Comparative leaf surface morphology and molecular data of five oaks of the subgenus Quercus Oerst (Fagaceae)

Abstract

Comparisons between different data sets are important in taxonomic studies, particularly for complex taxa such as the genus Quercus. However, there is scarce literature on comparative morphological and molecular analyses in oaks. Here we describe an experimental approach based on the correlation between micromorphological features and molecular data in order to characterize and discriminate among five closely related species: Q. robur, Q. petraea, Q. frainetto, Q. pubescens and Q. virgiliana.     

Species of the genus Quercus in Italy: Characterization by means of leaf surface observation

Abstract

We propose a study of the main species belonging to the genus Quercus in Italy, characterized and identified by means of leaf surface observation, with special attention devoted to waxes, trichomes and stomata. Comparing our results with the classification proposed by SCHWARZ (1984), we find that species belonging to Schwarz's subgenus Quercus are recognizable because their waxes are structured in vertical scales; the two other subgenera (Sclerophyllodrys and Cerris) present smooth wax structures, their distinctive feature being the shape of the stomatal rima, which is roundish in Sclerophyllodrys and elliptical in Cerris. The study characterizes Quercus pubescens Willd. and Quercus petraea Liebl. by analyzing some morphometric traits; but the authors feel that further research is needed on these critical taxonomic entities. Lastly, the study examines forms of was degeneration correlated to the phenomenon known as oak decline.     

Pollen grain morphology supports the taxonomical discrimination of Mediterranean oaks (Quercus,Fagaceae)

The exine architecture and some quantitative parameters (polar and equatorial axes and their ratio) of pollen grains allow a clear discrimination between groups of Mediterranean oak species. For the differentiation of species within groups only the quoted parameters prove useful.     

Role of micromorphological leaf traits and molecular data in taxonomy of three sympatric white oak species and their hybrids (Quercus L.)

Hybridisation and introgression occur with high frequency in the genus Quercus and interspecific hybrid individuals show patterns of morphological traits that might be influenced in different ways. Micromorphological leaf traits appear to be positive and stable in Quercus species, and by combining genetic and micromorphological analyses, it is possible to compare the patterns of variation in micromorphological leaf traits of pure and hybrid individuals. Trichomes and stomatal traits were examined using scanning electron microscopy at 150–2000 × magnification in sympatric oak species collected in a natural deciduous wood. Q. frainetto, Q. petraea and Q. pubescens appear to have a relatively predictable complement of trichome types. Both the pattern and quantitative values of each micromorphological trait examined (stomata and trichomes) have an important role in identifying hybrids and pure species; putative hybrids show a pattern of trichomes that is a combination of the parental types. These results, combined with the fact that micromorphological traits generally exhibit higher consistency, indicate that this source of information can be an excellent clue to hybridisation and introgression and useful in taxonomical, systematic and evolutionary studies on the European white oaks.     

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.     

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.     

Systematic leaf anatomy of Caladeniinae (Orchidaceae)

Leaf anatomy of 25 species in 15 genera of Caladeniinae (Diurideae, Orchidaceae), excluding Caladenia, was investigated to determine diagnostic characters to be used in forthcoming, broad-based cladistic analyses of the subtribe and to assess interspecific and intergeneric relationshipS. Of the characters examined, those that show the most variation among the study taxa are presence and types of trichomes, cuticular sculpturing, anticlinal walls of epidermal cells, heterogeneity of chlorenchyma, distribution and length: width ratios of stomata. Anatomical evidence supports the generic concept of Leptoceras Lindley but contradicts that of Drakonorchis Hopper & A.P. Brown. Paracaleana is not sufficiently distinct from Caleana to warrant generic status. Lyperanthus serratus and L.suaveolens are hyperstomatic, a rare condition in Orchidaceae. On the basis of these and other characters, Lyperanthus, as currently circumscribed, is polyphyletic. From leaf structure Caladeniinae as now conceived is polyphyletic and comprises seven groups: (1) Caladenia, Leptoceras, Elythranthera, Eriochilus, Glossodia, Aporostylis; (2) Adenochilus, Rimacola; (3) Arthrochilus, Chiloglottis, Spiculaea, Leporella; (4) Caleana (including Paracaleana); (5) Bumettia; (6) Lyperanthus suaveolens and L.serratus; (7) Lyperanthus nigricans.     

两种忍冬属植物叶表皮扫描电镜观察及其生态适应性的研究

利用扫描电镜观察2种忍冬属植物的叶表皮形态特征,观察指标包括气孔器、表皮毛、表皮细胞等。扫描电镜观察结果表明:气孔仅分布在下表皮,气孔器散生并为无规则型;下表皮密被表皮毛(包括腺状毛和非腺毛);上表皮细胞的垂周壁呈沟槽状下陷;同时在下表皮还分布有瘤状的草酸钙簇晶。为进一步研究这2种忍冬属植物叶的形态解剖特征,通过光学显微镜观察发现,叶均为异面叶,栅栏组织发达,细胞长柱形;海绵组织细胞类圆形,排列紧密;叶脉机械组织发达。以上这些构造特征,都有利于减少植物体水分的散失,以适应我国西南岩溶区干旱环境特征。通过进一步分析电镜观察结果,发现在忍冬叶的上表皮中脉处有呈线状分布的腺状表皮毛,而华南忍冬叶的上表皮则无表皮毛分布,这一微形态特征可以作为二者种间特异性差别,从而为二者的合理区分提供一定的依据。     

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.     

Two-year cycles of synchronous acorn and leaf production in biennial-fruiting evergreen oaks of subgenus Cyclobalanopsis (Quercus, Fagaceae)

Many masting species switch resources between vegetative growth and reproduction in mast and non-mast years. Although masting of oak species is well known, there have been few investigations of the relationship between vegetative growth and reproduction based on long-term monitoring data, especially in evergreen oaks of subgenus Cyclobalanopsis. We investigated annual variations over 13?years in acorn and leaf production of three evergreen oak species in subgenus Cyclobalanopsis, genus Quercus (Fagaceae)??Q. acuta, Q. salicina and Q. sessilifolia??in western Japan. In these species, the maturation of acorns occurs in the second autumn after flowering, which is known as a biennial-fruiting habit. We found a pattern of acorn production and masting in alternate years that was synchronized in all three species. Masting was not correlated with temperature and precipitation. Annual leaf-fall also showed 2-year cycle in the three oak species; peak years were synchronized between species and peak leaf-fall alternated with acorn production in all three species. Furthermore, there was a significant negative correlation between acorn and leaf production in all three species. Data showing 2-year cycles of acorn and leaf production and the negative correlation between them supports the hypothesis of resource switching between vegetative growth and reproduction. The 2-year cycle might be the basic, intrinsic rhythm of resource allocation in biennial-fruiting Cyclobalanopsis species.     

Foliar anatomy and micromorphology of eleven species of Eugenia L. (Myrtaceae)

The anatomy and micromorphology of eleven species of Eugenia found in the 'restinga' of Marica, Rio de Janeiro, Brazil, were studied and compared. The characteristics found to be of most use in distinguishing the species are: presence and types of trichomes persistent in the adult material, form and arrangement of overlying cells (epidermal cells over secretory cavities), pattern of anticlinal walls of epidermal cells (primary sculpturing), occurrence of epicuticular striae and folds (secondary sculpturing) and presence of peristomatal folds. An analytical key based on these characteristics is presented for the Eugenia species studied.     

中国栎属植物的数量分类研究

在全面衡量中国栎属49种4变种性状特征的基础上,观测了25个定性性状和18个数量性状。通过聚类分析,可以将中国栎属分为5个特征明显的表征群,即5组:麻栎组(Section Aegilops)、槲栎组(Section Quercus)、高山栎组(Section Brachylepides)、巴东栎组(Section Engleriana)和橿子栎组(Section Echinolepides)。在主成分分析中,虽然性状的累积贡献率增长不明显,前3个主成分累积贡献率仅有50.4%,但结果仍然显示出和聚类分析同样的结果。说明栎属的分类性状仍存在主要方面,只不过在演化过程中由于性状变异的多样性,产生了多个不同的表征类群。分析结果对我们解决个别有疑问种的分类地位也有启示作用。因此,数量分类方法对解决栎属组间及种间的分类问题有重要的意义。     

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.     

Geometric morphometrics and leaf phenotypic plasticity: assessing fluctuating asymmetry and allometry in European white oaks (Quercus)

Because plants are unable to move away from unfavourable habitats and environmental perturbations, leaf phenotypic plasticity facilitates light absorption and gas exchange. Oaks (Quercus spp.) are particularly known for their adaptability and plastic phenotypes, and leaf allometry and developmental instability may represent important mechanisms for their adaptation to environments and evolution. Because of its important role in the adaptation of plant populations to different environments, allometry can be involved in diversifying selection. Developmental instability is related to environmental perturbations and stresses by producing random deviations in structures characterized by bilateral symmetry, such as oak leaves. In addition, developmental instability can also arise from genetic bottlenecks or as a result of hybridization. The splitting of symmetric and asymmetric components of variation and their separate analysis allows the variability in leaf shape traits to be summarized, reducing the variation produced by developmental instability. The geometric morphometric approach is a useful method for the study of leaf asymmetry and allometric patterns. This method provides an important tool for the visualization of shape attributes that characterize species with highly variable leaf phenotypic patterns. In this study, leaf shape and size variability of three white oak species was investigated by means of a two‐dimensional landmark‐based method providing improved knowledge of variance partitioning, species discrimination, fluctuating asymmetry and allometric patterns of variation resulting from the different analyses. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 335–348.     

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.     

SEM study of surface structures of the spathe in Cryptocoryne and Lagenandra (Araceae: Aroideae: Cryptocoryneae)

SEM studies of the spathe structures in the two closely related genera Cryptocoryne and Lagenandra show differences between the inner and outer surfaces, as well as in cell structures in the various parts of the spathe. The cell structure reveals patterns mat makes it possible to depict homologous structures of the spathe, even though the spathes of the two genera look different. The basal part of the kettle has a mucilage covering of the cells, interpreted as a hitherto unnoticed food source. The cells of the inner surface of the kettle and tube have downward pointing trichomes. On the second day of flowering these collapse and sink into the cell lumen, which is suggested to create a unique lattice-like structure that enables the insects to climb out of the kettle and tube. The cell structure of the flap shows that it is a prolongation and continuation of the spathe tube margin.     

Application of plastid and nuclear markers to DNA barcoding of Euro‐Mediterranean oaks (Quercus,Fagaceae): problems,prospects and phylogenetic implications

DNA barcoding, a species identification system based on sequences from a short, standardized DNA region, has emerged recently as a new tool for taxonomists. We investigated the discriminatory power of a subset of highly variable proposed plant barcoding loci (matK, trnH‐psbA, ITS2) in Quercus, a taxonomically complex tree genus of global importance. The research included all currently recognized species and some major variants of the Mediterranean region and Europe (32 taxa) and 17 East Asian and North American species used for comparison. Based on sequence character state, we assigned unique plastid haplotypes to 40.8% of the investigated species; ITS2 increased the resolution up to 87.8% of total taxa. Nevertheless, unsuccessful genetic distance‐based discrimination questioned the potential efficiency of correct species identification for future studies. Most species appeared to be nonmonophyletic in parallel phylogenetic tests. Three subgeneric groups were outlined, with different rates of within‐group variability and geographical differentiation. Members of one of these groups (corresponding to the Eurasian Group Ilex) were paraphyletic to Group Quercus from the New and Old World and the Eurasian Group Cerris. The data gathered indicate that barcoding markers may help to identify closely related species clusters and contribute to the inference of major diversification and evolutionary patterns in oaks, but the methodology per se appears to be of limited efficacy in defining species limits, unless we make a profound revision of traditional Quercus taxonomic categories. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 172 , 478–499.     

Comparative morphology of the leaf epidermis in Fritillaria (Liliaceae) from China

The leaf epidermis of 16 species and one putative species of Fritillaria was examined using light microscopy (LM) and scanning electron microscopy (SEM). The results showed that the stomatal and other epidermal features were constant within species. Epidermal cells of Fritillaria under LM were usually polygonal and anticlinal cell walls were straight or curved. In a few species they were irregular, with sinuous anticlinal cell walls. The cuticular membrane of Fritillaria was usually striated, and the wax ornamentations were flaked, granular or concomitant. Based on leaf epidermal characteristics, the subdivision of Fritillaria is discussed, and the statistical t‐test method was used to ascertain the significance level of the differences in the stomata of each species. All orientations of the stomatal poles in Fritillaria were the same, and this phenomenon was named ‘stomatal orientation’. The stomatal characteristics support the origin of section Fritillaria in China from two floristic elements. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 160 , 93–109.     

Comparative leaf anatomical studies of some Sterculia L. species (Sterculiaceae)

A comparative study was undertaken on the leaves of 12 Sterculia species in order to assess anatomical variations which may be useful in species identification and to evaluate their significance in the taxonomy of the genus. All species have glandular and non-glandular trichomes, anomocytic stomata, multiple and mucilaginous epidermis, mucilaginous cavities and druses in mesophyll tissues. A small amount of variation was observed in the outline of transverse sections of midribs and petioles, the presence of central vascular bundles, and the type of trichomes. Some characters, such as the vertically divided adaxial epidermis in S. macrophylla , are unique to certain species and are therefore useful in diagnostic characters, while S. coccinea and S. elongata appear to be similar in the structure of the petiole and midrib. The results, however, do not show any groupings of species and do not support the observations of others on groups derived from wood anatomy.