Compartmentation of ions and organic compounds inSalicornia brachiata Roxb.

The actively growing stems ofSalicomia brachiata (60-d old) were dissected into three major tissue layers: vascular, spongy mesophyll and palisade. Each layer was analysed for chlorophyll, protein, amino acids, and sugar contents and the activities of ATPase and phosphatase (alkaline and acid). The differences in organic compounds and enzyme activities in these different tissues have been correlated with the ion content of the corresponding tissues.     

Increased vacuolar and plasma membrane H+-ATPase activities in Salicornia bigelovii Torr. in response to NaCl

The halophyte Salicornia bigelovii Torr. shows optimal growthand Na⁺ accumulation in 200 mM NaCl and reduced growth underlower salinity conditions. The ability to accumulate and compartmentalizeNa⁺ may result, in part, from stimulation of the H⁺ -ATPaseson the plasma membrane (PM-ATPase) and vacuolar membranes (V-ATPase).To determine if these two primary transport systems are involvedin salt tolerance, shoot fresh weight (FW) and activity of thePM- and V-ATPases from shoots in Salicornia grown in 5 and 200mM NaCI were compared. Higher PM-ATPase activity (60%) and FW(60%) were observed in plants grown in 200 mM NaCI and thesestimulations in growth and enzyme activity were specific forNa⁺ and not observed with Na⁺ added in vitro. V-ATPase activitywas significantly stimulated in vivo and in vitro (26% and 46%,respectively) after exposure to 200 mM NaCl, and stimulationwas Na⁺ -specific. Immunoblots indicated that the increasesin activity of the H⁺ -ATPases from plants grown in 200 mM NaCIwas not due to increases in protein expression. These studiessuggest that the H⁺-ATPases in Salicornia are important in salttolerance and provide a biochemical framework for understandingmechanisms of salt tolerance in plants. Key words: Salicornia, H⁺-ATPases, salt tolerance     

Light Environment within a Leaf I. Optical Properties of Paradermal Sections of Camellia Leaves with Special Reference to Differences in the Optical Properties of Palisade and Spongy Tissues

The attenuance (apparent absorbance), angular distribution oftransmitted light (scattering) and reflectance of paradermalsections of Camellia leaves were measured spectrophotometricallyand the data for the palisade and spongy tissues were compared. Attenuance in tissues could be expressed by Beer's law onlyfor wavelengths of strong absorption. At 680 nm, the apparentextinction coefficient of chlorophyll (e) for the spongy tissuewas about 1.4 times that for the palisade tissue. The largere for the spongy tissue is attributable mostly to the more effectiveincrease in the pathlength of light due to scattering at theinterfaces between the air space and cells because the differencebetween e for the two tissues was minimized by infiltrationof the air space in the tissues with a medium whose refractiveindex was similar to the index of the leaf cells. Scatteringwas larger for wavelengths of weak absorption, and the relativeincrease in attenuance caused by elongating the optical pathlengthwas even more prominent. Based on these data, we report an ecophysiological discussionof the internal light environment of a leaf and the meaningof the differentiation of mesophyll into the palisade and spongytissues. (Received May 18, 1983; Accepted September 21, 1983)     

Localization of Thermo-Osmotically Active Partitions in Young Leaves of Nuphar lutea

The submerged roots and rhizomes of the aquatic vascular macrophyteNuphar lutea (L.) Sm. are aerated, at least in part, by pressurizedventilation. Depending on temperature differences of up to 5K between the inside of young, just-emerged leaves and the surroundingair, pressure differences of 79 to 100 Pa higher than atmosphericare detectable inside the lacunuous spongy parenchyma of theleaf blades. The pressurization is a consequence of structuralfeatures of leaf tissues separating the air filled spaces ofthe spongy parenchyma from the atmosphere. These tissues areacting as thermo-osmotic partitions. Whereas the dimensionsof the stomatal openings (about 5·6 x 2·4 µm)and of the intercellular spaces of the palisade parenchyma (diametersabout 15 µm) are too large, those of the monolayers ofcells separating the palisade and the spongy parenchyma (diameters:0·7–1·2 µm) are small enough to impedefree gaseous diffusion. This inner non-homogeneous partitioninggives rise to the so-called Knudsen diffusion, a physical phenomenonleading to pressurization of the warmer air inside the spongyparenchyma. The rising pressure difference is strong enoughto establish an air flow through the aerenchyma of the wholeplant and out of the most porous older leaves in which a temperatureinduced pressurization is never detectable. These thermo-osmoticallyactive leaves enhance the influx of air to the rhizome and thediffusion path for oxygen to the roots is shortened to the distancebetween rhizome and root tips. Therefore, pressurized ventilationin Nuphar is seen to be of considerable ecological importancefor plant life in anaerobic environments. Key words: Aeration, leaf anatomy, thermo-osmosis of gases, Nuphar lutea     

Potential for the improvement of Salicornia bigelovii through selective breeding

Salicornia bigelovii Torr. is an annual salt marsh plant that produces seed oils, protein meal, fresh salad greens and forage on seawater irrigation. We compared S. bigelovii lines produced in two breeding programs with wild germplasm in greenhouse trials on brackish water (10 ppt NaCl) irrigation. S. bigelovii is an out-crossing species that is also capable of selfing, and the breeding programs showed it is possible to use both hybridization and pedigree breeding to improve the germplasm. Lines selected in a breeding program carried out in Eritrea, Africa, had smaller plant size and lower biomass yields than the starting germplasm, due to the need to compress the growth cycle within the cool months of the year in that hot climate, but seed yields and harvest index were improved. Lines produced from wild germplasm by mass selection and hybridization in Tucson, Arizona had higher biomass yield than starting germplasm. We conclude that S. bigelovii has sufficient genetic diversity among wild accessions and cultivars to support a crop improvement program.     

Compartmentation of Phenolic Compounds and Phenylalanine Ammonia-Lyase in Leaves of Phyllanthus tenellus Roxb. and their Induction by Copper Sulphate

The compartmentation of phenolic compounds in mature leavesof Phyllanthus tenellus and their induction by copper sulphatewere analysed at histological and subcellular levels. Lightand electron microscopy studies demonstrated that the vacuolesof spongy cells were the main sites of phenolic accumulation.Spraying plants with copper sulphate induced punctated lesionsformed by groups of necrotic cells which accumulated brownishsubstances. Histochemical tests and fluorescence microscopyanalysis of the sprayed leaves indicated that the phenolic compoundsincreased in spongy cells within the lesions. Ultrastructuralanalyses showed that 3 h after elicitation, the organelles ofthe cells within the lesion started to collapse and the contentof phenolic substances increased in the vacuole of spongy cells.Antibody against phenylalanine ammonia-lyase (PAL) from parsleycross-reacted with the crude extract of P. tenellus leaves.Two isoforms, one of 65 kD and the other of 66 kD, were identified.Immunocytochemical studies showed that PAL was synthesized inthe palisade and spongy cells, mainly in the cytoplasm and chloroplasts.The phytotoxicity of Cu²⁺ions induced the accumulation of PALin sub-cellular compartments of palisade cells. PAL accumulationstarted to increase 3 h after elicitation and reached a maximumafter 6 h, decreasing 12 h post-induction. The increase of PALwas more evident in cells within the necrotic punctated regionsthan in surrounding cells. Since the vacuole of palisade cellsdid not accumulate phenolic compounds, the in situ studies suggestedthat the end products of PAL synthesis play a role in palisadecell wall reinforcement or might accumulate in other tissues.The symptoms induced by copper sulphate suggest that this abioticelicitor may be a useful tool in the understanding of the regulationof biosynthetic phenolic pathways inP. tenellus . Copyright2000 Annals of Botany Company Cell death, copper sulphate, heavy metal, immunolabelling, phenolic compounds, phenylalanine ammonia-lyase,Phyllanthus , transmission electron microscopy, ultrastructure     

Paclobutrazol enhances photosynthesis and ajmalicine production in Catharanthus roseus

An investigation was carried out with paclobutrazol (PBZ) in order to study its effect on the photosynthetic and anatomical characteristics of Catharanthus roseus (L.) G. Don. The chlorophyll content and related parameters like net photosynthetic rate, transpiration rate and internal CO₂ concentration were measured from control and treated plant samples. The anatomical characteristics, viz., leaf thickness, epidermis and cuticle, palisade layer, spongy layer and vascular bundle were studied. The root alkaloid ajmalicine was extracted and estimated from both treated and control plant roots. Photosynthetic pigment increased under PBZ treatment. PBZ treatment increased the net photosynthetic rate and internal CO₂ concentration, but reduced transpiration. PBZ treatment increased the thickness of the leaf, epidermis and cuticle, palisade layer and spongy layer but reduced the diameter of xylem vessels significantly when compared to control, however the phloem elements had shown an increased diameter as compared to control in the PBZ treated plants. There was a significant enhancement in ajmalicine content under PBZ treatment when compared to control plants. The results suggested that the application of PBZ have significant effects on photosynthetic and anatomical responses thus can be used for improving productivity in medicinal plants.     

Anatomical considerations related to photosynthesis in cotton (Gossypium hirsutum L.) leaves, bracts, and the capsule wall

Light and electron microscopy was used to relate histologicaland ultrastructural differences of cotton (Gossypium hirsutumL.) leaves, bracts, and capsule walls to their different photosyntheticactivities. Light microscopy revealed that the leaf thicknesswas approximately 152µm, had a well-defined internal organizationwith elongated palisade mesophyll cells and loosely packed spongymesophyll cells. In contrast, the bract was thinner (111 µm),lacked a defined palisade layer, and was largely composed ofinternal air spaces. The capsule wall was very thick (1013µm)and composed of numerous tightly packed, paren-chymatous corticalcells with little or no intercellular air space. Chloroplastswith well-defined granal stacks and extensive stroma lamellaewere observed in each of these three tissues, however, theirdensity was always greater in the palisade cells of the leafcompared to spongy mesophyll cells of the bract and the parenchymatouscells of the capsule wall. The low rates of photosynthesis inthe bracts and the capsule wall were associated with the internalorganization of these tissues. Key words: Cotton, photosynthesis, anatomy, cuticle, tissues     

Habituation in Nicotiana bigelovii tissue cultures: Different behaviour of two varieties

The exceptionally high capacity for transformation to autotrophyfor hormones (habituation) discovered in Nicotiana bigeloviiled us to a comparison of the responses of 2 varieties (quadrivalvisand bigelovii) to different hormone treatments applied to theirseeds. Hormones used were: kinetin (0.8, 1.6, 3.2 ppm), 2,4-D(0.1, 0.4 ppm), IAA (2 ppm) and NAA (1, 2, 4 ppm). Callus formation and habituation were scored after transferon minimal medium (without hormones) 3 months after sowing.Var. quadrivalvis showed a much higher ability to form callusbut the callus formed was autonomous in a lower percentage ofcases when IAA or kinetin treatment was considered. Analysisof auxin-like substances in leaves of the 2 varieties showeda higher content in N. bigelovii var. quadrivalvis. Data arediscussed with particular regard to their relevance to tumorousphenomena in plants. ¹ Publication No 52 of the Laboratorio di Mutagenesi e Differenziamento,C. N. R. (Received March 30, 1971; )     

Effect of Phosphorus Nutrition on the Cellular Distribution of Acid Phosphatase in the Leaves of Lycopersicon esculentum L.

A histochemical study using light microscopy has been made ofthe distribution of acid phosphatase (EC 3.1.3.2 [EC] ) activity intransverse sections of fully expanded leaves of Lycopersiconesculentum grown in phosphate-deficient or sufficient media.Leaf tissues were prepared by two methods and were embeddedin paraffin wax. The location of acid phosphatase activity inleaf sections was determined by trapping orthophosphate releasedfrom p-nitrophenyl phosphate with lead acetate and subsequentlyconverting the lead phosphate to optically dense lead sulphide.In leaf sections from control tissue lead sulphide depositswere larpely confined to the spongy mesophyll cells. Whereasthe staining of the palisade cells was limited and of a granularnature, the staining of the spongy mesophyll cells was heavierand coincident with the outline of the individual cells. Moreover,the minor veins were more heavily stained than the surroundingmesophyll cells. Sections of phosphorus-deficient tissues wereheavily stained in both the palisade and spongy mesophyll layersand heavy deposits of lead sulphide were present in the regionsof the minor veins. It is suggested that the enhanced acid phosphataseactivity of the mesophyll cells in fully expanded leaves couldbe involved in the remobilization of phosphate within phosphorus-deficientplants, or be part of a phosphate transporting system, concentratingthe intracellular phosphate from the limiting supply in thesolution bathing the mesophyll cells. Lycopersicon esculentum L., tomato, acid phosphatase, phosphorus nutrition     

Vegetative Reproduction as Related to Biomechanics, Morphology and Anatomy of Four Cholla Cactus Species in the Sonoran Desert

Vegetative reproduction via the rooting of detached stem segments(joints) is well recognized for certain cylindropuntias (chollas).This mode of reproduction was characterized in the field forOpuntia acanthocarpa, O. bigelovii, O. echinocarpa and O. ramosissimain the northwestern Sonoran Desert and the southern Mojave Desertand related to the following: (1) morphology of terminal jointsand their junctions; (2) the biomechanics and anatomy of terminaljoint junctions; and (3) the rooting of detached terminal joints.Species that typically reproduce vegetatively were hypothesizedto possess mechanically weak terminal joint junctions and terminaljoints with high rooting abilities. In general, resistance tofailure of terminal joint junctions depended on the diameterof the junction, with larger diameters providing greater resistanceto mechanical failure. Junction strength also depended on thepresence or absence of fibres and the amount of parenchyma cellsper cross-sectional area. Rooting ability appeared to dependon joint diameter, which determines the amount of stored carbohydratesand water. Of the four species, only O. bigelovii showed evidenceof vegetative reproduction in the field and was also the onlyspecies that had both relatively weak junctions and joints witha high rooting ability. Furthermore, joints of O. bigeloviihad the most spines per tubercle area, which increases the chancethat their spines will catch on a passing vertebrate, allowingfor greater joint dispersal. Copyright 2001 Annals of BotanyCompany Biomechanics, Cactaceae, Cholla, libriform fibres, Opuntia, vascular tracheids, vessel elements     

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     

Comparative Photosynthetic Properties of Palisade Tissue Chloroplasts and Spongy Tissue Chloroplasts of Camellia japonica L.: Functional Adjustment of the Photosynthetic Apparatus to Light Environment within a Leaf

The photochemical properties of chloroplasts isolated separatelyfrom palisade and spongy tissues of Camellia leaves, were compared,and the following results were obtained: (1) The content ofthe light-harvesting Chi a/b-protein complex was higher in spongytissue chloroplasts (S-Chlts) than in palisade tissue chloroplasts(P-Chlts), while the contents of P700 and PS IT polypeptideswere higher in P-Chlts. (2) Fluorescence induction was slowerin P-Chlts, indicating that they had a larger plastoquinonepool than S-Chlts. (3) The quantum yield of PS II electron transportin S-Chlts was appreciably higher, while that of PS I electrontransport was higher in P-Chlts. (4) The maximal rates of bothPS I and PS IT electron transport under saturating light werehigher in P-Chlts than in S-Chlts. From these results, we concluded that the photochemical propertiesin P-Chlts are adjusted to high light intensity and those ofS-Chlts to low intensity enriched in green and far-red; bothare adjusted to their respective in situ light environments. (Received December 24, 1983; Accepted March 6, 1984)     

Effect of UV-B Radiation on Leaf Optical Properties Measured with Fibre Optics

Changes in the internal light microenvironment in leaves ofplants of Brassica campestris L. cv. Emma, B. carinata L., andMedicago saliva L. cv. Armour in response to exposure to UV-B(UV-B, 280–320 nm) radiation were measured using a fibreopticmicroprobe. Plants were exposed for 2 weeks either to high visiblelight or to supplemental ultraviolet-B radiation. The spectral regime (400–700 nm; PAR) was measured eithermidway through the leaf palisade or the spongy mesophyll. Afterexposure to UV-B radiation leaves of Brassica campesiris attenuatedtransmitted light more than the controls. At the same time bothforward and back scattered light increased in the palisade andspongy mesophylls. In contrast, UV-treatment of Medicago salivaleaves increased light transmission into the palisade, whilethe back scattered component showed little change. Leaves ofcariiwla showed little change in response to UV. Other responsesto UV-B radiation included increases in leaf thickness, decreasedtotal chlorophyll content, and changes in UV-B screening pigmentsand chlorophyll fluorescence induction kinetics. Brassica campestriswas most sensitive to exposure to enhanced levels of UV-B radiation,whereas leaves of B. carinata were the least sensitive. Ourdata indicate that exposure to UV-B radiation altered the lightmicroenvironment within leaves of the species different ways.These changes appeared to be caused by alterations in pigmentcontent and leaf anatomy. In turn, the altered distributionof PAR within the leaf could influence photosynthesis. Key words: Brassica campestris, Brassica carinata, fibre optics, light scattering, Medicago saliva, optical properties, ozone depletion, photosynthesis, ultraviolet radiation     

Calcium distribution in globoid crystals of cucurbita cotyledon protein bodies

Energy-dispersive x-ray analysis was used to investigate the location of globoid crystals with relatively high Ca levels within cotyledons of Cucurbita maxima, Cucurbita mixta, and Cucurbita andreana. The small globoid crystals in both upper and lower epidermal cells commonly contained Ca. Ca was present in globoid crystals of all provascular regions with the exception of the very small provascular regions of C. maxima. In C. maxima and C. mixta cotyledons, some cases were observed where Ca was found in the globoid crystals of the first layer of mesophyll cells surrounding the provascular region, but in general Ca was absent from globoid crystals of palisade and spongy mesophyll cells. In C. andreana, globoid crystals of palisade and spongy mesophyll cells commonly contained at least some Ca. Cell position and cell type are factors affecting the Ca content of globoid crystals in protein bodies.     

The paraveinal mesophyll of soybean leaves in relation to assimilate transfer and compartmentation

Nitrogen and carbohydrate assimilates were temporally and spatially compartmented among various cell types in soybean (Glycine max L., Merr.) leaves during seed filling. The paraveinal mesophyll (PVM), a unique cell layer found in soybean, was demonstrated to function in the synthesis, compartmentation and remobilization of nitrogen reserves prior to and during the seed-filling stages. At anthesis, the PVM vacuoles contain substantial protein which completely disappears by two weeks into the seed filling. Distinct changes in the PVM cytoplasm, tonoplast and organelles were correlated with the presence or absence of the vacuolar material. Microautoradiography following the accumulation of several radiolabeled sugars and amino acids demonstrated the glycoprotein nature of the vacuolar material. Incorporation of methionine, leucine, glucose, and glucosamine resulted in heavy labelling of the PVM vacuole, in contrast to galactose, proline, and mannose which resulted in a much reduced labelling pattern. In addition, starch is unequally compartmented and degraded among the various leaf cells during seed filling. At the end of the photoperiod at the flowering stage, the highest starch accumulation was in the second palisade layer followed by the spongy mesophyll and the first (uppermost) palisade layer. Starch in the first palisade layer was completely degraded during the dark whereas the starch in the second palisade and spongy mesophyll was not remobilized to any appreciable extent. By mid-podfilling (approximately five weeks postanthesis) starch was absent in the first palisade layer at the end of the photoperiod while the second palisade and spongy mesophyll layers contained substantial starch. Starch was remobilized from these latter cells during the remainder of seed filling when current photosynthetic production is low. Structural changes associated with cell senescence first appear in the upper palisade layer and then progress (excluding the PVM) to the second palisade and spongy mesophyll layer. The PVM and phloem appear to retain their structural integrity into the leaf yellowing stage. Reducing sink capacity by pod removal resulted in a continued accumulation of vacuolar protein, an increase in cytoplasmic volume, and fragmentation of the vacuole in the PVM. Pod removal also resulted in an increased amount of accumulated starch (which did not turn over) in all mesophyll layers, and an increase in cell size and cell-wall thickness.     

Possible Involvement of Greening in Cell Growth of Suaeda japonica under Salt Stress

Suaeda japonica, a member of the family Chenopodiaceae, is ahalophyte that grows on the shores of the Ariake Sea in Japan.Using yellowish and selected green callus tissues of S. japonica,we examined the correlation between cell growth and glycinebetainecontent, as well as the activity of betaine aldehyde dehydrogenase(BADH), under salt stress. Although the growth of yellowishcallus tissues was markedly inhibited by 0.1 M NaCl, that ofgreen callus tissues was not. However, in 0.3 M NaCl, the freshweight of green callus tissues was decreased by 15%. Both theendogenous level of glycinebetaine and the activity of BADHin yellowish callus tissues were very low, but green callustissues contained very high levels of glycinebetaine and highBADH activity. The activity in green callus increased significantlywith increases in the concentration of NaCl. The increase inlevels of Chl was also observed, but the increase occurred earlierthan that in the level of glycinebetaine. Functional developmentof chloroplasts (an increase in the level of Chl) seems to beinvolved in the increase in the activity of BADH, and the resultantincrease in levels of glycinebetaine seems to facilitate callusgrowth under salt stress. (Received October 2, 1996; Accepted November 18, 1996)     

A New Model for Leaf Photosynthesis Incorporating the Gradients of Light Environment and of Photosynthetic Properties of Chloroplasts within a Leaf

A leaf photosynthesis model was constructed based upon the notionthat the leaf photosynthesis is a summation of photosynthesisof each chloroplast under in situ micro-environmental conditions.Intra-leaf light environment was calculated using the valuesof transmittance and reflectance of leaf tissues reported previously.Simulations of light response curves of whole leaf photosynthesiswere carried out for the model leaves with different patternsof gradients in light environment and/or in photosynthetic activities.The results indicate that the higher absorption coefficientof chlorophyll in spongy tissue than in palisade tissue andintra-leaf vertical gradient in photosynthetic activity of thechloroplasts as reported for real dorsiventral leaves are bothadvantageous to the productivity of the leaf because these propertiesraise the efficiency of the light utilization. Intra-leaf light environment, leaf photosynthesis, light utilization, palisade tissue, photosynthetic productivity, spongy tissue     

Response of Melanthera biflora to Salinity and Water Stress

Melanthera biflora (Asteraceae) is a moderately salt-tolerantplant from the Indo-Pacific region. In laboratory studies itsgrowth was inhibited by salt above 50 mol m^–3, but itwas able to survive salinities approaching that of seawater,namely 400 mol m^–3. Shoot potassium concentrations weremaintained over a range of salinities up to 400 mol m^–3,while sodium and chloride accumulation followed closely theincrease in external osmotic pressure. In contrast, the increasein osmotic pressure of the leaf sap of Melanthera biflora, subjectedto water stress, was due mainly to a decrease in the ratio offresh weight/dry weight. 3-dimethylsulphoniopropionate (3-DMSP)and glycinebetaine were identified by fast atom bombardmentmass and ¹H -NMR spectroscopy, with 3-DMSP being the main oniumcompound and glycinebetaine absent in some accessions. Onium(quaternary ammonium and/or tertiary sulphonium) compounds andproline increased during salt and water stress due mainly toa decrease in the fresh weight/dry weight ratio of tissue, althoughpart of the increase in salt-stressed tissue was due to an increasein the accumulation of the onium compound. This salt-inducedincrease in 3-DMSP was inhibited in conditions of low sulphursupply and there was no compensatory increase in proline. Key words: Melanthera biflora, Asteraceae, salinity, glycinebetaine, 3-dimethylsulphonioproprionate