Salt-induced NO<Subscript>3</Subscript><Superscript>-</Superscript> uptake inhibition in cowpea roots is dependent on the ionic composition of the salt and its osmotic effect

Salinity remarkably inhibits NO₃ ^- uptake but the mechanisms are not well understood. This study was addressed to elucidate the role of ionic and osmotic components of salinity on NO₃ ^- influx and efflux employing classic kinetics involving a low affinity transport system (LATS) and a high affinity transport system (HATS). In the presence of KCl, NaCl, and Na₂SO₄ at 100 mM concentrations, in both LATS and HATS, Michaelis constant (K_m) was similar for the three salts and maximum rate (V_max) decreased as follows: KCl > NaCl > Na₂SO₄, compared to control indicating a non-competitive interaction with NO₃ ^-. Unexpectedly, iso-osmotic solutions (osmotic potential Ψ_π = -0.450) of polyethylene glycol (PEG, 17.84 %, v/v) and mannitol (100 mM) remarkably increased K_m in both the LATS and the HATS, but V_max did not change indicating a competitive inhibition. Under the PEG and mannitol treatments, K_m and V_max were higher than under the salt treatments. The salts increased slightly NO₃ ^- efflux in the following order KCl > NaCl > Na₂SO₄. In contrast, mannitol strongly stimulated and the PEG inhibited NO₃ ^- efflux. The obtained data reveal that salinity effects were not dependent on the anion type (Cl^- versus SO₄ ^2-) indicating a non-competitive inhibition mechanism between Cl^- and NO₃ ^-. In contrast, the cation types (K⁺ versus Na⁺) had a pronounced effect. The osmotic component is important to net NO₃ ^- uptake affecting remarkably the influx in both LATS and HATS components of cowpea roots.     

Intracellular position of mitochondria in mesophyll cells differs between C<Subscript>3</Subscript> and C<Subscript>4</Subscript> grasses

In C₃ plants, part of the CO₂ fixed during photosynthesis in chloroplasts is released from mitochondria during photorespiration by decarboxylation of glycine via glycine decarboxylase (GDC), thereby reducing photosynthetic efficiency. The apparent positioning of most mitochondria in the interior (vacuole side of chloroplasts) of mesophyll cells in C₃ grasses would increase the efficiency of refixation of CO₂ released from mitochondria by ribulose 1,5-bisphosphate carboxylase/?oxygenase (Rubisco) in chloroplasts. Therefore, in mesophyll cells of C₄ grasses, which lack both GDC and Rubisco, the mitochondria ought not to be positioned the same way as in C₃ mesophyll cells. To test this hypothesis, we investigated the intracellular position of mitochondria in mesophyll cells of 14 C₄ grasses of different C₄ subtypes and subfamilies (Chloridoideae, Micrairoideae, and Panicoideae) and a C₃–C₄ intermediate grass, Steinchisma hians, under an electron microscope. In C₄ mesophyll cells, most mitochondria were positioned adjacent to the cell wall, which clearly differs from the positioning in C₃ mesophyll cells. In S. hians mesophyll cells, the positioning was similar to that in C₃ cells. These results suggest that the mitochondrial positioning in C₄ mesophyll cells reflects the absence of both GDC and Rubisco in the mesophyll cells and the high activity of phosphoenolpyruvate carboxylase. In contrast, the relationship between the mitochondrial positioning and enzyme distribution in S. hians is complex, but the positioning may be related to the capture of respiratory CO₂ by Rubisco. Our study provides new possible insight into the physiological role of mitochondrial positioning in photosynthetic cells.     

Effects of NaCl, Na2SO4, and mannitol on storage lipid mobilization in the cotyledons and roots of purple alfalfa seedlings

The effects of NaCl, Na₂SO₄, and mannitol on mobilization of storage lipids in the cotyledon mesophyll and root meristem cells during the germination of purple alfalfa (Medicago sativa L.) seeds were studied using the transmission electron microscopy technique. The ultrastructural analysis showed that the number and shape of specific organelles, oleosomes, depended on the osmotic effects of a salt used. In control seedlings grown on distilled water, lipid mobilization in roots terminated by the 4th day, and in cotyledons, only by the 8th day. The disruptions in the storage lipid mobilization in the treated seedlings were specific and depended on the stress agent used, the osmotic pressure induced by this agent, as well as on the tissue type. The presence of a great number of oleosomes in the cytoplasm of cells subjected to osmotic stress, in contrast to the control ones, demonstrated the inhibition of mobilization processes. All concentrations of Na₂SO₄ and mannitol suppressed lipid mobilization in the cotyledons and roots. At the same time, NaCl markedly suppressed this process in the root cells only at a higher osmotic pressure of the solution. The results obtained suggest that the cytological analysis of cotyledon storage-lipid mobilization can tentatively assess the resistance of dicotyledonous plants to abiotic stresses.     

Effects of NaCl or Na<Subscript>2</Subscript>SO<Subscript>4</Subscript> salinity on plant growth,ion content and photosynthetic activity in <Emphasis Type="Italic">Ocimum basilicum</Emphasis> L.

Basil (Ocimum basilicum L., cultivar Genovese) plants were grown in Hoagland solution with or without 50 mM NaCl or 25 mM Na₂SO₄. After 15 days of treatment, Na₂SO₄ slowed growth of plants as indicated by root, stem and leaf dry weight, root length, shoot height and leaf area, and the effects were major of those induced by NaCl. Photosynthetic response was decreased more by chloride salinity than by sulphate. No effects in both treatments on leaf chlorophyll content, maximal efficiency of PSII photochemistry (F _v/F _m) and electron transport rate (ETR) were recorded. Therefore, an excess of energy following the limitation to CO₂ photoassimilation and a down regulation of PSII photochemistry was monitored under NaCl, which displays mechanisms that play a role in avoiding PSII photodamage able to dissipate this excess energy. Ionic composition (Na⁺, K⁺, Ca²⁺, and Mg²⁺) was affected to the same extent under both types of salinity, thus together with an increase in leaves Cl⁻, and roots SO₄ ²⁻ in NaCl and Na₂SO₄-treated plants, respectively, may have resulted in the observed growth retardation (for Na₂SO₄ treatment) and photosynthesis activity inhibition (for NaCl treatment), suggesting that those effects seem to have been due to the anionic component of the salts.     

Effects of salt-tolerant rootstock grafting on ultrastructure,photosynthetic capacity,and H<Subscript>2</Subscript>O<Subscript>2</Subscript>-scavenging system in chloroplasts of cucumber seedlings under NaCl stress

Plant growth, photosynthetic parameters, chloroplast ultrastructure, and the ascorbate-glutathione cycle system in chloroplasts of self-grafted and rootstock-grafted cucumber leaves were investigated. Grafted plants were grown hydroponically and were exposed to 0, 50, and 100 mM NaCl concentrations for 10 days. Under NaCl stress, the hydrogen peroxide (H₂O₂) content in cucumber chloroplasts increased, the chloroplast ultrastructure was damaged, and the gas stomatal conductance, intercellular CO₂ concentration, as well as shoot dry weight, plant height, stem diameter, leaf area, and leaf relative water content were inhibited, whereas these changes were less severe in rootstock-grafted plants. The activities of ascorbate peroxidase (APX; EC 1.11.1.11), glutathione reductase (GR; EC 1.6.4.2), and dehydroascorbate reductase (DHAR EC 1.8.5.1) were higher in the chloroplasts of rootstock-grafted plants compared with those of self-grafted plants under 50 and 100 mM NaCl. Similar trends were shown in leaf net CO₂ assimilation rate and transpiration rate, as well as reduced glutathione content under 100 mM NaCl. Results suggest that rootstock grafting enhances the H₂O₂-scavenging capacity of the ascorbate–glutathione cycle in cucumber chloroplasts under NaCl stress, thereby protecting the chloroplast structure and improving the photosynthetic performance of cucumber leaves. As a result, cucumber growth is promoted.     

Growth stimulation and inhibition by salt in relation to Na<Superscript>+</Superscript> manipulating genes in xero-halophyte <Emphasis Type="Italic">Atriplex halimus</Emphasis> L.

In the present study, Na⁺ manipulating genes could contribute not only to ion homeostasis but also to growth stimulation with exposing the halophyte Atriplex halimus L. to moderate NaCl concentration. The stimulation of growth was attributed to Na⁺ accumulation inside the vacuole leading to increase leaf cell size as well as accelerate leaf cell division. Increasing the assimilatory surface could result in enhancing the photosynthetic rate. The reduction of A. halimus growth compared to optimum growth at 50 and 200 mM NaCl could be attributed to osmotic effect rather than the ionic one of salt stress. The inhibition of photosynthesis seemed to be resulted from limitation of CO₂ due to the osmotic effect on stomatal conductance rather than the activity loss of photosynthetic machinery. The depletion of starch content along with the increase in sucrose content could imply that photosynthesis may be a limiting for A. halimus growth. The fast coordinate induction of Na⁺ manipulating genes could reveal that the tolerance of A. halimus to high concentrations evolved from its ability to regulate and control Na⁺ influx and efflux. V-H ⁺-PPase may play a vital role in A. halimus tolerance to osmotic and/or ionic stress due to its kinetics of induction. It seemed that H⁺-ATPase plays a pivotal role in A. halimus tolerance to stress due to the increase in its protein level was detected with all NaCl concentrations as well as with PEG treatments. Both of these genes might be useful in improving stress tolerance in transgenic crops.     

Changes in the levels of jasmonates and free polyamines induced by Na<Subscript>2</Subscript>SO<Subscript>4</Subscript> and NaCl in roots and leaves of the halophyte <Emphasis Type="Italic">Prosopis strombulifera</Emphasis>

Prosopis strombulifera, a common legume in high-salinity soils of Argentina, is a useful model for elucidation of salt tolerance mechanisms and specific biochemical pathways in halophytes, since its NaCl tolerance exceeds the limit described for most halophytic plants. We analyzed the effects of the increasing concentration of two main soil salts, Na₂SO₄ and NaCl, on growth parameters of P. strombulifera, chlorophyll levels, and content of jasmonates (JAs) and polyamines (PAs), which are key molecules involved in stress responses. P. strombulifera showed a halophytic response (growth promotion) to NaCl, but strong growth inhibition by iso-osmotic solutions of Na₂SO₄. Chlorophyll levels, number of leaves and leaf area were also differentially affected. An important finding was the partial alleviation of SO₄²⁻ toxicity by treatment with two-salt mixture. JAs are not directly involved in salt tolerance in this species since its levels decrease under all salt treatments. Beneficial effects of Putrescine (Put) accumulation in NaCl treated plants maybe inferred probably associated with the antioxidative defense system. Another novel finding is the accumulation of the uncommon PA cadaverine in roots under high Na₂SO₄, which may be related to SO₄²⁻ toxicity.     

Reorganization of interphase microtubules in root cells of <Emphasis Type="Italic">Medicago sativa</Emphasis> L. during acclimation to osmotic and salt stress

We examined the organization of microtubule system of interphase cells in roots of Medicago sativa L. during acclimation to salt and osmotic stress at different concentrations of NaCl, Na₂SO₄, and mannitol. We identified morphological changes of tubulin cytoskeleton in different root tissues during the acclimation to salt and osmotic stress: (1) decreased density of the cortical microtubule network, (2) random orientation of cortical microtubule bundles, (4) thickening of the bundles, (3) nonuniform density of the bundles, (4) fragmentation of the bundles, and (5) formation of microtubule converging centers. Network thinning and thickening of the bundles were observed both under osmotic and salt stress. Random orientation of cortical microtubules was visualized under osmotic stress but not during salt stress. Fragmentation of microtubule bundles took place under salt stress with a high concentration of mannitol. Formation of microtubule converging centers was common under prolonged action of sodium sulfate, less evident under sodium chloride, and not found after mannitol treatment. Our data show that, in alfalfa root cells, cortical microtubules rearrange not only in response to different ions, but also to osmotic pressure. Thus, the signaling pathways and molecular mechanisms inducing reorganization of the microtubule system may be triggered by sodium cations, as well as by sulfate and chloride anions at concentrations that do not cause irreversible cell damage.     

Photosynthetic limitations of a halophyte sea aster (<Emphasis Type="Italic">Aster tripolium</Emphasis> L) under water stress and NaCl stress

To understand the mechanisms of salt tolerance in a halophyte, sea aster (Aster tripolium L.), we studied the changes of water relation and the factors of photosynthetic limitation under water stress and 300 mM NaCl stress. The contents of Na⁺ and Cl^- were highest in NaCl-stressed leaves. Leaf osmotic potentials (Ψ _s) were decreased by both stress treatments, whereas leaf turgor pressure (Ψ _t) was maintained under NaCl stress. Decrease inΨ _s without any loss ofΨ _t accounted for osmotic adjustment using Na⁺ and Cl^- accumulated under NaCl stress. Stress treatments affected photosynthesis, and stomatal limitation was higher under water stress than under NaCl stress. Additionally, maximum CO₂ fixation rate and O₂ evolution rate decreased only under water stress, indicating irreversible damage to photosynthetic systems, mainly by dehydration. Water stress severely affected the water relation and photosynthetic capacity. On the other hand, turgid leaves under NaCl stress have dehydration tolerance due to maintenance of Ψ _t and photosynthetic activity. These results show that sea aster might not suffer from tissue dehydration in highly salinized environments. We conclude that the adaptation of sea aster to salinity may be accomplished by osmotic adjustment using accumulated Na⁺ and Cl^-, and that this plant has typical halophyte characteristics, but not drought tolerance. Electronic Publication     

Effects of Salt Stress on Photosynthetic Pigments and Activity of Ribulose-1,5-bisphosphate Carboxylase/Oxygenase in <Emphasis Type="Italic">Kalidium foliatum</Emphasis>

The effects of NaCl and Na₂SO₄ on photosynthetic pigments, malondialdehyde (MDA), Rubisco activity and superoxide dismutase (SOD) activity were investigated in Kalidium foliatum (Pall.) Moq., which is distributed in the saline soil of Hetao irrigation area in Inner Mongolia China. The K. foliatum plants were treated with NaCl (0, 100, 250, 400 and 500 mM), Na₂SO₄ (0, 100, 250, 400 and 500 mM) and NaCl + Na₂SO₄ (1: 1, v/v) (0, 100, 250, 400 and 500 mM of Na⁺ concentration, 0, 50, 125, 200 and 250 mM of Cl^– and SO ₄ ^2– concentration) for 10 days. Content of chlorophylls and carotenoids were significantly higher than control at increasing NaCl and Na₂SO₄ concentration, in contrast, were significantly reduced by higher concentration of NaCl + Na₂SO₄. Rubisco activity reduced steadily at 100 and 250 mM NaCl, while increased at 400 and 500 mM NaCl. Rubisco activity was significantly higher than control at 100 mM Na₂SO₄, and was no more change under NaCl + Na₂SO₄ treatment. The SOD activity increased with increasing NaCl and Na₂SO₄, and increased at moderate NaCl + Na₂SO₄ treatment. MDA content was lower than control at 250 mM salt concentration. On the basis of the data obtained, K. foliatum showed resistance to salt such as Na⁺, Cl^–and SO ₄ ^2– , Rubisco activity in K. foliatum might be more sensitive to salt.     

Some morphogenic effects of sodium sulfate on tobacco callus

Callus cultures of Nicotiana tabacum L cv. Wisconsin 38 were initiated and grown on shoot-forming (SF) and callus proliferation (CP) medium with or without Na₂SO₄. Two cultures were maintained on SF medium with 0, 0.75, 1 or 1.5% Na₂SO₄ for 2.5 and 3.5 years. In the older culture only callus grown on salt formed shoots throughout the maintenance period, while in the younger culture the control responded best and Na₂SO₄ was inhibitory. Callus from the older culture which had been grown on salt continued to form shoots in the absence of salt. Na₂SO₄ caused adventitious shoot formation in three cultures on CP medium. These shoots were present for 7 subcultures after removal of Na₂SO₄; but established, control callus, did not form shoots when transferred to Na₂SO₄. Callus initiated and maintained on NaCl or mannitol showed a slight increase in shoot initiation. On NaCl, Na₂SO₄ or mannitol, the tissue osmotic potential became more negative and proline concentration increased.     

Effect of sodium chloride on the response of the halophyte species <Emphasis Type="Italic">Sesuvium portulacastrum</Emphasis> grown in mannitol-induced water stress

Sesuvium portulacastrum is a halophytic species well adapted to salinity and drought. In order to evaluate the physiological impact of salt on water deficit-induced stress response, we cultivated seedlings for 12 days, in the presence or absence of 100 mmol l⁻¹ NaCl, on a nutrient solution containing either 0 mmol l⁻¹ or 25 mmol l⁻¹ mannitol. Mannitol-induced water stress reduced growth, increased the root/shoot ratio, and led to a significant decrease in water potential and leaf relative water content, whereas leaf Na⁺ and K⁺ concentrations remained unchanged. The addition of 100 mmol l⁻¹ NaCl to 25 mmol l⁻¹ mannitol-containing medium mitigated the deleterious impact of water stress on growth of S. portulacastrum, improved the relative water content, induced a significant decrease in leaf water potential and, concomitantly, resulted in enhancement of overall plant photosynthetic activity (i.e. CO₂ net assimilation rate, stomatal conductance). Presence of NaCl in the culture medium, together with mannitol, significantly increased the level of Na⁺ and proline in the leaves, but it had no effect on leaf soluble sugar content. These findings suggest that the ability of NaCl to improve plant performance under mannitol-induced water stress may be due to its effect on osmotic adjustment through Na⁺ and proline accumulation, which is coupled with an improvement in photosynthetic activity. A striking recovery in relative water content and growth of the seedlings was also recorded in the presence of NaCl on release of the water stress induced by mannitol.     

Characterization of Growth, Water Relations, and Proline Accumulation in Sodium Sulfate Tolerant Callus of Brassica napus L. cv Westar (Canola)

Unselected and sodium sulfate tolerant callus cultures of Brassica napus L. cv Westar were grown on media supplemented with mannitol, NaCl, or Na₂SO₄. In all cases, growth of tolerant callus, measured on a fresh weight or dry weight basis, was greater than that of unselected callus, which was also subject to necrosis on high levels of salt. Tissue water potential became more negative in both unselected and tolerant callus grown in the presence of mannitol or Na₂SO₄. Water potentials in unselected callus were more negative than those of the tolerant tissues; but over a range of Na₂SO₄ concentrations both cultures displayed osmotic adjustment, maintaining relatively constant turgor. Proline accumulation in both unselected and tolerant callus was low (15 to 20 micromoles per gram dry weight) in the absence of stress, but increased on media supplemented with mannitol, NaCl, or Na₂SO₄. Increases in proline concentration were approximately linear in tolerant callus, reaching a maximum of 130 to 175 micromoles per gram dry weight. In unselected callus, concentrations were higher, reaching 390 to 520 micromoles per gram dry weight. Proline accumulation was correlated with inhibition of growth, and there was a negative correlation between proline concentration and culture age for tolerant callus.     

Contributions of inorganic ions,soluble carbohydrates,and multiatomic alcohols to water homeostasis in <Emphasis Type="Italic">Artemisia lerchiana</Emphasis> and <Emphasis Type="Italic">A. pauciflora</Emphasis>

Two morphological forms of wormwood Artemisia lerchiana (f. erecta and f. nutans) and A. pauciflora Web. (morphological form erecta) were grown on sand culture at a range of NaCl concentrations in the nutrient medium and then assayed for Na⁺, K⁺, and Cl^? content in various organs. In addition, the content of mono-, di-, and trisaccharides and multiatomic alcohols (mannitol and glycerol); water content; and organ biomass were determined. All plants examined showed high NaCl tolerance, comparable to that of halophytes. They were able to maintain high tissue hydration under conditions of salinity-induced growth suppression. The intracellular osmotic pressure in wormwood organs was mainly determined by the presence of Na⁺, K⁺, and Cl^?, as well as by mono-, di-, and trisaccharides, mannitol, and glycerol. The high content of Na⁺ and Cl^? in wormwood organs was also observed in the absence of salinity, which implies the ability of these organs to absorb ions from diluted NaCl solutions and accumulate ions in cells of their tissues. With the increase in salinity, the content of Na⁺ and Cl^? in roots and leaves increased to even higher levels. It is concluded that the ability of wormwood plants to absorb and accumulate inorganic ions provides for sustainable high intracellular osmotic pressure and, accordingly, low water potential under drought and salinity conditions. Growing plants under high salinity lowered the content of monosaccharides in parallel with accumulation of the trisaccharide raffinose. It is supposed that soluble carbohydrates and multiatomic alcohols are not only significant for osmoregulation but also perform a protective function in wormwood plants. The lower osmotic pressure in root cells compared to that in leaf cells of all plants examined was mainly due to the gradient distribution of K⁺ and Cl^? between roots and leaves. The two Artemisia species and two morphological forms of A. lerchiana did not differ appreciably in the ways of water balance regulation. It is found that different morphologies of two A. lerchiana forms are unrelated to variations in intracellular osmotic and turgor pressures.     

Diffusional conductance to CO2 is the key limitation to photosynthesis in salt‐stressed leaves of rice (Oryza sativa)

Salinity significantly limits leaf photosynthesis but the factors causing the limitation in salt‐stressed leaves remain unclear. In the present work, photosynthetic and biochemical traits were investigated in four rice genotypes under two NaCl concentration (0 and 150 mM) to assess the stomatal, mesophyll and biochemical contributions to reduced photosynthetic rate (A) in salt‐stressed leaves. Our results indicated that salinity led to a decrease in A, leaf osmotic potential, electron transport rate and CO₂ concentrations in the chloroplasts (C_c) of rice leaves. Decreased A in salt‐stressed leaves was mainly attributable to low C_c, which was determined by stomatal and mesophyll conductance. The increased stomatal limitation was mainly related to the low leaf osmotic potential caused by soil salinity. However, the increased mesophyll limitation in salt‐stressed leaves was related to both osmotic stress and ion stress. These findings highlight the importance of considering mesophyll conductance when developing salinity‐tolerant rice cultivars.     

Plastid Structure and Development in Green Callus Tissues of Oxalis dispar

Cultured callus tissues derived from endosperm of Oxalis disparare shown to contain virescent amyloplasts. In darkness, proplastidsdevelop into typical amyloplasts, starch being deposited assingle or multiple grains. In light, amyloplasts are transformedinto chloroplasts. Thylakoid formation begins in spaces aroundand between existing starch grains. As thylakoids are assembledinto grana, starch slowly disappears; the plastids increasein size and the photosynthetic apparatus enlarges to fill thewhole of the plastid. Slight carotenoid synthesis takes placeas amyloplasts are laid down, but there is no chlorophyll synthesis.All pigments accumulate rapidly during the early stages of granaldevelopment, but slowly, and at a declining rate, during thelater stages. Treatment of the tissues with auxins suppressesthe development of thylakoid membranes, but has no effect uponthe development of amyloplast membranes. The possible significanceof this observation is discussed. Greening is accompanied by a marked decline in the rates ofboth cell division and cell expansion. This is attributed inpart to the diversion of nitrogen from the normal growth channelsinto the synthesis of thylakoid proteins.     

Disruption of <Emphasis Type="Italic">RCI2A</Emphasis> leads to over-accumulation of Na<Superscript>+</Superscript> and increased salt sensitivity in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> plants

For plant salt tolerance, it is important to regulate the uptake and accumulation of Na⁺ ions. The yeast pmp3 mutant which lacks PMP3 gene accumulates excess Na⁺ ions in the cell and shows increased Na⁺ sensitivity. Although the function of PMP3 is not fully understood, it is proposed that PMP3 contributes to the restriction of Na⁺ uptake and consequently salt tolerance in yeasts. In this paper, we have investigated whether the lack of RCI2A gene, homologous to PMP3 gene, causes a salt sensitive phenotype in Arabidopsis (Arabidopsis thaliana (L.) Heynh.) plants; and to thereby indicate the physiological role of RCI2A in higher plants. Two T-DNA insertional mutants of RCI2A were identified. Although the growth of rci2a mutants was comparable with that of wild type under normal conditions, high NaCl treatment caused increased accumulation of Na⁺ and more reduction of the growth of roots and shoots of rci2a mutants than that of wild type. Undifferentiated callus cultures regenerated from rci2a mutants also accumulated more Na⁺ than that from wild type under high NaCl treatment. Furthermore, when wild-type and rci2a plants were treated with NaCl, NaNO₃, Na₂SO₄, KCl, KNO₃, K₂SO₄ or LiCl, the rci2a mutants showed more reduction of shoot growth than wild type. Under treatments of tetramethylammonium chloride, CaCl₂, MgCl₂, mannitol or sorbitol, the growth reduction was comparable between wild-type and rci2a plants. These results suggested that RCI2A plays a role directly or indirectly for avoiding over-accumulation of excess Na⁺ and K⁺ ions in plants, and contributes to salt tolerance.     

C-4 Pathway in <Emphasis Type="Italic">Pennisetum purpureum</Emphasis>

HIGH rates of photosynthesis and low rates of photorespiration are found in several plant species in which the photosynthetic carbon reduction (PCR) cycle is augmented by a second carboxylation system, the C-4 pathway^1,2. This pathway does not result in a net fixation or reduction of carbon, but seems to operate as a mechanism for concentrating CO₂ at the site of reductive assimilation. It has been widely accepted that the carboxylation reaction of the C-4 pathway is associated with the chloroplasts of mesophyll -cells, while PCR cycle activity is restricted to the bundle sheath chloroplasts. Investigations^3–5 indicate, however that mesophyll chloroplasts do contain PCR cycle activity, arni that the C-4 pathway is located in the cytoplasm of non-green and mesophyll cells.     

Structural and biochemical bases of photorespiration in C<Subscript>4</Subscript> plants: quantification of organelles and glycine decarboxylase

In C₄ plants, photorespiration is decreased relative to C₃ plants. However, it remains unclear how much photorespiratory capacity C₄ leaf tissues actually have. We thoroughly investigated the quantitative distribution of photorespiratory organelles and the immunogold localization of the P protein of glycine decarboxylase (GDC) in mesophyll (M) and bundle sheath (BS) cells of various C₄ grass species. Specific differences occurred in the proportions of mitochondria and peroxisomes in the BS cells (relative to the M cells) in photosynthetic tissues surrounding a vein: lower in the NADP-malic enzyme (NADP-ME) species having poorly formed grana in the BS chloroplasts, and higher in the NAD-malic enzyme (NAD-ME) and phosphoenolpyruvate carboxykinase (PCK) species having well developed grana. In all C₄ species, GDC was localized mainly in the BS mitochondria. When the total amounts of GDC in the BS mitochondria per unit leaf width were estimated from the immunogold labeling density and the quantity of mitochondria, the BSs of NADP-ME species contained less GDC than those of NAD-ME or PCK species. This trend was also verified by immunoblot analysis of leaf soluble protein. There was a high positive correlation between the degree of granal development (granal index) in the BS chloroplasts and the total amount of GDC in the BS mitochondria. The variations in the structural and biochemical features involved in photorespiration found among C₄ species might reflect differences in the O₂/CO₂ partial pressure and in the potential photorespiratory capacity of the BS cells.Abbreviations BS Bundle sheath - GDC Glycine decarboxylase - M Mesophyll - NAD-ME NAD-malic enzyme - NADP-ME NADP-malic enzyme - PCK Phosphoenolpyruvate carboxykinase