Sucrose and Hexose Release by Excised Stem Segments of Vicia faba L.: The Sucrose-Specific Stimulating Influence of Cuscuta on Sugar Release and the Activity of Acid Invertase

By washing out solutes in 0.5 mM CaSO⁴at 25 °C during aperiod of 5–6 h, the release of sugars by excised stemsegments of Vicia faba L. was measured. The stem parasite Cuscutaeuropaea strongly stimulated the release of sucrose into theefflux medium; this effect was most marked during the last hoursof each experiment but this stimulating effect of the parasitecould not be detected for glucose and fructose. The fact thatparasitized stem segments released higher than normal hexoseamounts during the last hours of several experiments, couldbe explained as the result of extracellular hydrolysis of sucroseby free space invertase. A high free space acid invertase activitywas present in young stem segments of Vicia faba and in tissuesof Cuscuta. The stimulating influence of Cuscuta on sugar releaseby cells of stem segments appears sucrose-specific, supportingthe idea that the stimulating influence of Cuscuta on sugarrelease is restricted to the sieve-tube system. When metabolic inhibitors were added to the washing solutionor when segments were incubated at low temperature, no cleareffect of the parasite could be observed and for all segments(parasitized and non-parasitized) a strongly enhanced releaseof sucrose into the efflux medium was found during the lasthours of an experiment. These data support the idea that anintensive resorption of sucrose occurs within stem segments,after its release into the apoplast. Key words: Cuscuta europaea, Parasitic relationship, Phloem unloading     

Characteristics of Nectar Secretion by the Extrafloral Nectaries of Ricinus communis

The characteristics of nectar secretion by excised extrafloralnectaries of Ricinus have been examined. Secreted nectar wasfound to contain three sugars: sucrose, glucose and fructose,with glucose and fructose occurring in a 1: 1 ratio. All threesugars supported secretion when used in the culture medium andthe yield of nectar sugar was found to be concentration-dependent.Other sugar sources failed to support secretion. Experimentsusing ¹⁴C-sugars and ¹⁴CO₂ fed to intact plants allowed themovement of sugars through the nectary to be examined. Sucrosesynthesis occurs when excised glands are fed glucose and thisoccurs very early in the transport through the nectary. Themain sugar transported was sucrose, with little hydrolysis occurringuntil the final step of secretion. There was no evidence thatsucrose hydrolysis occurs either by invertase in the nectaror by a microbial flora. Inhibitors of respiration were foundto inhibit secretion as did anaerobiosis. Temperature also hada marked effect, with a temperature coefficient of 1.8. However,secretion of sucrose was not affected by anaerobic conditions,low temperatures or inhibitors of respiration as markedly asthat of glucose and fructose. Electron microscopy revealed the presence of a thickened andheavily stained wall at the inner border of the secretory epidermallayer. This wall contained numerous plasmodesmata at a frequencyof 14 per µm² and may represent an apoplastic barrier.Light microscope cytochemistry revealed that acid phosphataseis primarily located in the nectiferous tissue, while ATPaseis concentrated in the epidermis. The possibility that the nectarycontains two pathways for sucrose secretion, both apoplasticand symplastic, is discussed. Key words: Invertase, nectary, plasmodesmata, Ricinus communis, sucrose     

Influence of Different Carbon Sources on Invertase Activity and Growth of Sour Cherry (Prunus cerasus L.) Shoot Cultures

The influence of different sugars on shoot multiplication invitro and on the activity of invertase was studied with sourcherry (Prunus cerasus L.) cultures. The sugars sucrose, glucoseand fructose, and the sugar alcohol sorbitol, were investigatedat a wide range of concentrations. The optimum concentrationsof all carbon sources were 2% and 3% (w/v). Sucrose and glucosefavoured a similar rate of proliferation. However, in the presenceof fructose, proliferation was lowest but was coupled with thehighest frequency of formation of long shoots. The highest activityof total invertase was for tissues growing on a sucrose-containingmedium while, in a sugar-free medium, invertase activity wasmainly found in the ‘salt extracted’ fraction. Inthe remaining treatments, ‘soluble invertase’ dominated.For each sugar investigated, the activity of both forms of invertasewas significantly higher at 2% than at 3% (w/v) sugar. Key words: Tissue culture, sugars, invertase, shoot multiplication, sour cherry     

Development of Sink Capacity of the "Storage Root" in a Radish Variety with a Low Ratio of "Storage Root" to Shoot

Controling mechanisms of sink capacity are poorly understood.Previously we suggested that sucrose synthase (SuSy), but notinvertase, plays an important role for sink capacity of theradish "storage root" in a variety, Raphanus sativus L. (cv.White Cherish) [plant Cell Physiol. (1999) 40: 369]. With thisvariety about 50% of the total dry weight (DW) was in the "storageroot" at 21 d after sowing (DAS). We investigated the sink capacityof another radish variety, R. sativus L. (cv. Kosena) with alow ratio of "storage root" to shoot. With the latter varietyonly 3% of the total DW was in the "storage root" at 21 DAS.Sink activity (increase in DW of the "storage root" per unitof DW present per unit of time) of the "storage root" in Kosenaas well as White Cherish was strongly related to the level andactivity of SuSy but not to the activity of invertase. Theseresults confirmed that SuSy rather than invertase may be criticalfor the development of the sink activity of the radish "storageroot" and that the reaction products of UDP-glucose and fructoseare utilized for sink growth including biosynthesis of the cellwall. In Kosena photosynthates seemed to be partitioned mainlyinto developing leaves and fibrous roots. Differences in partitioningof photosynthates among various sinks with these two varietiesare discussed including anatomical considerations. (Received July 19, 1999; Accepted September 30, 1999)     

Functional Characterization of Two Ripening-related Sucrose Transporters from Grape Berries

The ripening of grape (Vitis vinifera L.) berries is associatedwith a large accumulation of glucose and fructose in the vacuolesof the fruit cells. These hexoses are derived from sucrose,which is released from the phloem and may be taken up by parenchymacells prior to hydrolysis. We have expressed two putative ripening-relatedsucrose transporters from grape berries, VvSUC11 (synonymouswith VvSUT1) and VvSUC12, in an invertase deficient yeast strainto characterize their transport activities. Sucrose was takenup by yeast transformed with either transporter at an optimumpH of <4.5 and with a Michaelis constant (K_m) of 0.9–1.4m M. The uptake of sucrose through VvSUC11 and VvSUC12 was inhibitedby protonophores and by vanadate. This is consistent with anactive uptake mechanism involving proton cotransport, typicalof sucrose/H⁺symporters. The transporters from grape berrieswere functionally similar to Scr1, a sucrose transporter fromRicinus cotyledons. It is likely that in grape berries VvSUC11and VvSUC12 facilitate the loading of sucrose from the apoplastinto the parenchyma cells. Copyright 2001 Annals of Botany Company Fruit, grape berries, plasma membrane, sugars, sucrose transporters, Vitis vinifera     

Characterization of wall-bound invertase isoforms of Picea abies cells and regulation by ectomycorrhizal fungi

In culture, the ectomycorrhiza-forming fungi Amanita muscaria (Pers. ex Fries) Hock. and Hebeloma crustuliniforme (Bull. ex Fries) Quel. only grow on media with glucose or fructose but not with sucrose as sole carbohydrate source. This is due to their lack of wall-bound invertase activity. Therefore, utilization of sucrose by the fungi within a mycorrhizal association is believed to depend on the wall-bound invertase activity of the host. This enzyme activity was studied in the apoplast of suspension cultured cells of Picea abies (L.) Karst. An ionically and a tightly wall-bound isoform of acid invertase were found that function as β-d -fructofuranoside-fructohydrolases (EC 3.2.1.26). The ionically bound enzyme could be easily released from walls of intact cells with buffer of high ionic strength. In its native form, the ionically bound invertase isoform is a monomeric protein with a molecular mass of 61 kDa, as determined by gel filtration and SDS-PAGE. Glycoprotein nature of the enzyme was demonstrated with antibodies directed against the digoxigenin-labeled protein. The K_m values of both enzymes for sucrose, their natural substrate, are relatively high (ionically bound invertase K_m= 16 mM, tightly bound invertase K_m= 8.6 mM). Activity of both wall-bound invertase isoforms strongly depends on the apoplastic pH. They have a narrow pH-optimum and exhibit highest activity at pH 4.5. with elevated activity between pH 4.5 and 6.0. Furthermore, fructose acts as competitive inhibitor of both isoforms, whereas glucose is not inhibitory. Unloading of sucrose from host cells to the apoplastic interface of the Hartig net in ectomycorrhizae appears to depend on the rate of hydrolysis by the wall-bound invertase of the host. Since the activity of the plant invertase depends on the actual pH value and the fructose concentration in the mycorrhizal interface, we suggest that the fungus can actively influence the activity of the plant invertase by acidification of the cell wall and by fructose uptake. Thus, the fungus itself can regulate its own supply of glucose and fructose.     

Invertase Development in Storage Tissue 4Beta vulgaris; its Nature, Extent, and Location

The development of invertase activity in storage tissue disksof Beta vulgaris during ageing under aseptic conditions hasbeen studied. The invertase is formed initially in the cellwall and subsequently appears in the cytoplasm. In disks upto 1.0 mm thick, invertase is formed throughout the tissue,but in thicker disks only limited activity appears in the interior.In disks up to 1.0 mm thick, treatment with ethyl acetate priorto enzyme assay increases the measured activity by rendering‘inaccessible’ soluble invertase available to sucrosein the assay medium. In thicker disks, pretreatment with ethylacetate also increases the measured invertase activity by facilitatingsucrose penetration to the centre of the tissue. Osmotic shockaffects invertase activity in the same manner as ethyl acetatetreatment. The significance of these results is discussed inrelation to the induction and development of invertase activityin the disk and in the cell.     

Invertase Activity and its Relation to Hexose Accumulation in Potato Tubers

Hexose accumulation was shown to occur in freshly harvestedmature potato tubers (Solanum tuberosum L.) both after storageat 10 ?C and when subsequently transferred to low temperature(3 ?C) storage. In general, changes in hexoses and sucrose werefound to be related to changes in acid invertase activity. Totalacid invertase activity (i.e. assayed after destroying the endogenousinvertase inhibitor present in the extracts) generally reflectedsugar changes more closely than did basal activity (i.e. assayedwith the inhibitor present). There was no evidence of a specificalkaline invertase. A comparison of the temperature responsesof cultivar Record with that of two SCRI² clones demonstrateddistinct genotypic variation in the extent of hexose accumulation.However, these differences were not always reflected by genotypicdifferences in total invertase activity. Key words: Invertase inhibitor, glucose, fructose, sucrose     

Import of Sucrose and its Partitioning in the Synthesis of Galactomannan and Raffinose-oligosaccharides in the Developing Guar (Cyamopsis tetragonolobus) Seed

Import of sucrose and its transformation to galactomannan andraffinose-oligosaccharides have been studied in the developingguar seed. The amount of galactomannan gradually increased withthe ageing of the seed. During the entire period of pod development,sucrose constituted the major portion of the free sugars inthe seed (both endosperm and cotyledons) as well as in the podwall. Besides myo-inositol, the free sugars detected in thedeveloping endosperm and cotyledons were glucose, fructose,raffinose and stachyose. Some compounds, possibly glycosides(RG values higher than that of fructose), were also detectedin the endosperm. In the later stages of seed development, therelative proportion of raffinose in the free sugars increased,reaching 50% of the total free sugars in 77-d-old cotyledons.With pod maturity, the activities of soluble acid and boundacid invertases in the pod wall increased manifold with a concomitantdecline in the non-reducing sugar content. These enzymes seemto be involved in the mobilization of sucrose from this fruitingstructure into the seed. An increased synthesis of raffinose-oligosaccharidesboth in the endosperm and cotyledons was associated with highactivities of soluble acid invertase (pH 4.8) and sucrose-UDPglucosyl transferase in these tissues. Feeding uniformly labelled¹⁴C-sugars to the detached intact pods as well as to the isolatedendosperm and cotyledons resulted in labelling of all endogenousfree sugars and galactomannan. The uptake and incorporationinto galactomannan of ¹⁴C was stimulated by Co²⁺, Mn²⁺ and Mg²⁺.Except for mannose, a major proportion of the ¹⁴C from glucose,fructose and sucrose appeared in sucrose in both endosperm andcotyledons indicating a fast reconstitution of sucrose in situ.Based on the present results, a possible mode of transformationof sucrose to galactomannan and raffinose-oligosaccharides hasbeen proposed. Key words: Sucrose, galactomannan, raffinose-oligosaccharides, invertase, sucrose-UDP glucosyl transferase, ¹⁴C-incorporation, guar seed     

Acid and alkaline invertases in suspension cultures of sugar beet cells

Alkaline invertase was induced during the initiation of suspension cultures of single cells from leaf explants of sugar beets in Murashige-Skoog liquid medium which contained benzyladenine. This activity was barely detectable in the leaves themselves. In suspension cultures, the presence of both acid and alkaline invertases was detected; alkaline invertase was only present in the cytoplasm of the cultured cells, whereas acid invertase was present in the cytoplasm and cell walls, and was also detected in the culture medium. The cell wall contained at least three types of acid invertase; two of these activities were solubilized by saline (saline-released) and EDTA (EDTA-released), respectively, and the third remained tightly associated with the cell wall. Saline-released and EDTA-released invertases from the cell wall showed the significant differences in their properties: the saline-released enzyme had the highest affinity for sucrose among the invertases tested, and was easily bound to cell walls, to DNA, and to a cation exchanger, unlike the EDTA-released enzyme. Sucrose is the source of carbon for plant cells in suspension culture and is probably degraded in the cell wall by the saline-released invertase, which had the highest activity and the highest affinity for sucrose. Hexose products of this degradation would be transported to cytoplasm. Soluble invertase, EDTA-released invertase from the cell wall, and one of two extracellular invertases behaved similarly upon chromatography on DEAE-cellulose. They had similar activity profiles with changing pH, and similar K_m values for sucrose. Thus it appears that they are identical. Two extracellular invertases found in the growth medium of the suspension cultures were probably identical with those in the soluble fraction of callus and seedlings of sugar beets, because they showed similar behaviors during chromatography on DEAE-cellulose, and had similar activity profiles with changing pH and K_m values for sucrose.     

Analysis of carbohydrate metabolism enzymes and cellular contents of sugars and proteins during spruce somatic embryogenesis suggests a regulatory role of exogenous sucrose in embryo development.

Carbohydrate metabolism was investigated during spruce somatic embryogenesis. During the period of maintenance corresponding to the active phase of embryogenic tissue growth, activities of soluble acid invertase and alkaline invertase increased together with cellular glucose and fructose levels. During the same time, sucrose phosphate synthase (SPS) activity increased while sucrose synthase (SuSy) activity stayed constant together with the cellular sucrose level. Therefore, during maintenance, invertases were thought to generate the hexoses necessary for embryogenic tissue growth while SuSy and SPS would allow cellular sucrose to be kept at a constant level. During maturation on sucrose-containing medium, SuSy and SPS activities stayed constant whereas invertase activities were high during the early stage of maturation before declining markedly from the second to the fifth week. This decrease of invertase activities resulted in a decreased hexose:sucrose ratio accompanied by starch and protein deposition. Additionally, carbohydrate metabolism was strongly modified when sucrose in the maturation medium was replaced by equimolar concentrations of glucose and fructose. Essentially, during the first 2 weeks, invertase activities were low in tissues growing on hexose-containing medium while cellular glucose and fructose levels increased. During the same period, SuSy activity increased while the SPS activity stayed constant together with the cellular sucrose level. This metabolism reorganization on hexose-containing medium affected cellular protein and starch levels resulting in a decrease of embryo number and quality. These results provide new knowledge on carbohydrate metabolism during spruce somatic embryogenesis and suggest a regulatory role of exogenous sucrose in embryo development.     

A novel sucrose synthase pathway for sucrose degradation in cultured sycamore cells

Enzymes of sucrose degradation and glycolysis in cultured sycamore (Acer pseudoplatanus L.) cells were assayed and characterized in crude extracts and after partial purification, in an attempt to identify pathways for sucrose catabolism. Desalted cell extracts contained similar activities (20-40 nanomoles per milligram protein per minute) of sucrose synthase, neutral invertase, glucokinase, fructokinase, phosphofructokinase, and UDPglucose pyrophosphorylase (assayed with 2 micromolar pyrophosphate (PPi). PPi-linked phosphofructokinase activity was virtually dependent upon fructose 2,6-bisphosphate, and the maximum activity exceeded that of ATP-linked phosphofructokinase. Hexokinase activity, with glucose as substrate, was highly specific for ATP, whereas fructokinase activity was relatively nonspecific. At 1 millimolar nucleoside triphosphate, fructokinase activity decreased in the order: UTP > ATP > CTP > GTP. We propose two pathways for sucrose degradation. One involves invertase action, followed by classical glycolysis of hexose sugars, and the other is a novel pathway initiated by sucrose synthase. The K_m for sucrose of sucrose synthase was severalfold lower than that of neutral invertase (15 versus 65 millimolar), which may determine carbon partitioning between the two pathways. The sucrose synthase pathway proposed involves cycling of uridylates and PPi. UDPglucose pyrophosphorylase, which is shown to be an effective `PPi-scavenger,' would consume PPi and form UTP. The UTP could be then utilized in the UTP-linked fructokinase reaction, thereby forming UDP for sucrose synthase. The source of PPi is postulated to arise from the back reaction of PPi-linked phosphofructokinase. Sycamore cells contained a substantial endogenous pool of PPi (about 3 nanomoles per gram fresh weight, roughly 1/10 the amount of ATP in these cells), and sufficient fructose 2,6-bisphosphate (0.09 nanomole per gram fresh weight) to activate the PPi-linked phosphofructokinase. Possible regulation and energetic differences between the sucrose synthase and invertase pathways are discussed.     

Sucrose Utilization of the Ectomycorrhizal Fungi Amanita muscaria and Hebeloma crustuliniforme Depends on the Cell Wall-bound Invertase Activity of their Host Picea abies

The role of apoplastic invertase (β-d -fructofuranoside — fructohydrolase, EC 3.2.1.26) of the host Picea abies for carbohydrate uptake and growth of two of its natural ectomycorrhiza partners was studied. For that purpose, hyphae of Amanita muscaria (Pers. ex Fries) Hock. and Hebeloma crustuliniforme (Bull. ex Fries) Quell., as well as roots and suspension cultured cells of Picea abies (L.) Karst. were used. Apoplastic invertase activity was demonstrated on roots and suspension cultured cells of spruce (in the latter case with 21.7 nkat (g fresh weight)^?1). Inhibition of the root cell wall invertase activity (pH optimum 4.5) by increasing the apoplastic pH allowed determination of the permanent release of sucrose from the root. However, under in vivo conditions at a lower cell wall pH the hydrolysation products glucose and fructose were predominantly found. In contrast to spruce cells and certain fungi, such as Saccharomyces (Novick et al., 1981) or Phycomyces (Ruiz-Herrera et al., 1989) invertase activity of the mycorrhizal fungi Hebeloma and Amanita was negligibly low. Furthermore, sucrose could not be consumed by Amanita and Hebeloma. As a consequence, cultures of these mycorrhizal fungi starved when kept on media with sucrose as sole carbohydrate source. But addition of invertase initiated hyphal growth immediately. Studies on carbohydrate uptake of host and fungal cells confirmed that the monosaccharides glucose and fructose were readily incorporated by spruce and fungal cells, with a clear preference for glucose. From these results it is suggested that apoplastic invertase activity of the host Picea abies is a precondition for the utilization of sucrose by the studied mycorrhizal fungi during the nutritional interaction of the symbiotic partners.     

Differential sugar uptake by cell suspension cultures of <Emphasis Type="Italic">Rudgea jasminoides</Emphasis>, a tropical woody Rubiaceae

The primary utilization of carbohydrates by cell suspension cultures of Rudgea jasminoides, a native woody Rubiaceae from tropical forests, was investigated. Sucrose, glucose + fructose, glucose, or fructose were supplied as carbon sources. The growth curves of R. jasminoides cultured in glucose + fructose, glucose, or fructose showed similar patterns to that observed when sucrose was supplied to the cells, except that an increase in dry mass was observed at the beginning of the stationary growth phase in the media containing only one monosaccharide. The increase in hexose levels in the media during the early stages of the cultures indicated extracellular hydrolysis of sucrose, which was further supported by the increase in the activity of acid invertase bound to the cell wall. Glucose was preferentially taken up, whereas uptake of fructose was delayed until glucose was nearly depleted from the medium. Measurements of intracellular sucrose content and cytoplasmatic and vacuolar invertases indicate that the enzymatic activity seems to be correlated with a decrease in the hexose flux into the cells of R. jasminoides. Our results indicate that the behavior of cell suspension cultures of R. jasminoides regarding sugar utilization seems to be similar to other dicotyledonous undifferentiated cell suspension cultures.     

Sucrose metabolism during endosperm development in wheat (Triticum aestivum)

This work reports changes in sucrose synthase and invertase activities throughout endosperm development in wheat, together with the associated substrates and metabolites, sucrose, UDP, glucose, fructose and UDP-glucose. Throughout endosperm development, sucrose synthase had consistently higher activity than invertase and indeed invertase activity did not change appreciably. The observed variation in pattern and amounts of glucose and fructose present during the mid- and late stages of endosperm development confirmed the suggestion that invertase was not the preferred pathway of sucrose catabolism. Kinetic parameters for sucrose synthase were determined in crude extracts. Estimates of UDP and sucrose concentrations suggest that sucrose synthase is unlikely to achieve its potential maximum velocity. This limitation may however be overcome in part by the apparent excess catalytic activity measured during endosperm development.     

The Relationship between Sucrose Supply, Sucrose-cleaving Enzymes and Flower Abortion in Pepper

Abortion of pepper flowers depends on the light intensity perceivedby the plant and on the amounts of sucrose taken up by the flower(Aloni B, Karni L, Zaidman Z, Schaffer AA. 1996.Annals of Botany78: 163–168). We hypothesize that changes in the activityof sucrose-cleaving enzymes within the flower ovary might beresponsible for the changes in flower abortion under differentlight conditions. In the present study we report that the activityof sucrose synthase, but not of cytosolic acid invertase, increasesin flowers of pepper plants which were exposed, for 2 d, toincreasing photosynthetically active radiation (PAR) in therange of 85–400 µmol m^-2s^-1at midday. Sucrose synthaseactivity increased in parallel with the increasing concentrationsof starch in the flower ovary. Feeding flower explants, preparedfrom 3-d-predarkened plants, with 100 mM sucrose for 24 h, causeda 23% increase in reducing sugars and a 2.5-fold increase instarch concentration, compared with explants fed with buffer.Likewise, feeding explants of pepper flowers with sucrose, glucose,fructose and also mannitol increased the sucrose synthase activityin the ovaries. Concomitantly, sucrose, glucose and fructose,but not mannitol, reduced the abortion of flower explants. Itis suggested that sucrose entry into the flower increases theflower sink activity by inhibiting abscission and inducing metabolicchanges, thus enhancing flower set. Pepper; Capsicum annuum L.; abscission; light; pepper flowers; sucrose; glucose; fructose; starch; acid invertase; sucrose synthase     

Sucrose utilization during somatic embryo development in black spruce: involvement of apoplastic invertase in the tissue and of extracellular invertase in the medium

The involvement of apoplastic invertase (Ap Inv) and sucrose synthase (SuSy) in the somatic embryo development of black spruce (Picea mariana) was investigated under different maturation conditions. Replacing 6% sucrose with 3% or 1% sucrose in the maturation medium drastically decreased Ap Inv activity and amount in embryogenic tissues. This was accompanied by a decrease in the hexose pool that resulted in a lower starch deposition and protein amount in embryogenic tissues together with a lower embryo production. Conversely, SuSy activity was stable during maturation regardless of the sucrose concentration used in the medium. The presence of an extracellular enzyme responsible for sucrose hydrolysis in the maturation medium was also verified. An immunodetection experiment with anti-acid invertase antibodies revealed the presence of an active 53 kDa polypeptide in the medium, which had a similar molecular mass to that of the Ap Inv polypeptide found in embryogenic tissues. Utilization of sucrose from the medium by the tissues was also studied using labelled 14C-sucrose. Distribution of the radioactivity between tissular sucrose, glucose, and fructose showed that sucrose was diffused into the cell wall of embryogenic tissues and partly hydrolyzed by Ap Inv. These results show that the utilization of sucrose from the medium, the Ap Inv activity in embryogenic tissues, and the release of an active invertase into the medium operate together for the utilization of the carbohydrates during somatic embryo development in black spruce.     

Uptake and Metabolism of Sugars by Suspension Cultured Catharanthus roseus Cells

The uptake and metabolism of sugars by suspension-cultured Catharanthusroseus cells were investigated. Substantially all the sucrosein the culture medium was hydrolyzed to glucose and fructosebefore being taken up by the cells. The activity of invertasebound to cell walls, determined in situ, was high at the earlystage of culture. Glucose was more easily taken up by the cellsthan was fructose. Tracer experiments using [U-¹⁴C]glucose and[U-¹⁴C]fructose indicated that glucose is a better precursorfor respiration than fructose, while fructose is preferentiallyutilized for the synthesis of sucrose, especially in the earlyphase of cell growth. Possible metabolic routes of sugar insuspension-cultured Catharanthus roseus cells are discussedin the context of these results. Catharanthus roseus, Madagascar periwinkle, suspension culture, sucrose, glucose, fructose, metabolism, glycolysis     

Physiological Aspects of Sugar Exchange between the Gametophyte and the Sporophyte of Polytrichum formosum

The sporophyte of bryophytes is dependent on the gametophyte for its carbon nutrition. This is especially true of the sporophytes of Polytrichum species, and it was generally thought that sucrose was the main form of sugar for long distance transport in the leptom. In Polytrichum formosum, sucrose was the main soluble sugar of the sporophyte and gametophyte tissues, and the highest concentration (about 230 mm) was found in the haustorium. In contrast, sugars collected from the vaginula apoplast were mainly hexoses, with traces of sucrose and trehalose. p-Chloromercuribenzene sulfonate, a nonpermeant inhibitor of the cell wall invertase, strongly reduced the hexose to sucrose ratio. The highest cell wall invertase activity (pH 4.5) was located in the vaginula, whereas the highest activity of a soluble invertase (pH 7.0) was found in both the vaginula and the haustorium. Glucose uptake was carrier-mediated but only weakly dependent on the external pH and the transmembrane electrical gradient, in contrast to amino acid uptake (S. Renault, C. Despeghel-Caussin, J.L. Bonnemain, S. Delrot [1989] Plant Physiol 90: 913-920). Furthermore, addition of 5 or 50 mm glucose to the incubation medium induced a marginal depolarization of the transmembrane potential difference of the transfer cells and had no effect on the pH of this medium. Glucose was converted to sucrose after its absorption into the haustorium. These results demonstrate the noncontinuity of sucrose at the gametophyte/sporophyte interface. They suggest that its conversion to glucose and fructose at this interface, and the subsequent reconversion to sucrose after hexose absorption by haustorium cells, mainly governs sugar accumulation in this latter organ.     

Sucrose and Nitrogen Supplies Regulate Growth of Maize Kernels

The growth of maize (Zea mays L.) kernels depends on the availabilityof carbon (C) and nitrogen (N) assimilates supplied by the motherplant and the capacity of the kernel to use them. Our objectiveswere to study the effects of N and sucrose supply levels ongrowth and metabolism of maize kernels. Kernel explants of Pioneer34RO6 were culturedin vitro with varying combinations of N (5to 30 m M) and sucrose (117 to 467 m M). Maximum kernel growthwas obtained with 10 m M N and 292 m M sucrose in the medium,and a deficiency of one assimilate could not be overcome bya sufficiency of the other. Increasing the N supply led to increasesin the kernel sink capacity (number of cells and starch granulesin the endosperm), activity of certain enzymes (soluble andbound invertases, sucrose synthase, and aspartate aminotransaminase),starch, and the levels of N compounds (total-N, soluble protein,and free amino acids), and decreased the levels of C metabolites(sucrose and reducing sugars). Conversely, increasing the sucrosesupply increased the level of endosperm C metabolites, freeamino acids, and ADPG-PPase and alanine transaminase activities,but decreased the activity of soluble invertase and concentrationsof soluble protein and total-N. Thus, while C and N are interdependentand essential for accumulation of maximum kernel weight, theyappear to regulate growth by different means. Nitrogen supplyaids the establishment of kernel sink capacity, and promotesactivity of enzymes relating to sucrose and nitrogen uptake,while sucrose regulates the activities of invertase and ADPG-PPase.Copyright 1999 Annals of Botany Company Zea mays, maize,, invertase, ADPG-PPase, media composition, sucrose, nitrogen, C/N.