Evolution ofSedum seriesRupestria (Crassulaceae): Evidence from chloroplast DNA and biosystematic data

Evolutionary relationships within the compariumSedum seriesRupestria (Crassulaceae) were investigated by phylogenetic analyses of restriction site variation of the chloroplast genome and morphological variation. DNAs were digested with 21 restriction enzymes and hybridized withS. album cpDNA probes covering the entire genome. A total of 141 ingroup variable restriction sites was identified. Morphological variation was observed in a total of 66 characters. Both datasets were analysed using parsimony methods and compared with evidence from cytology, artificial hybridization and biogeography to infer evolutionary relationships. The results 1) indicate a relatively high level of nucleotide sequence divergence withinS. ser.Rupestria; 2) indicate three main lines of plastome differentiation, which are in agreement with morphology, basic chromosome numbers, and centres of diversity; 3) do not provide a compelling solution for basal relationships; 4) support an earlier hypothesis on reticulation and provide evidence for a third case of allopolyploidy in the series; 5) suggest that interspecific gene flow inS. ser.Rupestria is funnelled through the presumably allopolyploidS. rupestre subsp.erectum.     

Geographical variation and systematics of the tetraploid marsh orchid Dactylorhiza majalis subsp. sphagnicola (Orchidaceae) and closely related taxa

Dactylorhiza majalis subsp. sphagnicola is an allotetraploid marsh orchid derived from parents closely similar to present‐day D. incarnata and the western European form of D. maculata subsp. maculata, suggesting that it has a postglacial origin. It extends from northwestern continental Europe into areas formerly covered by the Weichselian ice sheet in mid‐Scandinavia. Here, we studied the variation at both the plastid and nuclear marker systems to describe the geographical variation in subsp. sphagnicola and its evolutionary history. We investigated whether subsp. sphagnicola is affected by secondary hybridization and gene flow from its parental lineages or from other allotetraploid marsh orchids, and we also compared subsp. sphagnicola with other allotetraploids of similar origins. We analysed 492 plants from 50 populations. Thirty‐seven populations were collected as potential Dactylorhiza majalis subsp. sphagnicola, five as subsp. sesquipedalis (D. elata), one as D. elata subsp. brennensis, one as subsp. calcifugiens, one as subsp. occidentalis and the remaining five as populations with some affinity to subsp. lapponica (including D. traunsteineri). All populations were analysed for plastid haplotypes and nuclear internal transcribed spacer (ITS) allele frequencies, and a subset of 43 populations was analysed for five nuclear microsatellite loci. Dactylorhiza majalis subsp. sphagnicola was dominated by a single plastid haplotype that was also dominant in western European D. maculata subsp. maculata, and most of the alternative haplotypes differed by only one mutation from the dominant one. There was more variation in nuclear microsatellites and ITS, and the variation was geographically structured in these markers. Subspecies occidentalis and calcifugiens shared haplotypes with subsp. sphagnicola, whereas subsp. sesquipedalis and brennensis had other haplotypes. Dactylorhiza majalis subsp. sphagnicola may have a postglacial origin within its present continental distribution. It has incorporated genetic material from D. maculata subsp. maculata by secondary hybridization and introgression, and some northern populations have assimilated strongly divergent haplotypes from the northeastern form of D. maculata subsp. maculata. Subspecies sphagnicola has also evolved morphologically divergent local populations in the north that do not differ from the typical populations in genetic markers. It may form mixed populations with other allotetraploid subspecies of D. majalis and, at least at one site, it has become integrated with subsp. lapponica, demonstrating that independently derived allotetraploids may contribute to a common gene pool. Subspecies calcifugiens seems to be derived from subsp. sphagnicola, and further studies based on a larger sample may confirm that it is better recognized as a variety. The so‐called D. elata subsp. brennensis is of hybrid origin and combines markers from subsp. sesquipedalis with markers from the D. majalis core complex, possibly subsp. majalis. The new combination Dactylorhiza majalis subsp. sesquipedalis (Willd.) H.A.Pedersen & Hedrén comb. nov. is provided. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 168 , 174–193.     

ALLOPOLYPLOIDY IN NATURAL AND CULTIVATED POPULATIONS OF PORPHYRA (BANGIALES,RHODOPHYTA)1

To confirm whether allopolyploidy occurs in samples of previously identified Porphyra yezoensis Ueda, P. tenera Kjellm., and P. yezoensis × P. tenera from natural and cultivated populations, we examined these samples by using PCR‐RFLP and microsatellite analyses of multiple nuclear and chloroplast regions [nuclear regions: type II DNA topoisomerase gene (TOP2), actin‐related protein 4 gene (ARP4), internal transcribed spacer (ITS) rDNA and three microsatellite loci; chloroplast region: RUBISCO spacer]. Except for the ITS region, these multiple nuclear markers indicated that the wild strain MT‐1 and the cultivated strain 90‐02 (previously identified as P. yezoensis × P. tenera and cultivated P. tenera, respectively) are heterozygous and possess both genotypes of P. tenera and P. yezoensis in the conchocelis phase. Furthermore, gametophytic blades of two pure lines, HG‐TY1 and HG‐TY2 (F₁ strains of MT‐1 and 90‐02, respectively), were also heterozygous, and six chromosomes per single cell could be observed in each blade of the two pure lines. These results demonstrate that allopolyploidy occurs in Porphyra strains derived from both natural and cultivated populations, even though ITS genotypes of these strains showed homogenization toward one parental ITS.     

Molecular evidence for the hybrid origin of a new endemic species of Stylosanthes Sw. (Fabaceae) from the Mexican Yucatán Peninsula

Stylosanthes aff. calcicola is a formally undescribed tetraploid species from the Mexican Yucatán Peninsula, showing morphological similarities to the diploid species S. calcicola , but distinct in a number of characters. We used uni- and biparentally inherited molecular markers to infer the hybrid origin of this species in relation to known diploid species of Stylosanthes . Molecular characterization was based on length and/or DNA sequence variation of nuclear sequence-tagged site (STS) markers, the internal transcribed spacer (ITS) region of nuclear rDNA and the trnL intron of chloroplast DNA (cpDNA). Stylosanthes aff. calcicola contains a distinct cpDNA haplotype and nuclear DNA fragment, with closest relationship to the diploid species S. calcicola . In contrast, the DNA sequences of two nuclear loci reveal a closer relationship to the diploid species S. angustifolia , S. hispida , S. humilis , S. leiocarpa and S. viscosa . The majority of the STS markers showed additivity of PCR fragments in S. aff. calcicola , representing the combination of two genetically different genomes. We postulate that S. aff. calcicola is a distinct species of allotetraploid origin that appears to have originated once from hybridization between two divergent genomes, of which the maternal and paternal parent are closely related to, or derived from, a member of the lineages represented by S. calcicola and S. viscosa , respectively.  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 140 , 1–13.     

Endo-allopolyploidy of autopolyploids and recurrent hybridization—A possible mechanism to explain the unresolved Y-genome donor in polyploid Elymus species (Triticeae: Poaceae)

The genus Elymus L. in the tribe Triticeae (Poaceae) includes economically and ecologically important forage grasses. The genus contains the pivotal St genome from Pseudoroegneria in combination with other genomes in the tribe. Many Elymus species are tetraploids containing the StY genomes. It is thought that polyploidization characterizes the speciation of the genus in which the Y is considered as another key genome. Based on data from cytological, genome in situ hybridization, and molecular studies, we hypothesized an endo-allopolyploidy origin of the StY-genome species from the autotetraploid Pseudoroegneria species. To test this hypothesis, we amplified, cloned, and sequenced five single-copy nuclear genes (i.e., alcohol dehydrogenase 1–3, Adh1–Adh3, RNA polymerase II, Rpb2; and Waxy) from Elymus, Pseudoroegneria, and Hordeum species. The phylogenetic trees constructed based on the sequencing analyses of all genes indicated that diploid and autotetraploid Pseudoroegneria species were closely related, although with considerable genetic variation in tetraploids. In addition, the StY-genome Elymus species tended to have a close relationship with the diploid and autotetraploid Pseudoroegneria species, although different phylogenetic relationships among the gene trees were detected. These results indicated that the StY-genome species may have an autotetraploid origin and experienced recurrent hybridization. The complex St genomes in Pseudoroegneria in the polyploid state may gain more opportunities for within-species differentiation and recurrent hybridization. As a result, series modified versions of St genomes evolved into the StY genomes in some Elymus species.