Genetic diversity and hybrid formation in Central European club-mosses (Diphasiastrum, Lycopodiaceae) - New insights from cp microsatellites, two nuclear markers and AFLP.

In Europe, the genus Diphasiastrum (Lycopodiophyta) forms a reticulate network of six diploid taxa, including three parent species (D. alpinum, D. complanatum and D. tristachyum) and three hybrids (D. × issleri, D. × oellgaardii and D. × zeilleri). It was not clear if the hybrids arose once or repeatedly, if they have reproductive competence and if backcrossing occurs. We addressed these questions by analysing 209 accessions for chloroplast microsatellites (cp), two nuclear markers (introns of the RPB and LFY genes) and AFLP. For D. complanatum we show a sexual life cycle with alternation of generations: the gametophytic DNA amount is half of that of the sporophyte. With the exception of a single accession all hybrids display one of the two parental cp haplotypes; their frequencies do not differ significantly from a 1:1 ratio. Genotypes of nuclear markers are species-specific, displaying 2/4/1 (RPB) and 1/8/1 alleles (LFY) for the three parents mentioned above; all hybrids have one allele from each parent. All three hybrid taxa apparently represent independent F1 crosses. Hybridisation occurs bidirectional; no evidence for recent backcrossing was found. Asexual reproduction via agamospory is at least rare, since AFLP showed all hybrid plants to be different.

Flow cytometry confirms reticulate evolution and reveals triploidy in Central European Diphasiastrum taxa (Lycopodiaceae, Lycophyta).

BACKGROUND AND AIMS: Interspecific Diphasiastrum hybrids have been assumed to be homoploid and to produce well-formed spores serving sexual reproduction. If this were the case, forms intermediate between hybrids and parents or hybrid swarms should be expected. The purpose of this study was: (1) to check whether homoploidy consistently applies to the three hybrids throughout their Central European range; (2) to examine whether their genome sizes confirm their parentage as assumed by morphology; and (3) to perform a screening for detection of ploidy levels other than diploid and variation in DNA content due to backcrossing. METHODS: Flow cytometry was used first to measure the relative DNA values [with 4',6-diamidino-2-phenylindole (DAPI) staining] and ploidy level as a general screening, and secondly to determine the absolute DNA 2C values [with propidium iodide (PI) staining] in a number of selected samples with the main focus on the hybrids. KEY RESULTS: A considerable variation of DNA 2C values (5·26-7·52 pg) was detected between the three European Diphasiastrum species. The values of the diploid hybrids are highly constant without significant variation between regions. They are also intermediate between their assumed parents and agree closely with those calculated from their putative parents. This confirms their hybrid origin, assumed parentage and homoploid status. Considerably higher DNA amounts (9·48-10·30 pg) were obtained for three populations, suggesting that these represent triploid hybrids, an interpretation that is strongly supported by their morphology. CONCLUSIONS: Diploid hybrids have retained their genetic and morphological identites throughout their Central European range, and thus no indications for diploid backcrossing were found. The triploid hybrids have probably originated from backcrossing between a diploid gametophyte of a hybrid (derived from a diplospore) and a haploid gametophyte of a diploid parental species. By repeated crossing events, reticulate evolution patterns arise that are similar to those known for a number of ferns.

On the osmoregulation in Atriplex hymenelytra (Torr.) Wats. (Chenopodiaceae).

Aspects of osmoregulation were studied in leaves of irrigated and nonirrigated plants of Atriplex hymenelytra (Torr.) Wats. (Chenopodiaceae) from their natural habitat in Death Valley, California. Using a set of several data concentrations of inorganic electrolytes (Na+, K+, Cl-) and of oxalate in the mesophyll of this salt secreting species were calculated. The osmotic potential resulting from these solutes (under consideration of an empirically estimated osmotic coefficient) is in good agreement with field measurements of the overall osmotic potential in the leaf mesophyll as determined by pressure-volume curves. This indicates that these 4 electrolytes are the main osmotically active solutes. Oxalate is present in comparably high concentrations and is used to achieve ion balance.Organic solutes analyzed include soluble carbohydrates (mono-, di- and oligosaccharides), amino- and organic acids as well as glycinebetaine. Of these, organic- and amino acids (including proline) contribute only little to osmoregulation. Soluble carbohydrates and especially glycinebetaine exhibit concentrations high enough for generating considerable osmotic potentials, at least if these compounds are regarded to be restricted to the cytoplasm acting as compatible solutes.