Temporal and Spatial Development of the Cells of the Expanding First Leaf of Arabidopsis thaliana (L.) Heynh

The three-dimensional quantitative leaf anatomy in developingyoung (9–22 d) first leaves of wild type Arabidopsis thalianacv. Landsberg erecta from mitosis through cell and leaf expansionto the cessation of lamina growth has been studied. The domainsof cell division, the relative proportion of the cell typespresent during development and the production of intercellularspace in the developing leaf have been determined by image analysisof entire leaves sectioned in three planes. Mitotic activityoccurs throughout the youngest leaves prior to unfolding andcell expansion is initiated firstly at the leaf tip with a persistentzone of mitotic cells at the leaf base resulting in a gradientof development along the leaf axis, which persists in the olderleaves. Major anatomical changes which occur during the developmentare, a rapid increase in mesophyll volume, an increase in thevein network, and expansion of the intercellular spaces. Thepattern of cell expansion results in a 10-fold variation inmesophyll cell size in mature leaves. In the youngest leavesthe plan area of mesophyll cells varies between 100 µm²and 400 µm² whereas in mature leaves mesophyll cells rangein plan area from 800 µm² to 9500 µm². The volumesof mesophyll tissue and airspace under unit leaf area increase3-fold and 35-fold, respectively, during leaf expansion. Thevolume proportions of tissue types mesophyll:airspace:epiderrnal:vascularin the mature leaf are 61:26:12:1, respectively. This studyprovides comparative information for future identification andanalysis of leaf development mutants of Arabidopsis thaliana. Key words: Arabidopsis, quantitative leaf anatomy, leaf expansion, image analysis     

Genome Expression during Normal Leaf Development : I. CELLULAR AND CHLOROPLAST NUMBERS AND DNA, RNA, AND PROTEIN LEVELS IN TISSUES OF DIFFERENT AGES WITHIN A SEVEN-DAY-OLD WHEAT LEAF

Changes in genome expression during normal cellular and plastid development in the first leaf of young (7-day-old) wheat (Triticum aestivum var. Maris Dove) were investigated by examining homogeneous populations of leaf cells and plastids of several developmental ages present in the same leaf. The cells were characterized over a period immediately following the last cell division. All of the leaf cells had cytoplasmic contents and nuclei, and between 44% (young tissue) and 54% (older tissue) of the leaf cells were mesophyll cells. Chloroplast development was complete 36 hours after the chloroplasts had ceased dividing. Extremely large changes occurred in cellular constituents over a very short period of leaf development. Maximum rates of accumulation of ribulose bisphosphate carboxylase per mesophyll cell (80 picograms/hour), chlorophyll per mesophyll cell (9 picograms/hour), and 70S ribosomes per mesophyll cell (19 × 10⁵/hour) were recorded.     

Ability of leaf mesophyll to retain potassium correlates with salinity tolerance in wheat and barley

This work investigated the importance of the ability of leaf mesophyll cells to control K⁺ flux across the plasma membrane as a trait conferring tissue tolerance mechanism in plants grown under saline conditions. Four wheat (Triticum aestivum and Triticum turgidum) and four barley (Hordeum vulgare) genotypes contrasting in their salinity tolerance were grown under glasshouse conditions. Seven to 10‐day‐old leaves were excised, and net K⁺ and H⁺ fluxes were measured from either epidermal or mesophyll cells upon acute 100 mM treatment (mimicking plant failure to restrict Na⁺ delivery to the shoot) using non‐invasive microelectrode ion flux estimation (the MIFE) system. To enable net ion flux measurements from leaf epidermal cells, removal of epicuticular waxes was trialed with organic solvents. A series of methodological experiments was conducted to test the efficiency of different methods of wax removal, and the impact of experimental procedures on cell viability, in order to optimize the method. A strong positive correlation was found between plants' ability to retain K⁺ in salt‐treated leaves and their salinity tolerance, in both wheat and especially barley. The observed effects were related to the ionic but not osmotic component of salt stress. Pharmacological experiments have suggested that voltage‐gated K⁺‐permeable channels mediate K⁺ retention in leaf mesophyll upon elevated NaCl levels in the apoplast. It is concluded that MIFE measurements of NaCl‐induced K⁺ fluxes from leaf mesophyll may be used as an efficient screening tool for breeding in cereals for salinity tissue tolerance.     

Microtubule rearrangements accompanying dedifferentiation in mesophyll cultures ofZinnia elegans L.

Summary To determine the orientation of cortical microtubule arrays in mesophyll cells ofZinnia, a new technique designed to increase the rate of fixation of excised leaf tissue and subsequent permeabilization of mesophyll cell walls was developed. This procedure resulted in immunolabeling of high percentages of mesophyll cells, making it possible to quantify cells with different types of cortical microtubule arrays. When developing palisade mesophyll cells were fixed in situ, most of the cells had cortical microtubules organized in parallel arrays oriented transverse to the long axis. Delay in the transfer of leaf tissue to fixative resulted in increased numbers of cells with random cortical microtubule orientations, indicating that arrays may become reoriented rapidly during leaf excision and cell isolation procedures. The role of wound-induced microtubule reorientation in mesophyll dedifferentiation and tracheary element development is discussed.Abbreviations BSA bovine serum albumin - CMT cortical microtubule - TE tracheary element - TBS tris-buffered saline     

Recent stereological methods for measuring leaf anatomical characteristics: estimation of the number and sizes of stomata and mesophyll cells

Recent design-based stereological methods for measuring stomatalfrequency, mean epidermal cell length, number of mesophyll cellsand mean cell volume are described in an easy-to-follow manner.Two leaf types are considered: grass leaf and bifacial leaf.The presented methods are discussed and compared with alternativemethods. Key words: Disector, leaf, mesophyll, stereology, stomata     

青藏高原草地植物叶解剖特征

运用常规石蜡制片技术对我国青藏高原66种草地植物优势种的叶解剖特征进行研究,并分析了叶解剖特征与海拔、生长季降水及生长季均温之间的关系.结果表明:青藏高原草地植物叶片具有很多适应高寒环境的结构特征,如表皮层厚且表皮细胞大小差异显著,表皮毛等表皮附属物发达,异细胞丰富,通气组织普遍发达等;叶片各组成部分厚度的变异程度不同,其中海绵组织厚度变异最大,其次为上角质层、下表皮层、下角质层、上表皮层、栅栏组织,叶片厚度的变异最小;青藏高原草地植物叶片各组成部分的厚度存在协同进化,上下角质层厚度呈强烈正相关,海绵组织厚度与叶片厚度相关性最强;青藏高原草地植物叶片各组成部分的厚度与海拔、生长季降水、生长季均温3个重要环境变量呈较弱的相关性,总体表现为随海拔升高叶片各组成部分的厚度减小,而随生长季降水和生长季均温的增加叶片厚度增加.     

A comparative study on the anatomy and development of different shapes of domatia in Cinnamomum camphora (Lauraceae)

BACKGROUND AND AIMS: Domatia are small organs usually found in the axils of major veins on the underside of leaves and, although they have received wide attention from ecologists, few detailed reports exist on their anatomy or development. This study is focused on the domatia of Cinnamomum camphora (Lauraceae) and is the first comparative study on the anatomy and development of the different shapes of domatia within a single plant. METHODS: Four types of domatia in C. camphora leaves were observed on paraffin sections under a microscope. KEY RESULTS: The domatia consisted of six histological parts: the upper epidermis, the upper mesophyll tissue, spongy tissue, the lower mesophyll tissue, the tissue filling the rim opening, and the lower epidermis. They differed from the non-domatial lamina mainly in the cell structure of the upper and lower mesophyll tissue and the rim tissue. Differences in domatium shapes were mainly associated with differences in the structure of the upper mesophyll and in the number and size of the rim tissue cells. Differences in the development of domatium types were observed in terms of initiation timing, differentiation of the upper mesophyll cells and degree of rim tissue development. CONCLUSIONS: In domatia, active anticlinal division in the lower mesophyll cells, as compared with the upper mesophyll cells, was coordinated with dynamic growth of rim tissue cells and resulted in cavity formation. The anatomical or developmental differences among the four types of domatia were related to the positions of the domatia within a leaf. In terms of the ecological implications, the major anatomical difference between the domatia used by herbivorous and carnivorous mites was in the development of the rim tissue.     

Models for mesophyll cell arrangement in leaves of ryegrass (Lolium perenne L.)

Summary Three models for mesophyll cell arrangement in leaves of Lolium perenne L. are described. It is shown that mesophyll cells may be represented as spheres packed in a tetragonal lattice. The need to quantify mesophyll cell arrangement is discussed in relation to leaf photosynthesis.The variation on measurements of some anatomical features of leaves is examined and discussed.     

Structure of the Photosynthetic Apparatus in Leaves of Freshwater Hydrophytes: 2. Quantitative Characterization of Leaf Mesophyll and the Functional Activity of Leaves with Different Degrees of Submersion

The structure of leaf photosynthetic elements was investigated on 42 boreal plant species characterized by different degrees of submergence (helophytes, neustophytes, and hydatophytes). Six main types of mesophyll structures were identified. Quantitative characteristics for the mesostructure of the photosynthetic apparatus in these groups were determined, such as the size and abundance of cells and chloroplasts in the mesophyll and epidermis, the number of plastids per cell in each tissue, the total surface area of the mesophyll cells, epidermal cells, and chloroplasts per unit leaf area. Analysis showed that quantitative characteristics of the photosynthetic apparatus in hydrophytes are determined by two factors: (a) the degree of leaf submergence and (b) the type of mesophyll structure. With an increasing degree of immersion in water, the mesophyll types change in a sequence isopalisade dorsoventral homogeneous. The leaves become thinner, their weight per unit area diminishes, cells and chloroplasts become less numerous (on a per unit leaf area basis), but their dimensions become larger. Adaptation to aquatic medium is also manifested in the increasing contribution of the epidermal tissue to the overall photosynthesis: in submerged leaves, the epidermis accounts for more than 50% of the photosynthetic activity. The occurrence of six structural types of leaves contrasting in their characteristics was confirmed by discriminatory analysis according to the qualitative parameters of mesophyll.     

榉树叶片解剖构造和叶肉细胞超微结构的观察

对取自不同季节的榉树(Zelkova schneideriana hand.-Mazz.)叶的解剖构造和叶肉细胞超微结构进行了观察。榉树叶表面被密集锥状单细胞毛,幼叶则还有头状腺毛。气孔多分布在叶下表面,为不规则型。上表皮细胞外壁具有明显的角质层加厚,局部上表皮由2层细胞组成。叶肉细胞分化为栅栏组织和海绵组织;栅栏组织1—2层细胞;海绵组织发达,常与气孔相连。叶肉有含晶细胞存在。叶脉的维管束鞘明显,是厚壁细胞。成熟叶肉细胞细胞器丰富,有8-9个周壁叶绿体,为巨大的长椭圆形,基粒片层清晰,有淀粉粒,少量脂滴。叶片开始转变颜色时,叶绿体由长椭圆形变得趋于圆形;基粒片层逐渐变得杂乱至模糊不清;脂滴数目增多。秋季叶色不同的榉树叶片解剖构造存在一定差异。     

Curvature Correction Factors in the Measurement of Cell Surface Areas in Plant Tissues1

The simplest and most reliable method for the estimation ofthe surface area of cells in plant tissues involves the measurementof the total length of the perimeters of cell profiles in thinsections of the tissue. However, the method suffers from errorsdue to neglect of curvature of the cell surfaces. The effectsof the curvature can be compensated for by multiplying the measuredlength of profile perimeter by appropriate curvature correctionfactors. This paper presents values for these curvature correctionfactors, which are shown to be significantly large. The applicationof these correction factors to the measurement of cell surfaceareas in plant tissues is discussed with particular referenceto leaf mesophyll tissue. The choice of appropriate orientationsof the sections in a tissue is also discussed. Key words: Cell, Surface, Area, Curvature correction     

Comparative analyses of leaf anatomy of dicotyledonous species in Tibetan and Inner Mongolian grasslands

Knowledge of the leaf anatomy of grassland plants is crucial for understanding how these plants adapt to the environment. Tibetan alpine grasslands and Inner Mongolian temperate grasslands are two major grassland types in northern China. Tibetan alpine grasslands occur in high-altitude regions where the low temperatures limit plant growth. Inner Mongolian temperate grasslands are found in arid regions where moisture is the limiting factor. Few comparative studies concerning the leaf anatomy of grassland plants of the Tibetan Plateau and Inner Mongolian Plateau have been conducted. We examined leaf characteristics at 71 sites and among 65 species, across the alpine grasslands of the Tibetan Plateau and the temperate grasslands of the Inner Mongolian Plateau. We compared the leaf structures of plants with different life forms and taxonomies, and their adaptation to arid or cold environments. We explored relationships among leaf features and the effects of climatic factors (i.e., growing season temperature and precipitation) on leaf characteristics. Our results showed that (i) there were significant differences in leaf anatomy between Tibetan alpine and Inner Mongolian temperate grasslands. Except for mesophyll cell density, the values obtained for thickness of leaf tissue, surface area and volume of mesophyll cells were larger on the Tibetan Plateau than on the Inner Mongolian Plateau. (ii) Within the same family or genus, leaf anatomy showed significant differences between two regions, and trends were consistent with those of whole species. (iii) Leaf anatomy of woody and herbaceous plants also showed significant differences between the regions. Except for mesophyll cell density, the values obtained for the thickness of leaf tissue, and the surface area and volume of mesophyll cells were larger in herbaceous than in woody plants. (iv) Leaf anatomical traits changed accordingly. Total leaf thickness, thicknesses of lower and upper epidermal cells, and surface area and volume of mesophyll cells were positively correlated, while mesophyll cell density was negatively associated with those traits. (v) Growing season temperature had stronger effects on leaf anatomy than growing season precipitation. Although the communities in Tibetan and Inner Mongolian grasslands were similar in appearance, leaf anatomy differed; this was probably due to the combined effects of evolutionary adaptation of plants to environment and environmental stress induced by climatic factors.     

番荔枝科蚁花属和澄广花属叶的比较解剖学研究

利用扫描电镜技术,叶片离析方法和石蜡切片法对蚁花属1种和澄广花属9种植物叶的形态结构进行比较研究。结果表明,两属植物有许多相似之处,但又有以下一些显著不同;蚁花属植物叶表皮细胞均具一晶族,叶肉组织中具1-2层栅栏组织细胞,油细胞均匀分布在栅栏组织和海绵组织中,栅栏组织在主脉处不连续,而澄广花属植物叶的表皮细胞内具一单斜晶,叶肉组织中具1层栅栏组织细胞,油细胞仅分布在海绵组织中,栅栏组织在主脉处连续,结果为蚁花属和澄广花属的分类学处理提供了新证据。     

Basic Types of Structural Changes in the Leaf Mesophyll during Adaptation of Eastern Pamir Plants to Mountain Conditions

Quantitative characteristics of mesophyll structure were compared in leaves of eleven alpine plant species grown under natural conditions in the Eastern Pamirs at various altitudes, from 3800 to 4750 m. Basic types of changes in mesophyll structure, associated with plant adaptation to mountain conditions, were characterized. These changes manifested themselves in different numbers of cell layers and cell sizes in the palisade tissue and, as a consequence, in changed leaf thickness and cell number per unit of leaf area. Three plant groups were identified by the changes in the leaf structural characteristics depending on the type of mesophyll structure, ecological group of plant species, and altitude of plant habitat. The first group comprised alpine xerophytes with an isopalisade structure, in which the volume of palisade cells decreased and their number per unit of leaf area increased with the altitude of plant habitat. The number of mesophyll layers and leaf thickness decreased or did not change with altitude. The second group comprised subalpine plant species with a dorsoventral structure of mesophyll; these species occur below the line of continuous night frost. In these plant species, the number of mesophyll layers, leaf thickness, and cell number per unit of leaf area increased with altitude. The third group comprised mesophyte plants with a dorsoventral and homogenous mesophyll structure, which are encountered in a wide range of habitats, including the nival belt (from 4700 to 5000 m). In this group, cell volume increased and cell number per unit of leaf area decreased with altitude. We present a general scheme of leaf structural changes, which explains the changes in the quantitative characteristics of mesophyll as a function of altitude and highland environmental conditions.     

The responses of guard and mesophyll cell photosynthesis to CO2, O2, light,and water stress in a range of species are similar

High resolution chlorophyll a fluorescence imaging was used to compare the photosynthetic efficiency of PSII electron transport (estimated by Fq'/Fm') in guard cell chloroplasts and the underlying mesophyll in intact leaves of six different species: Commelina communis, Vicia faba, Amaranthus caudatus, Polypodium vulgare, Nicotiana tabacum, and Tradescantia albifora. While photosynthetic efficiency varied between the species, the efficiencies of guard cells and mesophyll cells were always closely matched. As measurement light intensity was increased, guard cells from the lower leaf surfaces of C. communis and V. faba showed larger reductions in photosynthetic efficiency than those from the upper surfaces. In these two species, guard cell photosynthetic efficiency responded similarly to that of the mesophyll when either light intensity or CO2 concentration during either measurement or growth was changed. In all six species, reducing the O2 concentration from 21% to 2% reduced guard cell photosynthetic efficiency, even for the C4 species A. caudatus, although the mesophyll of the C4 species did not show any O2 modulation of photosynthetic efficiency. This suggests that Rubisco activity is significant in the guard cells of these six species. When C. communis plants were water-stressed, the guard cell photosynthetic efficiency declined in parallel with that of the mesophyll. It was concluded that the photosynthetic efficiency in guard cells is determined by the same factors that determine it in the mesophyll.     

STRUCTURE OF THE LEAF OF PYROSSIA LONGIFOLIA—A FERN EXHIBITING CRASSULACEAN ACID METABOLISM

The leaf of Pyrossia longifolia (Burm.) Morton, an epiphytic fern known to exhibit CAM, was examined by light and electron microscopy. The relatively thick leaf contains a single-layered epidermis, “water-storage” tissue, and a reticulate vascular system embedded in mesophyll tissue not differentiated into palisade and spongy layers. Mesophyll is composed of large, slightly elongate cells each with a thin, parietal layer of cytoplasm and a large central vacuole. The chloroplast-microbody ratio in mesophyll cells indicates that Pyrossia may be a high photorespirer and thus similar in that sense to C₃ plants. Mesophyll is separated from the vascular tissue by a tightly-arranged layer of endodermal cells with Casparian strips. The inner layer of mesophyll cells and the endodermal cells lack suberin lamellae. The collateral veins contain sieve elements, tracheary elements, pericycle and vascular parenchyma cells, the latter conspicuously larger than the sieve elements. The vascular parenchyma is the only cell type in the leaf which contains plastids with a peripheral reticulum. The parenchymatic elements of the leaf are connected by plasmodesmata, all of which lack neck constrictions and sphincters, or sphincter-like structures. The connections between sieve elements and adjacent parenchymatic elements are pore-plasmodesmata characterized by prominent wall thickenings on the parenchymatic-element side of the wall. The distribution and relative frequencies of plasmodesmata between the various cell types of the leaf indicate photoassimilates may move either symplastically or by a combination of symplast and apoplast from the mesophyll to the site of phloem loading in the veins.     

Leaf structure and specific leaf mass: the alpine desert plants of the Eastern Pamirs, Tadjikistan

This study examines interrelationships between eight leaf attributes (specific leaf mass, area, dry mass, lamina thickness, mesophyll cell number per cm², mesophyll cell volume, chloroplast volume, and number of chloroplasts per mesophyll cell) in field-grown plants of 94 species from the Eastern Pamir Mountains, at elevations between 3800 and 4750 m. Unlike most other mountain areas, the Eastern Pamirs, Karakorum system, Tadjikistan provide localities where low temperatures and radiation combine with moisture stress at high altitudes. For all the attributes measured, significant differences were found between plants with different mesophyll types. Leaves with dorsiventral palisade structure (dorsal palisade, ventral spongy mesophyll cells) had thicker leaves with larger but fewer mesophyll cells, containing more and larger chloroplasts. These differences in mesophyll type are reflected in differences in the total surface of mesophyll cells per unit leaf area ( A _mes/ A ) or volume ( A _mes/ V ). Plants with isopalisade leaf structure (palisade cells under both dorsal and ventral surfaces) are more commonly xerophytes and their increased values of A _mes/ A and A _mes/ V decrease CO₂ mesophyll resistance, which is an important adaptation to drought. Path analysis shows the critical importance of mesophyll cell volume in leading to the covariance between the different leaf attributes and hence to specific leaf mass (SLM), even though mesophyll cell volume is not itself strongly correlated with SLM. This is because mesophyll cell volume increases SLM through its effects on leaf thickness and chloroplast number per cell, but decreases SLM through its negative effect on mesophyll cell density.     

Profiles of photosynthetic oxygen-evolution within leaves of Spinacia oleracea

Oxygen evolution was measured from mesophyll tissues in spinach leaves using a photoacoustic technique. The photosynthetic capacity of individual cell layers was measured by directing microscopic beams of light, 40 μm wide, to cells exposed within a leaf cross section. The resulting profile for oxygen-evolution potential was relatively flat, indicating a uniform capacity for photosynthesis in leaf mesophyll tissues. Two experimental approaches were used to estimate the photosynthetic performance of individual mesophyll cell layers when white light was applied to the adaxial leaf surface. These experiments indicated that oxygen was produced relatively uniformly across the mesophyll and that oxygen evolution increased with irradiance of the white light applied to the leaf surface. The measured profiles for oxygen evolution and capacity are flatter than previous measurements of profiles of fixed carbon and estimates of profiles for absorbed light within spinach leaves.     

Variation in Mesophyll Cell Number and Size in Wheat Leaves

The numbers of mesophyll cells in wheat leaves were determinedin a variety of wheat species differing in ploidy level andin leaves from different positions on the wheat plant. Leafsize and mesophyll cell number are linearly related in bothcases but differences were observed in mesophyll cell numberper unit leaf area with changing leaf size. Where changes incell size are caused either by nuclear ploidy or leaf position,differences in mesophyll cell number per unit leaf are negativelycorrelated with mesophyll cell plan area. The decrease in cellsize with increasing leaf position also results in a greaternumber of chloroplasts per unit leaf area. These results arediscussed in relation to anatomical variation of the wheat leaf. Mesophyll cell, cell numbers, leaf size, Triticum     

Leptosphaeria maculans elicits apoptosis coincident with leaf lesion formation and hyphal advance in Brassica napus

Programmed cell death, with many of the morphological markers of apoptosis, is increasingly recognized as an important process in plant disease. We have investigated the involvement and potential role of apoptosis during the formation of leaf lesions by the fungus Leptosphaeria maculans on susceptible Brassica napus cv. Westar. There were no signs of host cell damage until 7 to 8 days postinoculation (dpi), when trypan-blue-stained leaf mesophyll cells were first detected. Hyphae were visible in the intercellular spaces of the inoculated area from 5 dpi and were associated with trypan-blue-stained cells at 8 to 9 dpi. Hallmarks of apoptosis, observed coincident with or immediately prior to the formation of leaf lesions at 8 to 10 dpi, included membrane shrinkage of the mesophyll cell cytoplasm, loss of cell to cell contact in mesophyll cells, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling of nuclei in apparently "healthy" tissue immediately adjacent to dead areas. Hyphae were highly branched and prolific in the "healthy" tissue immediately adjacent to dead areas 9 to 10 dpi, and formed pycnidia inside dead areas 11 to 12 dpi. Coinfiltration of the tetrapeptide caspase inhibitor Ac-DEVD-CHO with spores of the pathogen significantly suppressed development of leaf lesions but did not affect fungus viability. We hypothesize that L. maculans elicits apoptosis as a dependent component of pathogenesis in susceptible B. napus, and that the fungus uses apoptotic cells as a source of nutrition for reproduction and further growth.