A rapid means of sex identification inSilene latifolia by use of flow cytometry

Sex identification in dioecious plants using nonflowering material would have broad applications in both basic and applied research. We present a method using flow cytometry for diagnosing the sex of the dioecious speciesSilene latifolia Poiret (Caryophyllaceae) by means of sexual differences in nuclear DNA content and base-pair composition. Males have a significantly larger genome, attributable to the known sex-chromosome heteromorphism. Males and females also differ in the AT/GC composition, attributable to differences in non-recombining portions of the sex chromosomes. The two measures enable assignment of individuals to sex with a combined error rate of 9%. These results forS. latifolia indicate useful directions for future research into sex diagnostics for other dioecious species.     

Plant population dynamics and species turnover on small islands near Karpathos (South Aegean,Greece)

The vascular flora of small offshore islets around Karpathos (Dodecanesos, Greece) has been surveyed and monitored during six consecutive years. The present study focuses on four islets that are inhabited by the islet specialist Silene holzmannii (Caryophyllaceae), a species that has a widely scattered distribution in the Aegean where it exclusively occurs on very small islands. Repeated census of its populations, partly following controlled extirpation, reveals extreme variation in number of individuals. Sowing experiments involving various species demonstrate the low probability of successful new colonizations. Differences in the amount of apparent turnover of species as well as the occurrence of both stable micro-populations and mass-populations are discussed.     

IDENTICAL MAJOR GENE LOCI FOR HEAVY METAL TOLERANCES THAT HAVE INDEPENDENTLY EVOLVED IN DIFFERENT LOCAL POPULATIONS AND SUBSPECIES OF SILENE VULGARIS

Heavy metal tolerant Silene vulgaris plants, originating from different metalliferous sites in Germany and one in Ireland, were crossed to each other and to nontolerant plants from a nonmetalliferous site in The Netherlands. Analysis of the crosses suggested that there were two distinct major gene loci for zinc tolerance among a total of five tolerant populations. The tolerance loci for zinc, copper, and cadmium in the Irish plants were shown to be identical with those in the German populations. It is argued that the occurrence of common major genes for tolerance among different geographically isolated populations must have resulted from independent parallel evolution in local nontolerant ancestral populations. Each of the tolerances studied seems to be controlled by only a few specific major genes.     

Heavy metal-induced accumulation of free proline in a metal-tolerant and a nontolerant ecotype of Silene vulgaris

Accumulation of free proline in response to Cu, Cd and Zn was studied in nontolerant and metal-tolerant Silene vulgaris (Moench) Garcke. In the nontolerant ecotype these metals induced a massive accumulation of proline, especially in the leaves. When compared at equimolar concentrations in the nutrient solution, Cu was the most effective inducer, followed by Cd and Zn, respectively. However, when compared at equal toxic strength, as estimated from the degree of root growth inhibition, proline accumulation decreased in the order Cd > Zn > Cu. The threshold exposure levels for proline accumulation coincided with the highest no-effect-concentrations for root growth. In the metal-tolerant ecotype the constitutive proline concentration in the leaves was 5 to 6 times higher than in the nontolerant ecotype. Exposure to Cu and Zn, however, was without any effect on the leaf proline concentration, even at exposure levels that caused a 50% root growth inhibition. Only Cd, when present at concentrations above the highest no-effect-concentration for root growth, induced a further increase of the leaf proline content. Reducing transpiration by placing the plants under a transparent polyethylene cover almost completely inhibited proline accumulation, even at metal accumulation rates in the leaves that caused a 10-fold increase of the proline level in leaves of uncovered plants. The results demonstrate that metal-induced proline accumulation depends on the development of a metal-induced water deficit in the leaves. Differential metal-induced proline accumulation in distinctly metal-tolerant ecotypes is a consequence, rather than a cause of differential metal tolerance.     

Reproduction and Pollination in Central European Populations of Silene and Saponaria Species

Flower morphology, flowering phenology, flower visitors, reproductive systems, and stigmatic receptivity of six species of Silene and Saponaria (Silene alba, S. dioica, S. vulgaris, S. nutans, S. noctiflora. Saponaria officinalis) were studied from April to October 1993 and from April to June 1994 in natural populations around Giessen in Hessen/Central Germany and, additionally, in individuals grown from seeds in the Botanical Garden of the University of Giessen. With the exception of Saponaria officinalis and S. noctiflora, all species were regularly visited and pollinated by crepuscular and nocturnal moths and hawkmoths, but only one species, S. alba, was exclusively pollinated by these night-active insects. The other species showed mixed pollination syndromes in which nocturnal and diurnal insects both promoted pollen transfer. Geitonogamy or even autogamy occurred in the gynodioecious and hermaphrodite species S. vulgaris, S. nutans, S. noctiflora, and Saponaria officinalis. Silene noctiflora, the only annual species, is pseudocleistogamous; the majority of its flowers did not open, and fruit set occurred after selfing in bud.     

Ageing of theSilene coeli-rosa L. shoot apex under non-inductive conditions: Changes in morphology,mitotic index,and polypeptide composition

Summary Ageing was studied in the shoot apex of the long day plant,Silene coeli-rosa by maintaining it in non-inductive short day conditions for 170 days. The dimensions, the zonation, and the polypeptidic pattern of the shoot apex, and the rate of leaf initiation were altered in 170-day-old plants compared with young plants grown under the same conditions (28 days). In aged plants, the number of cells increased in all shoot apical zones and, notably, the mitotic index increased in the axial zone; however, the rate of leaf initiation slowed down. These changes showed some similarities to those during the intermediate phase in quantitative photoperiodic species. The two-dimensional mini-gel electrophoretic study of protein extracts from shoot apices of young and ageing plants maintained in non-inductive conditions revealed 489 common polypeptidic spots, 13 unique to the young state and 24 new ones specific to aged plants. The spots characteristic for each state represented only 3.6% of the total identified polypeptides, but apical development under non-inductive conditions was characterized by qualitative changes in the polypeptide complement.Abbreviations C nuclear DNA complement - 1D gel first dimension gel - LD long day - NEPHGE non equilibrium pH gradient electrophoresis - SD short day - SDS sodium dodecyl sulfate - TCA trichloracetic acid     

Increased resistance to copper-induced damage of the root cell plasmalemma in copper tolerant Silene cucubalus

The relation between copper tolerance and the sensitivity of plants with respect to the effect of copper on the plasmalemma of root cells was studied using plants from one copper sensitive and two copper tolerant populations of Silene cucubalus Wib. In each population, the external copper concentration needed to induce ion leakage (a measure of damage to the permeability barrier) was similar to the highest no-effect-concentration of copper for root growth in that population. At higher concentrations, the degree of root growth inhibition paralleled the rate of ion leakage, the degree of trypan blue staining (a measure of plasmalemma integrity) and the accumulation of lipid peroxidation products. The amount of copper taken up by the plants was inversely related to their level of copper tolerance. Compared to copper sensitive plants, copper tolerant plants showed no increased resistance to either the sulfhydryl reagent N-ethylmaleimide or the free radical-producing compound cumene hydroperoxide.
These results indicate that damage to the permeability barrier of root cells constitutes the primary effect of copper toxicity in both sensitive and tolerant plants, and that copper tolerance is coupled to the ability of the plants to prevent such damage. This ability might depend on exclusion of copper by the root cell plasmalemma.     

Can the preservation of historical relicts permit the conservation of endangered plant species? The case of <Emphasis Type="Italic">Silene sennenii</Emphasis> (Caryophyllaceae)

Allozyme electrophoresis was used to measure levels and distribution of genetic diversity in the critically endangered (CR) Silene sennenii (Caryophyllaceae), a narrow endemic plant species found in northeastern Catalonia (Spain). At present, only 5 populations remain, containing no more than 5,000 individuals, which are subjected to several human pressures such as habitat fragmentation and land use changes. Interestingly, the largest population is located in and around a fortress built in the XVIII century, a circumstance which might facilitated its preservation until now. From the 21 satisfactorily interpreted loci, low levels of genetic variation were detected (P = 20.9, A = 1.31 and H _e = 0.071), which may be related to small population size, isolation, and fragmentation of extant populations. Moderate to high levels of inbreeding were also found, probably as consequence of the population’s genetic structuring. The conservation of the population located in the fortress would allow the preservation of all the alleles detected at species level; nevertheless, conservation of other populations, coupled with the inclusion of S. sennenii in the Spanish Catalogue of Endangered Species, would also be desirable in order to ensure the long-term survival of the species.