{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.     

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     

The Incorporation of C14-labelled Substrates into the Amino-acids of Groundnut Plants (Arachis hypogaea)

The amino-acid metabolism of groundnut plants has been studiedwith special reference to -methyleneglutamic acid (-MGA) andy-methyleneglutamine (-MG), constituents not found in the greatmajority of plant species. The sequence in which C¹⁴ from radioactive C¹⁴-carbon dioxideenters the amino-acids of leaves was determined. The patternof labelling was very similar to that found for leaves of otherspecies. The metabolic relationships existing between photosynthesisand amino-acid synthesis therefore do not seem to be affectedby the large quantities of -MGA and -MG present in the leaves.-MGA and -MG only gained traces of radioactivity. Experiments designed to study the incorporation of C¹⁴ fromuniformly labelled C¹⁴-alanine into the amino-acids of roots,immature leaves and cotyledons of seedlings and young plantsindicated that the main site of synthesis of -MGA was the cotyledons. Various C^I4-labelled substrates were fed to germinating seedsand, after a period of growth, the specific activities of theamino-acids of seedlings receiving different treatments weredetermined. Comparison of the specific activities enabled certaindeductions to be made concerning the probable biosynthetic pathwaysleading to -MGA and -MG. The results were consistent with theintact incorporation of pyruvate molecules, or another related3-carbon-atom containing compound, into -MGA.     

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     

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     

Morphogenetic Responses of Browallia Leaf Sections and Callus

Leaf sections of Browallia viscosa and B. speciosa were placedon Murashige and Skoog (1962) salts and vitamins medium (MS)containing auxins and cytokinins, singly or in combination,to elicit morphogenetic responses. B. viscosa developed extensiveroots in 4 weeks on media supplemented with indolebutyric acid(IBA), indol-3-yl acetic acid (IAA) or naphthalene acetic acid(NAA) (0·01, 0·1, 1·0, 5·0 and 10·0mg^–1), but with 2, 4-D (0·1 mg^–1) only lightyellow friable callus was obtained. Shoot initiation and elongationoccurred consistently in 4–6 weeks on leaf sections inthe presence of 6---dimethylallyl amino purine (2iP). Similarly,shoot regeneration from leaf-derived callus, initiated and sub-culturedon MS + benzyladenine (BA) + NAA only induced callus on leafexplants of both species. B. speciosa did not respond exceptfor moderate and prolific callus formation on MS + BA + NAAand Uchimiya and Murashige (1974) media respectively. Browallia viscosa, Browallia speciosa, tissue culture, regeneration, morphogenetic potential     

Water Relations of Sitka Spruce Seedlings After Root Damage

Sitka spruce[Picea sitchensis(Bong.) Carr] seedlings were subjectedto varying degrees of root damage in a growth room, rangingfrom careful transplanting to exposure of the root system toair for up to 3 h. After replanting, transpiration (E), leafwater potential (₁) and growth of the shoot and root were measuredand observations made on plant survival. Some plants in the root exposure treatments died 20–85days after planting. In plants which eventually died, E wasdepressed directly after treatment, but ₁ showed a variableresponse. In some plants ₁ decreased from —8·0x 10⁵ to —30 x 10⁵ Pa after only 10 days but in othersthere was little change in ₁ for 50 days. In spite of the maintenanceof a high water potential in some of the latter plants for longperiods, no root or shoot growth occurred. In plants which lived, the root damage reduced root and shootgrowth relative to untreated controls, and most treatments stronglydepressed E but had little or no effect on ₁. The changes of E and ₁ in treated plants suggest that the suppressionof E was often independent of ₁ although water stress eventuallydeveloped in some of the severely treated plants. Sitka spruce, Picea sitchensis (Bong.)Carr, water relations, root damage, transpiration, leaf water potential     

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.     

Vegetable Plant Part Relationships. II. A Quantitative Hypothesis for Shoot/Storage Root Development

A simple quantitative formulation of the concept of the controlof partitioning of assimilated carbon by the behaviour of plantcomponents as competing sinks is developed. An equation, In s = + In r—t, relating shoot (s) and storage root (r) d. wts, and the lengthof growth period (t), is constructed by considering possiblefates of imported assimilates into different plant parts. Thevalues of the equations' parameters depend on the relative sinkactivities of the plant parts, tissue respiration rates andinitial weights of plant components. The equation closely fitteddata collected from a number of carrot and beet experimentsin which planting density had been varied. Estimates of shootand storage root maintenance respiration rates, derived fromthe parameter , were of the correct order of magnitude. Othersets of experimental data are also discussed in the light ofpredictions of the theory and possible uses and extensions ofthis approach to assimilate partitioning are briefly discussed. Daucus carota L., Beta vulgaris, carrot, red beet, partition of assimilated carbon, maintenance respiration, storage root     

Nitrogenous Compounds and Nitrogen Metabolism in the Liliaceae: II. The Nitrogenous Compounds of Leaves of the Genus Tulipa: Environmental Effects on the Composition of Tulipa gesneriana

Because of the predominance of -methyleneglutamine in Tulipagesneriana a quantitative, chromatographic investigation hasbeen made to show the factors which determine the compositionof the alcohol-soluble nitrogen fraction in Tulipa A comparative study of the amino-acids that occur free in theleaves of fourteen different species of Tulipa has been made,-methyleneglutamine varied from as much as 60–70 per cent,of the amino-acid nitrogen down to levels below those whichpermit its chromatographic detection. The effects of temperature during growth and of diurnal variationon the nitrogenous compounds of the leaves of T. getnerianaare described. Excised shoots of T. gesneriana were exposed to light and darknessto affect the balance of protein synthesis and breakdown sothat the concomitant effects on the -methyleneglutamine acidcompounds and other main components of the soluble nitrogenfraction could be determined. Lastly, the relative ease of entry of C¹⁴ from C¹⁴O₂ into thesugars and certain other compounds of T. gesneriana is contrastedwith the relative difficulty with which it enters the -methyleneglutamineof the mature leaves on excised shoots. The occurrence and metabolism of -methyleneglutamine in T. gesnerianaleaves may, therefore, be regarded as additional to and superimposedupon an otherwise typical pattern of nitrogen metabolism: inthese leaves -methylene-glutamine does not assume the role usuallydischarged by glutamine.     

{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.     

Isolation of Subunits of Coupling Factor 1 from Maize and Spinach Chloroplasts and Properties of Combinations of Subunits with ATPase Activity

Subunits (, ß, ) and mixtures of subunits ( ß, , ß , ß ) were isolated without denaturationfrom a chloroform extract of chloroplast coupling factor 1 (CF₁)from maize (Zea mays var. Ushiku 5-4) and from spinach by fastprotein liquid chromatography (FPLC), on an anion-exchange columnof Mono-Q in the presence of n-octylglucoside (OG) and on achromatofocusing column of Mono-P. The ß -subunitcomplex (CF¹ ß ) was the minimum unit required forATPase activity, as was confirmed by the reconstituted complexof ß and subunits. An subunit isolated from maizeinhibited the ATPase activity of CF₁ ß from bothmaize and spinach. CF₁ ß was found to contain anOG-dependent Mg²⁺-ATPase. The ATPase activity of CF₁ ß required divalent cations, such as Mg²⁺ or Mn²⁺, for its expressionin the presence of OG; its optimum pH was 8.0 and it was markedlyinhibited by NaN₃. The enzyme hydrolyzed ATP in prefernece toGTP but not CTP, UTP, ADP, AMP or pNPP. Lineweaver-Burk plotsof its activity were curvilinear in the range of 0.6–0.7mM ATP.Mg²⁺. ¹Present address: Department of Biology, School of Education,Waseda University, Shinjuku-ku, Tokyo, 160 Japan. (Received February 15, 1989; Accepted April 20, 1989)     

Pumpkin (Cucurbita sp.) seed globulin III. Comparison of subunit structures among seed globulins of various Cucurbita species and characterization of peptide components

Various Cucurbita seed globulins showed patterns similar toone another on SDS-gel electrophoresis, and ß bandsfor unreduced globulins and , ', and ' bands for reduced ones.On gel electrophoresis in 6 M urea, reduced globulin gave twoacidic and two basic bands. These corresponded to and ' chainsand ₁ and ₂ chains, respectively, identified by two-dimensionalurea-SDS gel electrophoresis. The compositions of the and ßsubunits were proposed. (Received September 8, 1977; )     

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’.     

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     

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     

The Role of Glutamate Decarboxylase and {gamma}-Aminobutyric Acid in Germinating Barley

The variations in glutamate decarboxylase activity and in glutamicacid and -ABA concentration have been measured in barley embryosduring the uptake of water and in the roots and shoots for upto 6 days of growth. Glutamate decarboxylase activity was relativelysteady in the embryos during soaking but rose rapidly once growthbegan. This development paralleled an increase in the concentrationof glutamic acid in both roots and shoots at a time when theconcentration of -ABA was falling. During soaking in aeratedwater, the -ABA content of the embryos rose for 36 h, at whichpoint it accounted for 35 per cent of the soluble amino acids.-ABA was found to be a major free amino acid in roots but notin shoots. Experiments in vivo involving ¹⁴C-labelled glutamicacid and -ABA indicated that carbon from -ABA passed very rapidlyinto the citric-acid cycle intermediates and also that, throughoutthe period studied, -ABA was formed from glutamic acid despitethe alterations in relative concentrations of these amino acidsin the growing tissue.     

Keto-acids in the Alga Chlorella

The intracellular concentration of pyruvic and -ketoglutaricacids in Chlorella when grown under various conditions has beeninvestigated. The keto-acids were assayed after quantitativeelution of their 2,4: dinitrophenylhydrazones, chromatographicallyseparated. Very small amounts of keto-acids are found in Chlorella whengrown auto-trophically. The level was much higher under heterotrophicconditions and in nitrogen starvation. Further, with nitratethe level was considerably higher than if ammonia was supplied.In the presence of glucose this differential effect occurredwith pyruvic acid, but the level of -ketoglutaric acid appearedto be stabilized. Possible interpretations of these facts arediscussed.     

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.