Comparative Characterization of the Pigment Complex in Emergent, Floating, and Submerged Leaves of Hydrophytes

The content of chlorophylls (Chls) and carotenoids was studied in the leaves of 42 species of boreal aquatic plants with different degree of submergence (emergent, floating, and submerged) and isopalisade, dorsoventral, and homogenous types of mesophyll structure. Hydrophytes were shown to have a low Chl content (1–2 mg/g fr wt) and low Chls/carotenoids ratio (2.3–3.5) as compared to terrestrial plants. The pigment content per dry wt unit and unit leaf area was dependent on the type of mesophyll structure. It was a consequence of the changes in the parameters of leaf mesophyll structure characterizing the density of photosynthetic elements. In a sequence emergent floating submerged forms, the content of Chls and carotenoids decreased, and the photosynthetic capacity decreased due to a reduction in the chloroplast number per unit leaf area. Adaptation of submerged leaves to low illumination and slow CO₂ diffusion changed the functional properties of chloroplasts. An increase in the pigment content in the chloroplasts of submerged leaves (7 × 10^–9 mg Chl, 2 × 10^–9 mg carotenoids) as compared to emergent and floating leaves was accompanied by a decline in the photosynthetic capacity per Chl comprising 1.6 mg CO₂/(mg Chl h) versus 3.9 and 3.8 mg CO₂/(mg Chl h) in emergent and floating leaves, respectively.     

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.     

Leaf anatomy and photosynthetic acclimation in <Emphasis Type="Italic">Valeriana jatamansi</Emphasis> L. grown under high and low irradiance

Relationship of leaf anatomy with photosynthetic acclimation of Valeriana jatamansi was studied under full irradiance [FI, 1 600 mol(PPFD) m^–2 s^–1] and net-shade [NS, 650 mol(PPFD) m^–2 s^–1]. FI plants had thicker leaves with higher respiration rate (R _D), nitrogen content per unit leaf area, chlorophyll a/b ratio, high leaf mass per leaf area unit (LMA), and surface area of mesophyll cell (S _mes) and chloroplasts (S _c) facing intercellular space than NS plants. The difference between leaf thickness of FI and NS leaves was about 28 % but difference in photon-saturated rate of photosynthesis per unit leaf area (P _Nmax) was 50 %. This indicates that P _Nmax can increase to a larger extent than the leaf thickness with increasing irradiance in V. jatamansi. Anatomical studies showed that the mesophyll cells of FI plants had no open spaces along the mesophyll cell walls (higher S _c), but in NS plants wide open spaces along the mesophyll cell wall (lower S _c) were found. Positive correlation between S _c and P _Nmax explained the higher P _Nmax in FI plants. Increase in mesophyll thickness increased the availability of space along the mesophyll cell wall for chloroplasts (increased S _c) and hence P _Nmax was higher in FI plants. Thus this Himalayan species can acclimate to full sunlight by altering leaf anatomy and therefore may be cultivated in open fields.     

Photosynthetic Metabolism and Activity of Carboxylating Enzymes in Emergent,Floating, and Submersed Leaves of Hydrophytes

Radioisotope techniques were used to compare photosynthetic CO₂ fixation, activities of carboxylating enzymes, and the composition of photosynthates in 42 species of aquatic plants (emergent, floating, and submersed hydrophytes) collected from rivers Sysert' and Iset' in Sverdlovsk oblast (Russia). The submersed leaves, in comparison with the emergent and floating leaves, featured lower rates of potential photosynthesis (by 2.2 mg CO²/(dm² h) on average), low content of the fraction I protein, and low activity of Rubisco and phosphoenolpyruvate carboxylase (PEPC). The averaged activities of Rubisco and PEPC were diminished in submersed leaves by 10 and 1 mg/(dm² h), respectively. Different hydrophyte groups showed similar composition of assimilates accumulated after 5-min photosynthesis and did not differ in this respect from terrestrial plants. However, the incorporation of ¹⁴C into sucrose and starch in submersed leaves (30 and 9% of total labeling, respectively) was lower than in emergent and floating leaves (45 and 15%, respectively). At the same time, the incorporation of ¹⁴C into C₄ acids (malate and aspartate) was 1.5 times higher in submersed leaves than in other leaf types. Analysis of leaf differentiation, the Rubisco/PEPC activity ratio, the PEPC activity, and the composition of primary photosynthates in the pulse–chase experiments revealed no evidence of the C₄ effect in the submersed hydrophytes examined. The adaptation of hydatophytes to specific conditions of an aquatic environment was structurally manifested in the reduction (by a factor of 3–5) in the number of chloroplasts per 1 cm² leaf area. This small number of chloroplasts was responsible for low photosynthetic rates in submersed leaves, although metabolic activities of individual chloroplasts were similar for all three hydrophyte groups.     

Construction costs and mesostructure of leaves in hydrophytes

Construction costs (CC) and parameters of leaf structure (specific leaf weight, dry matter content, volume of photosynthesizing cells, and the number of cells per leaf area unit) were determined for 19 species of aquatic higher plants. The CC of 1 g dry matter varied from 0.98 g glucose in Lemna gibba L. to 1.48 g glucose in Nuphar pumila (Timm) DC. and Potamogeton natans L. The CC of leaf area unit varied to a greater extent than the CC of 1 g dry wt (from 10 to 97 g glucose/m²) and depended on the type of mesophyll structure. In leaves of hydrophytes with dorsoventral mesophyll structure, the CC of 1 m² leaf area was 3–9 times larger than in leaves with homogeneous structure. Variations in CC of 1 m² leaf area in hydrophytes were affected insignificantly (by 2% only) by variations of CC per 1 g dry wt and were mainly determined (by 82%) by changes in specific leaf weight. Two-factor analysis of variance has shown that the CC of 1 g dry wt in hydrophytes depended on the attachment of plants to the sediment: the CC was 1.2 times larger in rooted hydrophytes than in free floating plants. The second factor (the extent of submergence) potentiated the effect of rooting on CC. Reliable differences were found between the leaf CC for hydrophytes belonging to four groups distinguished by the extent of their contact with water and sediment. In a group series: rooted hydrophytes with floating leaves → submerged rooted hydrophytes → free floating submerged hydrophytes → free floating surface inhabiting hydrophytes, the CC of 1 g dry wt decreased by 1.3 times. Path analysis has shown that this trend was due to the increase in photosynthesizing cell volume and to reduction in number of cells per leaf area unit, which caused the decrease in dry matter content. The decrease in the content of leaf dry matter was accompanied by changes in its chemical composition: the content of carbon and nitrogen decreased. This led to a consistent decrease in leaf CC expressed per 1 g dry wt upon the increase in extent of plant hydrophilicity.     

Structural Adaptation of the Leaf Mesophyll to Shading

Structural characteristics of the mesophyll were studied in five boreal grass species experiencing a wide range of light and water supply conditions. Quantitative indices of the palisade and spongy mesophyll tissues (cell and chloroplast sizes, the number of chloroplasts per cell, the total cell and chloroplast surface area per unit leaf surface area) were determined in leaves of each of the species. The cell surface area and the cell volume in spongy mesophyll were determined with a novel method based on stereological analysis of cell projections. An important role of spongy parenchyma in the photosynthetic apparatus was demonstrated. In leaves of the species studied, the spongy parenchyma constituted about 50% of the total volume and 40% of the total surface area of mesophyll cells. The proportion of the palisade to spongy mesophyll tissues varied with plant species and growth conditions. In a xerophyte Genista tinctoria, the total cell volume, cell abundance, and the total surface area of cells and chloroplasts were 30–40% larger in the palisade than in the spongy mesophyll. In contrast, in a shade-loving species Veronica chamaedris, the spongy mesophyll was 1.5–2 times more developed than the palisade mesophyll. In mesophyte species grown under high light conditions, the cell abundance and the total cell surface area were 10–20% greater in the palisade mesophyll than in the spongy parenchyma. In shaded habitats, these indices were similar in the palisade and spongy mesophyll or were 10–20% lower in the palisade mesophyll. In mesophytes, CO₂ conductance of the spongy mesophyll accounted for about 50% of the total mesophyll conductance, as calculated from the structural characteristics, with the mesophyll CO₂ conductance increasing with leaf shading.     

Degradation of chloroplast DNA in second leaves of rice (Oryza sativa) before leaf yellowing

Summary The second leaf ofOryza sativa develops, grows and ages within the 10 days that follow imbibition under our controlled continuous-light conditions. Proplastids in the leaf cells develop, mature to become chloroplasts and then age and disintegrate. In an examination of this life process, we studied first the behavior and the number of copies of plastid DNA and levels of chlorophyll by epifluorescence microscopy after staining with 4,6-diamidino-2-phenylindole (DAPI), and by fluorimetry with a video-intensified microscope photon-counting system (VIMPCS). The results indicated that the number of copies of the plastid DNA per plastid increased and reached to plateau value of approximately 100 at the time when the elongation of the mesophyll cells and the enlargement of chloroplasts ceased 96 h after imbibition. However, 24 h later, the number of copies of plastid DNA per chloroplast began to decrease and fell rapidly to approximately 30 copies within 168 h after imbibition. Our examination of the number of chloroplasts per mesophyll cell indicated that no division of chloroplasts occurred more than 72 h after imbibition. The results suggest that the decrease in number of copies of plastid DNA per chloroplast was not due to an increase in the number of chloroplasts, but that this decrease was caused by degradation by unidentified enzymes. Since visible senescence of leaves, which was characterized by development of a yellowish color, began 168 h after imbibition, the degradation of plastid DNA seemed to occur 48 h before the visible leaf senescence. When we tested the nucleolytic activities in the second leaves after imbibition by digestion of plasmids in vitro and DNA-SDS polyacrylamide gel electrophoresis, five Ca²⁺–, four Zn²⁺–, and four Mn²⁺–dependent nucleases were detected in the leaf blades, and one of the Ca²⁺–, two of the Zn²⁺–, and two of the Mn²⁺–dependent nucleases were also identified in a purified preparation of intact chloroplasts. When the activity of the Zn²⁺–dependent nucleases (51 kDa and 13 kDa) increased markedly, degradation of the plastid DNA occurred. These results suggest that the destruction of chloroplast DNA, which occurs approximately 48 h before leaf yellowing, could be due to the activation of some metallo-nucleases and, furthermore, this enzymatic degradation propels the leaf towards senescence.     

Photosynthetic Response to Irradiance in Valeriana jatamansi Jones,a Threatened Understorey Medicinal Herb of Western Himalaya

Net photosynthetic rate (P _N) of Valeriana jatamansi plants, grown under nylon net shade or under different tree canopies, was saturated with photons at 1 000 mol m^–2 s^–1 photosynthetic photon-flux-density (PPFD), whereas open-grown plants were able to photosynthesise even at higher PPFD, e.g. of 2 000 mol m^–2 s^–1. Plants grown under net shade had higher total chlorophyll (Chl) content per unit area of leaf surface. However, Chl a/b ratio was maximal in open-grown plants, but remained unchanged in plants grown in nylon net shade and under different tree canopies. Sun-grown plants had thicker leaves (higher leaf mass per leaf area unit), higher wax content, and higher P _N than shade grown plants. Thus V. jatamansi is able to acclimate to high PPFD and therefore this Himalayan species may be cultivated in open habitat to meet the ever-increasing industrial demand.     

Variation of leaf traits and pigment content in three species of steppe plants depending on the climate aridity

Mesophyll structure and content of photosynthetic pigments in the leaves of three species of steppe plants, Centaurea scabiosa L., Euphorbia virgata Waldst. et Kit., Helichrysum arenarium (L.) Moench, were investigated in four geographical sites of the Volga region and the Urals located in the forest-steppe and steppe zones. Variations of the studied parameters between geographical points depended both on the species and on the structural organization of the leaf. The highest level of variation was observed for leaf area and pigment content per unit leaf area, the size and the number of chloroplasts in the cell changed to a lesser extent. The leaf thickness, leaf area and mesophyll cell sizes mostly depended on the plant species. C. scabiosa had large leaves (40–50 cm²) with large thickness (280–290 μm) and large mesophyll cells (up to 15000 μm³). The leaves of H. arenarium and E. virgata were ten times smaller and characterized by 1.5 times smaller thickness and 2?3 times smaller cell size. Geographical location and climate of the region affected leaf density, proportion of partial tissue volume, and the ratio of the photosynthetic pigments. In the southern point of Volga region with the highest climate aridity, all studied species were characterized by maximum values of volumetric leaf density (LD), due to the high proportion of sclerenchyma and vascular bundles, and specificity of the mesophyll structure. With the decline in latitude, chlorophyll (Chl) and carotenoid (Car) contents in leaf area were reduced, the ratio Chl/Car was increased, and the ratio Chl a/b was declined. The reduction of the pigment content in the leaf in all species was associated with a reduction in the amount of Chl per chloroplast, and for C. scabiosa and H. arenarium it was associated also with the reduction of chloroplast amount in the leaf area. In turn, chloroplast number per leaf area and the total cell area (A_mes/A) depended on the ratio of the number and size of mesophyll cells inherent to this plant species. At the same time, we found a similar mechanism of spatial organization of leaf restructuring for all studied species—decrease in A_mes/A was accompanied by increasing in the proportion of intercellular air spaces in the leaf. It is concluded that variations in structural and functional parameters of the photosynthetic apparatus of steppe plants were associated with plant adaptation to climate features. General direction of the changes of leaf parameters of the studied species with aridity was the increase of LD and the decrease of pigment content per leaf area however the cellular mechanisms of changes in the pigment content and integral parameters of mesophyll were determined by the plant species properties.     

Selection of soybean plant leaves which yield mesophyll cell isolates with maximal rates of CO2 and NO inf2 sup− photoassimilation

A problem often encountered when assaying mesophyll cell isolates prepared from mature soybean leaves, was that of poor reproducibility in rates of net ¹⁴CO₂ photoassimilation and NO₂ ^– photoreduction. It was known that soybean source leaves repeatedly displayed their most active net CO₂ photoassimilation in the period from attainment of maximal leaf area to approximately two to five days subsequent to that point. Advantage was taken of the fact that when soybean leaflets of each leaf reach their maximal area they also have reached their maximal leaf length from base to tip. This facilitates a more rapid determination of the point in time in which leaflet areas had reached A_max. Soybean plants (Glycine max cv. Williams) were propagated in the growth chamber with a 12 h light-12 h dark cycle, 25C, 65% RH, and 700 microeinsteins per meter squared per second. At 24 d post-emergence, the third leaf (numbered acropetally from the unifoliates) of each plant had just attained maximum leaflet areas (110 cm²) and lengths (13 cm). For this study, leaf mesophyll cells were enzymatically isolated, using commercially prepared pectinase, from leaflet sets of leaves selected from each of the second, third, and fourth leaf positions. Maximal rates of net ¹⁴CO₂ photoassimilation (with 5 mM HCO₃ ^–) for the second, third and fourth leaf (leaflet) isolates were, respectively, 27.0, 57.0, and 41.7 mol ¹⁴CO₂ assimilated per milligram chlorophyll per hour; simultaneously maximal rates of NO _inf2 ^sup– photoreduction (1 mM NO _inf2 ^sup– ) were, respectively, 4.4, 8.1, and 0.0 mol NO _inf2 ^sup– reduced per milligram chlorophyll per hour. These studies made it clear that in order repeatedly to attain reproducible maximal rates of leaf cell isolate net ¹⁴CO₂ photoassimilation and NO _inf2 ^sup– photoreduction, it always was necessary to select the newest, fully expanded leaves (e.g. leaf number 3) for cell isolation. Leaves from several plants only were pooled if they were excised from identically the same node on each of the plants.Abbreviations A_max - maximum leaflet (trifoliolate) area attained during ontogeny - CO₂ - CO₂ gas dissolved in solution - HCO _inf3 ^sup– - bicarbonate - L_max - maximum leaf blade length (midvein) attained during ontogeny - NiRase - chloroplast nitrite reductase (reduced ferredoxin) - NiPR - nitrite photoreduction - PE - post-emergence - Pn - net CO₂ photoassimilation (for leaflets and mesophyll cell isolates) - PPRC - pentose phosphate reductive cycle     

Comparative effects of growth irradiance on photosynthesis and leaf anatomy of Flaveria brownii (C4-like), Flaveria linearis (C3–C4) and their F1 hybrid

Photosynthetic rates and related anatomical characteristics of leaves developed at three levels of irradiance (1200, 300 and 80 umol · m^–2 · s^–1) were determined in the C₄-like species Flaveria brownii A.M. Powell, the C₃–C₄-intermediate species F. linearis Lag., and the F₁ hybrid between them (F. brownii × F. linearis). In the C₃–C₄ and F₁ plants, increases in photosynthetic capacity per unit leaf area were strongly correlated with changes in mesophyll area per unit leaf area. The C₄-like plant F. brownii, however, showed a much lower correlation between photosynthetic capacity and mesophyll area per unit leaf area. Plants of F. brownii developed at high irradiance showed photosynthetic rates per unit of mesophyll cell area 50% higher than those plants developed at medium irradiance. These results along with an increase in water-use efficiency are consistent with an increase of C₄ photosynthesis in high-irradiance-grown F. brownii plants, whereas in the other two genotypes such plasticity seems to be absent. Photosynthetic discrimination against ¹³C in the three genotypes was less at high than at low irradiance, with the greatest change occurring in F. brownii. Discrimination against ¹³C expressed as ¹³C was linearly correlated (r ² = 0.81; P<0.001) with the ratio of bundle-sheath volume to mesophyll cell area when all samples from the three genotypes were combined. This tissue ratio increased for F. brownii and the F₁ hybrid as growth irradiance increased, indicating a greater tendency towards Kranz anatomy. The results indicated that F. brownii had plasticity in its C₄-related anatomical and physiological characteristics as a function of growth irradiance, whereas plasticity was less evident in the F₁ hybrid and absent in F. linearis.Abbreviations A leaf surface area - A_ma, A_mn, A_lm total ma, mn or lm cell surface area - bs vascular bundle sheath - lm large spongy-mesophyll cells - ma mesophyll cells adjacent to bundle sheath - mn mesophyll cells not adjacent to bundle sheath - P_n net photosynthesis - (H, M, L) PPFD (high, medium, low) photosynthetic photon flux density - SLDW specific leaf dry wight - V_bs bs volume - V_{(ma + mn + bs)} total photosynthetic tissue volume - ¹³C ¹³C discrimination We thank Mrs. Lisa Smith for technical assistance in light microscopy and Dr. Ned Friedman (Department of Botany, University of Georgia, Athens, GA, USA) for the use of digitizing equipment. Participation of Dr. J.L. Araus in this work was supported by a grant Beca de Especialización para Doctores y Tecnólogos en el Extranjero, from Ministerio de Educatión y Ciencia, Spain.     

Shading-induced changes in the leaf mesophyll of plants of different functional types

Changes in the structural characteristics of mesophyll induced by shading were investigated in ten species of wild plants of diverse functional types. In all plant types, shading reduced leaf thickness and density by 30–50% and total surface of mesophyll, by 30–70%. The extent and mechanisms of mesophyll structural rearrangement depended on the plant functional type. In the ruderal plants, integral parameters of mesophyll, such as the surface of cells and chloroplasts and mesophyll resistance, changed threefold predominantly because of changes in the dimensions of the cells and chloroplasts. In these plants, shading reduced the volume of chloroplasts by 30%, and the chloroplast numbers per cell declined. The competitor plants showed a twofold increase in mesophyll resistance due to a decrease in the number of photosynthesizing cells per leaf area unit. Moreover, these plants maintained constant dimensions of mesophyll cells, ratios mesophyll surface/mesophyll volume and chloroplast surface/cell surface. In stress-tolerant plants, diffusion resistance of mesophyll remained the same irrespective of the growing conditions, and mesophyll rearrangement was associated with inversely proportional changes in the dimensions of the cells and cell volume per chloroplast. Noteworthy of these plants were relatively constant chloroplasts number per cell, per leaf area unit and total surface area of chloroplasts. The nature of relationship between the mesophyll diffusion resistance and structural parameters of leaf mesophyll differed in plants of diverse functional types.     

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.     

Differential positioning of chloroplasts in C4 mesophyll and bundle sheath cells

Chloroplast photorelocation movement is extensively studied in C₃ but not C₄ plants. C₄ plants have two types of photosynthetic cells: mesophyll and bundle sheath cells. Mesophyll chloroplasts are randomly distributed along cell walls, whereas bundle sheath chloroplasts are located close to the vascular tissues or mesophyll cells depending on the plant species. The cell-specific C₄ chloroplast arrangement is established during cell maturation, and is maintained throughout the life of the cell. However, only mesophyll chloroplasts can change their positions in response to environmental stresses. The migration pattern is unique to C₄ plants and differs from that of C₃ chloroplasts. in this mini-review, we highlight the cell-specific disposition of chloroplasts in C₄ plants and discuss the possible physiological significances.Key words: abscisic acid, aggregative movement, avoidance movement, blue light, bundle sheath cell, C₄ plant, chloroplast, cytoskeleton, environmental stress, mesophyll cellChloroplasts can change their intracellular positions to optimize photosynthetic activity and/or reduce photodamage occurring in response to light irradiation. On treating with high-intensity light, the chloroplasts move away from the light to minimize photodamage (avoidance response). Meanwhile, on irradiating with low-intensity light, they move toward the light source to maximize photosynthesis (accumulation response). These chloroplast-photorelocation movements are observed in a wide variety of plant species from green algae to seed plants,^1–3 although little attention has been paid to C₄ plants. There is a report stating that monocotyledonous C₄ plants showed changes in the light transmission of leaves in response to blue light,⁴ although the direction of migration of the chloroplasts is not described.C₄ plants have two types of photosynthetic cells: mesophyll (M) cells and bundle sheath (BS) cells, which have numerous well-developed chloroplasts. BS cells surround the vascular tissues, while M cells encircle the cylinders of the BS cells (Fig. 1). The C₄ dicarboxylate cycle of photosynthetic carbon assimilation is distributed between the two cell types, and acts as a CO₂ pump to concentrate CO₂ in the BS chloroplasts.^5,6 C₄ plants are divided into three subtypes on the basis of decarboxylating enzymes: NADP-malic enzyme (ME), NAD-ME and phosphoenolpyruvate carboxykinase. Although the M chloroplasts of all C₄ species are randomly distributed along the cell walls, BS chloroplasts are located either in a centripetal (close to the vascular tissue) or in a centrifugal (close to M cells) position, depending on the species (Fig. 1A).⁷ Thus, C₄ M and BS cells have different systems for chloroplast positioning: an M cell-specific system for dispersing chloroplasts and a BS cell-specific system for holding chloroplasts in a centripetal or centrifugal disposition.Open in a separate windowFigure 1The intracellular arrangement of chloroplasts in finger millet (Eleusine coracana), an NAD-ME-type C₄ plant. (A) Light micrograph of a transverse section of a leaf blade from a control plant. Bundle sheath (BS) cells surround the vascular tissues, while mesophyll (M) cells encircle the cylinders of the BS cells. BS chloroplasts are well developed, and are located in a centripetal position, whereas M chloroplasts are randomly distributed along the cell walls. B, bundle sheath cell; M, mesophyll cell; V, vascular bundle. (B) Transverse section of a leaf blade from a drought-stressed plant. Most M chloroplasts are aggregatively distributed toward the BS side, while the centripetal arrangement of BS chloroplasts is unchanged. (C and D) Transverse sections of leaf segments irradiated with blue light of intensity 500 µmol m⁻² s⁻¹ with or without 30 µM ABA for 8 h (C and D, respectively). The adaxial side of each leaf section (upper side in the photograph) was illuminated. In the absence of ABA, M chloroplasts exhibited avoidance movement on the illuminated side and aggregative movement on the opposite side. In the presence of ABA, aggregative movement was observed on both sides. Scale bars = 50 µm.     

Selenium modulates the activities of antioxidant enzymes,osmotic homeostasis and promotes the growth of sorrel seedlings under salt stress

Using physiological assays coupled with ultrathin tissue sections, we investigated the impacts of exogenous selenium (Se) on the growth, antioxidant enzymes, osmotic regulation and ultrastructural modifications of leaf mesophyll and root tip cells of 100 mM NaCl-stressed sorrel (Rumex patientia × R. tianshanicus) seedlings. At low concentrations (1–5 M), Se tended to stimulate the growth, the activities of superoxide dismutase and peroxidase enzymes, as well as the accumulation of water-soluble sugar in leaves of sorrel seedlings. At higher concentrations (10–30 M), Se exerted diminished beneficial effects on growth and enzyme activities. CAT activity did not change with Se addition (1–30 M). Electrolyte leakage of leaf cells declined, and K⁺ and Na⁺ ions increased in leaves with Se treatment, notably at 5 M of Se. TEM observations revealed that treatment with 5 M of Se positively promoted the integrity of membrane systems and cellular organelles, such as chloroplasts and mitochondria in leaf mesophyll and root tip cells. These results strongly suggest that an appropriate concentration of exogenous Se functions positively to promote the antioxidative and osmoregulatory capacity, and enhance the salt-resistance in sorrel seedlings.     

Light quality affects photosynthesis and leaf anatomy of birch plantlets in vitro

Cultures in vitro of Betula pendula Roth were subjected to light of different spectral qualities. Photosynthetic capacity was highest when the plantlets were exposed to blue light (max recorded photosynthesis, 82 mol CO₂ dm^–2 h^–1) and lowest when irradiated with light high in red and/or far-red wave lengths (max recorded photosynthesis, 40 mol CO₂ dm^–2 h^–1). Highest chlorophyll content (2.2 mg dm^–2 leaf area) was found in cultures irradiated with blue light, which also enhanced the leaf area. Morphometric analysis of light micrographs showed that the epidermal cell areas were largest in plantlets subjected to blue light and smallest in those subjected to red light. Morphometric analysis of electron micrographs of palisade cells, showed that the functional chloroplast area was largest in chloroplasts of leaves subjected to blue light and smallest in those exposed to red light. We suggest that light quality affects photosynthesis both through effects on the composition of the photosynthetic apparatus and on translocation of carbohydrates from chloroplasts.     

Climatic adaptability of populations ofDiplotaxis erucoides D.C. (Brassicaceae) from Sicily,based on leaf morphology,leaf anatomy and δ13 C studies

The morphological and anatomical variability ofDiplotaxis erucoides populations from Sicily was investigated. Populations growing during the summer months exhibit distinct xeromorphic features. Leaf area is strongly reduced and leaf thickness is increased when compared with winter populations. Cell size, as well as cell arrangement and mesophyll cell surface area differ significantly between summer and winter populations. Leaf thickness is almost three times higher in summer populations andA (cell)/A, i.e. the mesophyll cell surface area per unit leaf area changes from about 16 for winter populations to almost 52 for summer populations. These differences are partly due to differences in intercellular volume and partly due to alterations in mesophyll cell sizes. The organic materal of the summer populations exhibits ¹³C values in the order of –27%. to –28%., while the corresponding values for the winter populations are in the order of –31%. to –33%.. Analysis ofD. erucoides populations from the transition period revealed intermediate ¹³C values. Anatomical variations such as reductions or increases ofA (cells)/A and changes of intercellular volume correlate with the corresponding ¹³C data. The ¹³C data are discussed in conjunction with the differences in leaf anatomy.     

Leaf acclimation to experimental climate warming in meadow plants of different functional types

The use of open-top chambers (OTCs) installed in natural plant cover is one of the approaches to study plant responses to climate change. Three OTCs made from polyethylene film were installed on a herbgrass meadow in the subzone of the southern taiga before the beginning of the growing season. A significant increase in the average daily temperature values (by 0.5°C) and the relative humidity (by 10%) compared to control conditions was observed inside the chambers. Plant height, leaf parameters, and the pigments content were studied for six species of meadow plants during the growing season in two variants—inside the chamber and outside the chamber (control); more than 20 quantitative parameters of the mesophyll were studied for four of the species. It was found that the differences in microclimatic conditions had no effect on plant height and leaf area. A slight decrease in the thickness and density of the leaves and an increased water content were noted inside the OTCs. In contrast to weak changes in external leaf parameters, the internal leaf structure and the content of photosynthetic pigments varied considerably. Warming caused the reduction of the content of chlorophyll and carotenoids per unit leaf area in the majority of studied species, except for Veronica chamaedrys L., but the ratio of pigment forms did not change. Changes in the pigments content in the leaf were associated with some structural rearrangements in the mesophyll, whose mechanism depended on the functional properties of the species. Increased size of palisade cells and the number of chloroplasts per cell was noted in the ruderal species (R/CSR-strategist) Taraxacum officinale Wigg. s. l.; the reduction of chlorophyll content per leaf area occurred due to the decrease in chlorophyll content per a single chloroplast. Decreased number of cells and chloroplasts per leaf unit area without any changes in their size was marked for the species with S/CSR strategy Alchemilla vulgaris L. s. 1. and V. chamaedrys L. in a chamber, but the content of chlorophyll per a chloroplast increased. An increase in the number of cells and a simultaneous decrease in their size was observed in CR-strategist Cirsium setosum (Willd.) Bess. inside the OTC; the chlorophyll content per chloroplast did not change. It was concluded that the acclimation of plants to short-term climate warming was associated with the restructuring of leaf mesophyll, whose mechanism depended on the functional properties of the species.     

Leaf Structure of Tobacco In Vitro Grown Plantlets as Affected by Saccharose and Irradiance

Tobacco plantlets were cultured in vitro under high (200 µmol m^–2 s^–1) or low (60 µmol m^–2 s^–1) irradiance with or without saccharose in the medium. Light microscopy and image analysis were used to evaluate the effect of these culture conditions on leaf anatomy. Addition of saccharose resulted in thicker leaves (all leaf layers) and larger mesophyll cells under both growth irradiances. Various irradiance affected leaf anatomy differently when plantlets had been cultivated in presence or absence of saccharose in the medium. While under high irradiance in presence of saccharose leaf thickness and number of chloroplasts per cell section were increased, plantlets grown under high irradiance in absence of saccharose had thinner leaves and less chloroplasts per cell section. The changes were more pronounced in palisade parenchyma layer.     

Effects of water stress on cellular ultrastructure and on concentrations of endogenous abscisic acid and indole-3-acetic acid in Fatsia japonica leaves

Ultrastructural alterations in mesophyll cells as well as variations in bulk leaf endogenous ABA and IAA concentrations were studied in water-stressed field-grown plants of Fatsia japonica. Under water deficit cellular membranes were modified and an increase in vesicles was observed. The main damage to the chloroplasts included thylakoid swelling and disruption of the chloroplast envelope. Concomitant variations in abscisic acid and indole-3-acetic acid were observed. Despite the expected increased in endogenous ABA concentration in relation to water stress, after the highest concentration of ABA, observed at predawn in severely stressed plants (29-1), there was a sharp decline from 2768 pmol g fw^–1 to 145 pmol g fw^–1; thus in severely stressed plants ABA levels were not related to changes in bulk leaf ABA contents. Water stress did not influence the concentrations of indole-3-acetic acid, although the increase in the endogenous abscisic acid concentration could be related with the ultrastructural changes.Abbreviations ABA abscisic acid - IAA indole-3-acetic acid - leaf water potential