Reduced density due to logging and its consequences on mating system and pollen flow in the African mahogany Entandrophragma cylindricum

In tropical forests, selective logging removes large trees that are often the main contributors to pollination. We studied pollination patterns of the African mahogany, Entandrophragma cylindricum (Sapelli). We investigated two plots in Cameroon corresponding to three tree densities: unlogged forest (Ndama 2002), a mildly logged forest 1 year after logging (Ndama 2003) and a severely logged forest 30 years after logging (Dimako). We used four microsatellite markers to perform paternity analysis. Selfing remained below 2% in all treatments. Pollen flow was mainly long distance but with some proximity effects. Average observed within-plot pollination distances were 338, 266 and 385 m, and pollination by trees outside the plots was 70% (Ndama 2002), 74% (Ndama 2003) and 66% (Dimako). Despite sampling a limited number of seeds from a limited number of mother trees, we obtained seeds sired by 35.6-38.3% of the potential within-plot pollen donors. While trees 20 cm in diameter contributed to pollination, results in Dimako suggest that individual larger trees contribute more to pollination than small ones. This effect was not detected in the other treatments. The results suggest extensive pollen flow in Sapelli. Hence, in Sapelli, the main limiting factor for regeneration after logging may be a reduction in the number of trees capable of producing seeds rather genetic effects due to limits to pollen dispersal.     

Genetic variation within a fragmented population of swietenia humilis zucc

With large tracts of once continuous forest now modified by human use to leave reduced and highly fragmented stands of trees, the determination of the genetic consequences of forest fragmentation is a priority for ascertaining the conservation value of resultant stands, and in formulating sustainable management strategies. The levels and distribution of genetic diversity over 10 microsatellite loci were investigated within a fragmented population of the neotropical tree Swietenia humilis Zucc. High levels of genetic variation, typical of a highly outcrossing species, were found in all fragments at all loci (mean HE = 0.548). The majority of the variation was within rather than between fragments (RST = 0.032), giving high indirect estimates of gene flow (Nm = 8.9), probably reflecting the genetic structure of the trees present under more continuous forest. A high proportion of loci also showed significant departures from Hardy-Weinberg equilibrium with associated significant levels of FIS. The initial effects of fragmentation were, however, seen in the fragments through the loss of low-frequency alleles present in the continuous 'control' stand. The percentage of this allelic loss increased with a decrease in fragment size.     

Impacts of forest fragmentation on the mating system and genetic diversity of white spruce (Picea glauca) at the landscape level

We studied the mating system of white spruce (Picea glauca) in a landscape fragmented by agriculture in northern Ontario, Canada. We sampled 23 stands that ranged in size from 1 to >500 trees isolated by 250-3000 m from the nearest other stand. Six polymorphic allozyme loci from four enzyme systems were used to genotype approximately 10 000 embryos from 104 families. We detected no allele frequency heterogeneity in the pollen pool among stands or families (Phi(FT)=-0.025). Overall, estimates of outcrossing were high (t(m)=94% and mean t(s)=91%) but significantly different from unity. Bi-parental inbreeding (t(m)-t(s)=3.2%) was low but significantly different from zero. Allozyme-based outcrossing estimates did not differ significantly among three stand-size classes (SSCs): small (<10 trees), medium (10-100 trees) and large (> or =100 trees). The number of effective pollen donors was high in all SSCs, but was significantly lower in small stands (N(ep)=62.5) than in medium-sized and large stands (both N(ep)=143). The primary selfing rate was significantly higher in medium stands than in large stands. We found no significant difference in genetic diversity measures in the filial (seed) population among SSCs. Overall, these results indicate that white spruce stands in this fragmented landscape are resistant to genetic diversity losses, primarily through high pollen-mediated gene-flow and early selection against inbred embryos. We discuss the importance of using seed data, in conjunction with genetic data, to evaluate the impacts of fragmentation on natural populations.     

Allozyme variation of seed embryos and mating system in relict populations of Scots pine (Pinus sylvestris L.) from the Kremenets Hill Ridge and Maloe Poles'e

Allozyme variation at ten polymorphic loci and mating system was studied in three small isolated relict populations (4.4 to 22 ha) and in three artificial stands of Pinus sylvestris from the Kremenets Hill Ridge and Maloe Poles'e. It was established that the mean heterozygosity of 130 to 140 year-old trees from natural populations (H(O) = 0.288; H(E) = 0.277) was substantially lower, compared to 30 to 40 year-old trees from artificial stands (H(O) = 0.358; H(E) = 0.330). The observed heterozygosity of seed embryos (H(O) = 0.169 and 0.180) was substantially lower than of the mature trees from populations and artificial stands, respectively. In the embryo samples, irrespectively of the forest stand origin, substantial hetedrozygote deficiency was observed (at six to eight loci), compared to the Hardy-Weinberg expectations. The proportion of cross pollination in the populations and artificial stands was low, t(m) = 0.588 to 0.721; and t(m) = 0.455 to 0.837, respectively.     

Genetic diversity impacts of forest fires,forest harvesting,and alternative reforestation practices in black spruce (Picea mariana)

Benchmarks were established for genetic diversity inherent in natural mature populations, and genetic diversity impacts of forest fires, clearcut harvesting and alternative natural and artificial silvicultural regeneration practices were determined in black spruce (Picea mariana). Allozymes of 32 loci were used to determine and compare genetic diversity and genetic relationships of adjacent or nearby four stand types: post-fire natural mature (FNM), post-fire natural young (FNR), post-harvest natural young (HNR) and post-harvest plantation (PLT), of black spruce at each of the four study sites located in two ecoregions in Manitoba: Ecoregion 90-Lac Seul Upland (Eastern) and Ecoregion 158 - Mid-Boreal Lowland (Northern). Both allelic- and genotypic-based genetic diversity parameters, as well as latent genetic potential, were determined. Black spruce populations showed typical moderate to high levels of allozyme genetic diversity. The mean genetic diversity parameters over the 16 black spruce populations sampled were as follows: percent loci polymorphic - 67%, mean number of alleles per locus - 2.52, effective number of alleles per locus - 1.70, observed heterozygosity - 0.222, expected heterozygosity - 0.308, mean number of observed genotypes per locus - 3.65, mean number of expected genotypes per locus - 5.03, genotype additivity (observed) - 116.8, genotype additivity (expected) - 161, genotype multiplicity (observed) - 6.16 x 10(15), genotype multiplicity (expected) - 2.06 x 10(19) and latent genetic potential - 26.12. The four stand types (FNM, FNR, HNR and PLT) had comparable and statistically similar genetic diversity levels at each of the four study sites as well as overall. No significant differences in black spruce genetic diversity levels were observed between the two ecoregions in Manitoba, as well as between the post-fire and post-harvest regenerated stands. No particular order of genetic relatedness among the four stand types was observed. Black spruce populations showed some sort of site-related differentiation in their genetic constitution. Allelic heterogeneity and genetic distances among populations within stand types and among four stand types suggest that the genetic diversity was maintained at the landscape level in black spruce. The results of our study demonstrate that forest fires and currently used clearcut harvesting, and alternative natural and artificial silvicultural regeneration practices, do not adversely affect genetic diversity in black spruce, and that the genetic diversity effects of clearcut harvesting are not significantly different from those due to forest fires in black spruce.     

Molecular analysis of natural root grafting in jack pine (Pinus banksiana) trees: how does genetic proximity influence anastomosis occurrence?

Natural root grafting has been observed in more than 150 tree species where up to 90 % of trees could be interconnected within a stand. Intraspecific root grafting was previously found in Pinus banksiana stands, ranging from 21 to 71 % of trees grafted with one another. It is not known why root grafting is frequent in some species and not in others, or why not all roots that cross form root grafts. We investigated genetic diversity of grafted and non-grafted trees to determine if there was a relationship between genetic distance and the probability of forming natural root grafts. Seven plots were hydraulically excavated in four natural forest stands and three plantations of P. banksiana in the western boreal forest of Quebec, Canada. At pairs scale, we studied the effect of geographic and genetic distances on root grafting occurrence. At stand level, we analysed the effect of tree density, soil type, stand type and mean pairwise relatedness on the mean number of grafts per tree and on the percentage of grafted trees per plot. At pairs scale, our analysis revealed that root grafting presence was influenced by spatial distance between trees and less importantly, by genetic distance between individuals. At stand level, root grafting frequency was correlated with stand type (greater in naturally regenerated stands), but not with genetic diversity between individuals. In conclusion, root grafting appears to be principally linked to tree proximity and slightly to genetic proximity between individuals.     

Population structure, genetic diversity, and clone formation in Quercus chrysolepis (Fagaceae)

Stands of canyon live oak (Quercus chrysolepis, Fagaceae) are maintained for fuelwood, fire management, recreation, and as habitat for wildlife. Information about the link between the oak's reproductive ecology and its extent of genetic diversity is important in developing land management policies that will maintain the long-term viability of populations. Basal sprouting is the primary means of reproduction following fire or cutting, and stands frequently include groups of visibly connected trees in a clustered distribution that suggests cloning. We determined the extent to which clusters of trees were clonal and defined the spatial pattern and diversity of genotypes for six populations across nearly the entire east-west extent of the San Bernardino Mountains in southern California. We mapped over 100 trees at each of five sites and genotyped each tree for allozymes at seven polymorphic loci. We identified clones using these multilocus genotypes and detected an average of 34.4 ± 7.3 (SD) clones per site, most of which had unique genotypes. In general, clustered trees belong to single clones and most clones consist of few trees (mean = 3.4 ± 0.6 trees per clone). However, clone size increased significantly with increased individual heterozygosity, suggesting that selection may favor highly heterozygous clones. Clonal diversity and evenness were high relative to reports for most other clonal species; an average of 97% of clones had distinct genotypes, and Simpson's index of diversity averaged 0.95 ± 0.02. Population genetic analyses of 319 clones from six sites revealed high genetic diversity within sites (mean HS = 0.443). Only a small proportion of the total genetic diversity was explained by variation among sites (mean GST = 0.018), which is consistent with high gene flow among sites (Nm = 9.5). We found no significant substructure among plots within sites, and fixation indices within sites were generally small, suggesting that either little inbreeding occurs, and/or few inbred progeny survive. However, spatial autocorrelation analysis of clones indicated fine-scale genetic structure at distances under 4 m, possibly due to limited seed dispersal. Our data suggest that guidelines for seed collection of canyon live oak for use in restoration can be specified in a manner similar to that recommended for conifer species within the region studied.     

Genetic Variability of Wild Cherry (Prunus avium L.) Seed Stands in Slovenia as Revealed by Nuclear Microsatellite Loci

Microsatellite markers were used to describe the genetic variability of four seed stands of wild cherry (Prunus avium L.). One hundred and thirty one individuals were genotyped at ten nuclear microsatellite loci. Total genetic diversity was high (H _E = 0.704), while differences between stands were small but significant (F _ST = 0.053, G′ _ST = 0.234). There was a significant amount of clonal reproduction in one stand, with only 11 genotypes identified among 36 trees. One stand showed a significant excess (F _IS = −0.044) of heterozygosity, and one showed a deficit (F _IS = 0.044). Our results demonstrate the importance of taking into account the biological and genetic characteristics of species in forest management, especially when determining a new seed stand. The small genetic differences found between seed stands indicate that a large number of stands are not required. However, they should be carefully selected and should possess adequate genetic variability to ensure low relatedness between seed trees.     

Effects of Logging on the Genetic Diversity of Quercus tiaoloshanica Chun et Ko in a Tropical Montane Forest of Hainan Island, Southern China

Quercus tiaoloshanica Chun et Ko, which has a narrow range of distribution, is one of the important endemic species of the tropical montane rain forest on Hainan Island, southern China. Long-term logging and habitat destruction have resulted in population decline and distribution retreat of Q. tiaoloshanica. To determine the impact of logging on the genetic diversity of Q. tiaoloshanica, the authors investigated the genetic structures using amplified fragment length polymorphism markers in four regenerated stands after logging and in one unlogged stand. Compared with the unlogged stand, the effective number of alleles per locus dropped by 1.0% in selective logging stands and by 2.0% in clear logging stands, corresponding to reductions of 3.8% and 5.2%, respectively, in mean Nei＇s gene diversity and 2.9% and 3.5%, respectively, in mean Shannon diversity index. No substantial genetic erosion was detected in any of the regenerated stands owing to the high tree density and high heterogeneity of the Q. tiaoloshanica stands investigated. Meanwhile, there was no natural regeneration of the species observed in a Dacrydium pierrei Hickel plantation 700 m away from the regenerated stands, suggesting the limited ability of seed dispersal of Q. tiaoloshanica. Clear logging should be undertaken cautiously because the total number of plant species dropped by 15.2% in the clear-logged stands compared with the unlogged stand. To conserve the genetic diversity of this species, as well as the plant biodiversity of tropical forests, the habitats of Q. tiaoloshanica should be protected against exploitation in terms of agricultural or other forms of land use, and some mature trees should be preserved as seed sources to maintain an adequate regeneration base for this species in the management of logging.     

Reproductive and genetic characteristics of rare,disjunct pitch pine populations at the northern limits of its range in Canada

Pitch pine, Pinus rigida Mill., is a rare species in Canada, existing as a disjunct population in the St. Lawrence River Valley in eastern Ontario and two northern outlier stands in southern Quebec along Canada's southern border with the United States. Reproductive and genetic characteristics of these small, scattered stands were investigated to develop a foundation for management and restoration in the event of range expansion northwards under anticipated climate warming. Seed yields and seed quality appear to be comparable to other eastern conifers, and to pitch pine at the center of its geographic range. For seed and seedling growth traits, most of the variation was attributable to differences among trees within stands and, to a lesser extent, among stands within a population; whereas the population effect was non-significant. For reproductive traits, such as numbers of filled and empty seeds per cone, reproductive efficiency, and inbreeding estimates, high levels of variation (ranging from 26% to 33%) were found among stands, suggesting that stand structural features, such as stand size and tree density within stands, play an important role in pollination environment and overall reproductive success. Estimates of genetic diversity at 32 allozyme gene loci indicate that these small, isolated stands have maintained relatively high levels of genetic diversity compared with populations at the center of its geographic range, and also relative to other widely dispersed eastern conifers. The relatively high levels of viable seed production and genetic diversity in native pitch pine populations indicate that native Canadian populations may be suitable seed sources for species restoration and range expansion in Canada.     

SHORT COMMUNICATION: Do farmers reduce genetic diversity when they domesticate tropical trees? A case study from Amazonia

Agroforestry ecosystems may be an important resource for conservation and sustainable use of tropical trees, but little is known of the genetic diversity they contain. Inga edulis, a widespread indigenous fruit tree in South America, is used as a model to assess the maintenance of genetic diversity in five planted vs. five natural stands in the Peruvian Amazon. Analysis of five SSR (simple sequence repeat) loci indicated lower allelic variation in planted stands [mean corrected allelic richness 31.3 (planted) and 39.3 (natural), P = 0.009]. Concerns regarding genetic erosion in planted Amazonian tree stands appear valid, although allelic variation on-farm is still relatively high.     

Genetic analysis of forest species <Emphasis Type="Italic">Eugenia uniflora</Emphasis> L. through of newly developed SSR markers

Nine microsatellite loci for genetic analysis of three populations of the tropical tree Eugenia uniflora L. (pitanga or Brazilian cherry) from fragments of semideciduous forest were developed. We used the technique of building a (GA) _n and (CA) _n microsatellite-enriched library by capture with streptavidin-coated magnetic beads. We assessed the polymorphism of seven microsatellites in 84 mature trees found in three areas (Ribeirão Preto, Tambaú and São José do Rio Pardo), highly impacted by the agricultural practices, in a large region among Pardo river and Mogi-Guaçu river basins, in state of São Paulo, Brazil. All loci were polymorphic, and the number of alleles was high, ranging from 6 to 24, with a mean of 14.4. All stands showed the same high level of genetic diversity (mean H _E  = 0.83) and a low genetic differentiation (mean F _ST = 0.031), indicating that genetic diversity was higher within rather than among populations. Seven of the nine loci were highly variable, and sufficiently informative for E. uniflora. It was concluded that these new SSR markers can be efficiently used for gene flow studies.     

Limited Reintroduction Does Not Always Lead to Rapid Loss of Genetic Diversity: An Example from the American Chestnut (Castanea dentata; Fagaceae)

In restoring species, reasons for introducing limited numbers of individuals at different locations include costs of introduction and maintenance, limited founder supply, and risk “bet hedging.” However, populations initiated from few founders may experience increased genetic drift, inbreeding, and diversity loss. We examined the genetic diversity of an isolated stand of more than 5,000 American chestnut trees relative to that of the 9 surviving stand founders (out of 10 total) planted in the 1880s. We used minisatellite DNA probes to reveal 84 genetic markers (circa 24 loci) among the nine founders, and their genetic diversity was compared with three separate plots of descendant trees, as well as with two natural stands. The descendants were circa 7.3% more heterozygous than the founders (mean estimated H= 0.556 vs. 0.518, respectively; p < 0.0001). Genetic differentiation was not pronounced (F_ST < 0.031), and no markers, including those at low frequency among the founders, were lost in the descendants. The founders and natural transects were not significantly different in H or similarity (mean proportion of bands shared). Special planting or mating protocols for establishment of a vigorous American chestnut population from a low number of founders may not be required to avoid strong effects of genetic drift and inbreeding. These results demonstrate that loss of genetic diversity following reintroduction of a limited number of founders is not always inevitable, such as this case where the species is highly outcrossing, expression of heterozygous advantage may occur, the original founders remain as gene contributors over generations, and the establishing population expands constantly and rapidly.     

Regeneration mode affects spatial genetic structure of Nothofagus dombeyi forests

Disturbance may generate population bottlenecks by reducing population size and the number of founders establishing a new colony. We tested the hypothesis that the scale of disturbance affects the levels of genetic diversity and the spatial distribution of genotypes in naturally regenerating stands of Nothofagus dombeyi, an evergreen angiosperm tree, in northwestern Patagonia. At similar spatial scales, we predicted that old-growth stands characterized by fine-scale gap phase dynamics would be genetically diverse due to restricted gene flow among temporal and spatially isolated gaps. In contrast, young massively regenerated postfire cohorts resulting from coarse-scale disturbances would be genetically more homogeneous. At each of three paired old-growth and postfire stands a minimum of 50 trees were mapped and sampled within 1 ha. Fresh tissue was collected for isozyme analysis from a total of 361 trees along with tree cores and diameters. Tree age distributions reflected the dominant modes of regeneration. Six out of nine analysed loci were polymorphic. Mean genetic diversity parameters were greater but not significant in mature stands. Fixation indices suggested significant heterozygous deficit at two-thirds of possible tests indicating a Wahlund effect due to local recruitment of related seeds. F(ST) indicated moderate between-stand divergence. Mature stands concentrated half of positively like joins and yielded significant (P < 0.05) autocorrelation coefficients at small distance classes (< 20 m). Fine-scale patch dynamics within mature stands favours the maintenance of fine-scale genetic structure as a result of shade intolerance and local seed dispersal. Conversely, postfire stands suffer the effects of genetic drift given that a few reproductive trees produce a somewhat impoverished and genetically uniform progeny. Bottleneck effects will depend upon the density of remnant trees which could also be a function of the severity of fire.     

Temporal and spatial genetic structure in Vitellaria paradoxa (shea tree) in an agroforestry system in southern Mali

Ten microsatellite loci were used to investigate the impact of human activity on the spatial and temporal genetic structure of Vitellaria paradoxa (Sapotaceae), a parkland tree species in agroforestry systems in southern Mali. Two stands (forest and fallow) and three cohorts (adults, juveniles and natural regeneration) in each stand were studied to: (i) compare their levels of genetic diversity (gene diversity, HE; allelic richness, Rs; and inbreeding, FIS); (ii) assess their genetic differentiation (FST); and (iii) compare their levels of spatial genetic structuring. Gene diversity parameters did not vary substantially among stands or cohorts, and tests for bottleneck events were nonsignificant. The inbreeding coefficients were not significantly different from zero in most cases (FIS = -0.025 in forest and 0.045 in fallow), suggesting that the species is probably outbreeding. There was a weak decrease in F(IS) with age, suggesting inbreeding depression. Differentiation of stands within each cohort was weak (FST = 0.026, 0.0005, 0.010 for adults, juveniles and regeneration, respectively), suggesting extensive gene flow. Cohorts within each stand were little differentiated (FST = -0.001 and 0.001 in forest and fallow, respectively). The spatial genetic structure was more pronounced in fallow than in forest where adults showed no spatial structuring. In conclusion, despite the huge influence of human activity on the life cycle of Vitellaria paradoxa growing in parkland systems, the impact on the pattern of genetic variation at microsatellite loci appears rather limited, possibly due to the buffering effect of extensive gene flow between unmanaged and managed populations.     

The effects of genetic diversity,climate and defoliation events on trembling aspen growth performance across Canada

Tree genetic makeup may provide an important control of growth dynamics; however, no studies have previously attempted to evaluate its effects in natural trembling aspen stands. In this study, we examined the relative contribution of genetics (i.e. clonal diversity, observed heterozygosity) and environmental conditions (i.e. insects, climate) on aspen growth as represented by mean inter-tree correlation (RBAR), tree basal area increment (TBAI) and inter-annual growth variability (MS). We sampled 440 trees in 22 even-aged natural stands dominated by aspen along an east-west continental gradient of decreasing annual precipitation in the Canadian boreal forest. Linear and mixed-effect models tested the relationships between tree growth, genetics and environmental factors. We showed that clonal diversity and number of years with forest tent caterpillar (FTC) defoliation (N_FTC) reduced and increased the level of growth synchronicity (RBAR), respectively. Clonal diversity explained 30 % of variation in RBAR among sites. TBAI was positively influenced by high moisture conditions while N_FTC and climate explained the variation in MS among trees for each site. No genetic effect could explain either TBAI or the MS variation. Climate and N_FTC drive annual growth variability in trembling aspen at stand and subcontinental scales. Tree genetic makeup contributed to these dynamics, the annual growth dynamics of multi-clonal stands being less homogeneous than those of monoclonal stands. Maintaining diverse aspen stands may ensure a wider range of growth responses to environmental variability, which in turn may help maintain resilience of aspen stands under future climate.     

Microsatellite DNA analysis of genetic effects of harvesting in old-growth eastern white pine (Pinus strobus) in Ontario, Canada

Microsatellite DNA markers from 13 simple sequence repeat (SSR) loci were used to compare genetic diversity between preharvest pristine and postharvest residual gene pools of two adjacent virgin, old-growth ( approximately 250 years) stands of eastern white pine (Pinus strobus L.) in Ontario. There was concurrence in genetic diversity changes in the postharvest gene pools of the two stands. The total and mean numbers of alleles detected in each stand were reduced by approximately 26% after tree density reductions of approximately 75%. Approximately 18 and 21% of the low-frequency (0. 25 > P > or = 0.01) alleles and 76 and 92% of the rare (P < 0.01) alleles were lost from residual stands A and B, respectively, after harvesting. Multilocus gametic diversity was reduced by 38 and 85% and genotype additivity by approximately 50% in the residual stands after harvesting. Latent genetic potential of each stand was reduced by approximately 40%. Although heterozygosity was reduced (1-5%) in the postharvest residual stands, the reductions were not substantial and not comparable to those using other genetic diversity measures. The reductions in genetic diversity measures were slightly higher than those theoretically expected in postbottleneck populations according to drift theory. In the absence of substantial gene migration that could ameliorate the genetic losses, the ability of the postharvest white pine gene pools to adapt to changing environmental and disease conditions may have been compromised. The microsatellite DNA results for genetic effects of harvesting in old-growth eastern white pine stands were similar to those that we reported earlier from allozyme analysis (Buchert et al. 1997). The results indicate that silvicultural practices should ensure that the gene pools of remaining pristine old-growth stands are reconstituted in the regenerating stands.     

Effects of reforestation methods on genetic diversity of lodgepole pine: an assessment using microsatellite and randomly amplified polymorphic DNA markers

 We examined the effects of different methods of forest regeneration on the genetic diversity of lodgepole pine (Pinus contorta var ‘latifolia’) using two different DNA-based molecular markers [randomly amplified polymorphic DNA (RAPDs) and microsatellites or simple sequence repeats (SSRs)]. Genetic diversity was estimated for 30 individuals in each of four populations for the following three stand types: (1) mature lodgepole pine (>100 years); (2) 20- to 30-year-old harvested stands left for natural regeneration; (3) 20- to 30-year-old planted stands (4 stands of each type); and one group of 30 operationally produced seedlings. There was no significant effect of stand type on expected heterozygosity, although allelic richness and diversity were much higher for SSRs than for RAPDs. Expected heterozygosity ranged from 0.39 to 0.47 based on RAPDs and from 0.67 to 0.77 based on SSRs. The number of alleles per locus for SSRs ranged from 3 to 34 (mean 21.0), and there was a significant relationship between sequence repeat length and the number of alleles at a locus. Both marker types showed that over 94% of the variation was contained within the populations and that the naturally regenerated stands sampled had lower (not significant) expected heterozygosity than the planted or unharvested stands. The group of seedlings (assessed by RAPDs only) had expected heterozygosity and allele frequencies similar to those of the unharvested stands. Genetic distance measures were higher than obtained previously in the species using isozyme markers. There was no correlation between the two marker types for pair-wise genetic distances based on populations analyzed by both methods. Pair-wise genetic distance measures and an ordination of allele frequencies for both marker types showed little effect of geographic location or stand type on genetic similarity. Received: 18 August 1998 / Accepted: 17 September 1998     

Allozyme variation of seed embryos and mating system in relict populations of scots pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) from the Kremenets Hill Ridge and Maloe Poles’e

Allozyme variation at ten polymorphic loci and mating system was studied in three small isolated relict populations (4.4 to 22 ha) and in three artificial stands of Pinus sylvestris from the Kremenets Hill Ridge and Maloe Poles’e. It was established that the mean heterozygosity of 130 to 140 year-old trees from natural populations (H _O = 0.288; H _E = 0.277) was substantially lower, compared to 30 to 40 year-old trees from artificial stands (H _O = 0.358; H _E = 0.330). The observed heterozygosity of seed embryos (H _O = 0.169 and 0.180) was substantially lower than of the mature trees from populations and artificial stands, respectively. In the embryo samples, irrespectively of the forest stand origin, substantial hetedrozygote deficiency was observed (at six to eight loci), compared to the Hardy-Weinberg expectations. The proportion of cross pollination in the populations and artificial stands was low, t _m = 0.588 to 0.721; and t _m = 0.455 to 0.837, respectively.     

Extensive pollen immigration and no evidence of disrupted mating patterns or reproduction in a highly fragmented holm oak stand

Aims Forest fragmentation and reduced tree population densities can potentially have negative impacts on mating patterns, offspring genetic diversity and reproductive performance. The aim of the present study is to test these hypotheses comparing an extremely fragmented, low tree density (~0.02 trees/ha) holm oak (Quercus ilex L.) stand from Central Spain with a nearby high tree density stand (~50 trees/ha).Methods We genotyped adult trees and seeds from the low-density stand (436 seeds from 15 families) and the high-density stand (404 seeds from 11 families) using nine microsatellite markers. With these data, we performed paternity analyses, determined pollen flow, mating patterns and pollen pool structure, and estimated progeny genetic diversity in both stands. We also studied seed set and production and performed a pollen supplementation experiment to determine whether reduced tree density has limited foreign pollen availability.Important findings We have found extensive pollen immigration (>75%) into the low tree density stand and Monte Carlo simulations revealed that pollen moves larger distances than expected from null models of random dispersal. Mating patterns and differentiation of pollen pools were similar in the high-density stand and the low-density stand but we found higher inter-annual differentiation of pollen pools in the former. Progeny genetic diversity and self-fertilization rates did not differ between the low-density stand and the high-density stand. Seed set rates were significantly lower in the low-density stand than in the high-density stand and experimental cross-pollen supplementation evidenced that foreign pollen availability is indeed a limiting factor in the former. However, seed crops did not differ between the low-density stand and the high-density stand, indicating that limitation of foreign pollen is not likely to be of great concern in terms of reduced seed production and potential recruitment. Poor forest regeneration due to other ecological and human factors is probably a more important threat for the persistence of fragmented and low tree density stands than reduced pollen flow and only extremely small and isolated tree populations would be expected to suffer severe loss of genetic diversity in the long term.