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.     

近一世纪黄土高原区植物气孔密度变化规律

以黄土高原地区 4种典型 C3植物辽东栎 (Quercus liaotungensis)、虎榛子 (Ostryopsis davidiana)、酸枣 (Zizyphus jujubavar.spinosa)和狼牙刺 (Sophora viciifolia)植物腊叶样品标本为材料 ,利用数码图像显微镜处理系统 ,研究了从 2 0世纪 30年代至 2 0 0 2年中不同年代植物气孔密度的变化情况。结果表明 ,在近百年中 ,4种植物气孔密度随环境变化的演变规律不同 ,辽东栎气孔密度升高率为 1.31% ,几乎没有变化 ,虎榛子气孔密度降低率为 9.79% ,下降也不十分明显 ,而酸枣和狼牙刺的气孔密度降低率分别为 32 .35 %、4 6 .85 % ,下降非常明显。 4种植物气孔密度变化率比较 :狼牙刺 >酸枣 >虎榛子 >辽东栎。说明不同植物对气候环境因子的敏感性不同 ,具有不同的适应环境变化的策略 ,环境变化对气孔密度的影响 ,其效应因植物种类而异     

The effect of PlantformTM bioreactor on micropropagation of Quercus robur in comparison to a conventional in vitro culture system on gelled medium,and assessment of the microenvironment influence on leaf structure

Quercus robur L. was micropropagated by axillary bud proliferation testing two different shoot culture systems: (i) on gelled medium in Microbox (plastic vessel with a strip for ventilation) and (ii) in liquid culture in Plantform^TM bioreactor (a temporary immersion system). Two different conditions of temporary immersion were assessed: 12 min/8 h (Plantform 1) and 8 min/16 h (Plantform 2). The effect of the two culture systems was evaluated also during subsequent rooting phase, carried out on gelled medium. Finally, the influence of the different culture conditions on leaf structure was considered, taking also into consideration the micromorphological characters of young leaves from in-field-grown oaks. Nodal segments, excised from established in vitro shoots and cultured on modified Woody Plant Medium, showed a higher Relative Growth Rate in Plantform than in Microbox, but culture conditions provided in Plantform 1 favored shoot and leaf hyperhydricity. Shoots cultured in Microbox or Plantform 2 presented the same percentage of rooting after their transfer on gelled rooting medium. Leaves developed in the two different microenvironments had large stomata with elliptical shape, which indicates good functionality, and formed hairs, and epicuticolar waxes. These leaf features are considered to provide a good adaptability to ex vitro conditions.     

Abnormal germling development by brown rust and powdery mildew on cer barley mutants

The barley leaf rust fungus forms appressoria over host leaf stomata and penetrates via the stomatal pore. High levels of avoidance to leaf rust fungi have been described in some wild accessions of Hordeum species where a prominent wax layer on the stomata inhibits triggering of fungal appressorium differentiation. Leaf rust avoidance has not yet been found in H. vulgare. Since cuticular leaf waxes are implicated in the avoidance trait, we screened 27 eceriferum (cer) mutant lines of H. vulgare for avoidance to barley leaf rust. These mutations affect leaf waxes. Reduction in numbers of germ tubes forming appressoria over stomata was found in some lines, but the greatest reduction (ca 30%) was less than previously found in wild barley spp. or in an accession of H. chilense used here as a check. In one line (cer-zh654), avoidance was due to a combination of factors. Firstly, fewer germ tubes oriented towards stomata and so failed to contact them. Secondly, some germ tubes that encountered stomata did not form appressoria but over-grew them. In this line, therefore, the fungus tended to fail both to locate and to respond to stomata. The appressoria of barley powdery mildew form on leaf epidermal cells that they penetrate directly. On certain cer lines, a proportion of germlings of the barley powdery mildew fungus developed abnormally, suggesting that germlings failed to recognise and/or respond to the leaf surface waxes on these mutants.     

Leaf epidermis of Quercus subgen. Cyclobalanopsis (Oerst.) Schneid. (Fagaceae)

The characters of leaf epidermis (mainly epidermal cells, stomata, trichomes) have been examined in 48 species of Quercus subgen. Cyclobalanopsis of the Fagaceae. The adaxial leaf epidermis was investigated under light microscope. The epidermal cells of this surface are of two types in terms of the outline of their anticlinal wall. One type is characterized by having straight wall, and this type has been observed in most of the species studied, while the other is characterized by having sinuate or sinuolate wall, and this type has been found only in eight species, such as Q. blakei and Q. hui. There exist two types of trichome-bases on this surface: in one type the bases are singlecelled, and this type has been found in most of the species studied, such as Q. oxyodon and Q. augustinii; while in the other the bases consist of a group of cells, and this type has been found only in nine species, such as Q. thorelii and Q. chungii. The abaxial leaf epidermis was investigated under light microscope and scanning electron microscope. The epidermal cells and the trichome-bases on this surface under LM are in morphology essentially similar to those on the adaxial surface. The stomata under LM include three types: cyclocytic, subcyclocytic and anomocytic. The stomatal density is of some diagnostic value to distinguish between closely related species, e.g. the stomatal density in Q. glauca is higher than that in Q. schottkyana. Under SEM the stomata are elliptical or roundish. The most noteworthy feature of the adaxial leaf epidermis is the trichomes. Eight types of trichomes have been observed under SEM: simple-uniseriate, solitary, papillae, stellate, fused-stellate, stipitate-fasciculate, multiradiate and jellyfish-like. The first type, i.e. the simple-uniseriate trichome, is glandular, other six types, i.e. the solitary, papillae, stellate, fused-stellate, stipitate-fasciculate and multiradiate trichomes, are non-glandular, and the last one, i.e. the jellyfish-like trichome, may represent an intermediate type in structure between the glandular and the non-glandular trichomes. Papillae are very common in subgen. Cyclobalanopsis although in the Fagaceae they were previously reported only in a few species of Lithocarpus. The jellyfish-like trichome, observed only in Q. sichouensis, is reported in the Fagaceae for the first time. The evolutionary trends of trichomes are discussed. The trichomes seem to evolve toward the direction of increasing complexity in structure. Papillae are considered as the pedomorphism of trichomes. From papillae the solitary trichomes and then the stellate ones have evolved. The complicated trichome types with more arms may be produced from relatively simple ones with fewer arms by means of varions fusion: several stellate trichomes are fused to form the fused-stellate ones by basal fusion, and by the same way the fused-stellate ones to form the stipitate-fasciculate ones; the multiradiate trichomes are formed by the stellate ones, in which the arms emerge in a variety of seemingly random directions from a typically rounded common base. Based on the above results, it is considered that the trichome features are of important significance for a better understanding of the infrageneric division of Quercus and the phylogenetic relationships between this genus and the other genera in the Fagaceae. The very common presence of papillae in Quercus subgen. Cyclobalanopsis and in a few species of Lithocarpus may indicate that subgen. Cyclobalanopsis is a quite natural group on the one hand, and may have close relationship with Lithocarpus on the other hand. The thin-walled, jellyfish-like trichomes found in Q. sichouensis are in structure similar to the “glandular” peltate trichomes in Trigonobalanus, a genus generally regarded as the most primitive in subfam. Quercoideae, indicating that the affinity between subgen. Cyclobalanopsis and Trigonobalanus may becloser than that between subgen. Quercus and Trigonobalanus.     

栎属青冈亚属（壳斗科）的叶表皮研究

利用光镜和扫描电镜观察了栎属青冈亚属Quercus subgen. Cyclobalanopsis 48种植物的叶表皮,尤以对叶下表皮的毛被特征观察较为仔细。共观察到8种不同类型的叶表皮毛：单列毛、单毛、乳突、星状毛、溶和星状毛、具柄束毛、多出毛和水母状毛。其中乳突在青冈亚属中较常见,而在壳斗科其他属中仅在石栎属 Lithocarpus 少数种类中有报道;水母状毛首次在壳斗科中发现。毛被可能遵循以下的演化规律：乳突→单毛→星状毛;星状毛依照从简单→复杂的演化途径,分化出各种形态各异和结构复杂的毛系。初步讨论了毛被以及叶表皮其他特征(如毛基细胞和表皮细胞的形态、气孔的类型和密度等)的分类和系统学意义。     

Pre-exposure to ozone predisposes oak leaves to attacks by Diplodia corticola and Biscogniauxia mediterranea

One-year-old cork oak (Quercus suber) and turkey oak (Q. cerris) seedlings were exposed to ozone (110 ppb, 5 h day(-1), for 30 days) and were inoculated with Diplodia corticola and Biscogniauxia mediterranea, respectively, by spraying a suspension of spores on the leaves. Both fungi are endophytic and may act as weak parasites, contributing to oak decline. Ozone exposure stimulated leaf attacks after inoculation, although the physiological, visible, and structural responses of both oaks to O3 exposure were weak. In fact, steady-state gas exchange, leaf waxes, and wettability were not significantly affected by O3. In Q. cerris, O3 altered the structure of stomata, as observed by scanning microscopy, and reduced the leaf relative water content. No hyphal entry through stomata or growth towards stomata was, however, observed. Inoculations were performed in a humid chamber at low light; stomata were likely to be closed. When Q. cerris was inoculated in natural conditions, i.e., in a forest infected by B. mediterranea, seedlings pre-exposed to the enhanced O3 regime had a higher number of B. mediterranea isolates than the controls. This suggests that pre-exposure to O3 predisposed Q. cerris leaves to attacks by B. mediterranea independent of stomata. The hyphae of both fungi were able to enter the leaf through the cuticle, either by gradual in-growth into the cuticle or erosion of a hollow in the cuticle at the point of contact. The primary cause of increased leaf injury in O3-exposed seedlings appeared to be higher germination of spores than on control leaves.     

Systematic anatomical characters of the leaves of Dimorphandra and Mora (Leguminosae: Caesalpinioideae)

A systematic study of the leaf anatomy of Dimorphandra and Mora showed that the leaf epidermis provides taxonomic characters for the separation of the two sections of Dimorphandra, Pocillum and Dimorphandra , viz. the structure of the guard cells and the degree of sunkenness of the stomata. The diverse arrangements of the neighbouring cells around the stomata do not provide a clear distinction between the sections: in section Pocillum the stomata are paracytic but in section Dimorphandra several types are found. Papillae of diverse shape, size and disposition are present in the different species of Dimorphandra , and with all these variations in the epidermal characters it is possible to establish series of increasing complexity in the two sections.
Mora presents several characters which distinguish it from Dimorphandra (e.g. thickenings in the walls of the stomata and other epidermal cells, granular silica in the lumen of some epidermal cells, and sinuous anticlinal walls in the cells of both epidermides), thus supporting its recognition as a separate genus.     

Automatic measurement of stomatal density from microphotographs

Key message

An automated process using a cascade classifier allowed the rapid assessment of the density and distribution of stomata on microphotographs from leaves of two oak species.

Abstract

Stomatal density is the number of stomata per unit area, an intensively studied trait, involved in the control of CO₂ and H₂O exchange between leaf and atmosphere. This trait is usually estimated by counting manually each stoma on a given surface (e.g., a microphotograph), usually repeating the procedure with images from different parts of the leaf. To improve this procedure, we tested the performance of a cascade classifier to automatically detect stomata on microphotographs from two oak species: Quercus afares Pomel and Quercus suber L. The two species are phylogenetically close with similar stomatal morphology, which allowed testing the reuse of the cascade classifier on stomata with similar shape. The results showed that a cascade classifier trained on only 100 sample views of stomata from Q. afares was able to rapidly detect stomata in Q. afares as well as in Q. suber with a very low number of false positives (5 %/1.9 %) and a small number of undetected stomata (14.8 %/0.74 %), when partial stomata near the edge of the microphotographs were ignored. The remaining undetected stomata were due to obstacles such as trichomes. As an example of further applications, we used the positions detected by the cascade classifier to assess the spatial distribution of stomata and group them on the leaf surface. To our knowledge this is the first time that a cascade classifier has been applied to plant microphotographs, and we were able to show that it can dramatically decrease the time needed to estimate stomatal density and spatial distribution.     

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.     

The potential of stomata analysis in conifers to estimate presence of conifer trees: examples from the Alps

To estimate whether or not a plant taxon found in the fossil record was locally present may be difficult if only pollen is analyzed. Plant macrofossils, in contrast, provide a clear indication of a taxon’s local presence, although in some lake sediments or peats, macrofossils may be rare or degraded. For conifers, the stomata found on pollen slides are derived from needles and thus provide a valuable proxy for local presence and they can be identified to genus level. From previously published studies, a transect across the Alps based on 13 sites is presented. For basal samples in sandy silt above the till with high pollen values of Pinus, for example, we may distinguish pine pollen from distant sources (samples with no stomata), from reworked pollen (samples with stomata present). The first apparent local presence of most conifer genera based on stomata often but not always occurs together with the phase of rapid pollen increase (rational limit). An exception is Larix, with its annual deposition of needles and heavy poorly dispersed pollen, for it often shows the first stomata earlier, at the empirical pollen limit. The decline and potential local extinction of a conifer can sometimes be shown in the stomata record. The decline may have been caused by climatic change, competition, or human impact. In situations where conifers form the timberline, the stomata record may indicate timberline fluctuations. In the discussion of immigration or migration of taxa we advocate the use of the cautious term “apparent local presence” to include some uncertainties. Absence of a taxon is impossible to prove.     

中国西北地区松科和柏科气孔器形态

现代针叶树气孔器的研究为鉴定化石气孔器奠定了基础,对第四纪植被变化和古气候的研究起着很重要的作用,是第四纪孢粉学的一个重要补充。本文运用常规的标准的孢粉分析方法,对中国西北地区常见的松科3属8种和柏科3属4种植物气孔器进行了观察分析。结果表明凭气孔器大小可以区分松科与柏科。利用气孔器大小、T型结构的形状?上部木质片与气孔器茎之间的夹角进行属的鉴别,再结合茎的长度、宽度、中间木质部的宽、T型外角度和上部木质片外缘形状等可以进行种的鉴别,并编制了一个初步的检索表。     

北京山区落叶阔叶林优势种叶片特点及其生理生态特性

荆条、山杏、辽东栎、北京丁香、大叶白蜡、核桃楸等６种植物在不同光照条件下的气孔导度,不同生境下叶片形态参数和叶片解剖特征进行了研究。扫描电子显微镜图片显示这片植物的气孔全着生在远轴面,气孔密度大小是：辽东栎〉山杏〉北京丁香〉核桃楸〉大叶白蜡,方差分析结果显示这５种植物叶片上气孔密度存在极显著差异,且叶片外表微观上差异比较明显。生境不仅影响叶片的气孔密度,而且也导气孔导度的差异,全光照条件下,山杏、     

Genetic diversity and phylogeny of the family Osteoglossidae by the nuclear 18S ribosomal RNA and implications for its conservation

The genetic structure of the family Osteoglossidae from different geographical regions was examined using the nuclear 18S ribosomal RNA (18S rDNA) sequence. The results showed that the partial length of 18S rDNA was 1277 bp, and they were relatively conserved in the species of the family Osteoglossidae. A total of 62 (4.83%) polymorphic sites were observed, and 28 haplotypes were defined, which were characterized by different haplotype diversity (0.00105–0.900) and nucleotide diversity (0.00321–3.000). Analysis of molecular variance (AMOVA) showed genetic divergence of these species (79.76%). Phylogenetic analysis revealed that 18S rDNA was a suitable genetic marker and was able to distinguish phylogenetic relationships among different Osteoglossidae species, and it also supported current taxonomy. In addition, low genetic diversity of several species provides genetic assessment information and should be taken into consideration to implement a conservation management planning for some species, such as Scleropages formosus green arowana.     

植物角质层蜡质的化学组成研究综述

角质层是植物与外界的第一接触面,而角质层蜡质则是由位于角质层外的外层蜡质和深嵌在角质层中的内层蜡质两部分构成。植物角质层蜡质成分极其复杂,具有重要的生理功能。综述了有关植物角质层蜡质的化学组成信息,探讨了目前植物角质层蜡质化学成分研究中存在的一些问题,展望了角质层蜡质成分的研究前景。     

No heat-stimulated germination found in herbaceous species from burned subtropical grassland

The inverse relationship between numbers of stomata (stomatal frequency) on tree leaves and ambient CO₂ concentration is increasingly applied for reconstructing past atmospheric CO₂ levels. The abundance of leaf remains of Quercus robur in Holocene peat and lake deposits in Europe makes this species potentially suitable for high-resolution stomatal frequency analysis. In order to quantify the CO₂ responsiveness of the species, the behavior of the stomatal index for Q. robur during the current anthropogenic CO₂ increase is determined on the basis of buried, herbarium and modern leaf material from the Netherlands. The stomatal index (SI), expressing the ratio of the number of stomata in a given area divided by the total number of stomata and other epidermal cells in that same area, is used in order to minimize influences on stomatal frequency of environmental conditions other than CO₂. The sigmoid SI response pattern recorded for Q. robur resembles that of the closely related species Q. petraea, although there is a difference in the timing of the response limitation of the two species to increasing atmospheric CO₂. For calibration purposes only the linear phase of the sigmoidal response curve is taken into consideration in the presented CO₂ response model, which allows confident combination of Q. robur and Q. petraea over the interval from 290 to 325 ppmv CO₂. The model is conservative in reconstructing past CO₂ mixing ratios outside the range of monitored response. As a result of the observed SI response limit, the model predicts CO₂ levels below 325 ppmv with a mean error of 10.2 ppmv, whereas higher CO₂ levels are underestimated.     

6种松科植物叶表皮的扫描电镜观察

应用扫描电子显微镜,观察了松科(Pinaceae)3个属6种植物叶表皮特征。金钱松气孔带位下表面,气孔区与非气孔区表皮细胞形态差异很大,气孔细缝状;雪松针叶四面都具气孔线,气孔下陷,孔口光滑,针叶表面具较厚的角质层,表皮细胞形状观察不清;五针松针叶仅两腹面有气孔线,气孔有隆起的圆环围绕,表面角质层呈条纹状排列,形成一种沟槽;黑松、湿地松、马尾松针叶腹背面都有气孔线,气孔呈蜂窝状排列成行,气孔下陷,孔口角质化强烈。结果表明松科3个属植物叶表皮的形态结构及气孔器差异显著,一定程度上证实了3属是自然的分类群。     

Stomatocysts of Mallomonas hamata and M. heterospina (Mallomonadaceae, Synurophyceae) from South Swedish lakes

The stomato cysts of Mallomonas hamata and M. heterospina are described from lakes in South Sweden. These Mallomonas species develop cysts of similar appearance, size and form. Cyst formation for Mallomonas hamata was recorded at temperature 10–11°C and pH 5.9 and for M. heterospina at temperatures 4.2–11°C and pH 7.4. The cysts of M. hamata and M. heterospina are ovoid and easy to recognize, even with light microscopy, but difficult to distinguish from another. Thus their value as paleoindicators is limited, even if the two species have preference for waters of different nutrient and pH status.     

大别山北坡落叶阔叶林种间相关的研究

应用联结系数AC,点相关系数PCC,x~2检验和pearson相关系数r公式测定了大别山北坡落叶阔叶林的种间关系,依据种间联结程度将乔木层划分出3个生态种组,即栓皮栎──枫香种组,化香──短柄抱种组和锐齿栎──灯台树种组。