{gamma}-Amiobutyric Acid Metabolism in Plants: Part 1. Metabolism in Yeasts

The metabolism of -aminobutyric acid (AB) by two yeasts, Saccharomycescerevisiae and Torulopsis utilis, was investigated. Both yeastsgrew well upon AB as a sole source of nitrogen (N), and thelag phase for Torulopsis was shorter than when provided the N-source. The metabolism of AB by Torulopsis, whichwas associated with an increased O₂ uptake, was adaptive incharacter. The enzyme whose formation was induced by the supplyof AB was a transaminase, which was apparently specific forAB as the amino donor. Small amounts of transaminase were presentin unadapted, -grown cells. The optimum pH, equilibrium constant, Michaelis' constant, and coenzyme requirementwere investigated for the transamination reaction involving-ketoglutaric acid (KG) as amino group acceptor. Succinic semi-aldehyde(SSA) was a product of this transamination reaction.The possibility;that some AB was converted into SSA by a direct oxidative deaminationremained unconfirmed. The further conversion of SSA into succinic acid was establishedusing intact. cells for both yeasts. This oxidation processwas shown to be linked to the reduction of pyridine nucleotidesvising extracts of Saccharomyces as a source of SSA dehydrogenase.Dehydrogenase activity could be ascribed to two separate enzymes,one linked to DPN, and the other utilizing TPN and requiringMg⁺⁺ as an activator. The properties of the former enzyme, whichwas more important quantitatively, were investigated and comparedwith those described in the literature for an aldehyde dehydrogenaseof baker's yeast and for SSA dehydro-genases of Pseudomonas.Torulopsis extracts could catalyse the reduction of SSA to -hydroxybutyricacid (OHB); the OHB dehydrogenase involved required TPNH asa coenzyme. Certain other properties of this enzyme are recorded. The possibility is discussed that AB and SSA act as intermediatesin a metabolic pathway that may form a by-pass of the KG-succinatestage of the tricarboxylic acid cycle.     

{gamma}-Aminobutyric Acid Metabolism in Plants: Part 2. Metabolism in Higher Plants

The metabolism of -aminobutyric acid (AB) has been studied inhigher plants, particularly in peas and peanuts. Transaminationappeared to form the first step in AB degradation although transaminaseactivities were very low. The relatively active AB transaminaseassociated with whole pea plants possessing nodulated rootsappears to reside almost entirely within the nodules. AB transaminationwas demonstrated conclusively in extracts of mitochondria fromcotyledons of peanut seedlings; pyruvic acid acted as a betteramino-group acceptor than -ketoglutaric acid (KG). AB transaminaseactivity present in the microsomal and soluble cytoplasmic fractionsof the cells was very low AB was not metabolized perceptibly by intact mitochondria frompeanut, but when various organic acids were supplied simultaneously,an extra uptake of oxygen occurred and was associated with ABdisappearance. Aspartate, alanine, and ammonia were formed usingthe nitrogen atom of AB. The metabolic pathway followed by the carbon skeleton of ABwas traced by supplying C¹⁴-labelled material to leaf discsof peas and to mitochondria from peanut cotyledons. Radioactivitywas incorporated into organic acids, amino-acids, and respiratorycarbon dioxide in a manner suggesting that AB was convertedinto succinate which was then metabolized by the enzymes ofthe Krebs cycle present in the plant mitochondria. Glutamic decarboxylase was shown to be present largely in thenon-particulate (soluble) cytoplasm of cells. The enzymes responsiblefor AB synthesis and degradation, glutamic decarboxylase, andAB transaminase, respectively, therefore largely reside in differentsub-cellular fractions.     

Genotypic Differences in Leaf Water Relations between Brassica juncea and B. napus

Various kinds of measurement of tissue water status were madeseveral times during water stress and recovery in Brassica juncea(cv Canadian Black) and B napus (cv Drakkar) Unstressed plantsof the two species had similar leaf water potentials (_w), solute(_s) and turgor potentials (_p) Values of relative water content(RWC) and the slope of the linear relationship between _p andRWC (_p/RWC) were greater in B napus than in B juncea Statistical correlations of pooled data for the watered andstressed treatments differentiated the relationships among RWC,_w and its components in the two species The major statisticaldifference was that _p/RWC was related to RWC in B napus andto _w and _s in B juncea A decline in _p/RWC with decreasing _sin B juncea may be a mechanism for maintaining _p at low soilwater potentials through maintenance of more elastic cell walls. Brassica juncea, Brassica napus, osmotic adjustment, tissue elasticity, water relations     

Abscisic Acid and Water Relations of Rice (Oryza sativa L.): Sequential Responses to Water Stress in the Leaf

Water stress was imposed by withholding water at an early vegetativestage from plants of two rice cultivars (IR20 and 63–83)grown in pots. As stress intensified the following sequenceof responses of the leaves was observed: (i) rise in abscisicacid (ABA) content, (ii) closure of stomata, (iii) initiationof leaf rolling. In both cultivars, turgor (_p) declined linearly with total waterpotential () of the leaf. Bulk leaf ABA content increased linearlyas _p declined, and attained twice the control (unstressed) levelfollowing a reduction in _p of about 0.12 MPa. Stomatal conductance exhibited a sigmoidal relationship to _p,declining abruptly when a particular ‘critical’_p was reached (threshold response). The critical potentialsvaried considerably between experiments, but were closely correlatedwith control potentials and with the potentials at which ABAconcentration doubled relative to controls. Leaf rolling was initiated at s near to zero _p. Increases inthe ratio of adaxial to abaxial conductance were associatedwith rolling. Variations in the above responses could be accounted for byvariations in the rate of stress development, which in termsof reduction ranged from 0.38 to 0.86 MPa day^–1. Fastdrying rates resulted in: (a) reduced osmotic adjustment, (b)increased amounts of ABA in the leaf at a given level of or_p, (c) an increase in the ABA concentration present at 50 percent stomatal closure, and (d) initiation of leaf rolling ata higher . Oryza sativa L., rice, water stress, stomata, leaf rolling, abscisic acid     

Activity of Wall-bound Enzymes in Callus Cultures of Gossypium hirsutum L. During Growth

Activities of - and ß-glucosidase, - and ß-galactosidase,-mannosidase, ß-1,3-glucanase, acid and neutral invertaseswere detected in the cytoplasmic fraction as well as in cellwalls isolated from callus cultures of cotton. Activity of ß-mannosidase,however, could not be detected in the cell walls. Transfer ofcallus to a fresh medium did not immediately influence the activitiesof -glucosidase and ß-galactosidase but increasedsignificantly ß-glucosidase, -mannosidase, acid andneutral invertases. Addition of cycloheximide (1 and 100 mgl^–1) further stimulated acid and neutral invertases butnot other enzymes tested. Sodium chloride (NaCl) was effectivein extracting a-glucosidase, ß-glucosidase, ß-galactosidase,acid and neutral invertases. EDTA extracted most of the -galactosidase,-mannosidase, ß-1,3-glucanase and some -glucosidase.But, NaCl and EDTA could not extract some of the - and ß-glucosidasesand also acid and neutral invertases as evidenced from the residualand extra cellular activity. Studies with whole cells as a sourceof enzyme revealed that some of these enzymes were associatedwith the cell surface. Callus, glycosidases, glucanase, growth, Gossypium hirsutum     

Wilting of Leaves in Vicia faba L.

Wilting of leaves of Vicia faba L., which occurs when the pressurepotential (_p) is zero, and the leaf-water potential () at wiltingboth depend entirely upon the solute potential at incipientplasmolysis (_so) and not on soil-water status. Wilting in V.faba is acropetal; this is consistent with the hypothesis thatthere is a gradient of decreasing _so up the plant and that wateris transferred from the lower to the upper leaves, hasteningthe overall water loss from the lower leaves to the point when_p is zero. The gradient in _so up the plant is of the order of3–8 bar. It is proposed that wilting when _p>0 (i.e. > _so) shouldbe ‘apparent wilting’ and that when _p0 (i.e. _so),‘true wilting’.     

Seasonal Distribution of Carbohydrates in Nodules and Stem Exudate from Field-grown Soya Bean Plants

The concentration of carbohydrates in tap root nodules fromfield-grown soya bean [Glycine max (L.) Merr.] plants was verysimilar to the concentration of compounds previously reportedin greenhouse-grown nodules during vegetative growth of seedlings.The concentration of D-pinitol, sucrose and starch in nodulesdeclined during rapid fruit growth, but the concentration ofother compounds did not decline. The availability of carbohydratein nodules during fruit growth did not seem likely to be thecause of the decline in nitrogen-fixing activity of noduleswhich has been reported by others. All compounds except glucoseand , -trehalose declined to concentrations near zero duringa 10-day period of nodule decay. However, the decline in carbohydratedid not appear to cause nodule senescence because it did notprecede the period of decay and because decayed nodules containedsubstantial quantities of glucose and , -trehalose. Seasonalmean concentrations (72 samples from 24 dates) of compounds,in mg carbohydrate per g f. wt of nodule, were: sucrose, 2.84;D-pinitol, 1.14; D-chiro-inositol, 1.27; glucose, 1.40; , -trehalose,1.34; myo-inositol, 0.65; maltose, 0.31; and fructose, 0.21. Quantities of sugars and cyclitols in stem exudate collectedin the field on 13 dates were small (< 10 percent) relativeto the quantity of nitrogenous compounds transported from rootsto shoots. The seasonal pattern of pinitol transport in thexylem was very similar to the seasonal pinitol concentrationin nodules. A large increase in sugar concentration in stemexudate subsequent to 80 days after planting supports the viewthat lack of carbohydrate was not a cause of nodule senescence. Glycine max (L.) Merr, soya bean, cyclitols, , -trehalose, starch, D-pinitol, carbohydrates, root nodules, senescence     

Water-Relations Parameters of the Phloem. Determinations of Volumetric Elastic Modulus and Membrane Conductivity using an Applied Force Method and Shrinkage and Swelling of Tissues in Solutions at Differing Osmotic Pressure

Water-relations parameters were measured on sections of secondaryphloem from red oak (Quercus borealis michx. f.) and white ash(Fraxinus americana var. biltmoreana [Beadle] J. Wright) usinga linear displacement transducer. Changes in tissue thicknessin response to changes in the osmotic pressure of the bathingsolution were used to calculate the volumetric elastic modulusplus osmotic pressure (_v + ) of the tissue, and an applied forcemethod was used to estimate the time constant for water equilibration(T). The hydraulic conductivity of the cell membranes (L_p) wascalculated utilizing _v + and r values. The time-dependent behaviour of the tissue was much more complexthan originally expected. A correction for a time-dependentprocess that we call ‘drift’ was required to obtainnumbers for _v + . Furthermore, _v + was calculated on two assumptionsin order to relate changes in tissue dimensions to sieve elementparameters. In the first case, a lower limit for _v + of thesieve elements was determined by attributing all changes intissue dimensions to these cells. For red oak the average _v+ on this assumption is 72 bars. Assuming that all cell typeswere equally responsible for the changes in tissue dimensionsresulted in an _v + value of 192 bars for oak. If _v + and rare the same for all cells in the tissue, L_p for the sieve elementsof oak is 9.6 x 10^–8 cm s^–1 bar^–1. Exudationfrom the sieve elements of white ash during excision of thephloem led to artificially high values of _v + for that species. Quercus borealis michx. f., Fraxinus americana var, biltmoreana (Beadle) J. Wright, red oak, white ash, water relations, phloem, volumetric elastic modulus, membrane hydraulic conductivity     

The Nitrogen Metabolism of Groundnut Plants: the Role of {gamma}-methyleneglutamine and {gamma}-methyleneglutamic Acid

The changes occurring in the nitrogenous compounds during thegrowth of groundnut seedlings in the dark and light were compared,particular attention being centred on the levels of -methyleneglutamine,the principal amide of these plants, and -methyleneglutamicacid. The distribution of amino acids and amides in the mainorgans of normal young and mature plants was also examined.Suggestions are made concerning the possible pathways of synthesisand the functions of -methyleneglutamic acid and -methyleneglutaminein groundnut plants.     

Osmotic Properties of Drought Stressed Periwinkle (Catharanthus roseus) Genotypes

The pressure-volume technique was employed to compare waterrelations and moisture stress-induced osmotic adjustment ofPeriwinkle (Catharanthus roseus) cv. Pink (PC), Oscillatus (REC)and White (WC). Leaf water potential (_w), osmotic potential(_s), turgor potential (_p), bulk modulus of elasticity (), boundwater (RWC_w) and leaf hydration (H), were estimated by exposingthe plants to a drying cycle during which well watered plantswere dehydrated to zero turgor, and then irrigated. Osmoticadjustment (_w ¹⁰⁰) was calculated by comparing _a at full hydration(_a ¹⁰⁰) in stressed plants after recovery, with _a ¹⁰⁰ in controlplants. Values of ₂¹⁰⁰ were 0.76, 0.33 and 0.11 MPa in cv. PC,REC and WC, respectively. Maintenance of _p at lower ₃ and relativeleaf water content (RWC) in prestressed PC was attributableto a higher alkaloid content and greater leaf cell wall elasticity.RWCW was plotted against _p to determine its contribution tohydration maintenance at lower _p. Genotype PC showed greaterRWC_w at lower _p compared with REC and WC. The present studyhas demonstrated that there are cultivar differences in alkaloidaccumulation and water relations in acclimated plants and thatthe relative ranking for drought resistance within periwinkleappeared to correspond with the changes in osmotic properties. Medicinal plant, drought resistance, alkaloids, periwinkle [Catharanthus roseus (L.) G. Don]     

Occurrence of Multiple Forms of {alpha}-Amylase and Absence of Starch Phosphorylase in Soya Bean Seeds

Soya bean cultivars ‘Altona’ and ‘Chestnut’have active but quite low levels of -amylase. Activity was assayedwith specific substrates, oxidized amylose and ß-limitdextrin, which were resistant to attack by ß-amylase.During seed development -amylase activity increased to a maximumin both cultivars and then declined towards maturity. Matureand germinating seeds retain low activities of -amylase. Gelelectrophoresis separated the -amylase activity into six majorbands which occurred in both cultivars. The isozyme patternwas quite similar for developing, mature and germinating seed.although the relative proportion of activity in the variousbands changed somewhat. Starch phosphorylase was not detectedin any soya bean seed samples tested, but good activity wasfound in potato tuber extracts used as a control. Mixing experimentsusing soya bean and potato extracts indicated there were noinhibiting factors in soya bean seed extracts. Soya bean seedextracts probably do not contain starch phosphorylase. Glycine max (L.), Merr, soya bean, -amylase, isozymes, phosphorylase     

Natural Abundance of {delta}15N Confirms Insectivorous Habit ofRoridula gorgonias, Despite it Having No Proteolytic Enzymes

Natural abundance values of plant ¹⁵N give an indication asto the source of nitrogen. In particular, carnivorous plantsare expected to be relatively enriched due to trophic enrichmentof their prey. Values of ¹⁵N for adultRoridula gorgonias(mean+3.02)are 4–9 greater than co-occurring non-carnivorous plantspecies and 5.24 greater than juvenileR. gorgoniasplants. Theyare also 3.5–4.26 greater than co-occurringDroseraspecieswhich, being sundews, are considered to be carnivorous. Thesehigh levels of ¹⁵N in adult plants are best explained as beingdue to access to trophically enriched N from insects. As isthe case for other carnivorous plants, leaves and stems ofR.gorgoniasare highly ultraviolet reflective and are thereforeprobably attractive to potential insect prey. This is furthersupport for this plant species being insectivorous.Copyright1998 Annals of Botany Company Nitrogen isotopes, carnivorous plants, insectivorous plant, ultraviolet,Roridula gorgoniasL.     

Water Relations of Potato Leaves. II. Pressure--Volume Analysis and Inferences About the Constancy of the Apoplastic Fraction

Water relations characteristics of potato (Solanum tuberosumL. cv. Bintje) leaves were determined from pressure—volumeanalysis using a pressure chamber. Turgor was 077 MPa and thebulk volumetric modulus of elasticity 81 MPa at full turgidity;turgor loss occurred when water potential () had declined to–087 MPa at a relative water content (RWC) of 0912;the apoplastic water fraction (A) was 0235. As is usually found,there was a linear relation between 1/ and RWC beyond turgorloss. This finding supports the assumptions of the constancyof A during leaf dehydration. Beyond turgor loss the difference between and [measured afterfreezing and thawing (_d)] was about 01 MPa. This differencedid not increase as the leaf water content decreased. This resultcontradicts the constancy of A. It was concluded from calculations with a simple model of leafdehydration that analysis of the relation between and _d providesmore insight in the changes in the apoplastic fraction thanthe relation between 1/ and RWC. Research on the size of theapoplastic fraction and its changes with water potential wouldcomplement current understanding of leaf water relations. Solanum tuberosum, L., water potential, pressure chamber, osmotic potential, pressure potential, relative water content, apoplast, symplast     

Carotenoid Pigment Changes in Ripening Momordica charantia Fruits

The carotenoid composition of Momordica charantia fruit (pericarp)at four levels of maturity was extensively investigated. Thenumber of carotenoids isolated increased from five in the immaturefruit to six at the mature-green and 14 at the partly-ripe andripe stages. Cryptoxanthin, which could not be isolated at theimmature and mature stages, accumulated rapidly at the onsetof ripening to become the principal pigment of the ripe fruit.Moderate increases were seen in ß-carotene, zeaxanthinand lycopene concentrations as ripening progressed. The reversetrend was observed with lutein and -carotene which were themajor pigments of the immature fruit. Prior to the colour break,only the hydroxy derivatives of -carotene (zeinoxanthin andlutein) could be detected; the ß-hydroxy compounds(cryptoxanthin and zeaxanthin) appeared and predominated thereafter.The hydroxy carotenoids of the ripe fruit were almost entirelyesterified in contrast to those of the unripe fruit which weremainly unesterified. Traces of flavochrome, 5, 6-monoepoxy-ß-carotene,mutatochrome, -carotene, -carotene, -carotene and rubixanthinwere detected in the partly-ripe and ripe fruits but not inthe immature and mature-green samples. Phytofluene was observedin trace levels at all stages.     

Starch Deposition and Carbohydrase Activities in Developing and Germinating Soya Bean Seeds

Immature soya bean seeds accumulate starch as a transient reservematerial which is utilized later in development. Germinatingseeds also accumulate starch reserves, probably as a resultof gluconeogenesis from storage lipid. Developing beans showa rapid increase in ß-amylase activity which continuesinto early germination before declining. Distribution of ß-amylaseactivity is not consistent with its supposed role in starchdegradation. Soya bean seeds also contain -amylase and -glucosidaseactivities which could be responsible for starch mobilization. Glycine max (L.) Merr., soya bean, starch, carbohydrase, amylase, -glucosidase     

Effects of {alpha}-Hydroxysulphonate on Respiration of Mung Bean Root Tips

Hydroxysulphonate (-HPMS) was found to have an inhibitory effecton dark respiration of mung bean root tips. Inhibition was greatestin regions nearest the apex. The inhibition of root respirationby -HPMS was not due to an inhibition of glycolic acid oxidase.It is suggested that the action of -HPMS was probably on acetylCOA synthetase.     

A Transient Stimulation of Net Photosynthesis of Rye Leaves by {alpha}-Hydroxy-2-pyridinemethanesulphonic Acid ({alpha}-HPMS) due to Inhibition of Photorespiratory CO2 Release

The effects of -hydroxy-2-pyridinemethanesulphonic acid (-HPMS)upon net photosynthesis (P_n, the CO₂ compensation point (),post-lower illumination burst of CO₂ (PLIB) and post-lower temperatureburst of CO₂ (PLTB) in detached rye (Secale cereale L.) leaveswere investigated. At low concentrations ( 0.5 mol m^–3),-HPMS initially stimulated P_n and decreased the magnitude ofboth PLIB and PLTB. The decreased at all concentrations of-HPMS (0.05–5.0 mol m^–3. The effects of -HPMS onP_n and were time-dependent and, after a few minutes, the P_nwas inhibited while values increased considerably. At a higherconcentration (5.0 mol m ^–3), the transient effects of-HPMS were shorter () or not observed at all (P_n. Both PLIBand PLTB, when expressed in relation to P_n, increased at higherlevels of this compound. Similar data with respect to the effectsof -HPMS on PLIB and PLTB were found for leaves of dandelion(Taraxacum officinale L.). The results suggest that -HPMS may stimulate P_n by inhibitingphotorespiration, as originally suggested by Zelitch (1966),but only at low concentrations and over a short time span. Thedecrease of PLIB and PLTB values at low -HPMS levels is consistentwith these processes being a residual activity of the glycolatepathway. Key words: CO₂ compensation point, -hydroxy-2-pyridinemethanesulphonic acid, photorespiration, photosynthesis     

Dry Matter Distribution between the Shoot and Storage Root of Carrot (Daucus carota L.): I. Comparison of Varieties

The distribution of dry matter between shoot and storage rootof a wild variety and cultivars of carrot was investigated ina glasshouse experiment and a field experiment. In both experimentsthe deliberate introduction of variation in plant size revealeda linear relationship between the log weights of these partsat any time throughout growth. In the glasshouse experiment,in which growth was examined from approximately 10–600mg mean dry wt per plant, the slope of the relationship betweenIn shoot and In storage root dry wts decreased markedly duringdevelopment to 100 mg. Subsequently the slope became reasonablyconstant and the intercept progressively more negative. Datafrom the field experiment covered a similar timescale (48–115days) to the glasshouse experiment but the plants grew morequickly and were much heavier (180 mg–6 g dry wt). Therelationship between shoot and storage root for these data wassimilar to that for the plants of comparable size in the glasshouseexperiment. Over the range of development for which the slopeof the relationship was constant, data from both experimentswere described fairly well by the following equations: In shoot = – time+ In storage root where , and are constants. Between cultivars the slope ()and time constant () were similar, but the intercept () increasedwith maturity time of the cultivars. For wild carrot the slopewas larger and the time constant smaller than estimates forthe cultivars. These results on dry matter distribution arediscussed in relation to a model for partitioning assimilatein an individual plant. Daucus carota L., carrot, assimilate distribution, dry matter distribution, storage root, partitioning model