Leaf morphometric variation in Quercus affinis and Q. laurina (Fagaceae), two hybridizing Mexican red oaks

Leaf variation was examined in two hybridizing Mexican red oaks, Quercus affinis and Q. laurina . Data of nine traits were obtained for ten randomly chosen leaves in each of 512 individuals from 16 populations sampled along a geographical gradient, including the distribution area of both species and a putative area of secondary contact and hybridization. A cluster analysis recognized two main groups of populations, which were congruent with the taxonomic assignment of the predominant morphological type within the populations and were thus labelled ' Q. affinis -like' and ' Q. laurina -like' population groups. A nested analysis of variance revealed that the largest proportion of the total variation was contained within populations, as among-tree variation (28–54%, depending on the trait), and as intra-individual variation (17–56%). However, differences between the two groups of populations (3–27%) and among populations within groups (5–21%) were also significant for the nine traits. A distinct pattern of change across populations was observed for each trait. Variation was particularly pronounced along the geographical gradient for petiole length and leaf-margin teeth number, possibly implying selection on these two traits. Results suggest that phenotypic plasticity, gene flow, hybridization and natural selection have shaped foliar variation in this oak complex. © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 147 , 427–435.     

Comparative analysis of micromorphological characters in two distantly related Mexican oaks, Quercus conzattii and Q. eduardii (Fagaceae), and their hybrids

Interspecific hybridization occurs with high frequency in the genusQuercus, but few studies have analyzed and compared micromorphological characters in putative parental species and their hybrids.Quercus eduardii andQ. conzattii are two Mexican black oak species that, although distantly related, have formed at least one population of hybrid origin, where individuals with intermediate macromorphology are present. The purpose of this investigation was to analyze the degree of differentiation in micromorphological characters between the two species and to assess the expression of these characters in individuals with intermediate macromorphology. Foliar trichomes, epicuticular waxes, stomata, and pollen grains, were examined using scanning electron microscopy (SEM) in the three types of individuals (Q. conzatti, Q. eduardii, and intermediates). Trichome density was quantified with light microscopy. Types of trichomes present, length of trichome arms, types of epicuticular wax on the leaf surfaces, and the position of stomata with respect to the foliar surface were characters useful to differentiate betweenQ. conzattii andQ. eduardii. Plants with intermediate macromorphology displayed a pattern of micromorphological characters that were identical to one parental species (Q. conzattii), or extreme or novel relative to both species.     

Geographic structure of genetic and phenotypic variation in the hybrid zone between Quercus affinis and Q. laurina in Mexico

Analyzing the structure of hybrid zones is important for inferring their origin, dynamics and evolutionary significance. We examined the geographic structure of phenotypic and genetic variation in the contact zone between two Mexican red oaks, Quercus affinis and Q. laurina. A total of 105 individuals from seven populations were sampled along a 600‐km latitudinal gradient representing the distribution area of the two species and their contact zone. Individuals were genotyped for nine nuclear and four chloroplast DNA microsatellite loci (ncSSR and cpSSR, respectively), and characterized for several leaf and acorn traits. The cpSSR data revealed extensive haplotype sharing among populations of the two species, while a Bayesian assignment analysis based on ncSSRs identified two main genetic groups, each corresponding to one of the species, and two populations in the contact zone showing evidence of admixture. The proportion of genetic ancestry in the populations was strongly associated with latitude and showed a pattern of variation with the shape of a narrow sigmoidal cline. The variation in three of the seven phenotypic traits was partially congruent with molecular variation, while the other traits did not conform to a geographic cline but instead were correlated with environmental variables. In conclusion, the hybrid zone between the two oak species has some of the characteristics of a tension zone, but heterogeneous variation across traits suggests differential introgression and the action of extrinsic selection.     

Genetic admixing of two evergreen oaks,Quercus acuta and Q. sessilifolia (subgenus Cyclobalanopsis), is the result of interspecific introgressive hybridization

In forests worldwide, Quercus is a major genus; however, the boundaries between the constituent species are relatively weak, and hybridization is reported frequently. In this study, we examined Quercus acuta and Quercus sessilifolia (subgenus Cyclobalanopsis), which have a putative hybrid—Q. x takaoyamensis. We investigated leaf morphological traits and microsatellites of Q. acuta and Q. sessilifolia in the area where the two species are both found. Although the leaf traits overlapped, the two species could be distinguished morphologically as demonstrated by principal component analysis based on a range of these traits. They were also genetically differentiated, with F _ST?=?0.104. However, they shared most of the alleles at all eight loci examined, and considerable genetic admixing was detected. Admixture analysis demonstrated that Q. acuta and Q. sessilifolia, respectively, contained 11 and 24 % of individuals with a probability of less than 0.9 of being correctly assigned to their species. Model-based testing showed that this admixing was created by not only shared ancestral polymorphism but also by hybridization. Effective population size and migration rate were estimated using the coalescent approach. We estimated 8.843 and 71.98 effective numbers of migrants per generation to Q. acuta and Q. sessilifolia, respectively. Theoretically, one to ten migrants per generation are required to prevent complete genetic differentiation. Based on the results of this study, it appears that genetic admixing, with sharing of most alleles, is probably common in the two species and is maintained by interspecific introgressive hybridization.     

Genetic evidence for hybridization in red oaks (Quercus sect. Lobatae, Fagaceae)

? Premise of the study: Hybridization is pervasive in many plant taxa, with consequences for species taxonomy, local adaptation, and management. Oaks (Quercus spp.) are thought to hybridize readily yet retain distinct traits, drawing into question the biological species concept for such taxa, but the true extent of gene flow is controversial. Genetic data are beginning to shed new light on this issue, but red oaks (section Lobatae), an important component of North American forests, have largely been neglected. Moreover, gene flow estimates may be sensitive to the choice of life stage, marker type, or genetic structure statistic. ? Methods: We coupled genetic structure data with parentage analyses for two mixed-species stands in North Carolina. Genetic structure analyses of adults (including F(ST), R(ST), G'(ST), and structure) reflect long-term patterns of gene flow, while the percentage of seedlings with parents of two different species reflect current levels of gene flow. ? Key results: Genetic structure analyses revealed low differentiation in microsatellite allele frequencies between co-occurring species, suggesting past gene flow. However, methods differed in their sensitivity to differentiation, indicating a need for caution when drawing conclusions from a single method. Parentage analyses identifed >20% of seedlings as potential hybrids. The species examined exhibit distinct morphologies, suggesting selection against intermediate phenotypes. ? Conclusions: Our results suggest that hybridization between co-occurring red oaks occurs, but that selection may limit introgression, especially at functional loci. However, by providing a source of genetic variation, hybridization could influence the response of oaks and other hybridizing taxa to environmental change.     

Clonal and genetic structure of two Mexican oaks: Quercus eduardii and Quercus potosina (Fagaceae)

Quercus eduardii and Q. potosina are dominant oak species in Sierra Fría, Aguascalientes, Mexico. These species have been exploited for multiple purposes since the 16th century. Both species produce clonal offspring through root suckering and acorns through sexual reproduction. To understand clonality for the implementation of the most adequate actions for the conservation of these species, we addressed the following questions: (a) what is the spatial clonal structure of both species? (b) How much clonal and genetic diversity is maintained in these species? Random Amplified Polymorphic DNAs (RAPDs) were used as molecular markers for these analyses. Genets of both species have few ramets and these grow close the parent tree. Autocorrelation analyses at the ramet level showed an aggregated distribution at short distances and a random spatial distribution at larger distances. Also, at the genet level the autocorrelation analyses showed a random distribution. Clonal diversity was high in both species (Q. eduardii: D=0.963, G/N=0.60; Q. potosina: D=0.985, G/N=0.65). Genetic diversity was high within populations (Q. eduardii: H _e =0.33±0.11; Q. potosina: H _e =0.35±0.11). Low levels of genetic differentiation among populations were observed (Q. eduardii ? _st =0.19, P < 0.002; Q. potosina ? _st =0.13, P < 0.002). Both species maintain high levels of clonal and genetic diversity, probably due to successful sexual reproduction, which allows gene flow among populations. Conservation and/or reforestation programs must include seed collections and germplasm banks. Due to the small genet size and the high clonal diversity of these species, seeds can be collected in any place in Sierra Fría, Aguascalientes.     

Morphometric variation in oaks of the Apostle Islands in Wisconsin: evidence of hybridization between Quercus rubra and Q. ellipsoidalis (Fagaceae)

The Apostle Islands in Lake Superior are populated by trees that are clearly related to Quercus rubra L. However, several islands have trees with morphological characteristics suggestive of hybridization with Q. ellipsoidalis Hill. Leaf specimens were collected from trees in five locations: the outermost island, an intermediate island, the nearest-shore island, the northeast shoreline, and an inland forest about 24 km from the shoreline. Seventeen landmarks were digitized for two to five leaves per tree. These landmarks were used to generate nine linear characters and three angles. These characters, along with the number of bristle tips per leaf, were used in various combinations for several principal component analyses. In addition, the landmark configurations were examined using rotational-fit methods. The patterns observed in both types of analysis indicate phenotypic variation coincident with a line connecting the two most distant sample sites. The location nearest the geographic center of this line is also nearest the center of the two-dimensional view of phenotypic variation. Trees at each site illustrate a distinctive pattern in the rotational-fit analyses, and patterns of co-variation in the morphometric characters are different for each site. The observed morphometric variation is consistent with the hypothesis that there is hybridization between these two species, most likely in the form of introgression from Q. ellipsoidalis into Q. rubra.     

Type designation of the oaks Quercus ballota and Q. rotundifolia (Fagaceae)

The names Quercus ballota and Q. rotundifolia (Fagaceae), referring to taxa sometimes considered conspecific with the widely distributed Q. ilex, are discussed and typified. A specimen preserved at MPU is designated as the lectotype of Desfontaines's name Q. ballota. Lamarck's name Q. rotundifolia is neotypified using a specimen preserved at VAL, with several duplicates in other European herbaria.     

Drought response of two Mexican oak species, Quercus laceyi and Q. sideroxyla (Fagaceae), in relation to elevational position

To investigate the relationship between the altitudinal distribution of Quercus laceyi and Q. sideroxyla and their physiological responses to drought, we measured relative water content (RWC), water potentials (Ψ(predawn) and Ψ(midday)), photosynthesis (A(max)), stomatal conductance (g), chlorophyll fluorescence (F(v)/F(m)), and spectral reflectance (400-1100 nm) five times during a 7 wk acute drought. Quercus laceyi was drought tolerant, while Q. sideroxyla was a drought avoider; Q. laceyi tolerated lower RWC (Q. sideroxyla = 54%, Q. laceyi = 44%), Ψ(pd) (Q. sideroxyla = -2.6 MPa, Q. laceyi = -3.3 MPa), and Ψ(md) (Q. sideroxyla = -4.5 MPa, Q. laceyi = -6.6 MPa). The F(v)/F(m) also declined first in Q. sideroxyla in wk 6, whereas F(v)/F(m) did not decline in Q. laceyi until wk 7. A(max) and g fell in wk 4, 6, and 7 in drought seedlings of both species, suggesting a decline in CO(2) assimilation during the drought. Leaf spectral reflectance increased with time in response to decreases in leaf photosynthetic pigment concentrations in latter weeks of the drought. The results suggest a close association between the altitudinal distributions of these species and their adaptation to water stress.     

In vitro multiplication of Quercus leucotrichophora and Q. glauca: Important Himalayan oaks

Multiple shoots of Quercus leucotrichophora L. and Q. glauca Thunb. were induced from the intact embryos (decoated seeds) as well as from the cotyledonary nodes (with attached cotyledons but without radicle and primary shoot) of 3-weeks old in vitro grown seedlings on Woody Plant (WP; Lloyd and McCown, 1980) and Murashige and Skoog (MS; 1962) media supplemented with 6-benzyladenine (BA), either alone or in combination with gibberellic acid (GA₃)/ indole-3-butyric acid (IBA). BA (22.19 M) was effective for induction of multiple shoots and addition of GA₃ to the medium further enhanced the shoot number and shoot height but resulted in shoot thinness. High frequency shoot multiplication was achieved using cotyledonary nodes. Shoots were further multiplied from the original explant on WP medium supplemented with BA (22.19 M). Nearly 78% and 67% rooting was obtained in Q. leucotrichophora and Q. glauca microshoots (3–4 cm high), respectively on 1/2 strength WP medium supplemented with IBA (14.76 M). However, this was associated with basal callus formation. Treatment with IBA (25–100 M) for 24 or 48 h followed by transfer to PGR free 1/2 strength WP medium not only improved the rooting percentage but also avoided basal callus formation. IBA at 100 M for 24 h was most effective (90% and 100% rooting in Q. leucotrichophora and Q. glauca, respectively). In vitro rooted plants were hardened and established in garden soil.Growth performance of 6-month-old in vitro raised plants was compared with ex vitro plants (seedlings) of the same age. The photosynthesis and transpiration rates of eight months old in vitro and ex vitro raised plants of both species were measured under different light (0, 600, 900, 1200, 1500 and 2000 mol m^–2s^–1) and temperature (20, 25, 30, 35 and 40 °C). Light optimum for photosynthesis was around 2000 mol m^–2s^–1 in Q. leucotrichophora and around 1500 mol m^–2s^–1 in Q. glauca whereas optimum temperature for photosynthesis was 25 °C in Q. leucotrichophora and 30 °C in Q. glauca. The rate of transpiration at different temperatures (20–40 °C), in the two species, increased with increase in the light intensity up to the highest level, i.e., 2000 mol m^–2s^–1. Temperatures beyond 35 °C adversely affected the rate of transpiration in in vitro raised as well as ex vitro plants of both the species. In vitro raised and hardened plants of both the species were comparable to ex vitro plants in terms of gas and water vapour exchange characteristics, within the limits of this study.     

Spatial and genetic structure of two sandhills oaks: Quercus laevis and Quercus margaretta (Fagaceae)

Quercus laevis Walt, (turkey oak) and Q. margaretta Ashe (scrubby post oak) are important scrub oaks in the sandhills forest communities of the Coastal Plain of the southeastern United States. We used allozyme loci and Ripley's L-statistics to examine clonal structure and spatial dispersion in these species. Q. laevis greater than 1.5 m in height were randomly dispersed on a scale of 0–40 m; smaller individuals (< 1.5 m) were slightly clustered on a scale of 0–12 m. Larger individuals separated by ≤1 m had 15% probability of being ramets of the same clone. Q. margaretta showed strong clustering on a scale of 0–20 m. Stems separated by ≤ 1 m had a 71% probability of being ramets of the same clone. Clonal offspring were strongly clustered about the presumed clonal parent: 50% fell within 0.50 m of this individual. Simulation modeling and direct comparison of adult and juvenile genotypes indicated that acorns are dispersed on a scale of tens of meters for both species, suggesting animal vectors such as squirrels or blue jays.     

Natural hybridisation between kermes (Quercus coccifera L.) and holm oaks (Q. ilex L.) revealed by microsatellite markers

Hybridisation between species of the genus Quercus is a common phenomenon as a result of weak reproductive isolation mechanisms between phylogenetically close species that frequently co-occur in mixed stands. In this study, we use microsatellite markers to analyse introgression between kermes ( Quercus coccifera L.) and holm ( Q. ilex L.) oak, two closely related taxa that frequently dominate the landscape in extensive areas in the Mediterranean region. All tested microsatellites amplified and were polymorphic in both kermes and holm oaks. Bayesian admixture analyses showed a good correspondence between each species and one of the two inferred genetic clusters. Five sampled individuals were a priori tentatively identified as hybrids on the basis of intermediate morphological characteristics, and it was confirmed that they also presented mixed genotypes. However, we also detected different levels of genetic introgression among morphologically pure individuals, suggesting that successful backcrossing and/or reduced phenotypic expression of genetic variance in certain individuals may have resulted in strong convergence towards a single species phenotype.     

Molecular differentiation and diversity among the California red oaks (Fagaceae; Quercus section Lobatae)

A recent epidemic of Phytopthora (Sudden Oak Death) in coastal woodlands of California is causing severe mortality in some oak species belonging to the red oak (Lobatae) group. To predict the risks of spread of this disease, an understanding of the relationships among California's red oak species and of their population genetic structure is needed. We focus here on relationships among the four species of red oak. Whereas morphological distinction of Quercus wislizeni and Quercus parvula can pose problems, Quercus kelloggii and Quercus agrifolia in pure forms are easily distinguishable from one another and from Q. wislizeni and Q. parvula in the field. However, hybrids among all species combinations are known to occur in nature and these can confound data from ecological studies. Our results revealed greatest differentiation of the deciduous Q. kelloggii, with only weak AFLP fragment differentiation of the three remaining evergreen species. The molecular data suggest a closer affinity of Q. agrifolia with Q. wislizeni and Q. parvula contrary to earlier suggestions that its origins are likely to have been with northern deciduous oaks probably through a common ancestor with Q. kelloggii. Interior and coastal populations of Q. wislizeni separated in dendrograms based on phenetic and genetic distances suggesting probable isolation in different glacial refugia. The position of Q. parvula remains ambiguous, having a closer affinity with interior populations of Q. wislizeni and with Q. agrifolia, than with coastal populations of Q. wislizeni. Mean population differentiation in Q. wislizeni was 0.18, which is somewhat higher than the average for other oak species, suggesting that range fragmentation has occurred in the past, resulting in a metapopulation structure. Our results provide evidence that introgression among these species may be causing reticulation, further confounding species separation. Whereas Phytopthora has been reported on Q. agrifolia, Q. parvula and Q. kelloggii, it has not yet been detected in natural populations of Q. wislizeni. The species relationships that our molecular data show suggest that this is more likely a result of escape due to ecological tolerances than to genetic differences.     

Early seedling growth of pine (Pinus densiflora) and oaks (Quercus serrata,Q. mongolica,Q. variabilis) in response to light intensity and soil moisture

In climatic chambers seed germination and seedling growth of Pinus densiflora Sieb. et Zucc., Quercus serrata Thunb., Quercus mongolica Fisch. ex Turcz. and Quercus variablilis Bl. were investigated as functions of light intensity and soil moisture. In Korea these tree species occur widely and form mixed forests with different distributions. Species clearly differed in the pattern of germination and early seedling growth between light and soil treatments. The germination of pine did not differ between the experimental treatments until the breaking of the primary buds. After that, light intensity was the deciding factor for further development. In the most moist treatment (approx. field capacity) growth of the pine seedlings was strongly inhibited. For the three oak species, differences between experimental treatments first occurred after complete formation of primary leaves. Seed development strongly correlated with the weight of the acorn. The large seeded Q. variabilis (acorns with mean weight of 4.7 g) developed faster and reached larger dimensions towards the end of the experiment than Q. mongolica (2.8 g per acorn) and Q. serrata (0.9 g per acorn). Regarding height and biomass growth, the oak species showed a higher shade tolerance than pine. The proleptic shoot growth was clearly influenced by the light intensity. Root formation was favoured by a high exposure to light. In case of the oak species reduction of soil moisture increased the length of primary roots and the number of secondary roots.     

The effects of seed size, cotyledon reserves, and herbivory on seedling survival and growth in Quercus rugosa and Q. laurina (Fagaceae)

In a greenhouse experiment, seedling survival of two oak species (Quercus rugosa and Q. laurina) was greatly affected by the excision of cotyledons 1 mo after germination, with a greater impact on Q. laurina. The effect of seed size was also significant for both species, with a positive correlation between seed mass and survival and growth. The effect of cotyledon excision on seedling growth persisted throughout the first growing season in Q. rugosa and was not analyzed for Q. laurina due to the low number of seedlings that survived cotyledon excision. Seed size significantly affected seedling height, diameter, leaf area, and biomass at 6 mo. Seed size and cotyledon retention affected the ability of Q. rugosa to recover from herbivory, as both factors had a significant effect on relative growth rates after aerial biomass removal. The results show that seedlings originating from large seeds can better endure loss of cotyledons and aerial biomass and thus are better equipped to confront stress early in their lives.     

Regeneration of oaks (Quercus robur/Q. petraea) and three other tree species during long-term succession after catastrophic disturbance (windthrow)

In broadleaved temperate forests in Europe, oak (Quercus robur/Q. petraea) regeneration is reported to be weak or absent. However, most work concern seedlings or saplings, studied relatively few years. We studied a Picea abies stand in Sweden, windthrown and logged (all stems harvested) in 1969, testing the hypothesis that oaks regenerate in the long term among competing tree species after catastrophic disturbance. In 2011, after 40 years/growth seasons, we recorded live and dead trees in the new stand and investigated the surroundings, competition, and succession. The following trees, up to 26 m tall, colonized: Sorbus aucuparia, Betula pendula/B. pubescens, Fagus sylvatica, Q. robur/Q. petraea, and Corylus avellana (a shrub). Betula dominated, and only Fagus was regenerating in 2011. Sorbus had produced most of the dead trees, mainly or partly through intraspecific competition. In the stand, compared to the surroundings, Quercus, Picea, and Alnus glutinosa were under-represented, and Sorbus, Betula, and Fagus were over-represented. Yet, the density of Quercus was far from negligible; 48 large trees/ha. Most of the oaks (74 %) were co-dominant trees and many grew near Sorbus. Thus, oaks can survive and grow fast among pioneer trees and browsing animals, a conclusion which is supported by the literature (nine studies identified). High mortality of Sorbus is part of a long-term succession, where Fagus might come to dominate. However, Quercus likely will persist, in low density. We propose three key traits contributing to long-term persistence of Q. robur/Q. petraea in European temperate forests: long life span, ecological plasticity, and resistance to disturbances.