Heterophylly in Ranunculus flabellaris: The Effect of Abscisic Acid on Leaf Anatomy

Ranunculus flabellaris Raf., the yellow water crowfoot, exhibitsstriking heterophylly between submerged and terrestrial leaves.Leaves produced under water are highly divided with numerousnarrow lobes and deep sinuses, whereas terrestrial leaves havefew broad lobes and shallow sinuses. When plants are submergedin a 25 µM solution of ABA, the typical transition fromterrestrial to submerged leaves is completely suppressed and,instead, terrestrial-like leaves are produced. Image analysistechniques show that, in addition to this modification of leafmorphology, leaves produced under ABA treatment possess surfaceand internal features characteristic of terrestrial leaf anatomy.This study provides evidence that the environmental factorsthat influence the morphological and anatomical expression ofheterophylly may act through endogenous ABA. Ranunculus flabellaris, yellow water crowfoot, ABA, heterophylly, leaf anatomy     

Heterophylly in Ranunculus flabellaris Raf.: The Effect of Abscisic Acid

Ranunculus flabellaris Raf., the Yellow Water-Crowfoot, is aheterophyllous semi-aquatic dicotyledonous plant, showing strikingmorphological and anatomical differences between terrestrialleaves and those formed underwater. After plants are submergedin a 25 µM solution of abscisic acid, leaves are producedat a normal rate but they exhibit many of the characteristicsof terrestrial leaves. Ranunculus flabellaris Raf., Yellow Water-Crowfoot, heterophylly, abscisic acid     

A Vertical Gradient of the Chloroplast Abundance among Leaves of Chenopodium album

The abundances of chloroplasts in leaves on the main stems ofChenopodium album at different height levels were investigatedin relation to the photosynthetic capacity and light environmentof the leaves. (1) The number of chloroplasts per mesophyllcell decreased with descending position of leaves, except foryoung developing leaves at the top of plants that had smallerchloroplast numbers per cell than matured leaves beneath them.Contents of chlorophyll and ribulose-1,5-bisphosphate carboxylase/oxygenaseper leaf area that were highest in the topmost young leavesand decreased with decreasing height level indicate that thereis a vertical gradient of chloroplast abundance per leaf areadecreasing from the top of the leaf canopy with depth. (2) Light-saturatingrate of photosynthetic oxygen evolution per leaf area of maturedleaves decreased more steeply with decreasing leaf positionthan the chloroplast number per cell. Gradients of chlorophylland the enzyme protein contents were also steeper than thatof the chloroplast number. Loss of photosynthesis in lower leavesis, therefore, ascribed partly to loss of whole chloroplastsand partly to reduced photosynthetic capacities of the remainingchloroplasts. (3) The chloroplast number per cell in newly expandedsecond leaves was comparable to those in leaves that have developedat later stages of the plant growth but decreased graduallyduring leaf senescence both in the dark and light. The formationof the vertical gradient of chloroplast abundance is, therefore,ascribed to loss of whole chloroplasts during senescence ofleaves. (4) Irradiance a leaf receives decreased sharply fromthe top of the canopy with depth. The physiological or ecophysiologicalsignificance of the vertical distribution of chloroplasts amongleaves was discussed taking light environments of leaves intoconsideration. (Received July 31, 1995; Accepted October 20, 1995)     

辣椒彩色斑叶突变体叶片显微结构及超微结构研究

以彩色斑叶辣椒突变体、紫叶辣椒、白色斑叶辣椒突变体功能叶为试验材料,利用光学显微镜和透射电子显微镜,通过观察叶片不同斑区的显微结构及超微结构,分析彩色斑叶的显色部位、显色特征、细胞器数量及形态变化,从细胞水平上探讨彩色斑叶辣椒复杂叶色的成因。结果表明：(1)彩色斑叶辣椒突变体子叶为紫色,自第一片真叶展开出现异色斑块,斑块位置、频率、色彩深度无明显规律。(2)叶肉细胞内叶绿体少甚至缺失形成白斑,花色素苷在叶肉细胞和保卫细胞均有分布,其在叶肉细胞不均匀分布是紫色深度不同的主因。(3)辣椒彩色斑叶突变体绿斑区内细胞形态良好,细胞器结构较好;紫色斑区和白色斑区细胞呈中度肿胀,细胞器明显异常。(4)叶肉细胞内叶绿体少甚至缺失、花色素苷不均匀分布是叶片呈现彩色的原因,该叶斑类型属于色素型。     

Cellular Structure and Light Absorbtion in Leaves of Frithia pulchra (Mesembryanthemaceae)

In order to quantify the structural differences between celltypes of leaves from a ‘ window’ plant, an ultrastructuralmorphometric analysis was made of the epidermal, window andchlorenchyma tissues of Frithia pulchra. Epidermal cells arethe largest cells found in Frithia leaves and are characterizedby the presence of a thick outer tangential cell wall and numerousvacuolar inclusions. Epidermal tissue has an optical densityof 0.30. The transparent window tissue (i.e. optical density= 0.08) has a uniform ultrastructure throughout the length ofthe leaf. The vacuome comprises aproximately 97 per cent ofthe protoplasmic volume of window cells. Chlorenchyma cellspossess thin cell walls and are surrounded by numerous intercellularspaces. Cells of the apical chlorenchyma tissue possess approximately30 plastids per cell. These chloroplasts have an average individualvolume of 220 µm². Cells of the basal chlorenchyma tissuecontain chloroplasts that are five to six times smaller andmore numerous than those in cells of the apical chlorenchyma.The increased volume of chloroplasts in the apical comparedwith basal chlorenchyma cells (i.e. 31.4 and 20.2 per cent ofthe protoplasm, respectively) is positively correlated withtheir optical densities of 1.46 and 0.97, respectively. Frithia pulchra, stereology, leaf, light absorption, window plant     

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     

Observations on the Senescence of a Mutant Non-yellowing Genotype of Phaseolus vulgaris L.

A non-yellowing mutant of Phaseolus vulgaris L. was used toinvestigate factors involved in chlorophyll breakdown duringfoliar senescence. The mutant showed physiological changes similarto those of the normal yellowing type during senescence exceptthat leaf chlorophyll did not decline. Transmission electronmicroscope studies did not reveal appreciable differences inchloroplast ultrastructure between the two genotypes, suggestingthat chloroplast membrane integrity was not the factor preventingchlorophyll degradation in the mutant. However, the lack ofplastoglobuli in senescent mutant chloroplasts suggested thatthe lipid environment may be different from that of senescentnormal chloroplasts. Banding patterns of total soluble protein,resolved by sodium dodecyl sulphate-poly aery lamide gel electrophoresisshowed few, if any, differences between mature non-senescentnormal and mutant leaves; however, bands at 14 kD and 58 kDdiminished in senescent normal leaves, but remained in senescentmutant non-yellowing leaves. Key words: Non-yellowing mutant, Phaseolus vulgaris, senescence, chlorophyll degradation     

Membrane-bound Plastid Inclusions and Chloroplast Thylakoid Formation in Sunflower (Helianthus annuus L.)

The ultrastructure, distribution and frequency of membrane-boundplastid inclusions present in the epidermal cells of leavesof intact sunflower plants (Helianthus annuus L.) and in theepidermal and mesophyll cells of sunflower leaf discs culturedin darkness have been studied. These inclusions appear to bedilated thylakoids containing a granular material which, undernormal conditions, is probably involved in chloroplast membraneformation. It is suggested that this material accumulates, andinclusions form, in the chloroplasts of sunflower leaves intwo specific situations. Firstly, in the completely differentiatedcells of the epidermis where the chloroplasts, although at arelatively immature stage, have nevertheless reached a terminalstage of development. Secondly, in the mesophyll cells of youngleaves when chloroplast development has been arrested at animmature stage by a 5-day dark period. In the latter situationthe material can be remobilized if plastid development is restimulated.The plastids of sunflower leaf discs cultured in darkness containboth membrane-bound inclusions and prolamellar bodies, indicatingthat they are separate and distinct structures possibly containingdifferent membrane components. Helianthus annuus L., sunflower, chloroplast, ultrastructure, plastid inclusions, thylakoid formation     

Tobacco-mosaic-virus-induced increase in abscisic-acid concentration in tobacco leaves:

The concentrations of free and bound abscisic acid (ABA and the presumed ABA glucose ester) increased three- to fourfold in leaves of White Burley tobacco (Nicotiana tabacum L.) systemically infected with tobacco mosaic virus. Infected leaves developed a distinct mosaic of light-green and dark-green areas. The largest increases in both free and bound ABA occurred in dark-green areas. In contrast, virus accumulated to a much higher concentration in light-green tissue. Free ABA in healthy leaves was contained predominantly within the chloroplasts while the majority of bound ABA was present in non-chloroplastic fractions. Chloroplasts from light-green or dark-green tissues were able to increase stromal pH on illumination by an amount similar to chloroplasts from healthy leaf. It is unlikely therefore that any virus-induced diminution of pH gradient is responsible for increased ABA accumulation. Tobacco mosaic virus infection had little effect on free ABA concentration in chloroplasts; the virus-induced increase in free ABA occurred predominantly out-side the chloroplast. The proportional distribution of bound ABA in the cell was not changed by infection. Treatment of healthy plants with ABA or water stress increased chlorophyll concentration by an amount similar to that induced by infection in dark-green areas of leaf. A role for increased ABA concentration in the development of mosaic symptoms is suggested.Abbreviations ABA abscisic acid - TMV tobacco mosaic virus     

Ultrastructural Changes in Chloroplasts of Quercus rotundifolia Lam. in Response to Evernic Acid

The amount of total chlorophyll, chlorophylls a and b as wellas the ratio of a to b decreased in chloroplasts isolated fromQuercus rotundifolia leaves, kept for 17 d in a solution of35.5 µM evernic acid in 1 mM Na HCO₃, when compared withthe chloroplasts of control leaves (kept in NaHCO₃). The chloroplastsin the spongy parenchyma were smaller and the amount of starchand plastoglobuli lower. The number of grana per chloroplastsection, the number of thylakoids per grana and the height ofgrana stacks were also less in the chloroplasts of leaves treatedwith evernic acid. Quantitative ultrastructural differenceswere determined by means of electron microscopy and image analysistechniques. Quercus rotundifolia Lam., chloroplasts, ultrastructure, lichens, evernic acid     

Effects of Copper Toxicity on Leaves of Oregano (Origanum vulgare subsp. hirtum)

The effects of increasing concentrations of soil copper on anumber of leaf structural parameters in oregano plants werestudied to determine the effect of copper toxicity. Copper-stressedleaves were small and chlorotic and underwent a thickening oftheir lamina, due principally to an increase in the number andvolume of mesophyll cells. The number of stomata and glandularand non-glandular hairs increased significantly. Chloroplastsof mesophyll cells declined dramatically in number and volume.Grana and stroma thylakoids of chloroplasts did not undergoany noticeable alterations, but starch grains disappeared, plastoglobulibecame larger and the double membrane limiting the chloroplastbecame dilated. Leaf chlorosis was determined by total chlorophyllanalysis and measurement of the leaf Cu, Fe and Mg content.The effects of copper toxicity on oregano leaves comprised significantstructural alterations which reflect reduced metabolic activity.Copyright 2001 Annals of Botany Company Origanum vulgare subsp, hirtum, oregano, copper toxicity, leaf, structure and ultrastructure, morphometry, inorganic elements, tissue relative volumes     

Senescence-Associated Changes during Long-day-induced Leaf Senescence and the Nature of the Graft-transmissible Senescence Substance in Kleinia articulata

Schwabe, W. W. and Kulkarni, V. J. 1987. Senescence-associatedchanges during long-day-induced leaf senescence and the natureof the graft-transmissible senescence substance in Kleinia articulata.— J. exp. Bot. 38: 1741–1755. The long-day-induced senescence in Kleinia articulata leaveswas characterized by a loss in fresh and dry weight, in therate of leaf expansion and progressive loss of chlorophyll inthe detached rooted leaves. Ultrastructural examination of mesophyllcells of leaves from plants grown in continuous light showedthat osmiophilic globules accumulating in the chloroplasts werethe first visible sign of senescence in the organdies. Thesefirst signs of senescence could be detected in very young leavesof plants in continuous light, even before the leaves had expanded.Attempts were made to study the cause of this photoperiodicsenescence which, from previous work, appeared to involve agraft-transmissible substance. Leaves in continuous light showed reduced stomatal opening andextracts from them had very much higher activity in the Commelinastomatal closure assay (ABA-like activity ?) compared with non-senescingleaves grown in short days (8 h). However, even if all the activitywere due to ABA, this on its own does not appear to be the senescencesubstance because a much longer exposure to continuous lightwas required to induce irreversible senescence than to reachmaximum stomatal closure promoting activity in the bioassay.Moreover, severe water stress (high ABA?) did not lead to senescenceunless combined with continuous light or ethylene treatment.It is postulated that while ABA may play an important role inKleinia leaf senescence its lethal effect may not be realizedunless ethylene-induced membrane changes may synergisticallyassist. Key words: Leaf senescence, ABA, Daylength, stomatal movement, Kleinia     

Ageing-related Anatomical and Ultrastructural Changes in Leaves of Birch (Betula pendula Roth.) Clones as Affected by Low Ozone Exposure

Effects of ozone on the leaf anatomy and ultrastructure of fivebirch (Betula pendula Roth.) clones were studied during onegrowing season in open-field conditions. Cumulative ozone exposurewas 1·5 times higher than ambient. Ozone exposure decreasedtotal leaf thickness in one, ozone sensitive, clone. The effecton palisade spongy mesophyll thickness was clone-specific, whilethe amount of palisade intercellular space was reduced in allclones. A second effect was a change in the relative amountsof adaxial and abaxial epidermis. In palisade and spongy parenchymacells of all clones, ozone increased the number of irregularand spherical shaped chloroplasts, the electron density of chloroplaststroma, swelling and curling of thylakoids, translucency ofthe mitochondrial matrix and also the amount of cytoplasmiclipids. In the sensitive clone shorter chloroplasts and reducedamount of starch were observed in ozone-exposed plants, whilst,in the tolerant clone, the size of chloroplasts and the amountof starch were unaffected. Ozone effects on number, size andelectron density of plastoglobuli and vacuolar tannin were clone-dependent.At the ultrastructural level, the normal leaf ageing processprogressed at different rates in the birch clones. Ozone acceleratedsenescence-related structural changes, in accordance with earlierobservations of deciduous species.Copyright 1995, 1999 AcademicPress Betula pendula Roth., birch, clones, ageing, ozone, leaf anatomy, ultrastructure     

Leaf anatomy and chloroplast ultrastructure of Mn-deficient orange plants

Leaf samples of Mn-deficient and Mn-sufficient (control) ‘Navelate’ orange plants grown in a greenhouse were taken to investigate the effects of Mn deficiency in leaf structure and chloroplast ultrastructure. Total leaf chlorophyll concentration was significantly lower in Mn-deficient plants than in control ones. Entire lamina thickness was not altered due to Mn deficiency. However, Mn deficiency resulted in disorganization of mesophyll cells, mainly of palisade parenchyma cells. The number of mesophyll chloroplasts per cellular area and their length were both affected negatively. The membranous system of chloroplasts was also disorganized. The percentages of starch grains and plastoglobuli per chloroplast of Mn-deficient leaves were significantly greater than those of control leaves.     

Effects of ethylene and abscisic acid upon heterophylly in<Emphasis Type="Italic"> Ludwigia arcuata</Emphasis> (Onagraceae)

In this study, we examined the effects of ethylene and abscisic acid (ABA) upon heterophyllous leaf formation of Ludwigia arcuata Walt. Treatment with ethylene gas resulted in the formation of submerged-type leaves on terrestrial shoots of L. arcuata, while treatments with ABA induced the formation of terrestrial-type leaves on submerged shoots. Measurement of the endogenous ethylene concentration of submerged shoots showed that it was higher than that of terrestrial ones. In contrast, the endogenous ABA concentration of terrestrial shoots was higher than that of submerged ones. To clarify interactions of ethylene and ABA, simultaneous additions of these two plant hormones were examined. When L. arcuata plants were treated with these two plant hormones, the effects of ABA dominated that of ethylene, resulting in the formation of terrestrial-type leaves. This suggests that ABA may be located downstream of ethylene in signal transduction chains for forming heterophyllous changes. Further, ethylene treatment induced the reduction of endogenous levels of ABA in tissues of L. arcuata, resulting in the formation of submerged-type leaves. Thus the effects of ethylene and ABA upon heterophyllous leaf formation are discussed in relationship to the cross-talk between signaling pathways of ethylene and ABA.Abbreviations ABA abscisic acid - ACC 1-aminocyclopropane-1-carboxylic acid - L/W ratio ratio of leaf length to width - LN leaf number - GAs gibberellins     

日光温室光温因子对黄瓜叶绿体超微结构及其功能的影响

在日光温室内,研究了光温因子对黄瓜叶绿体超微结构及其功能的影响．结果表明,因季节之间光、温条件不同,日光温室黄瓜叶片显微结构和叶绿体超微结构有一定差异,1月份光照弱叶肉细胞较大,而5月份光照强叶绿体数较多．在该试验条件下,未发现叶片光合速率与叶绿体超微结构之间有直接或密切的相关性．在各生长季节其光合速率均为第4叶>初展叶>基部叶,与叶龄及各叶位的受光量有关．如果将不同叶位叶放在相同的光照下,则差异明显减少．黄瓜叶片的叶肉细胞、叶绿体和淀粉粒的大小以及叶绿体数、基粒数、基粒厚度、基粒片层数都随叶位的下降而呈增加趋势。不同品种、同品种不同生长时期的叶片显微结构和叶绿体超微结构及其功能也有一定的差异．限制日光温室冬季黄瓜光合作用的主要因素是光照弱、有效光照时数少,而在晴天温度的限制作用相对较小。阴天因光照弱而导致的室内低温则是限制黄瓜生长的关键因素．     

Chloroplast Structure and Starch Grain Accumulation in Leaves That Received Different Red and Far-Red Levels during Development

An important step in understanding influence of growth environment on carbon metabolism in plants is to gain a better understanding of effects of light quality on the photosynthetic system. Electron microscopy was used to study chloroplast ultrastructure in developing and fully expanded leaves of tobacco (Nicotiana tabacum L. cv Burley 21). Brief exposures to red or far-red light at the end of each day during growth under controlled environments influenced granum size, granum number and starch grain accumulation in chloroplasts, and the concentration of sugars in leaf lamina. Far-red-treated leaves had chloroplasts with more but smaller grana than did red-treated leaves. Red light at the end of the photosynthetic period resulted in more and larger starch grains in the chloroplasts and a lower concentration of sugars in leaves. Chloroplast ultrastructure and starch grain accumulation patterns that were initiated in the expanding leaves were also evident in the fully expanded leaves that received the treatment during development. It appears that the phytochrome system in the developing leaves sensed the light environment and initiated events which influenced chloroplast development and partitioning of photosynthate to adapt the plant for better survival under those environmental conditions.     

Effects of soil resistance to root penetration on leaf expansion in wheat (Triticum aestivum L.): composition, number and size of epidermal cells in mature blades

Wheat seedlings {Triticum aestivum L.) were grown on soils withcontrasted resistances to root penetration (measured as penetrometerresistance, R_s. High R_s reduced the rates of leaf appearanceand expansion. Although the duration of expansion was increased,mature leaves were smaller. Underlying changes in leaf anatomywere investigated on cleared mature leaves, focusing on theepidermes. Three leaves were analysed: leaves 1 and 3 whichstarted their development in the embryo, and leaf 5 which wasinitiated on the seedling, after imposition of contrasted soilconditions. In all leaves, high R_s, caused a reduction in maturecell sizes, lengths and widths, and a shift in the relativeproportions of functionally different cell types, with a decreasein the relative proportions of stomata and associated cell types(interstomatal and sister cells) and an increase in the proportionsof unspecialized elongated epidermal cells and of trichomes.In leaves 3 and 5 the number of cellular files across the bladewas also reduced, while in leaf 1 it was similar at the twoR_s. These differences between leaves are attributed to differencesin their developmental stage when root stress was first perceived.Remarkably, R_s had no effect (leaf 1) or relatively small effects(leaves 3 and 5) on the total number of cells per file, suggestingthat this parameter is either largely insensitive to variationin root environment, or is programmed at the outset before stresswas perceived at the apex. Key words: Wheat, anatomy, mature epidermis, root impedance     

Structure and distribution of chloroplasts and other organelles in leaves with various rates of photosynthesis

The ultrastructure and distribution of chloroplasts, mitochondria, peroxisomes, and other cellular constituents have been examined in cross sections of leaves from plants with either high or low photosynthetic capacity. Photosynthetic capacity of a given plant cannot be correlated with the presence or absence of grana in bundle sheath cell chloroplasts, the presence or absence of starch grains in bundle sheath or mesophyll cell chloroplasts, the chloroplast size in bundle sheath or mesophyll cells, or the location of chloroplasts within bundle sheath cells. We conclude that the number and concentration of chloroplasts, mitochondria, and peroxisomes in bundle sheath cells is the most reliable anatomical criterion presently available for determining the photosynthetic capacity of a given plant.     

Changes of Anatomical Features, Photosynthesis and Ribulose Bisphosphate Carboxylase-Oxygenase Content of Mango Leaves

Changes in anatomical and physiological features, includingchanges in amount per unit area of anthocyanin and chlorophyll,in leaves of seedling mango (Mangifera indica L. cv. Irwin)trees were determined to understand what controls the rate ofphotosynthesis (Pn) at various stages of development. The youngleaves of seedling trees contained high concentrations of anthocyanin.During enlargement of leaves, the disappearance of anthocyaninand the accumulation of chlorophyll occurred concomitantly;the anthocyanin content began to decrease markedly once theleaf area had reached a maximum. During the early period ofleaf development, the thickness of mesophyll tissue decreasedtemporarily, but when the length of the leaf reached half thatof a mature leaf, the mesophyll began to thicken again. Smallstarch grains appeared in the chloroplasts of the young leavesand chloroplast nucleoids (ct-nuclei) were distributed throughoutthe chloroplasts. When leaves matured, ct-nuclei were displacedto the periphery of chloroplasts because of the accumulationof large starch grains. Compared with young leaves, green andmature leaves contained greater concentrations of ribulose bisphosphatecarboxylase-oxygenase (RuBisCO) protein. The results of immunocytochemicalexamination of RuBisCO under the light microscope reflectedthe results of electrophoresis measurements of RuBisCO. Pn waslow during the chocolate-coloured stage of early leaf development.In green and mature leaves Pn was higher; the average Pn was7·6 mg CO₂ dm^-2 h^-1 under light at intensities above500 µmol m^-2 s^-1.Copyright 1995, 1999 Academic Press Mangifera indica L., mango leaf, chloroplast nucleoids, chloroplast ultrastructure, starch accumulation, anthocyanin, chlorophyll, DAPI staining, SDS-PAGE, immunocytochemical technique