Showing papers in "Planta in 1976"

The presence of glutathione and glutathione reductase in chloroplasts: A proposed role in ascorbic acid metabolism.

Abstract: Both glutathione and an NADPH-dependent glutathione reductase are present in spinach (Spinacia oleracea L.) chloroplasts. It is proposed that glutathione functions to stabilise enzymes of the Calvin cycle, and it may also act to keep ascorbic acid in chloroplasts in the reduced form.

Purification and viability determinations of plant protoplasts.

Abstract: A method is described for purifying plant protoplasts from cellular and subcellular debris. The procedure utilizes a density buffer containing 9.6% sodium metrizoate and 5.6% Ficoll. The use of fluorescein diacetate for assessing the viability of plant protoplasts is also reported.

Formation of hydrogen peroxide by isolated cell walls from horseradish (Armoracia lapathifolia Gilib.).

Abstract: Isolated cell-wall suspensions from horseradish in the presence of 5×10(-4) M MnCl2 catalyze the production of hydrogen peroxide at the expense of either NADPH or NADH. This reaction is inhibited by scavengers of the superoxide free radical ion such as ascorbate or dihydroxyphenols or by superoxide dismutase, and stimulated by monophenols such as p-coumaric acid. On comparison with isolated (commercial) horseradish peroxidase it becomes evident that (a) cell-wall-bound peroxidase(s) is (are) responsible for the production of hydrogenperoxide, involving the superoxide free radical ion as an intermediate of the complex reaction chain.

Water permeability of isolated cuticular membranes: The effect of cuticular waxes on diffusion of water.

Abstract: The water permeability of astomatous cuticular membranes isolated from Citrus aurantium L. leaves, pear (Pyrus communis L.) leaves and onion (Allium cepa L.) bulb scales was determined before and after extraction of cuticular waxes with lipid solvents. In pear, the permeability coefficients for diffusion of tritiated water across cuticular membranes (CM) prior to extraction [Pd(CM)] decreased by a factor of four during leaf expansion. In all three species investigated Pd(CM) values of cuticular membranes from fully expanded leaves varied between 1 to 2×10-7 cm-3 s-1·Pd(CM) values were not affected by pH. Extraction of cuticular waxes from the membranes increased their water permeability by a factor of 300 to 500. Permeability coefficients for diffusion of THO across the cutin matrix (MX) after extraction [Pd(MX)] increased with increasing pH. Pdvalues were not inversely proportional to the thickness of cuticular membranes. By treating the cutin matrix and cuticular waxes as two resistances acting in series it was shown that the water permeability of cuticles is completely determined by the waxes. The lack of the Pd(CM) values to respond to pH appeared to be due to structural effects of waxes in the cutin matrix. Cuticular membranes from the submerse leaves of the aquatic plant Potamogeton lucens L. were three orders of magnitude more permeable to water than the cuticular membranes of the terrestrial species investigated.

Abscisic acid in developing wheat grains and its relationship to grain growth and maturation.

Abstract: During the later stages of growth of grains of wheat (Triticum aestivum L. cvs. WW15 and Gabo) there is a dramatic increase (up to 40fold) in the content of abscisic acid (ABA) to 4–6 ng per grain. This level remains high from 25 to 40 days after anthesis. Then, in association with natural or forced drying of the grain, there is a rapid drop (5–10 fold) in the ABA content and a brief increase in the content of bound ABA. The bulk of ABA in an ear was in the grain (95%) and although the embryo contributed 19% of this ABA it was less than 5% of the grain by weight. There was no clear relationship between ABA content and the growth of grains in various spikelet or floret positions. Application of (±)-ABA to the ear had no effect on grain growth rate but led to an earlier cessation of grain growth and hastened the drying of the grain. Isolated embryos and whole grains were capable of germinating during the mid grain growth period (15–25 days), but germination capacity declined subsequently as ABA accumulated. Later, still, with grain drying and loss of ABA, embryo and grain became germinable again. At this time there was also a dramatic increase in the ability of the grain to synthesize α-amylase. It is suggested that the accumulation of ABA at the later stages of grain growth prevents precocious germination and premature hydrolysis of starch reserves of the morphologically mature but still unripe grain. An inevitable consequence of such action may be in triggering grain maturation.

Chromium uptake and transport in barley seedlings (Hordeum vulgare L.).

Abstract: Potassium chromate is more toxic to the growth of barley in solution culture than chromic chloride, though apparent uptake of the latter is much faster. Inhibitor studies indicate that CrO42- uptake is “active” whereas Cr3+ uptake is passive, demonstrating that the two forms do not share a common uptake mechanism. Studies on the form of Cr inside root cells show that in plants fed CrO42- the Cr remains largely unchanged whereas in plants fed Cr3+ a little CrO42- (0.5 per cent) is produced. This conversion is dependent on the presence of living material and is probably enzymatic. Chromate uptake follows Michaelis-Menten kinetics at low concentration and is competitively inhibited by sulphate. Transport of chromium up the root is very slow, accounting for the low levels of Cr in the shoots. Chromate is transported better than Cr3+ though still to a very limited extent. These experiments provide a physiological basis for previous observations.

Water permeability of isolated cuticular membranes: The effect of pH and cations on diffusion, hydrodynamic permeability and size of polar pores in the cutin matrix

Abstract: The upper astomatous cuticle of Citrus aurantium L. leaves was isolated enzymatically or chemically, extracted with lipid solvents and used for the determination of water diffusion (Pd) and osmotic water permeability (Pf). The water permeability was strongly dependent on the pH value and the cations of the buffer solutions. In presence of monovalent alkali metal ions Pd increased almost five fold between pH 3 and 11. The shape of the plot Pd vs. pH suggests the presence of 3 different dissociable groups fixed to the membrane matrix. They are tentatively identified as two carboxyl groups dissociating between pH 3 to 6 and 6 to 9, respectively, and as phenolic hydroxyl groups dissociating above pH 9. The carboxyl group dissociating between pH 6 and 9 discriminated between alkali metal ions according to their ionic radius. Water permeability was lowest in the Li+ from and increased in the order Li+

The major growth current through lily pollen tubes enters as K(+) and leaves as H (+).

Abstract: Growing lily (Lilium longiflorum Thunb.) pollen always drive a current into their tubes and out of their grains. The only external ions needed for growth (and the growth current) are K+, H+, and Ca2+. Increases in K+ immediately stimulate the current; while decreases in K+ immediately inhibit it. Comparable changes in H+ have the opposite effect; while those in Ca2+ have very little effect. We infer that most of the steady growth current is carried in by a potassium leak and out by a proton pump; but other considerations indicate that a minor, but controlling, component of the inward current consists of calcium ions.

Hydrogen isotope discrimination in higher plants: Correlations with photosynthetic pathway and environment.

Abstract: The ratio of deuterium to hydrogen (expressed as δD) in hydrogen released as water during the combustion of dried plant material was examined. The δD value (metabolic hydrogen) determined on plant materials grown under controlled conditions is correlated with pathways of photosynthetic carbon metabolism. C3 plants show mean δD values of-132‰ for shoots and -117‰ for roots; C4 plants show mean δD values of -91‰ for shoots and-77‰ for roots and CAM plants a δD value of-75‰ for roots and shoots. The difference between the δD value of shoot material from C3 and C4 plants was confirmed in species growing under a range of glasshouse conditions. This difference in δD value between C3 and C4 species does not appear to be due to differences in the δD value (tissue water) in the plants as a result of physical fractionation of hydrogen isotopes during transpiration. In C3 and C4 plants the hydrogen isotope discrimination is in the same direction as the carbon isotope discrimination and factors contributing to the difference in δD values are discussed. In CAM plants grown in the laboratory or collected from the field δD values range from-75‰ to +50‰ and are correlated with δ(13)C values. When deprived of water, the δD value (metabolic hydrogen) in both soluble and insoluble material in leaves of Kalanchoe daigremontiana Hamet et Perr., becomes less negative. These changes may reflect the deuterium enrichment of tissue water during transpiration, or in field conditions, may reflect the different δD value of available water in areas of increasing aridity. Whatever the origin of the variable δD value in CAM plants, this parameter may be a useful index of the water relations of these plants under natural conditions.

Purification and characterization of peridinin-chlorophyll a -proteins from the marine dinoflagellates Glenodinium sp. and Gonyaulax polyedra

Abstract: A peridinin-chlorophyll a-protein complex (PCP) was obtained in large quantity from the marine dinoflagellates, Glenodinium sp. and Gonyaulax polyedra. The chromoproteins have similar molecular weights, 35,500 for Glenodinium sp. and 34,500 for G. polyedra. The proteins from the PCP complex of Glenodinium sp. dissociated from the chromophore on treatment with 1% sodium dodecyl sulfate (SDS) at room temperature. The protein component was a single subunit with a molecular weight of 15,500. Proteins from the PCP complex of G. polyedra were composed of a single polypeptide with a molecular weight of about 32,000. Two peridinin-chlorophyll a-proteins from Glenodinium sp. accounted for 70% of the PCP complex and had isoelectric points of 7.4 and 7.3. The PCP complex from G. polyedra was dominated by a single chromoprotein with an isoelectric point of 7.2 Chromophore analysis indicated the presence of only peridinin and chlorophyll a in a molar ratio approaching 4:1. Other pigments characteristically found in dinoflagellates were absent. Fluorescence excitation spectra of purified PCP indicated an efficient energy transfer from peridinin to chlorophyll a, an observation that lends support to the reported role of peridinin as an accessory pigment in photosynthetic oxygen evolution. In several other brown colored dinoflagellates examined, PCP representtd less than 20% of the total peridinin. However, no PCP could be isolated from cultures of Amphidinium carterae (PY-1). This study provides further evidence that PCP is a normal component of most peridinin-containing dinoflagellates, and functions as a light-harvesting component of the dinoflagellate chloroplast. No fucoxanthin-containing analog of PCP was detected in the chrysophyte, Cricosphera carterae and the dinoflagellate Glenodinium foliaceum.

Biochemical differentiation of plastids and other organelles in rye leaves with a high-temperature-induced deficiency of plastid ribosomes.

Abstract: 1. In developing rye (Secale cereale L.) leaves the formation of plastidic ribosomes was selectively prevented in light as well as in darkness, when the seedlings were grown at an elevated temperature of 32° instead of 22° where normal development ocurred. Plastid ribosome deficient parts of lightgrown leaves were chlorotic at 32°. — 2. At both temperatures the leaves contained under all conditions (light or dark, on H2O or nutrient solution) equal or very similar amounts of total amino nitrogen. In light, the contents of total protein and dry weight were lower at 32° than at 22°, especially when the plants were grown on nutrient solution. — 3. Mitochondrial marker enzymes had normal or even higher activities in 32°-grown leaves. Respiration rates were similar for segments of leaves grown on water in light either at 32° or at 22° but by 20–30% lower for 32°-grown plants when they had been raised in darkness or on nutrient solution. In contrast to 22°-grown tissue, respiration of 32°-grown leaf segments was rather insensitive to KCN. Comparative inhibitor studies indicated the presence of both the cyanide-sensitive and the cyanide-insensitive pathway of respiration in 32°-grown leaves. — 4. Leaf microbody marker enzymes were present in leaves grown at 32°. From chlorotic parts of 32°-light-grown leaves a typical microbody fraction was isolated on sucrose densitygradients. — 5. Leaves of seedlings grown at 32° contained only very low levels of ribulosediphosphate carboxylase activity and of fraction I protein. Photosynthetic 14CO2-fixation of such leaves was only a few per cent of that observed in normal leaves, and no photosynthetic oxygen evolution was observed in chlorotic leaf segments. However, ten other soluble enzymes which are exclusively or partially localized in chloroplasts reached high activities under all conditions at 32° (Table 4). — 6. From chlorotic parts of 32°-light-grown leaves as well as from etiolated 32°-grown leaves a fraction of intact plastids was isolated and purified by sucrose gradient centrifugation which contained several soluble chloroplast enzymes. From the results we conclude that cytoplasmic protein synthesis must contribute a functional chloroplast envelope including the mechanism for the recognition and uptake of chloroplast proteins which are synthesized on cytoplasmic ribosomes.

The regulation of ammonia assimilating enzymes in Lemma minor.

Abstract: Lemna minor has the potential to assimilate ammonia via either the glutamine or glutamate pathways. A 3-4 fold variation in the level of ferredoxindependent glutamate synthase may occur, when plants are grown on different nitrogen sources, but these changes show no simple relationship to changes in the endogenous pool of glutamate. High activities of glutamate synthase and glutamine synthetase at low ammonia availability suggests that these two enzymes function in the assimilation of low ammonia concentrations. Increasing ammonia availability leads to a reduction in level of glutamate synthase and glutamine synthetase and an increase in the level of glutamate dehydrogenase. Glutamine synthetase and glutamate dehydrogenase are subject to concurrent regulation, with glutamine rather than ammonia, exerting negative control on glutamine synthetase and positive control on glutamate dehydrogenase. The changes in the ratio of these two enzymes in response to the internal pool of glutamine could regulate the direction of the flow of ammonia into amino acids via the two alternative routes of assimilation.

The effects of water stress on abscisic-acid levels and metabolism in roots of Phaseolus vulgaris L. and other plants.

Abstract: Abscisic-acid (ABA) levels in roots of bean plants exposed to a—4 bar stress in the root medium increased ca. 10fold within 1 h and 16fold by the end of the 2nd h. Several types of experiments indicated that there is no transport requirement from the shoot for the increase to occur. ABA levels in roots from pea (Pisum sativum L.) and sunflower (Helianthus annuus L.) also increased in response to a—4 bar stress, although not as dramatically as in bean. When (S)-[2-14C]-ABA was fed to excised bean roots dihydrophaseic acid (DPA) was the major metabolite formed. The levels of endogenous DPA and phaseic acid increased markedly during a 27-h stress period. These results are consistent with a possible role for ABA in roots of water-stressed plants.

Cellulose microfibril orientation and cell shaping in developing guard cells of Allium: The role of microtubules and ion accumulation.

Abstract: The role of microtubules and ions in cell shaping was investigated in differentiating guard cells of Allium using light and electron microscopy and cytochemistry. Microtubules appear soon after cytokinesis in a discrete zone close to the plasmalemma adjacent to the common wall between guard cells. The microtubules fan out from this zone, which corresponds to the future pore site, towards the other sides of the cell. Soon new cellulose microfibrils are deposited on the wall adjacent to the microtubules and oriented parallel to them. As the wall thickens, the shape of the cell shifts from cylindrical to kidney-like. Studies with polarized light show that guard cells gradually assume a birefringence pattern during development characteristic of wall microfibrils radiating away from the pore site. Retardation increases from 10 A when cells just begin to take shape, to 80–100 A at maturity. Both microfibril and microtubule orientation remain constant during development. Observations on aberrant cells including those produced under the influence of drugs such as colchicine, which leads to loss of microtubules, abnormal wall thickenings and disruption of wall birefringence, further support the role of microtubules in cell shaping through their function in the localization of wall deposition and the orientation of cellulose microfibrils in the new wall layer. Potassium first appears in guard mother cells before division and rapidly accumulates afterwards during cell shaping, as judged by the cobaltinitrite reaction. Some chloride and perhaps organic acid anions also accumulate. Thus, these ions, which are known to play a role in the function of mature guard cells, also seem to be important in the early growth and shaping of these cells.

Mesurement of longitudinal ion profiles in single roots of Hordeum and Atriplex by use of flameless atomic absorption spectroscopy

Abstract: A method is described by which the Na+ and K+ content in 0.5 mm sections of single roots of Hordeum distichon L. and Atriplex hortensis L. can be determined by use of flameless atomic absorption spectroscopy. By this method the longitudinal profiles of K+ and Na+ along low salt roots and roots which had been equilibrated with or grown in K+-free 1 mM Na+-solution were determined. The profiles reveal that high K+/Na+ ratios in the cytoplasm are maintained also in K+-free solutions. In solutions containing 1 mM Na+ a high K+/Na+ selectivity was found to be dependent on sufficient aeration. From the ion profiles the cytoplasmic (110 mM) and vacuolar (20 mM) K+ concentration in low salt barley roots—values which are unobtainable by compartmental analysis—could be estimated.

The role of peridinin-chlorophyll a-proteins in the photosynthetic light adaption of the marine dinoflagellate, Glenodinium sp.

Abstract: The marine dinoflagellate, Glenodinium sp., was cultured at a series of light levels and growth, pigmentation, and photosynthetic rates were compared. Under decreasing light conditions, growth rates decreased, cellular chlorophyll a and peridinin content per cell increased, and maximum cellular photosynthetic rates remained unchanged. Pigmentation changes were related to alterations in cellular concentrations of a peridinin-chlorophyll a-protein and an unidentified chlorophyll a component of the chloroplast membrane. Maintenance of photosynthetic rates with decreased irradiance is interpreted as an increase in the number of pigment molecules in the light-harvesting antenna associated with the reaction centers of the photosynthetic apparatus, thus increasing the potential for photon capture for photosynthesis.

Nitrate influx and efflux by intact wheat seedlings: Effects of prior nitrate nutrition.

Abstract: Wheat (Triticum vulgare L., cv. Blueboy) seedlings, grown with 0.25, 1.0 and 15 mM nitrate in complete nutrient solutions, were transferred 10 days after germination to 1.0 mM K15NO3 (∼99 A% 15N) plus 0.1 mM CaSO4 at pH 6.0. The solutions were replaced periodically over a 6-h period (5 mW cm-2; 23°). Changes in the [15N]- and [14N]nitrate in the solution were determined by nitrate reductase and mass-spectrometric procedures and potassium by flame photometry. Influx of [15N]nitrate was depressed in plants grown at 1.0 mM nitrate relative to those grown at 0.25 mM, but there was no appreciably difference in [14N]nitrate efflux. Prior growth at 15 mM further restricted [15N]nitrate influx which, together with a substantial increase in [14N]nitrate efflux, resulted in no net nitrate uptake during the course of the experiment. Efflux of [14N]nitrate occurred to solutions containing no nitrate but it was significantly enhanced upon exposure to [15N]nitrate in the external solution. Influx of [15N]nitrate was more restricted at 5°, relative to 23°, than was [14N]nitrate efflux. The nitrate concentrations of the root tissue immediately before exposure to the K15NO3 solutions did not give a precise indication of the subsequent [15N]nitrate influx rates nor of the [14N]nitrate efflux rates. Net K+ uptake was related to the magnitude of the net nitrate uptake, not to the initial K+ concentration in the roots. The data are interpreted as indicating that [15N]nitrate influx and [14N]nitrate efflux are largely independent processes, subject to different controls, and that net nitrate uptake provides the driving force for net potassium uptake.

The de novo origin of the quiescent center regenerating root apices of Zea mays

Abstract: In roots from which the root cap and quiescent center have been removed new apical tissues regenerated in line with the main axis of the root. Regeneration of these tissues occurred from the region of the proximal meristem, which extends for no more than 350 μm from the cut surface. Accompanying the regeneration of new apical tissues is a change in the architecture of the root apex and initiation and enlargement of a new quiescent center. A possible “role” for the quiescent center in the establishment of pattern at the apex is considered. Regeneration of the original apex failed to occur in those roots from which the root cap, quiescent center and proximal meristem were excised.

Auxin binding to corn coleoptile membranes: Kinetics and specificity

Abstract: Detailed examination of binding over the range 10-7–10-6 M suggests that membrane preparations from coleoptiles of Zea mays L., cv Kelvedon 33 contain at least two sets of high affinity binding sites for 1-naphthylacetic acid (NAA), with dissociation constants of 1.8×10-7 M (site 1) and 14.5×10-7 M (site 2). Similar studies with 3-indolylacetic acid (IAA) also indicate two sets of binding sites, whose concentrations are closely comparable to those deduced for NAA. A substantial proportion of the total binding activity is retained in a detergent-solubilized preparation. Using [14C]NAA the interactions of a range of analogues with each of the binding sites have been examined with the aid of double reciprocal plots. The specificity of site 2 is compatible with that expected for an auxin receptor, in that only active auxins, antiauxin transport inhibitors are able to compete with [14C]NAA for the binding sites. Site 1 on the other hand is less specific, since it appears to bind all compounds tested, including physiologically inactive analogues.

Separation and localization of two classes of auxin binding sites in corn coleoptile membranes.

Abstract: Further evidence is presented for the discrete nature of the two classes of high affinity auxin binding sites in corn (Zea mays L.) coleoptile membranes, site 1 and site 2. Fractions can be obtained by differential centrifugation that exhibit binding kinetics characteristic of site 2, but not site 1. Membrane preparations containing both binding sites may be resolved on sucrose gradients into a light and a heavy band, whose binding kinetics and analogue binding specificities correspond to those deduced for site 1 and site 2 respectively in unfractionated membranes. Evidence from enzymic and chemical assays and from electron microscopy suggests that site 2, the auxin-specific binding site, is located in fractions enriched in plasma membrane, whereas site 1 is associated with Golgi membranes and/or endoplasmic reticulum.

Fusicoccin-induced growth and hydrogen ion excretion of Avena coleoptiles: Relation to auxin responses

Abstract: The fungal toxin fusicoccin (FC) induces both rapid cell elongation and H+-excretion in Avena coleoptiles. The rates for both responses are greater with FC than with optimal auxin, and in both cases the lag after addition of the hormone is less with FC. This provides additional support for the acid-growth theory. The FC responses resemble the auxin responses in that they are inhibited by a range of metabolic inhibitors, but the responses differ in three ways. First auxin, but not FC, requires continual protein synthesis for its action. The auxin-induced H+-excretion is inhibited by water stress or by low external pH, while the FC-induced H+-excretion is much less sensitive to either. It is concluded that auxin-induced and FC-induced H+-excretion may occur via different mechanisms.

Transmitting tissue in the pistil of tobacco: Light and electron microscopic observations.

Abstract: The pistil of tobacco (Nicotiana tabacum L. cv. Wisconsin 38) is comprised of two fused carpels. The stigma is bilobed, papillose, and at maturity is covered with a sticky exudate. The style is solid. Both stigma and style are made up of four tissue elements—epidermis, cortex, vascular, and transmitting tissue. Transmitting tissue in this species is chlorophyllous. Transmitting cells have thin primary walls and are separated by massive deposits of denselystaining amorphous material. The cells contain numerous mitochondria, dictyosomes, RER, amyloplasts, ribosomes, as well as crystal-containing microbodies and myelin-like formations. Observations are discussed in relation to other reports dealing with similar cell populations.

Suspensor, gibberellin and in vitro development of Phaseolus coccineus embryos.

Abstract: Embryos of Phaseolus coccineus in different stages of development (from 0.5 to 5 mm in length) were grown in vitro. Both intact embryos (with suspensor) and embryos deprived of suspensor were studied. It was found that removal of the suspensor has no effect on the development of embryos which have reached a length of 5 mm. With younger embryos, removal of the suspensor reduces embryo development, the negative effect being the greater the younger the embryo. It was shown that gibberellic acid (GA3) concentrations of 10(-8) to 10(-6)M can replace the suspensor in heart-shaped and early cotyledonary embryos (0.5 to 1.5 mm in length), whereas they reduce the development of suspensor-deprived embryos of later stages (embryos 2 to 3 mm in length) as compared with intact embryos of similar size grown on hormone-free medium. GA3 concentrations of 10(-5) and 10(-4)M are generally inhibitory and may stimulate callus formation in some embryos. The present data and those of Alpi et al. (1975) concur in ascribing a major role to gibberellins in characterizing the physiological function of the suspensor in early embryogenesis in Phaseolus coccineus.

Lipid composition of envelopes, prolamellar bodies and other plastid membranes in etiolated, green and greening wheat leaves.

Abstract: Comparative studies of lipid composition were made on prolamellar bodies, envelopes and other plastid membranes separately extracted from etiolated, green or greening (intermittent or continuous light) wheat (Triticum sativum L.) leaves. The different membrane fractions were examined by electron microscopy. The major lipid was digalactosyldiglyceride in the envelopes and prolamellar bodies and monogalactosyldiglyceride in stroma lamellae and grana. Phosphatidylcholine represented 60% of total phospholipids in the envelopes, 30% in prolamellar bodies and 14% in grana. All types of envelopes had the same lipid proportions. For all lipids the lowest fatty acid unsaturation was always found in the envelope membranes. The relative amount of {ie193-1} acid in the phosphatidylglycerol of envelopes increased from 4% (etioplasts) to an average of 15% (etiochloroplasts and chloroplasts).

Polyhedral bodies and ribulose 1,5-diphosphate carboxylase of the blue-green alga Anabaena cylindrica.

Abstract: The ribulose 1,5-diphosphate carboxylase (RUDP Case E.C. 4.1.1.39) activity of late log phase Anabaena cylindrica Lemm. was measured in vitro in fractions obtained by sucrose density gradient centrifugation. Two peaks of enzymic activity were obtained. One, accounting for about 80% of the total measurable activity occurred at the top of the gradient and appeared to be soluble activity; the second showed maximum activity in the 55% (w/w) sucrose fraction and represented 20% of the total activity. When the distribution of RUDP Case was assayed by immunoprecipitation using antiserum to RUDP Case from Euglena gracilis, the corresponding values were 59% and 41%. Electron microscopy of the various fractions showed that polyhedral bodies, which are sites of RUDP Case activity in other autotrophic prokaryotes, were also most abundant in the 55% (w/w) sucrose fraction.

Reversible inactivation of nitrate reductase in Chlorella vulgaris in vivo

Abstract: The NADH-nitrate oxidoreductase of Chlorella vulgaris has an inactive form which has previously been shown to be a cyanide complex of the reduced enzyme. This inactive enzyme can be reactivated by treatment with ferricyanide in vitro. In the present study, the activation state of the enzyme was determined after different prior in vivo programs involving environmental variations. Oxygen, nitrate, light and CO2 all affect the in vivo inactivation of the enzyme in an interdependent manner. In general, the inactivation is stimulated by O2 and inhibited by nitrate and CO2. Light may stimulate or inhibit, depending on conditions. Thus, the effects of CO2 and nitrate (inhibition of reversible inactivation) are clearly manifested only in the light. In contrast, light stimulates the inactivation in the presence of oxygen and the absence of CO2 and nitrate. Since the inactivation of the enzyme requires HCN and NADH, and it is improbable that O2 stimulates NADH formation, it is reasonable to conclude that HCN is formed as the result of an oxidation reaction (which is stimulated by light). The formation of HCN is probably stimulated by Mn(2+), since the formation of reversibly-inactivated enzyme is impaired in Mn(2+)-deficient cells. The prevention of enzyme inactivation by nitrate in vivo is in keeping with previous in vitro results showing that nitrate prevents inactivation by maintaining the enzyme in the oxidized form. A stimulation of nitrate uptake by CO2 and light could account for the effect of CO2 (prevention of inactivation) which is seen mainly in the presence of nitrate and light. Ammonia added in the presence of nitrate has the same effect on the enzyme as removing nitrate (promotion of reversible inactivation). Ammonia added in the absence of nitrate has little extra effect. It is therefore likely that ammonia acts by preventing nitrate uptake. The uncoupler, carbonylcyanide-m-chloro-phenylhydrazone, causes enzyme inactivation because it acts as a good HCN precursor, particularly in the light. Nitrite, arsenate and dinitrophenol cause an enzyme inactivation which can not be reversed by ferricyanide in crude extracts. This suggests that there are at least two different ways in which the enzyme can be inactivated rather rapidly in vivo.

Source, sink and hormonal control of translocation in wheat

Abstract: An analysis of the pattern of movement of 14C-labelled flag leaf assimilates in wheat (Triticum aestivum l. c.v. Gabo) during grain development, indicated that the greater the requirement for assimilates by the ear the more rapid was the speed of movement of these through the peduncle to the ear and also the lower their concentration. Experiments with [14C] indoleacetic acid ([14C]IAA) suggested that auxin production by the grains was not responsible for the control of assimilate translocation through the peduncle. Limiting the supply of available assimilates by shading the lower parts of the plant, did not significantly alter the speed of movement of 14C-photosynthate through the peduncle, while severing half of the vascular tissue in the peduncle altered the pattern of movement of 14C to the ear and enhanced the speed of movement of 14C through the remaining functional conducting tissue. These results are discussed in relation to the mechanism of translocation.

An enzyme to degrade lettuce endosperm cell walls. Appearance of a mannanase following phytochrome- and gibberellin-induced germination

Abstract: Lettuce seeds (Lactuca sativa L. cv. Grand Rapids) stimulated to germinate by gibberellin and red light produce large amounts of endo-β-mannanase. This enzyme increases markedly following radicle emergence and is capable of degrading mannose-containing polysaccharides, which are the major components of the endosperm cell wall. Non-germinated seeds contain little enzyme and under conditions where gibberellin- or red light-stimulated germination is prevented (eg. by abscisic acid or prolonged far red light) enzyme levels remain low. Cycloheximide inhibits the increase in enzyme levels when supplied to germinating seeds, but the enzyme once produced is stable in vivo in the presence of this inhibitor for at least 24h. The majority of the extractable mannanase activity is located in the endosperm and we propose that the function of this enzyme is to mobilise the endosperm cell wall polysaccharides as a nutrient source for the growing embryo.

A structural glycoprotein, containing hydroxyproline, isolated from the cell wall of Chlamydomonas reinhardii.

Abstract: A soluble extract from purified cell walls of C. reinhardii has been separated by gel filtration into three fractions which together account for 94% of the cell wall. The major fraction (accounting for 70% of the extract) is a glycoprotein, with a molecular wt. in sodium perchlorate of 298,000, which can be split into 4 electrophoretically distinct species. It contains 35% protein with high levels of hydroxyproline, arabinose and galactose, and is capable of self assembly into crystalline structures identical to those found within the cell wall. The second fraction (25% of the extract) is a similar glycoprotein, but contains 24% protein, a higher proportion of mannose, and is incapable of self assembly. The third fraction (3–6% of the extract) is shown to be an adsorbed impurity from the growth medium used.

Cell-wall formation in Pelvetia embryos. A freeze-fracture study.

Abstract: The cell-wall formation in the egg of Pelvetia fastigiata (J.G. Agardh) DeToni (Fucaceae) was studied with freeze-fracture. 1. The wall is lamellated with microfibrils approximately parallel in each lamella. The average orientation of microfibrils turns about 35° in each subsequent lamella. This slow turn gives rise to bow-shaped arcs when the wall is obliquely cross fractured. 2. The organization of the fibrils in the innermost lamellae is visualized by their imprints on the plasma membrane. These imprints are the result of both turgor pressure and adhesion of fibrils to the membrane. 3. Strings of membrane particles appear on the plasma membrane shortly after fertilization. They seem to be formed by a fertilization-induced aggregation of isolated membrane particles. Later each string comes to lie under a fibril and along its imprint. Peculiar lateral rips indicate that some strings are tightly bound to a fibril and may be involved in its orientation. 4. Wall formation in Pelvetia is marked by pronounced secretory activities. Following fertilization, the fusion of cortical vesicles and other vesicles make numerous loci in the plasma membrane. In older embryos, fibril-free patches in the plasma membrane mark the position of microfibril elongation centers in the wall matrix. Prior to germination, these elongation centers and their corresponding membrane patches reach a high density at the presumptive rhizoid end.