High nuclear genetic diversity, high levels of outcrossing and low differentiation among remnant populations of Quercus petraea at the margin of its range in Ireland

• Background and Aims Quercus petraea colonized Irelandafter the last glaciation from refugia on mainland Europe. Deforestation,however, beginning in Neolithic times, has resulted in small,scattered forest fragments, now covering less than 12 000 ha. • Methods Plastid (three fragments) and microsatellitevariation (13 loci) were characterized in seven Irish populationssampled along a north–south gradient. Using Bayesian approachesand Wright’s F-statistics, the effects of colonizationand fragmentation on the genetic structure and mating patternsof extant oak populations were investigated. • Key Results All populations possessed cytotypes commonto the Iberian Peninsula. Despite the distance from the refugialcore and the extensive deforestation in Ireland, nuclear geneticvariation was high and comparable to mainland Europe. Low populationdifferentiation was observed within Ireland and populationsshowed no evidence for isolation by distance. As expected ofa marker with an effective population size of one-quarter relativeto the nuclear genome, plastid variation indicated higher differentiation.Individual inbreeding coefficients indicated high levels ofoutcrossing. • Conclusions Consistent with a large effective populationsize in the historical migrant gene pool and/or with high geneflow among populations, high within-population diversity andlow population differentiation was observed within Ireland.It is proposed that native Q. petraea populations in Irelandshare a common phylogeographic history and that the presentgenetic structure does not reflect founder effects.     

Species distinction in Irish populations of Quercus petraea and Q. robur: morphological versus molecular analyses

BACKGROUND AND AIMS: Populations of oak (Quercus petraea and Q. robur) were investigated using morphological and molecular (AFLP) analyses to assess species distinction. The study aimed to describe species distinction in Irish oak populations and to situate this in a European context. METHODS: Populations were sampled from across the range of the island of Ireland. Leaf morphological characters were analysed through clustering and ordination methods. Putative neutral molecular markers (AFLPs) were used to analyse the molecular variation. Cluster and ordination analyses were also performed on the AFLP markers in addition to calculations of genetic diversity and F-statisitcs. KEY RESULTS: A notable divergence was uncovered between the morphological and molecular analyses. The morphological analysis clearly differentiated individuals into their respective species, whereas the molecular analysis did not. Twenty species-specific AFLP markers were observed from 123 plants in 24 populations but none of these was species-diagnostic. Principal Coordinate Analysis of the AFLP data revealed a clustering, across the first two axes, of individuals according to population rather than according to species. High F(ST) values calculated from AFLP markers also indicated population differentiation (F(ST) = 0.271). Species differentiation accounted for only 13 % of the variation in diversity compared with population differentiation, which accounted for 27 %. CONCLUSIONS: The results show that neutral molecular variation is partitioned more strongly between populations than between species. Although this could indicate that the populations of Q. petraea and Q. robur studied may not be distinct species at a molecular level, it is proposed that the difficulty in distinguishing the species in Irish oak populations using AFLP markers is due to population differentiation masking species differences. This could result from non-random mating in small, fragmented woodland populations. Hybridization and introgression between the species could also have a significant role.     

Natural hybridisation between Quercus petraea (Matt.) Liebl. and Quercus pubescens Willd. within an Italian stand as revealed by microsatellite fingerprinting

Interspecific gene flow is frequently reported in the genus Quercus . However, interfertile oak species often seem to remain distinct, even within areas of sympatry. This study employed molecular markers to verify, at a fine scale, the presence of interspecific gene flow in a natural population of Quercus petraea and Quercus pubescens . Within a delimited area of 6 ha, all adult trees belonging to the studied oak complex and seeds from a subsample of such trees were collected and analysed using molecular microsatellite markers. A low interspecific genetic differentiation and a high level of interspecific genetic admixture suggested past hybridisation. Paternity inference of seeds allowed the estimation of pollination frequencies from the three groups of pollen donors ( Q. petraea, Q. pubescens , intermediate). We also assayed pollen viability and germinability of each species group. We observed natural hybridisation between Q. petraea and Q. pubescens, with a predominant component in the direction Q. petraea → Q. pubescens : Q. pubescens displayed a higher level of heterospecific pollination by Q. petraea (25.8%) and intermediate morphotypes (14.7%), compared to Q. petraea acting as pollen receptor (with less than 5% heterospecific pollinations). Intermediate 'mother trees' were pollinated in similar proportions by Q. petraea (23.1%), Q. pubescens (37.8%) and intermediate morphotypes (39.1%). The asymmetrical introgression observed for the studied generation may be caused, among other factors, by the relative abundance of trees from each species group in the studied area.     

Ontogeny of the proventitious epicormic buds in Quercus petraea.

In the present work, we described the fate of proventitious epicormic buds on the trunks of 40-year-old Quercus petraea trees and in parallel the vascular trace they produced in the wood. Our results show that small and large individual epicormic buds can survive as buds for 40 years and that both are composed of a terminal meristem and scales. Meristematic areas are detected in the scale axils of small buds; in addition to these meristems the large buds also have secondary bud primordia. The small buds are connected to the pith of the main stem by a unique trace, whereas the large buds are connected by one or multiple traces. A single trace might imply that the whole bud is still alive and multiple traces might indicate that the terminal meristem has died. In the latter case, each trace is connected to a secondary bud of the large bud. The buds found in a cluster are composed of a terminal meristem and scales with axillary meristems in the scale axils. A cluster is connected to the pith of a stem either by a unique trace when it seems to be the result of partial abscission of an epicormic shoot or multiple traces when it might have originated from an epicormic bud in which the terminal meristem has died. Whatever the type of the bud, the vascular trace in the bark is composed of a cambium, secondary xylem and parenchyma cells and the trace present in the wood had parenchyma cells with vestiges of secondary xylem. Each year, the vascular trace should be produced in the bark by the cambium of the tree but not by the bud itself. On 40-year-old Q. petraea, we observed a proliferation of epicormic buds and in parallel a multiplication of the number of vascular traces in the trunk, but the knots caused by the traces of epicormic buds in the wood, either as individuals or in clusters, are minor since their colours are only slightly darker than those of woody rays and they are less than 2 mm in diameter. The knots will appear when epicormic buds develop into shoots. Received: 30 March 1999 / Accepted: 09 June 1999     

Comparison of radial increment and volume growth in stems and roots of Quercus petraea

Dendrochronological studies dealing with roots, stems and branches are very rare or often take the form of short notes. The difficulties of detecting rings and of quantifying the radial growth in roots have already been described for various species. In oak the anatomical root structure differs from stemwood. The roots are radial-porous or diffuse-porous, and there is often no clear distinction between individual rings. In our study visual and radiographic techniques were used to examine radial increment in roots of sessile oak (Quercus petraea L.) which was compared with radial growth in branches and along the stem. Coarse roots were cut from four 30- to 34-year-old trees that had been uprooted mechanically and disks were taken at different distances from the stem-root base. Ring widths were measured in the stem at height of 0.3 m, at breast height (1.3 m), beneath the crown, in branches of the crown, and in roots every 20 cm. The ring widths were cross-dated, and the heterogeneity of growth within a root and within the root system were analysed. Asymmetric growth frequently occurred in roots so that ovals, I-beam and T-beam shapes were developed. With the method used in our study the annual growth layers close to the central cylinder could be distinguished as well as beneath the bark. Pointer years were detected in all sections of the tree and permitted correction of ring widths in roots. Root system, stem and branch showed a basic similarity in their radial sequence of ring width. The annual biomass increment was weaker and more variable with several consecutive changes in the roots than in the stems. The root/shoot ratio reached a minimum rather early, beginning at the cambial age of 20 years. This revised version was published online in June 2006 with corrections to the Cover Date.     

海拔对高山栎光合气体交换和叶性状的影响

理解影响植物分布的式样及过程是生态学研究的中心内容之一,但对许多物种而言,限制其分布的原因还不清楚。为了认识高山栎分布与生理生态特性的关系,我们在不同海拔的4个观测点研究了帽斗栎的光合气体交换、叶氮含量、叶绿素含量和比叶重。由于高的水气压亏缺和气温,帽斗栎的光合作用和蒸腾作用在午间表现出明显的降低现象。帽斗栎的饱和光合速率、水分利用效率、最大羧化速率、最大电子传递速率和氮利用效率在海拔中部比低海拔或高海拔处的为高。不同海拔的叶氮含量在5月份有差异,8月份则没有明显不同。叶片厚度随海拔增加,但叶绿素含量及光合最适温度随海拔升高而降低。帽斗栎光合作用的海拔变化与叶片的生化效率和氮含量有关,而与比叶重无关。研究结果说明,温度的海拔变化对高山栎的光合作用和叶性状有明显影响,最适宜帽斗栎光合碳获取及生长的海拔范围是3180～3610m。     

Presence of triploids among oak species

BACKGROUND AND AIMS: Polyploids, organisms that have multiple sets of chromosomes, are common in certain plant and animal taxa. However, there are only a few reports of intraspecific ploidy variation within the genus Quercus. The aim of the study was to investigate the suspected ploidy level of two oaks that have unusual microsatellite banding patterns. METHODS: Polyploidy was investigated by using microsatellite analysis, stomata length measurements and nuclear DNA content estimation by flow cytometry. KEY RESULTS: Each putative triploid tree has patterns of microsatellite variation unexpected for diploid genomes, with up to three alleles at some loci, significantly longer stomata and 1.5 times more DNA per nucleus compared with diploids. CONCLUSIONS: To our knowledge, this report contains the first evidence for triploidy in Q. petraea and confirmation of this phenomenon in Q. robur. Regardless of the positive or negative aspects of the presence of triploid oaks in forest stands, it is of value to be able to screen for them. This study demonstrates that nuclear microsatellites and estimation of DNA content by flow cytometry can readily be used for this purpose.     

Water in the stems of sessile oak (Quercus petraea) assessed by computer tomography with concurrent measurements of sap velocity and ultrasound emission

Concurrent measurements of heat pulse velocity and ultrasound acoustic emission were performed on the trunks of adult Quercus petraea plants under different water stress conditions. On the trunk section of the plants the wood density was measured non-destructively using a mobile computer tomograph which measures the attenuation of a collimated beam of radiation which traverses the trunk. By scanning the trunk in different directions, it is possible to map wood density in the section. As wood density is strictly correlated with water content, this method allows evaluation of the water content in the trunk section and the water conditions in the different parts of the section. The computer-tomograph technique is non-invasive and is not influenced by climatic fluctuations. A close agreement was found between wood density and heat pulse velocity; the relationship between these two parameters and ultrasound acoustic emission is discussed. Trunks of sessile oak appear well suited as water storage reservoirs playing an important role in tree survival during extended periods of low soil water availability, especially in the context of global climatic change. Here the computer-tomograph methodology is described and suggestions arc made for further research development.     

The effects of liming and gypsum applications on a sessile oak (Quercus petraea (M.) Liebl.) stand at La Croix-Scaille (French Ardennes) I. Site characteristics, soil chemistry and aerial biomass

In a former 45 to 50 year old sessile oak ( Quercus petraea (M.) Liebl.) coppice mixed with birch (Betula pubescens Ehrh.) and rowan (Sorbus aucuparia L.) on a poor acidic forest soil at la Croix-Scaille in the French Ardennes, several liming amendments were applied in 1990 and 1994. Data on soil and soil solution composition, as well as stand growth and foliar composition were collected between 1994 and 1997. All treatments, containing 1.4 t ha-1 equivalent of CaO supplied as lime, gypsum or a mixture of the two, resulted in an increase of cation exchange capacity and base saturation down to 15 cm and for CaSO4 treatments down to 30 to 45 cm, increases of soil pH and Ca concentration at the surface and a decrease of Al concentration in the soil and soil solution in the surface layers. No negative effects like increased nitrate or cation leaching were observed. Although Mg nutrition was not improved by the treatments (not containing Mg), a relative and maintained gain of radial increment of sessile oak in the order of 40% for both lime and gypsum applied, was observed immediately from the first year on, after the application (1991).     

Genetic variation at allozyme and RAPD loci in sessile oak Quercus petraea (Matt.) Liebl.: the role of history and geography

The nuclear genetic variation within and among 21 populations of sessile oak was estimated at 31 RAPD loci in conjunction with previous estimates of variation at eight allozyme loci. The aim of the study was to assess the relative role of isolation-by-distance and postglacial history on patterns of nuclear variation. Because of its small effective population size and maternal transmission, the chloroplast genome is a good marker of population history. Both kinds of nuclear variation (RAPD and allozyme) were therefore compared, first, to the geographical distances among populations and, secondly, to chloroplast DNA restriction polymorphism in the same populations. Multiple Mantel tests were used for this purpose. Although RAPDs revealed less genetic diversity than allozymes, levels of genetic differentiation ( G _ST) were identical. The standard genetic distance calculated at all RAPD loci was correlated with geographical distances but not with the genetic distance calculated from chloroplast DNA data. Conversely, allozyme variation was correlated with chloroplast DNA variation, but not with geography. Possibly, divergent selection at two allozyme loci during the glacial period could explain this pattern. Because of its greater number of loci assayed, RAPDs probably provided a less biased picture of the relative role of geography and history.     

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.     

Investigation of genetic diversity in fragmented geometric tortoise (Psammobates geometricus) populations

Microsatellite DNA was used to investigatelevels of genetic variability in severelyfragmented populations of the geometrictortoise, Psammobates geometricus, themost endangered tortoise on mainland Africa.Eight microsatellite markers were used toassess genetic variability within and betweenthree naturally occurring populations of P. geometricus. These populations areseparated by the Cape Fold mountain ranges inthe western Cape region of South Africa. Levels of variability were quantified usingallelic diversity, genotypic frequencies andheterozygosity. Evidence for populationsub-structuring was examined using Fstatistics, R_st and ².High levels of variability were found in allthree populations. Low levels of populationdifferentiation were found suggestingsignificant gene flow between the populations.     

Ontogeny of proventitious epicormic buds in Quercus petraea. I. In the 5 years following initiation

 The persistence of large epicormic shoots is one of the main factors that reduces timber quality and value in Quercus petraea. The early phases of epicormic shoot formation, i.e. the initiation of the epicormic buds, their survival and their proliferation over the years, are not clearly understood. In the present work, we studied the initiation of the axillary buds giving rise to epicormic buds and shoots, and followed their behaviour during the first 5 years using both scanning electron microscopy and light microscopy. Two types of proventitious epicormic buds have been identified. The first type has small axillary buds associated with the rings of bud-scale scars which are found at the base and tip of each growth unit. These buds are made of a terminal meristem surrounded only by scales; no leaf primordium is detected. During the second and third years of epicormic life, meristematic areas appear in the scale axil. Progressively, the meristematic areas organize into secondary bud primordia composed solely of the terminal meristem surrounded by scales. The second type of epicormic bud has secondary buds produced by a large axillary bud when this large bud either developed into a shoot or partially abscised. The epicormic potential in Q. petraea is characterized by a balance between the epicormic buds in apparent rest, enclosing meristematic areas and secondary bud primordia, and their mortality over the years. Received: 22 January 1998 / Accepted: 8 May 1998     

A hierarchical analysis of minisatellite DNA diversity in Gambel oak (Quercus gambelii Nutt.; Fagaceae)

Gambel oak is a short shrub to medium-sized tree of southwest North America for which the potential to grow in large clonal stands has been proposed. Here we use three different multilocus VNTR DNA probes (synthesized via PCR) to demonstrate that clones growing together can be identified, but individual clones exceeding 50 m in diameter do not commonly occur in the study population of Quercus gambelii . Further, using a hierarchical sampling scheme (three circular transects with diameters of 9, 1100, and 28 000 m), we estimate that for nonclonal individuals: (i) mean number of bands analysed per individual = 22.92; (ii) mean similarity (band sharing) between individuals = 0.322; (iii) mean probability that two randomly chosen individuals share all bands = 4.93 × 10^–11; and (iv) mean estimated heterozygosity = 0.796. F _ST calculated for the two nonclonal levels of the sampling hierarchy was 0.023, indicating that little genetic differentiation exists between them. These results support previous findings that, due to life-history traits of the genus and this species (out-crossing, wind-pollinated, animal-dispersed, long-lived woody perennials), gene flow is high and genetic subdivision of populations is low in oaks. At this study site, the clonal nature demonstrated for Gambel oak appears to have little detectable effect on these population genetic characteristics.     

The effect of lime and gypsum applications on a sessile oak ( Quercus petraea (M.) Liebl.) stand at La Croix-Scaille (French Ardennes) II. Fine root dynamics

Fine root distribution, quantities, dynamics and composition were studied in a sessile oak coppice stand in the French Ardennes on an acidic soil (< pH-H2O 4.5), one to five years after lime or gypsum applications. Fine root biomass and length increased and specific root length decreased after lime or gypsum treatments. The treatment responses were strongest four to five years after the applications, but the tendencies after one year were similar. The effects were pronounced in the top 15 cm but also at 30–45 cm four to five years after liming. The latter effect suggests an indirect positive feedback from the aerial parts of the trees into the deeper soil layers. Sequential sampling for two years revealed large differences in total fine root length between the years, and also indicated that fine root turnover was lower after liming or gypsum applications than in the control. This seemed to be related to a lower fine root mortality and higher longevity rather than to increased fine root production. The improved nutrient status of the fine roots corroborates this and coincides with improved foliar nutrition and tree growth. Moderate doses of lime and gypsum appeared effective in enhancing root system uptake function, resulting in increased above ground growth.     

Pollen dispersal inferred from paternity analysis in a mixed oak stand of Quercus robur L. and Q. petraea (Matt.) Liebl.

Paternity analysis was used to determine the spatial distribution of male parents of 984 offspring collected from 13 identified mother trees in a natural stand of 5.76 ha and comprising 296 adult trees of Quercus petraea and Q . robur . For seven of the 13 maternal progeny arrays sampled, we found an excess of nearby matings and a preferential direction of pollination. For the remaining progeny arrays, no departure from random distribution of male parents was detected. A common trend among all families was a high percentage (averaging 65% for Q. robur and 69% for Q. petraea ) of offspring that were pollinated by male parents from outside the study site. By pooling the data over all families, the average pollen dispersal curve within the stand was inferred and fitted to a negative exponential distribution. This model extrapolated for distances over the spatial scale of the study stand was insufficient to explain the high level of gene flow detected by the paternity analysis, suggesting a substantial level of long-distance pollination events. The genetic composition of the pollen pools received by each maternal tree was compared and showed significant differentiation that could be attributed to differences in male reproductive success. By contrast, no significant differentiation between the pollen clouds from outside and inside the study stand was detected, suggesting genetic homogeneity between the surrounding forest and the study stand.     

Water relations and gas exchange in Fagus sylvatica L. and Quercus petraea (Mattuschka) Liebl. in a mixed stand at their southern limit of distribution in Europe

Water status and gas exchange of beech (Fagus sylvatica L.) and sessile oak [Quercus petraea (Mattuschka) Liebl.] were studied in a mixed stand in the Montejo de la Sierra forest (central Iberian Peninsula), one of the southernmost locations of both species in Europe. Gas exchange and water potential were measured in leaves at different canopy levels over several days in two growing seasons. The daily variation pattern was established with the measurements of three selected dates per year, representative of the soil moisture content situations in early, mid- and late summer. A similar daily time course of leaf water potential was found for the two species. Nevertheless, beech showed a most noticeable decrease of water potential at midmorning and maintained lower leaf water potential than oak in the early afternoon. In 1994 the sessile oak saplings showed higher values of predawn water potential (Ψ_pd) than beech at the end of summer, when soil moisture content was lowest (20 cm depth). Beech showed a significantly lower net assimilation rate (A) than sessile oak for leaves under the same PPFD. Maximum net photosynthesis values (A _max) for beech and sessile oak on sunny leaves were 10.1±0.4 μmol m^–2 s^–1 and 17.8±1.7 μmol m^–2 s^–1 respectively, and those for water vapour stomatal conductance (g _wv) were 265±31 mmol m^–2 s^–1 and 438±74 mmol m^–2 s^–1. Differences in A and g _wv between the two species were maintained throughout the day on all measurement dates. No clear relationship was found between water status of saplings and stomata performance; there was only a negative correlation between Ψ_pd and g _wvmid in beech. Nevertheless, a significant response to the air vapour pressure gradient between leaf and air was translated into stomata closure on an hourly basis, more intensively in beech. Received: 4 March 1999 / Accepted: 21 December 1999