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     

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     

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     

Changes in {alpha} and {beta}-Amylase Activities during Germination of Seeds of Alfalfa (Medicago sativa L.)

We examined the methods available for the assay of -amylasein alfalfa (Medicago sativa) and found the Phadebas test mostsuitable. The Phadebas assay and activity staining on ampholinegels after isoelectrofocusing revealed that an amylase is presentin the dry seeds of alfalfa and that its activity decreasesrapidly after the second day of seed germination. An amylasewas purified by affinity chromatography and gel filtration.The kinds of sugar generated from soluble starch by the purifiedamylase resembled those generated by other -amylases from plants,in particular those from mung bean (Vigna radiata). These resultsindicate that the amylase in alfalfa seeds belongs to the familyof -amylases. The molecular weight and isoelectric point ofthe -amylase were determined to be 43 kDa and 4.92, respectively. The Pantrac assay and activity staining on immobiline gels afterisoelectrofocusing revealed that the activities of ß-amylasesincreased during the initial 4 to 5 days of germination. Furthermore,treatment of whole seedlings with cycloheximide or actinomycinD inhibited the increase in activity of ß-amylasesbut did not affect the reduction in activity of -amylase. During germination of alfalfa seeds, -amylase activity decreaseswhile, in contrast, ß-amylase activity increases (inthe cotyledons of germinating seeds), changes that are specificto the germinating seeds of alfalfa. (Received September 8, 1990; Accepted February 20, 1991)     

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.     

Amylase Activity and Loss of Viability in Wheat

Changes in total and -amylase synthesis were compared in twowheat samples aged up to 70% viability level. The results showedthat differences in amylase levels can be detectable betweenunaged and aged wheats and also between the two aged samples.Thus seed viability loss is characterized by decreases in amylaseproduction. However, these changes appear to affect the germinationrate, but not the final germinability. Trticum durum L. cv. Appulo, wheat, seed ageing, amylase, -amylase     

Changes in Carotenoid Composition of Certain Roses with Age

The carotenoid composition of the anthers and styles and thefruits of four roses of different ages have been investigated.ß-carotene and its epoxides were present at all stagesstudied, as were the ß-carotene derived xanthophyllauroxanthin and the -carotene derived flavoxanthin and chrysanthemaxanthin.As the various parts aged, the control of carotenoid synthesiswas removed, oxidative processes took place with the resultthat very few members of the -carotene series were found, andepoxy-carotenoids and their derivatives were the main carotenoidspresent. Rubixanthin, 3-hydroxy--carotene characteristic ofrose hips was found in fairly large amounts in anthers and stylesand -carotene was probably the precursor of this pigment. Rosehips were also investigated for their vitamin-A contents whichwere not as high as those of All Gold flowers.     

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’.     

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     

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     

Pumpkin (Cucurbita sp.) seed globulin V. Proteolytic activities involved in globulin degradation in ungerminated seeds

Two proteolytic activities I and II involved in the globulindegradation were detected in pumpkin seeds. Activity I, hydrolyzing and ß subunits of the globulin to form F_ß,was found in both dry seeds and cycloheximide-treated cotyledons,and decreased during germination. Activity II, hydrolyzing F_ßto produce small peptides and amino acids, was not observedin dry seeds but found in cycloheximide-treated cotyledons,increased up to 4 days, and gradually decreased during germination. Activity I gave limited hydrolytic products from the globulinand the chain, but not from F_ß, the chain and some animal proteins. It was inhibitedby EDTA. On the other hand, activity II hydrolyzed F_ßand the chain faster than the globulin, the chain and some animal proteins. It was inhibitedby EDTA and p-chloromer-curibenzoate, and activated by ß-mercaptoethanol,dithiothreitol and CoCl₂. Optimum pH's were at about 6.8 andat 6.0 to 6.8 for activities I and II, respectively. The degradation process of the globulin can be divided intotwo steps: the first step is the conversion of globulin to F_ßand the second step, F_ß to small peptides and aminoacids. (Received November 9, 1979; )     

{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.     

{beta}-Amylase Activity as an Index for Germination Potential in Rice

Seeds of different vigour also differ in their germination ability.In rice (Oryza sativa), this difference was correlated withthe level of incorporation of ³⁵S-methionine into 25-60% ammoniumsulphate precipitable material that was rich in amylase proteins.This protein fraction, from dry seeds, contained no -amylaseactivity. In contrast, ß-amylase activity was presentin all seed stocks capable of 99% germination, although thelevel was lower in seeds that grew slowly when germinated. Inlow viability low vigour stock (i.e. extensively deterioratedseeds) ß-amylase activity was absent. Alpha-amylaseactivity in all stocks was detected only after 24 h from thestart of imbibition. These results indicate that ß-amylaseactivity is reliable indicator of the germination ability ofrice seed stocks and of their vigour during germination.Copyright1995, 1999 Academic Press Rice (Oryza sativa L.,), germination, ß-amylase, -amylase, seed vigour     

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)     

Molecular dynamics simulations of oligosaccharides and their conformation in the crystal structure of lectin-carbohydrate complex: importance of the torsion angle {psi} for the orientation of {alpha}1,6-arm

The conformation of the heptasaccharide Man-1,6-(Man-1,3)(Xyl-ß1,2)-Man-ß,4-GlcNAc₂-ß1,4-(L-Fuc-1,3)-GlcNAc₁,the carbohydrate moiety of Erythrina corallodendron lectin (EcorL),the hexasaccharide Man-1,6-(Man-1,3) (GlcNAc-ß1,4)-Man-ß1,4-GlcNAc-ß1,4-GlcNAcand their disaccharide fragments have been studied by moleculardynamics (MD) simulations for 1000 ps with different initialconformations. In the isolated heptasaccharide, the most frequentlyaccessed conformation during MD has a value of 180° aroundMan-1,6-Man linkage. This conformation is stabilized by theformation of a hydrogen bond between the carbonyl oxygen ofGlcNAc₂ with the O3/O4 hydroxyls of the 1,6-linked mannose residue.The conformation of the heptasaccharide found in the crystalstructure of the EcorL-lactose complex (Shaanan et al., Science,254, 862, 1991), that has a value of 76° around Man-1,6-Manlinkage, is accessed, although less frequently, during MD ofthe isolated oligosaccharide. The ,, = 58°,–134°,–60°conformation around Man-₁,6-Man fragment observed in the crystalstructure of the Lathyrus ochnrs lectin complexed with a biantennaryoctasaccharide (Table I in Homans,S.W., Glycobiology, 3, 551,1993) has also been accessed in the present MD simulations.These values for the 1,6-linkage, which are observed in theprotein-carbohydrate crystal structures and are accessed inthe MD simulations, though occasionally, have not been predictedfrom NMR studies. Furthermore, these different values of leadto significantly different orientations of the 1,6-arm for thesame value of . This contrasts with the earlier predictionsthat only different values of can bring about significant changesin the orientation of the ₁,6-arm. The MD simulations also showthat the effects of bisecting GlcNAc or ß1,2-xyloseare very similar on the ₁,3-arm and slightly different on the₁,6-arm. bisecting GlcNAc carbohydrates glycoprotein lectinsaccharide complex     

Carotenoids of Rowan Berries

The berries of Sorbus aucuparia have been investigated for theircarotenoid contents. The pigments identified were: phytofluene,-, ß-carotene, cryptoxanthin, monoepoxy--carotene,monoepoxy-ß-carotene, aurochrome, and mutatochrome.Usually when green fruits ripen the control of carotenoid synthesisis removed when the chlorophylls disappear, there is a rapidincrease of carotenoids in an over-all oxidative manner anddifferent carotenoids appear. However, the results obtainedsuggest that a different mechanism takes place in S. aucuparia.This may be the exception that proves the rule.     

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     

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     

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.     

Competition for Light in a Plant Monoculture Modelled as a Spatial Stochastic Process

In a glasshouse experiment closely spaced plants were regularlysub-irrigated with nutrient solution and it was assumed thatcompetition between individuals was for light. Interaction betweenneighbours is modelled as a spatial process in relation to differencesin plant height. The influence one plant has on another is calculatedas the ratio of the angle ß between their apices,and an angle which represents the intensity of competitionin the population as a whole. When ß < suppressionis considered to occur, i.e. tall plants suppress short plantsbut not vice versa, competition is one-sided. The status ofeach plant within the population is calculated by multiplicationof its individual plant neighbour interaction terms where ß< . This need not be restricted to nearest neighbours. Plantswith status = 1 are free from competition, those with status= 0 are totally suppressed. It is acknowledged that other processes than that modelled ascompetition may result in variation in relative growth rates.To account for this plant status is used to modify a relativegrowth rate drawn from a population of mean µ and standarddeviation . Plants for which status µ is less than anestimated are considered to die. Parameter values were estimatedfor a sequence of fortnightly harvests of glasshouse grown Tagetespatula. There was a decrease in µ and , increased andplants came to compete with their second nearest neighboursas well as their nearest as the community developed. The greatest was at an intermediate stage, suggesting that more frequentassessments of the competition process are required at somestages than at others. Model parameters fitted to one data setwere used in a Monte Carlo testing procedure with a second,independent data set. The importance of this technique is stressedbecause plants in a single community are not independent realizationof the competition process. Simulations with the model reproducedboth bimodality in the frequency distributions of plant sizeand an even spatial distribution of large or surviving plants,features which have been observed in a range of plant monocultures.To achieve these features it was essential that plant interactionbe modelled as a one-sided process. Tagetes patula L., competition, light, monoculture, growth model