30 pg in octoploid H. brachiatum. 1Cx-values ranged from 1.72 pg in H. pilosella to 2.16 pg in H. echioides (1.26-fold). The DNA content of (high) polyploids was usually proportional to the DNA values of their diploid/low polyploid counterparts, indicating lack of processes altering genome size (i.e. genome down-sizing). Most species showed constant nuclear DNA amounts, exceptions being three hybridogenous taxa, in which introgressive hybridization was suggested as a presumable trigger for genome size variation. Monoploid genome sizes of hybridogenous species were always between the corresponding values of their putative parents. In addition, there was a good congruency between actual DNA estimates and theoretical values inferred from putative parental combinations and between DNA values of experimental F(1) hybrids and corresponding established hybridogenous taxa. CONCLUSIONS: Significant differences in genome size between hawkweed species from hybridogenous lineages involving the small-genome H. pilosella document the usefulness of nuclear DNA content as a supportive marker for reliable delineation of several of the most problematic taxa in Hieracium sub-genus Pilosella (including classification of borderline morphotypes). In addition, genome size data were shown to have a good predictive value for inferring evolutionary relationships and genome constitution (i.e. putative parental combinations) in hybridogenous species.     

Interaction between Some Pasture Species and 2

Possible control options are investigated for the introduced Hieracium weeds, particular problems in South Island high country, New Zealand. In a pot experiment regression of input to output ratios of above ground biomass over successive harvests, from binary mixtures was used to determine the competitive interaction between 13 pasture species and two Hieracium species, H. pilosella and H. praealtum, in a low fertility soil. Treatments also included a factorial of presence or absence of compartments separating root and shoots of species. Species differed in their mean growth rate, relative to Hieracium species. The ranking of mean growth rate relative to H. pilosella was Trifolium repens (best), Bromus inermis, Sanguisorba minor, Festuca novaezelandiae, F. rubra, Arrhenatherum elatius, Anthoxanthum odoratum, Lotus corniculatus, Hypochoeris radicata, Trifolium repens, T. hybridum, T. medium and Astragalus cicer. Shoot partitions decreased Hieracium's interaction with white clover while root partitions increased interaction with B. inermis. However, the rates were not related to the proportion of Hieracium in the mixtures, indicating a general lack of specific competitive effects against Hieracium.     

Reproduction mode in the allopolyploid facultatively apomictic hawkweed Hieracium rubrum (Asteraceae, H. subgen. Pilosella)

The versatility of the breeding system in the hybridogenous hexaploid, Hieracium rubrum, was demonstrated in emasculation and crossing experiments. The flow-cytometric ploidy analysis of 1095 seedlings amongst its' progeny enabled the determination and quantification of the reproductive pathway (apospory, haploid parthenogenesis, cross-fertilization of both reduced and unreduced female gametes) responsible for each progeny class. The progeny profiles were stable between two successive years. The percentage of hybrids arisen from crosses with tetraploid sexual H. pilosella was 7.9%. The rate of trihaploids generated by H. rubrum when crossed to H. pilosella was 3.95%, similar to that of pentaploid (n + n) and octoploid (2n + n) hybrids (3.95% and 3.39%). Unreduced pollen from H. pilosella contributed to hybridization much less frequently (0.56% of all progeny), than unreduced eggs of H. rubrum. The increased frequency of trihaploids formed by crossed compared to emasculated plants suggests an interaction between the presence of pollen and the autonomous development of meiotic embryo sacs. Although the environmental conditions (garden compared to unheated glasshouse) influenced the progeny following the emasculation, no corresponding response to environment was detected amongst the autonomously derived progeny following pollination with H. pilosella. However, there was an influence of pollen parent on progeny which varied under the different environmental treatments. The importance of residual sexuality in reproduction of apomicts is evident. The non-maternal progeny was generated with highly variable frequency especially in the glasshouse, with some capitula reaching up to 50%. Having an autonomous endosperm development, Hieracium subgen. Pilosella is almost unique among other aposporous genera, in which such variable reproduction mode is connected with pseudogamy.     

Phylogeographic analysis of the brown alga Cutleria multifida (Tilopteridales,Phaeophyceae) suggests a complicated introduction history

In depth genetic comparisons of populations of Cutleria multifida (Tilopteridales, Phaeophyceae) collected from Europe, the northwestern Pacific Ocean, Australia and New Zealand using the DNA sequences of four gene regions (the mitochondrial cox2 and cox3 genes, the intergeneric spacer region adjacent to cox3, and the open reading frame) suggested that the northwestern European and Japanese populations were considerably greater in terms of their genetic divergence than Mediterranean, Australian or New Zealand populations. The haplotypes of the populations in northwestern European (distribution range including the type locality, seven haplotypes) and Japanese populations (seven haplotypes) were unique except for one shared haplotype. There were weak but positive correlations between the geographical distance and the genetic divergence among northwestern European and Japanese populations. Moreover, both female and male gametophytes occurred in eight of the nine Japanese localities, suggesting Japanese populations showed normal sexual heteromorphic life history of the species. In light of these results, it appears that Japanese populations were native to the area despite earlier hypothesis. In contrast, Australian and New Zealand populations were composed of only one haplotype that is very close to those found in northwestern Europe and Japan, suggesting a recent introduction history from Europe (or from northeastern Asia via Europe) by ship transport to Australia and New Zealand. The Mediterranean populations included two haplotypes identical to those found in northwestern Europe and Japan, and it is suggestive of transoceanic introductions of some populations between Mediterranean and Japanese coasts.     

Distribution and diet of chamois (

Changes in Hieracium abundance in Eastern Otago tussock grassland were examined by sampling 163 sites in 1982 and again in 1992. For Hieracium pilosella, H. praealtum and H. lepidulum, as well as Agrostis capillaris for comparison, colonisation of new sites was recorded, as well as extinction of species from sites over the 10 years, and changes in cover. H. pilosella colonised the majority of sites from which it had been absent in 1982; it disappeared from only a few sites where it had been present at very low cover. In sites where it remained over the decade, its cover increased by 50%. Its rate of increase was very similar to that recorded for North Canterbury, but with no indication of the 34% stabilisation point that has been hypothesised for the latter area. H. praealtum and H. lepidulum, in contrast, remained absent from most sites, and colonised only a few; in very few sites was their cover greater than 1%. Agrostis capillaris was widespread and abundant, but increased little over the period. The sites where H. pilosella colonised tended to be degraded, with a greater, and increasing, percentage of bare ground and low tussock cover. The few sites where it disappeared were ones where pasture improvement had occurred, as seen in the increase of species indicative of pasture development. Several explanations are considered for the increase in H. pilosella in recent decades. The increase of this species in Eastern Otago has been greater than that of most other exotic species. Changes in land management have contributed to its increase, but probably changes caused indirectly by government policy, not gradual degradation. Genetic changes in the species, and metapopulation dynamics, have not been investigated, but they could well have contributed to Hieracium invasion.     

Sporophytic ovule tissues modulate the initiation and progression of apomixis in Hieracium

Apomixis in Hieracium subgenus Pilosella initiates in ovules when sporophytic cells termed aposporous initial (AI) cells enlarge near sexual cells undergoing meiosis. AI cells displace the sexual structures and divide by mitosis to form unreduced embryo sac(s) without meiosis (apomeiosis) that initiate fertilization-independent embryo and endosperm development. In some Hieracium subgenus Pilosella species, these events are controlled by the dominant LOSS OF APOMEIOSIS (LOA) and LOSS OF PARTHENOGENESIS (LOP) loci. In H. praealtum and H. piloselloides, which both contain the same core LOA locus, the timing and frequency of AI cell formation is altered in derived mutants exhibiting abnormal funiculus growth and in transgenic plants expressing rolB which alters cellular sensitivity to auxin. The impact on apomictic and sexual reproduction was examined here when a chimeric RNAse gene was targeted to the funiculus and basal portions of the ovule, and also when polar auxin transport was inhibited during ovule development following N-1-naphthylphthalamic acid (NPA) application. Both treatments led to ovule deformity in the funiculus and distal parts of the ovule and LOA-dependent alterations in the timing, position, and frequency of AI cell formation. In the case of NPA treatment, this correlated with increased expression of DR5:GFP in the ovule, which marks the accumulation of the plant hormone auxin. Our results show that sporophytic information potentiated by funiculus growth and polar auxin transport influences ovule development, the initiation of apomixis, and the progression of embryo sac development in Hieracium. Signals associated with ovule pattern formation and auxin distribution or perception may influence the capacity of sporophytic ovule cells to respond to LOA.     

Changes in the Structure of Tall Tussock Grasslands and Infestation by Species of

A plant sociological survey of tall-tussock grasslands in the Mackenzie country was repeated after an interval of 26-28 years. Changes in physiognomy of the grasslands which have been inferred from earlier studies have been found to be continuing on many sites. A noteworthy feature of most sites has been a reduction in number of indigenous species found. An increase in abundance of Hieracium pilosella or H. praealtum has occurred at most sites. About 140 species and 9 variables from 53 sites were interrelated in a direct unimodal ordination (canonical correspondence analysis). Ordination results of the 1960s and the 1989 data were compared and interpreted with respect to spread of Hieracium pilosella and H. praealtum. The dynamics of Hieracium infestation was studied within changing community structure. A distinct trend is demonstrated of increasing infestation with increasing grassland degradation.     

rbcL sequences reveal multiple cryptic introductions of the Japanese red alga Polysiphonia harveyi

In Europe, the last 20 years have seen a spectacular increase in accidental introductions of marine species, but it has recently been suggested that both the actual number of invaders and their impacts have been seriously underestimated because of the prevalence of sibling species in marine habitats. The red alga Polysiphonia harveyi is regarded as an alien in the British Isles and Atlantic Europe, having appeared in various locations there during the past 170 years. Similar or conspecific populations are known from Atlantic North America and Japan. To choose between three competing hypotheses concerning the origin of P. harveyi in Europe, we employed rbcL sequence analysis in conjunction with karyological and interbreeding data for samples and isolates of P. harveyi and various congeners from the Pacific and North Atlantic Oceans. All cultured isolates of P. harveyi were completely interfertile, and there was no evidence of polyploidy or aneuploidy. Thus, this biological species is both morphologically and genetically variable: intraspecific rbcL divergences of up to 2.1% are high even for red algae. Seven rbcL haplotypes were identified. The four most divergent haplotypes were observed in Japanese samples from Hokkaido and south-central Honshu, which are linked by hypothetical 'missing' haplotypes that may be located in northern Honshu. These data are consistent with Japan being the centre of diversity and origin for P. harveyi. Two non-Japanese lineages were linked to Hokkaido and Honshu, respectively. A single haplotype was found in all North Atlantic and Mediterranean accessions, except for North Carolina, where the haplotype found was the same as that invading in New Zealand and California. The introduction of P. harveyi into New Zealand has gone unnoticed because P. strictissima is a morphologically indistinguishable native sibling species. The sequence divergence between them is 4-5%, greater than between some morphologically distinct red algal species. Two different types of cryptic invasions of P. harveyi have therefore occurred. In addition to its introduction as a cryptic sibling species in New Zealand, P. harveyi has been introduced at least twice into the North Atlantic from presumed different source populations. These two introductions are genetically and probably also physiologically divergent but completely interfertile.     

Changes in vegetation states in grazed and ungrazed Mackenzie Basin grasslands, New Zealand, 1990-2000

Changes in vegetation from 1990 to 2000 were examined at 10 high country localities, representing four grassland types: fescue tussock (Festuca novae-zelandiae), snow tussock (Chionochloa rigida), red tussock (C. rubra), and silver tussock (Poa cita). At each locality, three treatments were established: ambient sheep+rabbit grazing, rabbit grazing only, and no grazing. The mutivariate methods of classification and ordination were used on individual-quadrat cover data to define vegetation states and to examine transitions between them over time. Vegetation states in quadrats already dominated by Hieracium pilosella(> 50% cover) in 1990 showed little change in species composition regardless of grassland type and grazing treatment. In fescue tussock grassland, H. pilosellaincreased regardless of grazing treatment in states with low initial H. pilosellacover (< 5%), while the cover of Carex colensoi, Aira caryophyllea and Rumex acetosella decreased. In the single silver tussock locality, Poa citadecreased markedly in the ungrazed treatment as adventive species such as Dactylis glomerataand Echium vulgare increased. However, Poa citaalso decreased, probably due to drought, in the grazed treatment. Snow tussock and red tussock grassland states were more stable than those in short tussock grasslands, but there was also a general trend towards increasing H. pilosellacover in intertussock vegetation regardless of treatment. However, at one snow tussock locality, transitions from H. piloselladominated to C. rigida-dominated states occurred in ungrazed quadrats, while the reverse occurred in grazed vegetation. Implications for the management of tussock grasslands for conservation are discussed.     

Influence of pastoral management on plant biodiversity in a depleted short tussock grassland, Mackenzie Basin

This study investigated the effects of different management inputs (fertiliser and seed) and grazing patterns on plant biodiversity in a short tussock grassland with a strong Hieracium pilosella component. Cover abundance of vascular and non-vascular plants and environmental variables were measured in 32 10×10-m plots located in five blocks with different management treatments. Ordination of the floristic data separated the block with the highest management inputs from other blocks. Several adventive species were significantly more abundant in this block, while several native species were either absent or uncommon, but were significantly more abundant in other study blocks. H. pilosella was significantly more abundant in blocks with lower management inputs. Diversity was significantly higher in the block with the highest management inputs. The native tussock Poa colensoi had significantly greater cover abundance while Festuca novae-zelandiae tussocks were significantly taller in this block. Our results suggest that high management inputs reduce the abundance of H. pilosella and diversity of native species, but increase the abundance of other adventive species and the cover and vigour of native tussocks. Our results highlight an interesting management conundrum for short tussock grasslands. No-input management is likely to result in a decline in native biodiversity, as well as production values, as H. pilosella mats deplete soil nutrients and restrict regeneration of native species. However, input of fertiliser and adventive seeds to enhance production values, although resulting in an increase in the vigour and abundance of some native species (mainly tussocks) and a reduction in H. pilosella abundance, will also result in a decline in overall native species richness.     

Isolation of a diploid, apomictic plant of Hieracium aurantiacum

 As in most taxa that contain gametophytic apomicts, all of the apomictic biotypes studied so far in the genus Hieracium subgenus Pilosella have been recorded as polyploids. As part of a study of variation in apomictically derived populations of H. aurantiacum, a diploid plant was identified. Apospory, the mechanism of apomixis typically observed in this taxon, was observed in this plant. Unreduced megagametophytes at various developmental stages were commonly observed in the developing ovules, and endosperm formation was autonomous. The eventual seed set, however, was low. This appears to have been due to the proliferation of competing megagametophytes and embryos within each ovule. Pollen sterility was also observed, primarily resulting from the dysfunction of microgametogenesis at the uninucleate stage. Received: 3 September 1996 / Revision accepted: 21 March 1997     

Variation inHieracium subgen.Pilosella (Asteraceae): What do we know about its sources?

The present paper reviews mechanisms producing complicated patterns of variation withinHieracium subgen.Pilosella. The taxonomic complexity of this subgenus is due to highly variable basic species and intermediate (hybridogenous) species. The most important sources of variation are polyploidy, hybridization and (mostly) facultative apomixis of the aposporous type. The combination of hybridization, apomixis and clonal growth leads to the maintenance of various hybrids having originated from backcrossing and hybridization among more than two species, which is possible because of the fertile pollen of apomictic hybrids. Ever since Mendel’s experiments, some of F₁ hybrids have been found to be highly variable, probably reflecting the high heterozygosity of some of the basic species. Variable progeny can also result from unreduced gametes, or the rare parthenogenetic development of reduced gametes. While these processes were detected in experiments, their role within field populations remains unknown. However, multiple origins of intermediate species, and introgression within basic species are highly likely to result in high levels of variation. While few population level studies have been undertaken in Europe, several such studies have been carried out on adventive populations in New Zealand, and these show a different pattern. Aneuploid plants, rare in Europe, are common in New Zealand, and there is frequently more than one ploidy level within a population.     

Rapid short-tussock grassland decline with and without grazing, Marlborough, New Zealand

Species abundance, species richness, and ground cover were measured over 10 years on nine paired grazed and exclosure plots in short-tussock grassland in the early stages of invasion by Hieracium species. With and without grazing, H. pilosella and H. caespitosum increased markedly and H. lepidulum increased locally. In contrast, 50% of all other common species and species groups, and total, native, and exotic species richness declined significantly. Exclusion increased or had no effect on rates of increase in Hieracium species and rates of decline in short tussocks, and did not reduce rates of decline in other species. Exclusion had no effect on decline in native species richness, but mainly accelerated declines in total and exotic richness. Declines in 13 key vegetation variables were significantly predicted by increase in Hieracium abundance, suggesting competitive exclusion. With or without grazing, Hieracium species will become more dominant and other species will continue to decline. The effects of large herbivores on plant species diversity can often be predicted from site productivity. Our results indicate the need also to account for species origin, spatial scale, time, and exotic invasion.     

Chromosome numbers in Hieracium and Pilosella species (Asteraceae) from the Iberian Peninsula and the Balearic Islands

Mitotic chromosome numbers are reported from 16 Hieracium and two Pilosella species from the Iberian Peninsula and the Balearic Islands that are little known cytogenetically. The chromosome numbers of Hieracium aragonense Scheele (2 n  = 27, 36), H. compositum Lapeyr. (2 n  = 27), H. murcandidum G. Mateo (2 n  = 27), H. spathulatum Scheele (2 n  = 27), H. segurae Mateo (2 n  = 27), H. teruelanum Mateo (2 n  = 27), H. valentinum Pau (2 n  = 27), Pilosella pseudovahlii (De Retz) Mateo (2 n  = 18), and P. tardans (Peter) Sójak (2 n  = 36) were determined for the first time. New cytotypes were detected in H. cordifolium Lapeyr. (2 n  = 27) and H. loscosianum Scheele (2 n  = 36). The karyotype of the studied species was similar in overall morphology and comprised metacentric and submetacentric chromosomes; secondary constrictions (up to two) were detected in the chromosome complements of some species.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 311–320.     

Cytogeography of Pilosella officinarum (Compositae): altitudinal and longitudinal differences in ploidy level distribution in the Czech Republic and Slovakia and the general pattern in Europe

BACKGROUND AND AIMS: Pilosella officinarum (syn. Hieracium pilosella) is a highly structured species with respect to the ploidy level, with obvious cytogeographic trends. Previous non-collated data indicated a possible differentiation in the frequency of particular ploidy levels in the Czech Republic and Slovakia. Therefore, detailed sampling and ploidy level analyses were assessed to reveal a boundary of common occurrence of tetraploids on one hand and higher ploids on the other. For a better understanding of cytogeographic differentiation of P. officinarum in central Europe, a search was made for a general cytogeographic pattern in Europe based on published data. METHODS: DNA-ploidy level and/or chromosome number were identified for 1059 plants using flow cytometry and/or chromosome counting on root meristem preparations. Samples were collected from 336 localities in the Czech Republic, Slovakia and north-eastern Hungary. In addition, ploidy levels were determined for plants from 18 localities in Bulgaria, Georgia, Ireland, Italy, Romania and Ukraine. KEY RESULTS: Four ploidy levels were found in the studied area with a contrasting pattern of distribution. The most widespread cytotype in the western part of the Czech Republic is tetraploid (4x) reproducing sexually, while the apomictic pentaploids and mostly apomictic hexaploids (5x and 6x, respectively) clearly prevail in Slovakia and the eastern part of the Czech Republic. The boundary between common occurrence of tetraploids and higher ploids is very obvious and represents the geomorphologic boundary between the Bohemian Massif and the Western Carpathians with the adjacent part of Pannonia. Mixed populations consisting of two different ploidy levels were recorded in nearly 11% of localities. A statistically significant difference in a vertical distribution of penta- and hexaploids was observed in the Western Carpathians and the adjacent Pannonian Plain. Hexaploid populations tend to occur at lower elevations (usually below 500 m), while the pentaploid level is more or less evenly distributed up to 1000 m a.s.l. For the first time the heptaploid level (7x) was found on one site in Slovakia. In Europe, the sexual tetraploid level has clearly a sub-Atlantic character of distribution. The plants of higher ploidy level (penta- and hexa-) with mostly apomictic reproduction prevail in the northern part of Scandinavia and the British Isles, the Alps and the Western Carpathians with the adjacent part of Pannonia. A detailed overview of published data shows that extremely rare records on existence of diploid populations in the south-west Alps are with high probability erroneous and most probably refer to the closely related diploid species P. peleteriana. CONCLUSIONS: The recent distribution of P. officinarum in Europe is complex and probably reflects the climatic changes during the Pleistocene and consequent postglacial migrations. Probably both penta- and hexaploids arose independently in central Europe (Alps and Carpathian Mountains) and in northern Europe (Scandinavia, Great Britain, Ireland), where the apomictic plants colonized deglaciated areas. We suggest that P. officinarum is in fact an amphidiploid species with a basic tetraploid level, which probably originated from hybridizations of diploid taxa from the section Pilosellina.     

Factors predisposing short-tussock grasslands to

The effects of environment and management on the composition of short-tussock grasslands and the abundance of the invasive weed Hieracium pilosella were investigated in two small catchments. Species composition and site factors were recorded on a total of 182 plots and the management history of each catchment was reviewed. H. pilosella was present on >80% of all plots, but was at an early stage of invasion in one catchment (<5% cover) and dominant in the other (25% cover). Classification and ordination revealed strong between-catchment differences in community composition that reflected differences in environment (soil fertility and rainfall), disturbance history (animal populations and burning), and the stage of invasion by H. pilosella. In both catchments H. pilosella tended to be least abundant on the wettest, driest, and most fertile soils. However, such relationships were weak. Generalised additive models and regression showed that in the earlier stage of invasion individual site factors explained less than 20% of the variation in H. pilosella cover. Topographic position and slope (both indicative of soil moisture) were the most significant combined predictors, but together explained only 32% of the variation. In the later stage of invasion individual factors explained up to 33% of the variation. Topsoil sulphur, slope, and topsoil calcium were the most significant combined predictors, but together explained only 53% of the variation. Between-catchment comparisons highlighted the inter-related roles of environment, disturbance history, geographic location, availability of H. pilosella propagules, and stage of invasion in more fully explaining the abundance of H. pilosella. Of five models that have been proposed for Hieracium invasion, the: "grassland decline" model best incorporated the inter-related factors that influence spatial and temporal variation in H. pilosella abundance in the study area. This model concentrates on identifying predisposing and trigger factors that increase the likelihood of invasion and accounts for multiple causes and interactions by specifying five key factors that influence the ability of a plant species to invade existing vegetation: environment, disturbance, vegetation structure and composition, life history attributes of the invader, and the availability of invading propagules. The model potentially provides a comprehensive framework for evaluating the causes of Hieracium invasion, targeting research effort, and developing sustainable management strategies.