植物叶片解剖结构的量化及其在C4植物高粱中的应用北大核心CSCD

测量切片内所有细胞或细胞器的总周长,并乘以切片厚度和校正因子是一种量化植物叶片解剖结构的常用方法。前人在使用该方法时因忽视了细胞大小、切片厚度等影响估算精确度的因素,从而导致计数样本偏少并产生较大误差。通过分析该方法误差产生的原因及其与精确度的关系,明确了该方法在实际应用中不同精确度所要求对应的切片数量。在此基础上,将该方法用于C4植物高粱(Sorghum bicolor)叶片结构的量化,提出叶肉细胞与维管束鞘细胞接触面积(CABM)等参数,发展并完善了植物叶片解剖结构的量化方法。     

青藏高原高寒草地和内蒙古高原温带草地主要双子叶植物叶片解剖特征的比较研究

青藏高原高寒草地和内蒙古高原温带草地同属中国地带性的草地类型.然而,主导它们的环境因子有所不同.前者主要受生长季低温的影响,后者主要受干旱的影响.发育在这两类草地上的植物,在解剖特征上有何共性和差异,目前少有报道,但却是理解植物适应环境的关键所在.本文通过对青藏高原高寒草地和内蒙古高原温带草地71个样地65种双子叶植物叶片解剖特征的研究,对比分析了不同科、属及不同生活型植物叶片对干旱环境以及高寒环境的适应特征,探讨了叶片总厚度、上表皮和下表皮厚度、叶肉组织厚度、叶肉细胞密度、叶肉细胞表面积和体积等多个解剖特征之间的关系以及气候因子(生长季温度、降水)对它们的影响.结果表明:(1)总体上看,青藏高原与内蒙古高原草地植物的叶片解剖特征存在显著差异,除叶肉细胞密度外,青藏高原高寒草地植物的叶片各组成部分厚度、叶肉细胞表面积和体积均大于内蒙古高原温带草地植物;(2)同一科属植物的叶片解剖特征在两地存在显著差异,并显示出与所有物种总体分析一致的规律;(3)在调查范围内,两地草本植物除叶肉细胞密度外,叶片各组成部分厚度以及叶肉细胞表面积、体积均显著大于木本植物;(4)叶片的各解剖指标间存在显著的协同变化:叶片总厚度,上、下表皮厚度,叶肉细胞厚度,叶肉细胞表面积和体积之间均呈显著正相关,而叶肉细胞密度与这些指标显著负相关;(5)总体上看,青藏高原和内蒙古高原草地植物叶片的解剖特征与生长季温度的关系比与生长季降水密切.因此,尽管青藏高原高寒草地和内蒙古高原温带草地在外貌上有趋同性,但叶片解剖结构有着显著差异,这可能是由于植物对环境的适应进化及(或)环境的筛选作用共同作用的结果.     

暗罗属植物叶的比较解剖学研究

利用扫描电镜技术、叶片叶片离析方法和石蜡切片法对暗罗属12种植物叶和形态结构进行了比较研究。结果表明,叶表皮细胞形状、气孔器形态、表皮毛类型、表皮细胞中晶体类型、叶肉中油细胞分布位置、栅栏组织和海绵组织厚度的比值,以及主脉维管组织的结构特征等具有明显的种间差异。可以利用这些叶的解剖特征将暗罗属植物相互区别开来。     

蒙古莸叶片解剖结构的地理种源变异及其对环境变化响应的意义

长期受到生长环境影响而形成的遗传变异对植物生长发育有着显著的影响。叶片是植物对环境变化最敏感的器官, 了解叶片解剖结构在不同环境中产生的适应性变异是探索植物对环境适应的基础。同质园试验是研究遗传与环境因素对植物生长代谢等影响的一种有效方法, 该研究利用同质园试验排除了环境梯度的影响, 通过常规石蜡切片、多重比较、相关性分析、一般线性模型分析等方法, 对7个不同种源地的蒙古莸(Caryopteris mongholica)叶片解剖结构及其影响因素进行了定量比较。结果表明, 7个种源地的蒙古莸叶片均为等面叶, 无海绵组织分化, 其上表皮细胞较下表皮细胞厚, 上栅栏组织较下栅栏组织厚; 叶片各解剖结构参数间存在显著的自相关性, 不同种源叶片解剖结构存在显著差异: 随种源地年平均气温升高, 叶厚度、栅栏组织厚度呈增大趋势, 其中, 最西南部的阿左旗种源蒙古莸叶片的上下栅栏组织、叶厚度及叶片结构紧密度值均最大, 表现出明显的抗旱特征。种源地经纬度、气温、降水等对解剖结构指标有显著的影响, 其解释程度为34.09%-81.43%。同质园试验说明, 种源地气候差异驱动的遗传变异是引起不同种源叶片解剖结构差异的重要因素。     

蒙古莸叶片解剖结构的地理种源变异及其对环境变化响应的意义

长期受到生长环境影响而形成的遗传变异对植物生长发育有着显著的影响。叶片是植物对环境变化最敏感的器官,了解叶片解剖结构在不同环境中产生的适应性变异是探索植物对环境适应的基础。同质园试验是研究遗传与环境因素对植物生长代谢等影响的一种有效方法,该研究利用同质园试验排除了环境梯度的影响,通过常规石蜡切片、多重比较、相关性分析、一般线性模型分析等方法,对7个不同种源地的蒙古莸(Caryopteris mongholica)叶片解剖结构及其影响因素进行了定量比较。结果表明,7个种源地的蒙古莸叶片均为等面叶,无海绵组织分化,其上表皮细胞较下表皮细胞厚,上栅栏组织较下栅栏组织厚;叶片各解剖结构参数间存在显著的自相关性,不同种源叶片解剖结构存在显著差异:随种源地年平均气温升高,叶厚度、栅栏组织厚度呈增大趋势,其中,最西南部的阿左旗种源蒙古莸叶片的上下栅栏组织、叶厚度及叶片结构紧密度值均最大,表现出明显的抗旱特征。种源地经纬度、气温、降水等对解剖结构指标有显著的影响,其解释程度为34.09%–81.43%。同质园试验说明,种源地气候差异驱动的遗传变异是引起不同种源叶片解剖结构差异的重要因素。     

六种沉香属植物叶片解剖结构研究

为比较沉香属不同种植物间的叶片形态解剖特征,将不同来源的六种沉香属植物在海南省兴隆南药园种植,运用石蜡切片法和撕片法对其成熟叶片的解剖特征进行观察,并对叶片的上下表皮,叶脉和叶横切面等12项数量性状进行统计分析。结果表明:六种沉香属植物叶片解剖结构基本一致,均为典型的异面叶,由表皮、叶肉和叶脉组成,表现出典型的旱生形态特点。表皮细胞单层,气孔微下陷,仅分布在下表皮,上下表皮上零星分布着表皮毛。叶肉组织发达,栅栏组织由1~2层排列紧密地圆柱状细胞组成,其间分布着大量的长方晶体,海绵组织内有一层排列较整齐,染色较深的异细胞组成的下皮层。主脉维管束双韧型,呈圆环状,内含大量异细胞。方差分析表明,除栅海比外,叶片厚度、叶脉条数、主脉厚度等其余11项数量指标在六种植物间差异均达到显著水平。聚类分析将这六种植物聚成3类,Aquilaria sinensis(白木香),A.crassna和A.banaensis聚为一类,A.baillonii和A.malaccensis聚为一类;A.yunnanensis(云南沉香)单独为一类。该研究结果为沉香属植物的物种鉴定提供了解剖学依据,同时对沉香属植物合理开发利用具有重要意义。     

新疆四种补血草属植物叶片的解剖学研究

利用叶片离析法和石蜡切片法研究了补血草属4种植物叶片的形态结构。结果表明:(1)4种植物的叶片有许多共同的结构适应特征,表皮细胞排列紧密,表面有厚的角质层;气孔类型均为不等型,气孔位置为平置或略微下陷;上下表皮还具有多细胞构成的盐腺;栅栏组织发达,多为等面叶;存在粘液细胞和单宁细胞;机械组织和维管组织都不发达等。(2)不同种间有不同的结构适应特征,如表皮细胞的形状、大小、垂周壁饰样,气孔密度,盐腺密度,叶片厚度和栅栏组织厚度等。通过叶的结构特征反映出盐生植物与旱生植物的不同。     

狗尾草属5种植物叶片的形态特征

为了解狗尾草属(Setaria Beauv.)植物的叶片形态特征,采用石蜡切片法和徒手切片法观察5 种狗尾草属植物的叶片形态结构。结果表明,5 种狗尾草属植物叶表皮的解剖特征较一致,叶表皮细胞长度、气孔器大小、叶厚、角质层厚度等性状在种间存在显著差异,其中长细胞形状、垂周壁深浅、气孔器副卫细胞的形状和刺毛数量可以用来区分种类。叶片性状的相关分析表明,叶片气孔密度与气孔长度以及气孔宽度呈显著负相关,泡状细胞厚度与角质层厚度以及上表皮厚度呈显著正相关。根据禾本科叶表皮特征的演化趋势,狗尾草(S. viridis)最原始,金色狗尾草(S. glauca)和大狗尾草(S. faberii)较进化,皱叶狗尾草(S.plicata)和莩草(S. chondrachne)最高级。这些为狗尾草属植物分类学和系统演化关系提供资料。     

陕北黄土高原优势植物叶片解剖结构的生态适应性

以陕北黄土高原地区7种优势植物为材料,比较了它们沟间地和沟谷地植株叶片解剖结构的差异,以揭示该地区优势植物对干旱强光照环境的生态适应性.结果表明:(1)7种植物各自具有抵抗黄土高原干旱强光照环境的特殊解剖结构:白羊草(Bothriochloa ischaemum)叶上表皮具有发达的泡状细胞,叶内具有花环结构;长芒草(Stipa bungeana) 叶上表皮下陷形成气孔窝,表皮下具有2至多层的厚壁组织;猪毛蒿(Artemisia scoparia)叶具有贮水组织、分泌组织和环栅型叶肉细胞;铁杆蒿(Artemisia sacrorum)叶肉全特化为栅栏组织;茭蒿(Artemisia giraldii)具有双栅型叶肉细胞和分泌结构;达乌里胡枝子(Lespedeza daurica)叶具有发达的粘液细胞;杠柳(Periploca sepium)叶表皮具厚蜡质层.(2)与沟谷地植物叶片结构相比,干旱强光照的沟间地植物叶片厚度、叶上表皮角质层厚度、栅栏组织厚度、贮水组织厚度增加,上表皮细胞体积、韧皮部面积增大,而木质部面积、木质部面积/韧皮部面积缩小.(3)叶片变异系数可反映植物适应环境的潜在能力,7种植物综合变异系数由高到低依次为猪毛蒿、铁杆蒿、茭蒿、白羊草、达乌里胡枝子、长芒草、杠柳.潜在适应能力最强的猪毛蒿已成为陕北黄土高原地区植被生态恢复的先锋物种之一.     

兴安落叶松针叶解剖结构变化及其光合能力对气候变化的适应性

叶片易受环境因子影响,其形态解剖结构特征不但与叶片的生理功能密切相关,而且反映树木对环境变化的响应和适应。叶片结构的改变势必会改变树木的生理功能。同一树种长期生长在异质环境条件下,经过自然选择和适应,会在形态和生理特性等方面产生变异,形成特定的地理种群。另外,母体所经受的环境胁迫也会影响到其子代的生长、发育和生理等特征。因此,了解植物叶片形态结构对环境变化的响应与适应是探索植物对环境变化的响应适应机制的基础。兴安落叶松(Larix gmelinii Rupr.)是我国北方森林的优势树种,主要分布在我国东北地区,但日益加剧的气候变化可能会改变其现有的分布区。为了区分叶片对气候变化的可塑性和适应性,本研究采用同质园法比较测定了6个不同气候条件下的兴安落叶松种源的32年生树木的针叶解剖结构和光合生理相关因子,利用石蜡切片方法分析了针叶的解剖结构特征、光合能力（Pmax-a）、水分利用效率（WUE）之间的关系及其对气候变化的适应性。结果表明：表皮细胞厚度、叶肉细胞厚度、传输组织厚度、维管束厚度、内皮层厚度以及叶片总厚度均存在显著的种源间差异（P < 0.05）。叶肉细胞厚度与Pmax-a、气孔导度和WUE之间均存在显著的正相关关系（P < 0.05）。叶肉细胞厚度、表皮细胞厚度、叶片总厚度以及叶肉细胞厚度和表皮细胞厚度在叶片总厚度中所占比例均与种源地的干燥度指数（即年蒸发量与年降水量之比）呈正线性关系。这些结果说明：不同种源兴安落叶松针叶解剖结构因对种源原地气候条件的长期适应而产生显著的差异,从而引起其针叶光合作用、水分利用等生理功能发生相应的变化,从而有利于该树种在气候变化的情景下得以生存和繁衍。     

MEASUREMENT OF THICKNESS WITHIN SECTIONS BY QUANTITATIVE ELECTRON MICROSCOPY

To apply the method of quantitative electron microscopy to the measurement of mass in thin sections, the thickness of the section at or very near the structure to be studied must be known. Dowex anion exchange resin AG 1 x 2, stained with phosphotungstic acid (PTA) at pH 6.4, was used as a thickness standard which could be embedded and sectioned. The sectioned PTA-Dowex appeared uniformly stained and exhibited suitable electron opacity. The stoichiometry of the reaction between PTA and the Dowex resin was measured by three independent methods based on gravimetric, colorimetric, and nitrogen determinations whose results showed close agreement. From the PTA uptake, the density of the stained spheres was calculated. Mass of a defined area of PTA-Dowex was measured by quantitative electron microscopy, and from this mass and density, the volume and then the thickness were calculated. The values for thickness were compared to those obtained by interference microscopy on the embedding medium alone in the same sections.     

Roles of climate and functional traits in controlling toothed vs. untoothed leaf margins

? Premise of the study: Leaf-margin state (toothed vs. untoothed) forms the basis of several popular methods for reconstructing temperature. Some potential confounding factors have not been investigated with large data sets, limiting our understanding of the adaptive significance of leaf teeth and their reliability to reconstruct paleoclimate. Here we test the strength of correlations between leaf-margin state and deciduousness, leaf thickness, wood type (ring-porous vs. diffuse-porous), height within community, and several leaf economic variables. ? Methods: We assembled a trait database for 3549 species from six continents based on published and original data. The strength of associations between traits was quantified using correlational and principal axes approaches. ? Key results: Toothed species, independent of temperature, are more likely to be deciduous and to have thin leaves, a high leaf nitrogen concentration, a low leaf mass per area, and ring-porous wood. Canopy trees display the highest sensitivity between leaf-margin state and temperature; subcanopy plants, especially herbs, are less sensitive. ? Conclusions: Our data support hypotheses linking the adaptive significance of teeth to leaf thickness and deciduousness (in addition to temperature). Toothed species associate with the "fast-return" end of the leaf economic spectrum, providing another functional link to thin leaves and the deciduous habit. Accounting for these confounding factors should improve climate estimates from tooth-based methods.     

中国东部亚热带青冈种群叶片的生态解剖

运用生态解剖学的方法对我国亚热带东部分布的１０个青冈种群叶片的解剖特征进行了比较分析,结果如下：①青冈叶片结构在种群间存在着广泛的差异,这种差异是青冈适应不同环境条件的结构基础。青冈叶片各部分结构特征在种群间的变化不晃同步的,变异系数（ＣＶ）在６．０％～２０．５％之间;变异幅度最大的是栅栏组织和上角质膜厚度,最小的是下表皮厚度。②相关分析表明,温度和降雨是影响青冈叶片地理变异的主导因子,其中温度、     

A quantitative method for assessing co-localization in immunolabeled thin section electron micrographs

A method is introduced for the analysis of nearest neighbor distances between immunogold particles marking proteins on electron micrographs. Deviation from the distribution that is predicted by chance indicates co-localization of the labeled species, and the potential for productive interaction in vivo. Application of this method to the analysis of nearest neighbor distances in experiments with pea leaf thin sections and isozyme-directed antibodies indicates that glyceraldehyde-3-P dehydrogenase is located near P-glycerate kinase and near aldolase in the chloroplast stroma, consistent with the notion that these enzymes are part of a multi-enzyme photosynthetic CO(2)-fixation complex in situ.     

Reconstructing adhesion structures in tissues by cryo-electron tomography of vitrified frozen sections

Cryo-electron tomography enables three-dimensional insights into the macromolecular architecture of cells in a close-to-life state. However, it is limited to thin specimens, <1.0 μm in thickness, typically restricted to the peripheral areas of intact eukaryotic cells. Analysis of tissue ultrastructure, on the other hand, requires physical sectioning approaches, preferably cryo-sectioning, following which electron tomography (ET) may be performed. Nevertheless, cryo-electron microscopy of vitrified sections is a demanding technique and typically cannot be used to examine thick sections, >80-100 nm, due to surface crevasses. Here, we explore the potential use of cryo-ET of vitrified frozen sections (VFSs) for imaging cell adhesions in chicken smooth muscle and mouse epithelial tissues. By investigating 300-400 nm thick sections, which are collected on the EM grid and re-vitrified, we resolved fine 3D structural details of the membrane-associated dense plaques and flanking caveoli in smooth muscle tissue, and desmosomal adhesions in stratified epithelium. Technically, this method offers a simple approach for reconstructing thick volumes of hydrated frozen sections.     

Impact of elevated CO(2) and O(3) on gas exchange parameters and epidermal characteristics in potato (Solanum tuberosum L.)

Potato plants (Solanum tuberosum L. cv. Bintje) were grown in open-top chambers (OTCs) under three CO(2) levels (ambient and 24 h d(-1) seasonal mean concentrations of 550 and 680 micromol mol(-1)) and two O(3) levels (ambient and a seasonal mean 8 h d(-1) concentration of 50 nmol mol(-1)). The objectives were to determine the effects of season-long exposure to these key climate change gases on gas exchange, leaf thickness and epidermal characteristics. The experimental design also provided an ideal opportunity to examine within-leaf variation in epidermal characteristics at the whole-leaf level. Stomatal and epidermal cell density and stomatal index were measured at specific locations on the youngest fully expanded leaf (centre of lamina, mid-way between tip and base) and representative whole leaves from each treatment. Effects on leaf conductance, assimilation rate and instantaneous transpiration efficiency were determined by infrared gas analysis, while anatomical characteristics were examined using a combination of leaf impressions and thin sections. Exposure to elevated CO(2) or O(3) generally increased leaf thickness, leaf area, stomatal density, and assimilation rate, but reduced leaf conductance. The irregular stomatal distribution within leaves resulted from a combination of uneven differentiation and expansion of the epidermal cells. The results are discussed with reference to sampling protocols and the need to account for within-leaf variation when examining the impact of climate change or other environmental factors on epidermal characteristics.     

An efficient direct induction of protocorm-like bodies from leaf subepidermal cells of Doritaenopsis hybrid using thin-section culture

High-frequency protocorm-like body (PLB) formation directly from thin leaf sections of Doritaenopsis hybrid was achieved in order to develop a mass-scale propagation system. Concentrated efforts were made to study the effects of different cytokinins on in vitro PLB induction from thin leaf sections. Among the cytokinins tested, thidiazuron (TDZ) was found to be a more effective inducer of PLBs than benzyladenine and zeatin. A modified Murashige and Skoog medium supplemented with 9.0 µM TDZ was found to be the optimum concentration for PLB development from thin leaf sections of Doritaenopsis hybrid. Of the two different explant types used in the present experiment, the highest percentage of PLB formation (72.3%) and highest number of PLBs (18) per explant were observed on thin leaf sections (1 mm thick), while only 20% (4.3 per explant) of comparatively large leaf segments (5 mm thick) were able to produce PLBs under the same culture conditions. Light microscopy observations indicated that the initial cell divisions for PLB formation occurred on the region near the cut surface and that an intact epidermal layer appeared to play an important role in PLB formation. Proembryo initiation occurred from several cells just beneath the intact epidermal cell, and globular PLBs were clearly visible after 3 weeks of culture and subsequently developed into mature PLBs.     

Increased ethylene and decreased phenolic compounds stimulate somatic embryo regeneration in leaf thin section cultures ofDoritaenopsis hybrid

The effects of endogenous levels of ethylene and phenolic compounds on somatic embryogenesis, medium-browning, and peroxidase activity were evaluated in thin section cultures ofDoritaenopsis. Cultures were maintained for 8 weeks with four different treatments: i) thick leaf segment culture, ii) thin leaf section culture, iii) thin leaf section culture with ventilation, or iv) thin leaf section culture after expiants were first washed. Expiants cultured in closed vessels produced a larger number of somatic embryos than those reared in the ventilated vessels. This enhanced formation confirmed the greater involvement of accumulated ethylene under non-ventilated conditions, because wound-induced tissues from thin leaf sections normally release high level of ethylene. When expiants were washed in the liquid medium and inoculated on the same solid medium, somatic embryo production was 1.7 and 18.5 times higher than in the thin section cultures and thick segment cultures, respectively. Reducing the level of phenolics in expiants at the initial stage of culturing apparently stimulated this embryo regeneration.     

Intraspecific variation in fiber properties inYucca elata andHesperaloe funifera (Agavaceae)

Yucca elata andHesperaloe funifera possess long, thin fibers that have potential for making specialty papers. The objective of this study is to examine patterns ofintraspecific variation in fiber properties in these two species. InYucca elata most of the variation in fiber length is found within populations where fiber length is highly correlated with leaf length. In contrast, inHesperaloe funifera there is significant variation between populations and random variation in fiber lengths within most populations. Within-plant variation inHesperaloe was also examined. Fiber length does not vary between leaves of different ages but does vary within leaves. Fibers from the base of the leaf are shorter and wider than those from the middle and distal sections; fibers from distal sections are narrowest.     

Phyllotactic pattern is altered in the transition to flowering in the early ears of Zea mays landrace chapalote (Poaceae)

The origin of polystichy in the maize ear and central tassel spike continues to challenge our understanding of evolution in this important crop species. In this paper we tested the hypothesis that the change in phyllotaxy occurs in the region of husk leaf production before the transition to reproductive growth. Young ear or presumptive ear primordia were dissected to examine the transition from distichous husk leaves below the ear through spiral phyllotaxy to the polystichous arrangement of spikelet pair primordia in the young ear. Serial transverse sections were used to document the thickness of successive disks of insertion of lateral primordia and to reconstruct the path of procambial differentiation. The transition in phyllotaxy, though variable, typically occurs in the vegetative zone and is associated with periodic heterogeneity in the thickness of leaf bases and a delay in the development of waves of procambial differentiation into the base of the young ear.