The intriguing complexity of parthenogenesis inheritance in Pilosella rubra (Asteraceae, Lactuceae)

Neither the genetic basis nor the inheritance of apomixis is fully understood in plants. The present study is focused on the inheritance of parthenogenesis, one of the basic elements of apomixis, in Pilosella (Asteraceae). A complex pattern of inheritance was recorded in the segregating F(1) progeny recovered from reciprocal crosses between the facultatively apomictic hexaploid P. rubra and the sexual tetraploid P. officinarum. Although both female and male reduced gametes of P. rubra transmitted parthenogenesis at the same rate in the reciprocal crosses, the resulting segregating F(1) progeny inherited parthenogenesis at different rates. The actual transmission rates of parthenogenesis were significantly correlated with the mode of origin of the respective F(1) progeny class. The inheritance of parthenogenesis was significantly reduced in F(1) n?+?n hybrid progeny from the cross where parthenogenesis was transmitted by female gametes. In F(1) n?+?0 polyhaploid progeny from the same cross, however, the transmission rate of parthenogenesis was high; all fertile polyhaploids were parthenogenetic. It appeared that reduced female gametes transmitting parthenogenesis preferentially developed parthenogenetically and only rarely were fertilized in P. rubra. The fact that the determinant for parthenogenesis acts gametophytically in Pilosella and the precocious embryogenesis in parthenogenesis-transmitting megagametophytes was suggested as the most probable explanations for this observation. Furthermore, we observed the different expression of complete apomixis in the non-segregating F(1) 2n?+?n hybrids as compared to their apomictic maternal parent P. rubra. We suggest that this difference is a result of unspecified interactions between the parental genomes.     

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.     

The genusPicris (Asteraceae,Lactuceae) in Tropical Africa

Three species of the genusPicris L. are native in Tropical Africa:P. abyssinica Sch. Bip. (Ethiopia),P. xylopoda Lack, spec. nova (Nigeria, Ethiopia) andP. humilis DC. (Senegal, Mali). There are indications that the two perennial species,P. abyssinica andP. xylopoda, are related to and have evolved from a primitive Central Asiatic stock in a manner parallel to many African species ofCrepis L.P. humilis, on the other hand, is a small annual plant with a high number of derived characters. The introduced species of European origin growing south of the Sahara are briefly mentioned.     

Phenotypic Similarity and Genetic Relationship Among Populations of Microseris bigelovii (Asteraceae: Lactuceae)

Twelve inbred strains have been derived from 10 representative populations of the North American annual M. bigelovii. The distribution and correlation of 21 phenotypic characters distinguishing these strains has been studied. The genetic similarity of these strains has been previously determined using nu clear DNA polymorphisms. Genetically similar strains (always geographic near neighbors) share phenotypic character states but these involve different characters in different cases. A few character state differences are correlated across all strains via early rosette growth rate, timing of reproduction, or overall capitulum size. Most character states recombine freely. This is evidence for effective gene flow (probably by achene dispersal) among these isolated populations.     

Geographical distribution of wildLactuca species (Asteraceae, Lactuceae)

A broad survey of available world literature showed that at least 98 wildLactuca spp. (Asteraceae) have been described taxonomically. The distribution of the genusLactuca worldwide includes 17 species in Europe, 51 in Asia, 43 in Africa, and 12 in the Americas (mostly the North American subcontinent). Species originating in Asia, Africa, and the Americas form ca. 83% of knownLactuca spp. richness; however, they are very poorly documented from the viewpoint of taxonomic relationships, ecogeography, and variability. The phytogeography ofLactuca spp. regarding their distribution on different continents and in relation to the structure of the lettuce gene pool is discussed. A more detailed analysis of geographical distribution and habitats is given for some species (L. serriola, L. saligna, L. virosa, L. perennis, L. quercina, L. tatarica), which represent the primary, secondary, and tertiary gene pools of cultivated lettuce (L. sativa). Original and synanthropic distributions ofLactuca spp. and their occurrence in natural and secondary habitats are discussed, along with the representation of wildLactuca spp. in world gene-bank collections. Global biodiversity ofLactuca spp. and their representation in germplasm collections are poorly documented. Future studies of taxonomy, phytogeography, ecology, phylogenetic relationships, and genetic diversity are needed for a more complete understanding of this genus and taxonomically related genera.     

Hypervariable DNA Fingerprint Loci in Microseris pygmaea (Asteraceae,Lactuceae)

Oligo-GATA containing probes reveal DNA fingerprinting patterns in DNA of Microseris pygmaea after restriction enzyme digestion. There are only a few major restriction fragments. These are longer than 2 kb, and they differ completely among plants from different populations, among most members of a single local inbreeding population, and even among some parent and offspring plants. In the F2 of an interpopulation hybrid, major fingerprint bands could be assigned to 2 unlinked loci by 3: 1 and 1: 2: 1 segregations respectively, one further band segregated roughly 1: 1, one band appeared irregularly. The hypervariability of the GATA-fingerprint loci contrasts with a low variability of other genetic markers in M. pygmaea and with a more complex but less variable GATA-fingerprint pattern in related species.     

Substantial Genome Size Variation in Taraxacum stenocephalum (Asteraceae, Lactuceae)

There are only a few exceptions to the rule that polyploidy in Taraxacum is associated with agamospermy. One of them is the sexual, tetraploid species Taraxacum stenocephalum. Incidentally, remarkable variation in karyology was found in this species. The present study aims to confirm this variation by an extensive screen of nuclear DNA content. Individuals from two large populations in the Lesser and Greater Caucasus, Georgia were analyzed using flow cytometry to ascertain intraspecific nuclear DNA content variation. Across the whole data set comprising all 159 individuals, a 1.223-fold difference was detected based on propidium iodide (PI) analyses. To verify this finding, we compared flow-cytometric data obtained using DAPI (4′,6-diamidino-2-phenylindole) and PI staining using a representative subset of individuals. This comparison revealed a 1.194-fold difference in DNA content for DAPI and a 1.219-fold difference for PI. Mean nuclear genome size in absolute terms (2C value ± s.d.) was estimated at 4.38?±?0.21 pg, ranging from 4.01 pg to 4.89 pg, despite the invariable chromosome counts of 2n?=?32. A regression analysis comparing the datasets for DAPI and PI staining found a strong correlation between data obtained by the DAPI and PI dyes (R?=?0.976; P?=?0.0001). Simultaneous high-resolution flow-cytometric analyses also proved the accuracy of our findings. We discuss possible sources of these large differences in DNA content within Taraxacum stenocephalum. Further research is needed to identify the source of this remarkable variation.     

Karyotype evolution in South American species of Hypochaeris (Asteraceae, Lactuceae)

The genus Hypochaeris (Asteraceae, Lactuceae) contains ten species in Europe, three in Asia, and approximately 50 in South America. Previous cytotaxonomic studies have shown two groups of taxa: (1) European species with different basic chromosome numbers and differentiated karyotypes, and (2) South American species with x=4 and uniform asymmetric and bimodal karyotypes. Karyotypic data are synthesized for South American species of Hypochaeris with new information for six Chilean species: H. acaulis, H. apargioides, H. palustris, H. spathulata, H. tenuifolia and H. thrincioides. Four main groups can be distinguished based on presence and localization of secondary constrictions – SCs (bearing Nucleolar Organizer Regions – NORs) on chromosomes 2 and 3, and 18S–25S and 5S rDNA loci number, localization, and activity. We propose karyotypic evolution of South American Hypochaeris (x=4) from H. maculata-like (x=5) European ancestors. The original South American karyotype would have possessed two SCs, one on the long arm of chromosome 2, and the other on the short arm of chromosome 3 (in terminal position). Further evolution would have involved inversion within the short arm of chromosome 3 and inactivation/loss of the SC on chromosome 2.     

Molecular phylogenetics reveals Leontodon (Asteraceae, Lactuceae) to be diphyletic

The plastid matK gene, trnL/F spacer, and nuclear rDNA ITS were sequenced for 36 species of Leontodon and 29 taxa of related genera of tribe Lactuceae. Phylogenetic relationships inferred from the independent and combined data are largely congruent and reveal that Leontodon sensu lato (s.l.) as presently defined is diphyletic: L. subgenus Leontodon forms a clade with Helminthotheca, Picris and Hypochaeris as sister genera, whereas L. subgenus Oporinia appears as a separate clade with strong bootstrap support and is thus better treated as a separate genus. Previous sectional classifications of Leontodon s.l. are considered in the light of DNA and additional morphological and karyological data. Support is presented for a core group of Hypochaeridinae sensu stricto (s.s.) with the two clades of Leontodon s.l., Helminthotheca, Picris, and Hypochaeris, whereas Urospermum, Hyoseris, Aposeris, and Rhagadiolus appear to be positioned more distantly.     

Sesquiterpenoids as chemosystematic markers in the subtribe Hypochaeridinae (Lactuceae,Asteraceae)

Sesquiterpene lactones are well established as chemosystematic markers in the Asteraceae family. From the Lactuceae tribe of the Asteraceae family a large number of sesquiterpene lactones – mainly of the guaiane type – have been isolated. One of the 11 subtribes of the Lactuceae recognized by Bremer is the subtribe Hypochaeridinae, which encompasses 10 genera with approximately 170 species. The present communication summarizes the sesquiterpene derivatives reported from these 10 genera, points out, which constituents are characteristic for particular groups, and discusses the occurrence of these secondary metabolites in a chemosystematic context. To this end, each of the reported sesquiterpenoids reported for the Hypochaeridinae is classified into one of three main compound classes (MCCs; eudesmane-, germacrane-, guaiane-derivatives) and into one of a number of compound classes (CCs) within these principal groups. The distribution of sesquiterpenoids belonging to these particular classes of sesquiterpenoids generally follows the currently accepted generic limits. However, the genus Helminthoteca, which is included into Picris by many authors, possesses an array of compounds implying a closer relationship to the genus Hypochaeris than to the remainder of the genus Picris. Furthermore, Leontodon subgenus Oporinia shows closer similarity in secondary metabolite patterns to the genus Picris than to Leontodon subgenus Leontodon. On the other hand Leontodon subgenus Leontodon has more chemical characters in common with the genus Hedypnois than with Leontodon subgenus Oporinia. These findings are in-line with recent results of molecular analyses, which imply that current generic limits within the Hypochaeridinae might not reflect the phylogeny of the subtribe.     

Allozyme variation in sexual and apomictic Taraxacum and Pilosella (Asteraceae) populations

Allozyme spectra of peroxidase, esterase, superoxid dismutase, tyrosinase, alcohol dehydrogenase, lactate dehydrogenase, and acid phosphatase were examined in populations of sexual (Taraxacum serotinum and Pilosella echioides) and apomictic (T. officinalis and P. officinarum) plant species. The heterozygosity in these populations (0.455-0.620) proved to be considerably higher than the average level characteristic of plant populations (0.058-0.185). The populations examined did not differ in the mean phenotype number mu, i.e., they exhibited the same diversity (3.213-3.380). The proportion of rare phenotypes h also did not differ between the sexual and apomictic species of the same genus, whereas this parameter in the Pilosella populations (0.150-0.174) was significantly higher than in the Taraxacum ones (0.093-0.114). The populations were characterized by numerous isozyme spectra (more than 11 per populations) and displayed multiple allelism (the mean allele frequency was 3.63-4.38 per locus). They exhibited a high percentage of rare (occurring at a frequency lower than 5%) spectra (35-80%). This indicates that agamic complexes, to which these populations belong, may have a more complicated genetic structure of both apomictic and sexual populations than the species that do not belong to agamic complexes.     

A new Spanish species of Tragopogon (Asteraceae: Lactuceae)

Tragopogon cazorlanum (Asteraceae: Lactuceae) is proposed as a new Spanish species with distribution restricted to the Baetic mountains (south-eastern Spain). Its morphological, palynological, chromosomal, biogeographical and ecological features are discussed, as well as its main relationships and differences with reference to other Spanish species of the genus.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 146 , 505–511.     

Evolution of microsporangium numbers in Microseris (Asteraceae: Lactuceae)

The genus Microseris contains species with disporangiate stamens and species with tetrasporangiate stamens. We determined the number of microsporangia per stamen in serial sections of heads from all 13 species of Microseris, its close relative Uropappus lindleyi, and the two allopolyploid species of Slebbinsoseris. Four Microseris species, three diploid and one tetraploid, have two microsporangia per stamen; all other species investigated have four. The most parsimonious assumption is that the disporangiate condition is derived and arose once in the evolution of Microseris. The inheritance of the number of microsporangia per stamen in crosses between M. bigelovii (disporangiate) and M. douglasii (tetrasporangiate) was determined. Segregation of microsporangium number per stamen in F2s derived from these crosses is quantitative rather than Mendelian. The average number of microsporangia per stamen in the F2 plants ranges from 2.0 to 4.0. There is a predominance of tetrasporangiate stamens in the F1 and in most F2 plants. The observed pattern of inheritance suggests a major gene with dominance and quantitative modifiers.     

Genetic components of heterocarpy inMicroseris hybrid B87 (Asteraceae,Lactuceae)

Microseris B87 is derived from a single hybrid specimen betweenM. pygmaea with few, weakly hairy peripheral achenes and aM. bigelovii with many, strongly hairy peripheral achenes. Offspring through the F4 and F5 generations obtained by spontaneous selfing were analyzed for the segregation of quantitative and qualitative characters relating to achene dimorphism. The phenotypic effects of two previously identified unlinked genes determining the relative number of outer achenes are characterized in partially and completely homozygous sublines. We show that two morphological markers genetically linked to one of these genes are themselves regulated by the system inducing heterocarpy. Not more than two more unlinked genes are involved in the genetic basis of the heterocarpic response. The interaction of these genes in determining the heterocarpy phenotypes is discussed in the framework of a model postulating genes for a morphogen gradient across the capitulum and genes responding to this gradient.     

Allopatric distribution,ecology and conservation status of the Pilosella alpicola group (Asteraceae)

The Pilosella alpicola group comprises four morphologically distinct and geographically vicariant alpine taxa. We performed a thorough herbarium revision and literature survey to infer their distributional pattern(s). Pilosella alpicola s.s. occurs in the Alps in two disjunct areas: the Swiss Valais Alps and the Italian Dolomites. Historical records come also from the Austrian Alps (Gurktaler Alps and Hohe Tauern) and from one site from the Alpes Maritimes (Col de Larche), but the localities have not been recently confirmed. Pilosella rhodopea, a Balkan subendemic taxon, is quite widespread in Bulgaria (Stara planina Mts, Rila Mts and Pirin Mts), but is more rare in Albania, Greece and Macedonia. Interestingly, this species has also been recorded at two isolated sites in the Romanian southern Carpathians (the C?p??înii and Cozia Mts). This occurrence underlines the floristic affinities of this part of the Carpathians to the Balkan flora. Only two localities of P. serbica, based on voucher specimens, have been recorded so far; Kopaonik Mts in Serbia and the Prokletije Mts in Montenegro. The records from other ranges are related to P. rhodopea. Pilosella ullepitschii, the detailed distribution of which has already been published, is a Carpathian endemic with its core area of distribution in the western Carpathians (Slovakia and Poland). Three isolated localities are also known in the eastern Carpathians (Nemira Mts) and one locality in the southern Carpathians (Bucegi Mts). The possible causes of disjunctions between and within species ranges are briefly discussed. Based on the distributional data, population sizes and ecology, we evaluate the conservation status of the P. alpicola taxa and propose their inclusion in national Red Lists.     

Diplospory and Parthenogenesis in Sexual x Agamospermous (Apomictic ) Erigeron (Asteraceae) Hybrids

Segregation for asexual seed production was evaluated for 130 experimental F1 hybrids resulting from a cross between diploid (2n=18) sexual Erigeron strigosus and triploid (2n=27) agamospermous Erigeron annuus. Paternity of hybrids was documented using 13 RAPD markers. The distribution of F1 chromosome numbers is bimodal, centering on diploid and triploid ploidal levels but with underrepresentation of diploids. Diplosporous versus meiotic megagametophyte development was ascertained microscopically for >/=100 ovules per plant. Diplospory ranges from 0% to 100% among all progeny but is uniformly low (0%-3%) for 17 diploid hybrids. The inheritance of diplospory in Erigeron appears to be best explained by a one-locus-two-allele polysomic model with selection against gametes homozygous for diplospory. Parthenogenesis, estimated via seed counts, ranges from 0% to 60% and apparently is contingent upon diplospory, as seed production was absent or very low in predominantly meiotic hybrids. However, the absence of parthenogenesis in many highly diplosporous hybrids indicates that these two aspects of agamospermous development are not strictly associated. The segregation of both diplospory and parthenogenesis in this population will permit further genetic dissection of these traits with molecular marker-based analyses.     

Phylogeographic patterns in Hypochaeris section Hypochaeris (Asteraceae, Lactuceae) of the western Mediterranean

Aim  To analyse phylogeographic patterns in the four species of Hypochaeris sect. Hypochaeris , evaluating possible areas of origin and the microevolutionary processes that have shaped their morphology, genetics and distribution.
Location  Western Mediterranean area.
Methods  We applied amplified fragment length polymorphism (AFLP) markers to a total of 494 individuals belonging to 82 populations of Hypochaeris arachnoidea , H. glabra , H. radicata and H. salzmanniana to determine population structure.
Results  Populations with the largest proportion of private and rare AFLP fragments were found in Morocco. This region was consequently inferred to be the ancestral area for H. arachnoidea , H. glabra , H. radicata and H. salzmanniana . The Guadalquivir River (southern Spain) was inferred to be an effective dispersal barrier for H. glabra and H. radicata. The Strait of Gibraltar was inferred to be a somewhat weaker barrier than the Guadalquivir River for H. radicata and a much weaker barrier for H. glabra . The main barrier for H. salzmanniana coincides with the extension of the Rif Mountains to the Atlantic coast in Morocco, and the Strait of Gibraltar is a much weaker barrier for this species. Hypochaeris arachnoidea appears to have originated in the Atlas Mountains.
Main conclusions  The highest levels of genetic variation in La Mamora forest ( H. glabra and H. salzmanniana ) or the adjacent central Middle Atlas ( H. arachnoidea and H. radicata ) in Morocco suggest that these areas were a centre of origin of Hypochaeris sect. Hypochaeris . All three potential barriers – the Guadalquivir River, the Strait of Gibraltar, and the Rif Mountains – have been important in shaping genetic diversity in species of section Hypochaeris .     

Allozyme Variation in Diploid, Polyploid and Mixed-Ploidy Populations of the Pilosella alpicola Group (Asteraceae): Relation to Morphology, Origin of Polyploids and Breeding System

The Pilosella alpicola group includes four species (P. alpicola s.str., P. ullepitschii, P. rhodopea and P. serbica) with allopatric distributions (Alps, Balkans, Carpathians) and contrasting cytotype patterns (diploid, diploid-polyploid and polyploid species). Whereas diploid taxa (P. ullepitschii and P. serbica) reproduce sexually, the mode of reproduction of polyploid cytotypes reflects their origin: autopolyploids of P. rhodopea reproduce sexually, while allopolyploid cytotypes of P. alpicola s.str. apomictically. We used allozymes to elucidate overall genetic variation within the group and to test their utility for taxon discrimination, assessment of polyploid origin and possible correlations with breeding systems. Variation of five allozyme systems encoded by eight polymorphic loci and 29 alleles was studied in 20 populations and 298 plants representing all taxa. Allozymes were proved to be only of limited usefulness for the taxonomic classification within the P. alpicola group. The Western Carpathian populations of P. ullepitschii formed the only genetically well-differentiated group. The same allele suite shared by all cytotypes of P. rhodopea and presence of both balanced and unbalanced heterozygotes in tetraploids was consistent with autopolyploid origins of polyploids and provided further evidence for a primary contact zone. An isolated relic population of P. rhodopea from the Southern Carpathians exhibited lowered values of genetic diversity when compared to the core area. Pronounced fixed heterozygosity was found in P. alpicola s.str., supporting its allopolyploid origin. In accordance with assumptions, genotypic variability was significantly higher in sexually reproducing diploid and diploid-polyploid taxa than in apomictic P. alpicola s.str.     

Phenotypic Plasticity of Capitulum Morphogenesis in Microseris pygmaea (Asteraceae: Lactuceae)

Inbred populations of the annual Chilean species Microserispygmaea show phenotypic plasticity in the number of floretsper capitulum. In order to find out how this plastic variationmay arise during development, two inbred lines (A92 and C96)were grown under short day conditions. Groups of plants fromeach strain were transferred to long day conditions at about2 week intervals. In this way we introduced variation for plantsize (number of leaves per plant) at onset of flowering. Thenumber of florets per mature capitulum increased linearly withplant size. After transfer to long day conditions, plants wereharvested daily for light microscopic measurements of meristemwhole mounts. Only the first capitulum of each plant was analysed.All florets were formed after 12 d in strain C96 and after 14d in strain A92. In order to detect the effect of plant sizeon morphogenesis, we performed a multiple regression analysisof developmental parameters on time and number of leaves. Widthand height of the capitulum receptacle increased daily witha growth rate depending on the starting size. Differences inmeristem size were detected already in vegetative plants ofdifferent sizes. In contrast, there was no influence of plantsize on floret primordium size. We combined the multiple regressionmodels in one simple model for prediction of floret numbersfrom numbers of leaves per plant at onset of flowering. Predictionsof this model agree with observed relationships in both inbredlines.Copyright 1994, 1999 Academic Press Asteraceae, capitulum, inflorescence, meristem, meristic character, Microseris pygmaea, morphogenesis, phenotypic plasticity