Anna Krahulcová

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.

Introgressive hybridization between a native and an introduced species:Viola lutea subsp.Sudetica versusV. Tricolor

A hybrid swarm betweenViola lutea subsp.sudetica (2n=50, native species) andViola tricolor (2n=26, introduced species) originated in the 1970’s in the Krkonoše Mts. Analyses of chromosome numbers, reproductive biology, morphology, and ecology gave the following results: (1) Compared to the plants found in the 1970’s, the number of colour morphs have decreased and the types now prevailing in the field are morphologically closer toV. lutea subsp.sudetica, forming a continuum. (2) In the field, hybrids having approximately the same chromosome number as the primary hybrids are most common. Some plants of the hybrid swarm have certain characters unknown to their parents. (3) In the field, autogamous types similar toV. tricolor were found. (4) A limited number of plants from the field and culture have higher somatic chromosome numbers thanV. lutea subsp.sudetica; there was a tendency towards increasing chromosome numbers in their progeny (up to 2n=62). These plants have some new morphological characters (a small proportion of hexacolpate pollen) not found in the parents (nor in the other hybrids with prevailing tetracolpate and pentacolpate pollen grains) and higher pollen fertility in comparison to the other hybrids. These plants also have the highest germination rate. (5) There is a tendency for chromosome numbers to decline in the progeny of open pollinated hybrid plants in the lowland experimental graden. (6) The phenology of the plants in the mountain grasslands and the lowland garden is different; the parents behave in a totally contrasting manner. (7) In the field at least some hybrids extend outside the geographical and ecological ranges of the parental species, invading new communities. (8) Seeds ofV. tricolor do not show any dormancy and germinate in the year of production; most of theV. lutea subsp.sudetica seeds germinate during the spring of the following year. Hybrids with intermediate chromosome numbers had both types of germination strategy. The germination rate of intermediates with high chromosome numbers was even higher than that ofV. tricolor.

Enriching Ploidy Level Diversity: the Role of Apomictic and Sexual Biotypes of Hieracium subgen. Pilosella (Asteraceae) that Coexist in Polyploid Populations

The capacity to generate variation in ploidy and reproductive mode was compared in facultatively apomictic versus sexual maternal plants that coexist in two model populations. The population structure was studied in polyploid hybrid swarms comprised of Hieracium pilosella (usually sexual, less commonly apomictic), H. bauhini (apomictic), and their hybrids (sexual, apomictic, or sterile). Relationships among established biotypes were proposed on the basis of their DNA ploidy level/chromosome number, reproductive mode and morphology. Isozyme phenotypes and chloroplast DNA haplotypes were assayed in the population that was richer in hybrids. The reproductive origin of seed progeny was identified in both sexual and apomictic mothers, using alternative methods: the karyological, morphological and reproductive characters of the cultivated progeny were compared with those of respective mothers, or flow cytometric seed screening was used. In both populations, the progeny of sexual mothers mainly retained a rather narrow range of ploidy level/chromosome number, while the progeny of facultatively apomictic mothers was more variable. The high-polyploid hybrids, which had arisen from the fertilization of unreduced egg cells of apomicts, mainly produced aberrant non-maternal progeny (either sexually and/or via haploid parthenogenesis). Apparently, such versatile reproduction resulted in genomic instability of the recently formed high-polyploid hybrids. While the progeny produced by both true apomictic and sexual mothers mostly maintained the maternal reproductive mode, the progeny of those ‘versatile’ mothers was mainly sexual. Herein, we argue that polyploid facultative apomicts can considerably increase population diversity.

Ecology of two cytotypes ofButomus umbellatus I. Karyology and breeding behaviour

Chromosome numbers were counted inButomus umbellatus from 99 localities in both Czech and Slovak Republics and the karyotype morphology was studied. Basic chromosome sets are asymmetrical and uniform among diploids (2n=26) and triploids (2n=39). Diploids occur less frequently than triploids in this region. Their clonal populations are usually fertile owing to self-compatibility. The clonal populations of triploids are selfincompatible and usually sterile. Thus, the different self-compatibility is the main biological character distinguishing diploids from triploids. Pollen of triploids is viable in spite of meiotic irregularities in pollen mother cells (PMCs). Hybridization both between cytotypes and between the different triploid genets may take place, if they occur sympatrically. Offspring having diverse chromosome numbers between diploid and triploid level can originate as the consequence of such hybridization.

Autogamy inHieracium Subgen.Pilosella

The presence of autogamy inHieracium subgen.Pilosella is reported for diploidH. lactucella and tetraploidH. pilosella. Self-compatibility is induced under the influence of pollen from another species (mentor effects).

The biology of Butomus umbellatus in shallow waters with fluctuating water level

Butomus umbellatus L. is a plant species typical of littoral communities of river and stream shores. It can form continuous stands in shallow reservoirs with fluctuating water level. Their expansion is promoted by: (a) intensive vegetative reproduction of plants, (b) crowded sprouting from rhizome fragments on emerged pond bottom, (c) shallow water layer in the year following summer drainage. Expansion of B. umbellatus depends on ploidy level: two cytotypes were found in the Czech and Slovak Republics, differing in their reproductive ability. Seed production of triploids is strongly limited (they are self-incompatible within clones), while diploids can be fully fertile. Nevertheless, even in diploids, the efficiency of seed reproduction under natural conditions is low. Triploids spread by intensive vegetative reproduction, which is decisive for clonal growth of populations and their regeneration after scraping of bottom surface. During seasonal development, maximum of aboveground biomass is produced in early summer, while underground biomass increases till autumn. Growth of the plants is limited by cutting before maximum underground biomass is attained, or by duck grazing.

Variation inTaraxacum bessarabicum and allied taxa of the sectionPiesis (Compositae): allozyme diversity, karyotypes and breeding behaviour

Allozyme techniques, karyotype analyses and cultivation experiments were carried out on 20 population samples ofTaraxacum sect.Piesis (Compositae), viz.T. bessarabicum (17 samples from W. and C. Europe, Ukraine and Crimea, Central Asia and the Altai),T. salsum from Crimea,T. x mesohalobium from Crimea, andT. stenolepium from the Causasus. The taxa studied share a primitive, symmetrical karyotype. All taxa studied are sexual,T. bessarabicum mostly autogamous. Sexuality at the tetraploid level (T. stenolepium) and occasional male sterility in a sexual (T. bessarabicum in Moravia) were recorded for the first time in the genus. Selfing, diploidy and the highly predictable habitat may account for the mostly low level within population allozyme variation inT. bessarabicum. W. and C. European samples ofT. bessarabicum are almost invariable allozymically and, as a group, have no unique alleles. The Crimean and Asian group of populations shows higher levels of allozyme variation and has 15 alleles not found in the former group at 13 loci studied. Recent migration from one source region is suggested to account for the homogeneity of the western group, refugial persistence of alleles and possible introgression from sympatric species may have resulted in allele richness in the eastern group. Hybridization betweenT. bessarabicum andT. salsum was documented by allozyme patterns in a few plants in Crimea. Some aberrant allozyme or karyotypic features of two populations are discussed as well.

Selected chromosome counts of the Czechoslovak flora III

Chromosome numbers compared with as yet published data are given for the following 12 Phanerogams (both native species and aliens) from Czechoslovakia:Ambrosia trifida L.,Cardamine chelidonia L.,Dephne cneorum L.,Epipactis albensisNováková etRydlo,Linum flavum L.subsp flavum, Lunaria rediviva L.,Nepeta grandiflora M.BIEB.,Reseda luteola L.,Thlaspi montanum L.,Tithymalus salicifolius (Host)Klotzsch etGarcke,Tithymalus virgultosus (Klokov) Holub andVerbascum speciosumSchrad. subsp.speciosum. The chromosome number 2n=40 is presented for the first time in autogamousEpipactis albensisNováková etRydlo. New chromosome numbers were found inCardamine chelidonia L. (2n=32) and inTithymalus salicifolius (Host) Klotzch etGarcke (2n=40). Known but less frequent cytotypes are reported inLinum flavum L. subsp.flavum (2n=28) and inVerbascum speciosumSchrad. subsp.speciosum (2n=30).

Evolution of apomixis loci in Pilosella and Hieracium (Asteraceae) inferred from the conservation of apomixis-linked markers in natural and experimental populations

The Hieracium and Pilosella (Lactuceae, Asteraceae) genera of closely related hawkweeds contain species with two different modes of gametophytic apomixis (asexual seed formation). Both genera contain polyploid species, and in wild populations, sexual and apomictic species co-exist. Apomixis is known to co-exist with sexuality in apomictic Pilosella individuals, however, apomictic Hieracium have been regarded as obligate apomicts. Here, a developmental analysis of apomixis within 16 Hieracium species revealed meiosis and megaspore tetrad formation in 1 to 7% of ovules, for the first time indicating residual sexuality in this genus. Molecular markers linked to the two independent, dominant loci LOSS OF APOMEIOSIS (LOA) and LOSS OF PARTHENOGENESIS (LOP) controlling apomixis in Pilosella piloselloides subsp. praealta were screened across 20 phenotyped Hieracium individuals from natural populations, and 65 phenotyped Pilosella individuals from natural and experimental cross populations, to examine their conservation, inheritance and association with reproductive modes. All of the tested LOA and LOP-linked markers were absent in the 20 Hieracium samples irrespective of their reproductive mode. Within Pilosella, LOA and LOP-linked markers were essentially absent within the sexual plants, although they were not conserved in all apomictic individuals. Both loci appeared to be inherited independently, and evidence for additional genetic factors influencing quantitative expression of LOA and LOP was obtained. Collectively, these data suggest independent evolution of apomixis in Hieracium and Pilosella and are discussed with respect to current knowledge of the evolution of apomixis.

Ploidy Level Selection During Germination and Early Stages of Seedling Growth in the Progeny of Allohexaploid Facultative Apomict, Hieracium rubrum (asteraceae)

Selection within progeny of a facultative apomict,Hieracium rubrum, was studied using flow cytometry of embryos in seeds (a modified method of Flow Cytometric Seed Screen) and seedlings. Flow cytometric screening of particular progeny classes was based on distinct ploidy categories, reflecting the way of their origin.The results of both estimations of progeny composition significantly differed, which makes direct comparison of proportions detected in seeds or seedling stage impossible. The results suggest that progeny originating from reduced egg cells have higher mortality during germination and the early establishment phase than that from unreduced egg cells. Within the progeny of emasculated plants, the proportion of polyhaploid progeny decreased in favor of apomictically derived plants. Within the progeny of plants pollinated byH. pilosella, the proportion of polyhaploid progeny decreased significantly in favor of apomictically derived plants and 2n + n hybrids.It is argued that at least a proportion of the sexually derived progeny of this facultatively apomictic maternal parent has a lower survival rate than apomictically derived progeny.