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.     

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.     

Chloroplast DNA diversity of Hieracium Pilosella (Asteraceae) introduced to New Zealand: reticulation, hybridization, and invasion

The European hawkweed Hieracium pilosella is a successful invader and a troublesome weed in New Zealand. The systematics of the genus Hieracium is extremely complex and contentious, probably due to recent speciation, hybridization, polyploidy, and diverse reproductive strategies. In the first chloroplast DNA survey of the group, we sequenced 285 plants (including H. pilosella and 12 other species of subgenus Pilosella) from New Zealand and Europe for 900 bp of trnL-trnF. Eleven haplotypes were identified with much sharing among species. Three haplotypes (A, D, G) were found in seven, three, and four species, respectively, but two species (H. lactucella and H. auricula) had single, private haplotypes. Our cpDNA data for subgenus Pilosella are consistent with the group's having incomplete lineage sorting and/or recent reticulate evolution. Six haplotypes were identified in H. pilosella, four of these unique to this taxon in our sample. In New Zealand, haplotype A was common and occurred in plants of different ploidy (i.e., 4×, 5×, 6×), whereas haplotypes C, B, and M were restricted to 4×, 5×, and 6× plants, respectively. The distribution of haplotype variation suggests that some or all of the H. pilosella seeds accidentally introduced into New Zealand probably came from east Europe rather than the United Kingdom and that a minimum of four lineages were introduced. Within New Zealand, hybridization of H. pilosella with a related taxon (probably H. praealtum) has occurred at least three times, involving both obligate sexual tetraploids and facultative apomictic pentaploids of H. pilosella.     

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 in Bulgarian Centaurea s.l. (Asteraceae)

Thirty-nine species and subspecies of the genera Centaurea, Colymbada, Psephellus and Cyanus (all included in Centaurea s.l.) including many rare and endemic taxa of preponderantly Bulgarian distribution have been investigated with Feulgen DNA image densitometry for holoploid and monoploid genome size (C- and Cx-values). Cyanus varies gradually 2.17-fold between 0.74 pg and 1.56 pg (1Cx). In the remaining taxa two major genome size groups are found, which differ about 1.8-fold in Cx-value. Low values occur in Centaurea subgenera Acrolophus, Solstitiaria, Phalolepis (0.77 pg to 0.90 pg, 1Cx) and Jacea (0.95 pg to 1.09 pg, 1Cx), high values in the genera Colymbada (1.65 pg to 1.93 pg, 1Cx) and Psephellus (1.79 pg, 1Cx, in P. marschallianus). Cx-values support a distinction of Colymbada from Centaurea. Genome size variation is discussed with regard to phylogeny, life form (annual versus perennial), polyploidy, chromosome basic numbers, altitude of occurrence and climate, endemism, and rarity.     

Genome size variation in the genus Carthamus (Asteraceae, Cardueae): systematic implications and additive changes during allopolyploidization

BACKGROUND AND AIMS: Plant genome size is an important biological characteristic, with relationships to systematics, ecology and distribution. Currently, there is no information regarding nuclear DNA content for any Carthamus species. In addition to improving the knowledge base, this research focuses on interspecific variation and its implications for the infrageneric classification of this genus. Genome size variation in the process of allopolyploid formation is also addressed. METHODS: Nuclear DNA samples from 34 populations of 16 species of the genus Carthamus were assessed by flow cytometry using propidium iodide. KEY RESULTS: The 2C values ranged from 2.26 pg for C. leucocaulos to 7.46 pg for C. turkestanicus, and monoploid genome size (1Cx-value) ranged from 1.13 pg in C. leucocaulos to 1.53 pg in C. alexandrinus. Mean genome sizes differed significantly, based on sectional classification. Both allopolyploid species (C. creticus and C. turkestanicus) exhibited nuclear DNA contents in accordance with the sum of the putative parental C-values (in one case with a slight reduction, frequent in polyploids), supporting their hybrid origin. CONCLUSIONS: Genome size represents a useful tool in elucidating systematic relationships between closely related species. A considerable reduction in monoploid genome size, possibly due to the hybrid formation, is also reported within these taxa.     

Genome size in Hieracium subgenus Hieracium (Asteraceae) is strongly correlated with major phylogenetic groups

Background and Aims

Hieracium subgenus Hieracium is one of the taxonomically most intricate groups of vascular plants, due to polyploidy and a diversity of breeeding systems (sexuality vs. apomixis). The aim of the present study was to analyse nuclear genome size in a phylogenetic framework and to assess relationships between genome size and ploidy, breeding system and selected ecogeographic features.

Methods

Holoploid and monoploid genome sizes (C- and Cx-values) of 215 cultivated plants from 89 field populations of 42 so-called ‘basic’ Hieracium species were determined using propidium iodide flow cytometry. Chromosome counts were available for all analysed plants, and all plants were tested experimentally for their mode of reproduction (sexuality vs. apomixis). For constructing molecular phylogenetic trees, the external transcribed spacer region of nuclear ribosomal DNA was used.

Key Results

The mean 2C values differed up to 2·37-fold among different species (from 7·03 pg in diploid to 16·67 in tetraploid accessions). The 1Cx values varied 1·22-fold (between 3·51 and 4·34 pg). Variation in 1Cx values between conspecific (species in a broad sense) accessions ranged from 0·24% to 7·2%. Little variation (not exceeding the approximate measurement inaccurracy threshold of 3·5%) was found in 33 species, whereas variation higher than 3·5% was detected in seven species. Most of the latter may have a polytopic origin. Mean 1Cx values of the three cytotypes (2n, 3n and 4n) differed significantly (average of 3·93 pg in diploids, 3·82 pg in triploids and 3·78 pg in tetraploids) indicating downsizing of genomes in polyploids. The pattern of genome size variation correlated well with two major phylogenetic clades which were composed of species with western or eastern European origin. The monoploid genome size in the ‘western’ species was significantly lower than in the ‘eastern’ ones. Correlation of genome size with latitude, altitude and selected ecological characters (light and temperature) was not significant. A longitudinal component was only apparent for the whole data set, but absent within the major lineages.

Conclusions

Phylogeny was the most important factor explaining the pattern of genome size variation in Hieracium sensu stricto, species of western European origin having significantly lower genome size in comparison with those of eastern European origin. Any correlation with ecogeographic variables, including longitude, was outweighed by the divergence of the genus into two major phylogenetic lineages.Key words: Apomixis, chromosome numbers, Compositae, genome size, hawkweeds, Hieracium subgenus Hieracium, mode of reproduction, nuclear DNA content, phylogeny, polyploidy     

Genome size variation in Central European species of Cirsium (Compositae) and their natural hybrids

BACKGROUND AND AIMS: Nuclear DNA amounts of 12 diploid and one tetraploid taxa and 12 natural interspecific hybrids of Cirsium from 102 populations in the Czech Republic, Austria, Slovakia and Hungary were estimated. METHODS: DAPI and PI flow cytometry were used. KEY RESULTS: 2C-values of diploid (2n = 34) species varied from 2.14 pg in C. heterophyllum to 3.60 pg in C. eriophorum (1.68-fold difference); the 2C value for the tetraploid C. vulgare was estimated at 5.54 pg. The DNA contents of hybrids were located between the values of their putative parents, although usually closer to the species with the smaller genome. Biennial species of Cirsium possessed larger nuclear DNA amounts than their perennial relatives. Genome size was negatively correlated with Ellenberg's indicator values for continentality and moisture and with eastern limits of distribution. A negative relationship was also detected between the genome size and the tendency to form natural interspecific hybrids. On the contrary, C-values positively corresponded with the spinyness (degree of spinosity). AT frequency ranged from 48.38 % in C. eriophorum to 51.75 % in C. arvense. Significant intraspecific DNA content variation in DAPI sessions was detected in C. acaule (probably due to the presence of B-chromosomes), and in tetraploid C. vulgare. Only the diploid level was confirmed for the Pannonian C. brachycephalum, generally considered to be tetraploid. In addition, triploidy was discovered for the first time in C. rivulare. CONCLUSIONS: Considerable differences in nuclear DNA content exist among Central European species of Cirsium on the diploid level. Perennial soft spiny Cirsium species of wet habitats and continental distributions generally have smaller genomes. The hybrids of diploid species remain diploid, and their DNA content is smaller than the mean of the parents. Species with smaller genomes produce interspecific hybrids more frequently.     

Natural hybridization and hybrid zones between Quercus crassifolia and Quercus crassipes (Fagaceae) in Mexico: morphological and molecular evidence

Hybrid zones provide interesting systems to study genetic and ecological interaction between different species. The correct identification of hybrids is necessary to understand the evolutionary process involved in hybridization. An oak species complex occurring in Mexico formed by two parental species, Quercus crassifolia H. & B. and Q. crassipes H. & B., and their putative hybrid species, Q. dysophylla, was analyzed with molecular markers (random amplified polymorphic DNA [RAPDs]) and morphological tools in seven hybrid zones (10 trees per taxa in each hybrid zone) and two pure sites for each parental species (20 trees per site). We tested whether geographic proximity of hybrid plants to the allopatric site of a parental species increases its morphological and genetic similarity with its parent. Seventeen morphological traits were measured in 8700 leaves from 290 trees. Total DNA of 250 individuals was analyzed with six diagnostic RAPD primers. Quercus crassifolia differed significantly from Q. crassipes in all the examined characters. Molecular markers and morphological characters were highly coincident and support the hypothesis of hybridization in this complex, although both species remain distinct in mixed stands. Clusters and a hybrid index (for molecular and morphological data) showed that individuals from the same parental species were more similar among themselves than to putative hybrids, indicating occasional hybridization with segregation in hybrid types or backcrossing to parents. Evidence does not indicate a unidirectional pattern of gene flow.     

Nuclear DNA content in some species of Lessingianthus (Vernonieae, Asteraceae) by flow cytometry

The nuclear DNA content was determined for the first time in 25 species of the South American genus Lessingianthus H.Rob. (Vernonieae, Asteraceae) by flow cytometry. This analysis constitutes the first estimation of the genome size for the Vernonieae tribe. The 2C- and 1Cx-values were calculated in all the species. The 2C-value ranged from 2.04 to 14.34 pg. The 1Cx-value ranged from 0.995 to 1.43 pg. The general tendency indicated a decrease in the 1Cx-value with increasing ploidy level, with some exceptions, in some species the 1Cx-value increased with the ploidy increase. The measuring of DNA content allowed reporting a new cytotype for L. polyphyllus (Sch.Bip.) H.Rob.     

Nuclear DNA content in allopolyploid species and synthetic hybrids in the grass genus Paspalum

DNA content was estimated by flow cytometry in seventeen taxa from the Dilatata, Quadrifaria and Paniculata groups of Paspalum and five synthetic hybrids. Results were compared to known genome constitutions and phylogenetic relationships. DNA 2C-values ranged from 1.24 pg in diploid P. juergensii to 3.79 pg in a hexaploid biotype of P. dilatatum. The I genome of three Quadrifaria diploids is 1.2 to 1.5-fold larger than the J genome of P. juergensii (Paniculata). The 2C-values of the IIJJ tetraploids of the Dilatata group are lower than expected based on putative genome donors. Reduction of genome sizes could have occurred after the formation of the allopolyploids of the Dilatata group. The DNA content of all synthetic hybrids is in accordance with the sum of parental C-values. The interactions driving genome downsizing may operate differently during the transition from diploidy to polyploidy than on subsequent increases in ploidy level.     

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.     

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.     

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.     

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.     

Relative DNA content in diploid,polyploid, and multiploid species of <Emphasis Type="Italic">Paspalum</Emphasis> (Poaceae) with relation to reproductive mode and taxonomy

It is generally accepted that polyploids have downsized basic genomes rather than additive values with respect to their related diploids. Changes in genome size have been reported in correlation with several biological characteristics. About 75 % of around 350 species recognized for Paspalum (Poaceae) are polyploid and most polyploids are apomictic. Multiploid species are common with most of them bearing sexual diploid and apomictic tetraploid or other ploidy levels. DNA content in the embryo and the endosperm was measured by flow cytometry in a seed-by-seed analysis of 47 species including 77 different entities. The relative DNA content of the embryo informed the genome size of the accession while the embryo:endosperm ratio of DNA content revealed its reproductive mode. The genome sizes (2C-value) varied from 0.5 to 6.5 pg and for 29 species were measured for the first time. Flow cytometry provided new information on the reproductive mode for 12 species and one botanical variety and supplied new data for 10 species concerning cytotypes reported for the first time. There was no significant difference between the mean basic genome sizes (1Cx-values) of 32 sexual and 45 apomictic entities. Seventeen entities were diploid and 60 were polyploids with different degrees. There were no clear patterns of changes in 1Cx-values due to polyploidy or reproductive systems, and the existing variations are in concordance with subgeneric taxonomical grouping.     

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.     

Sexual and vegetative reproduction of Hieracium pilosella L. under competition and disturbance: a grid-based simulation model

We used a spatially explicit simulation model to examine the relative importance of vegetative and sexual reproduction in Hieracium pilosella L. Based on an understanding of the complex life cycle of this species and on data from in situ population dynamics in a calcareous grassland in NW Switzerland, we simulated growth and the relative contribution of clonal reproduction by stolons and reproduction by seeds across a gradient of increasing soil fertility. Competition by a clonal grass resulted in nearly complete exclusion of H. pilosella from the more fertile part of the simulation plot. Under low soil fertility, when grass could not survive, H. pilosella largely persisted by vegetatively produced rosettes. This pattern of a sharp separation of both species was shifted slightly in favour of H. pilosella by introducing random disturbances. Only by adding: (1) long-distance seed dispersal, and (2) facilitation of seedling establishment in the vicinity of grass tussocks in vegetation gaps was a more realistic representation of field observations realised, with rosettes of H. pilosella grown from seeds occasionally distributed within dense grass vegetation. Phenotypic plasticity of stolon length was a decisive factor for the maintenance of H. pilosella populations. We conclude that a mixed strategy of clonal growth and reproduction by seeds in H. pilosella is necessary to maintain populations of this species in the presence of high interspecific competition and a shortage of open space.     

Genetic relationships among Hylocereus and Selenicereus vine cacti (Cactaceae): evidence from hybridization and cytological studies

BACKGROUND AND AIMS: Hylocereus and Selenicereus are native to tropical and sub-tropical America. Based on its taxonomic status and crossability relations it was postulated that H. megalanthus (syn. S. megalanthus) is an allotetraploid (2n = 4x = 44) derived from natural hybridization between two closely related diploid taxa. The present work aimed at elucidating the genetic relationships between species of the two genera. METHODS: Crosses were performed and the putative hybrids were analysed by chromosome counts and morphological traits. The ploidy level of hybrids was confirmed by fluorescent in situ hybridization (FISH) of rDNA sites. Genomic in situ hybridization (GISH) was used in an attempt to identify the putative diploid genome donors of H. megalanthus and an artificial interploid hybrid. KEY RESULTS: Reciprocal crosses among four diploid Hylocereus species (H. costaricensis, H. monacanthus (syn. H. polyrhizus), H. undatus and Hylocereus sp.) yielded viable diploid hybrids, with regular chromosome pairing. Reciprocal crosses between these Hylocereus spp. and H. megalanthus yielded viable triploid, pentaploid, hexaploid and aneuploid hybrids. Morphological and phenological traits confirm the hybrid origin. In situ detection of rDNA sites was in accord with the ploidy status of the species and hybrid studied. GISH results indicated that overall sequence composition of H. megalanthus is similar to that of H. ocamponis and S. grandiflorus. High sequence similarity was also found between the parental genomes of H. monacanthus and H. megalanthus in one triploid hybrid. CONCLUSIONS: The ease of obtaining partially fertile F1 hybrids and the relative sequence similarity (in GISH study) suggest close genetic relationships among the taxa analysed.