Bioregulator enhancement of sink activity in sugar beet

The sink mobilizing abillity is partially determined by sugar uptake rates of storage cells. Two synthetic growth regulators (Pix and BAS 106W) were tested for their effect on sucrose uptake in root tissue discs or glucose uptake in cell cultures of sugar beet. In tissue discs, uptake at the plasmalemma was determined by incubating the discs for 1 h in the presence of 5 mM sucrose and at the tonoplast for 4 h in the presence of 40 mM sucrose. Cell cultures were incubated for 1 h in the presence of 1 mM glucose. Pix (10 mg l^–1) caused a 20% stimulation of active sucrose uptake at the plasmalemma. Active sucrose uptake at the tonoplast was increased 67% by 100 mg l^–1 Pix. No effect of BAS 106W was observed on sucrose uptake in tissue discs. In cell cultures, a 65% enhancement of active glucose uptake was observed with both Pix and BAs 106W. When the bioregulators were applied to the root medium of seedlings, Pix but not BAS 106W resulted in increased root/shoot ratio, translocation of ¹⁴C-assimilates, and allocation of more biomass to the root sink. The data suggested that sugar transport and translocation may be used as biochemical criteria for rapid screening of effective yield enhancing bioregulators.     

Effects of decreased net carbon exchange on carbohydrate metabolism in sugar beet source leaves

The relationship between CO₂ concentration and starch synthesis and degradation was studied by measuring leaf starch content and disappearance of ¹⁴C-starch. At a concentration of 340 microliters CO₂ per liter, starch accumulated without degradation of previously synthesized starch. Degradation of starch began when CO₂ concentration was lowered, but its synthesis continued. At 120 microliters CO₂ per liter rates of synthesis and degradation were equal. Even at the CO₂ compensation point, synthesis of starch continued. Concomitant starch synthesis and mobilization supported export from the leaf. Changes in starch metabolism that occur when photosynthesis is CO₂-limited provide a means to study regulation of starch metabolism and carbon allocation in translocating leaves.     

Effects of boron on nitrogen metabolism and sugar levels of sugar beet

Summary The effects of deficient and toxic levels of boron on various aspects of nitrogen metabolism in sugar beet are studied. Plant analysis shows a nitrate ion accumulation, a decrease in the activity of the nitrate reductase enzyme and a lower molybdenum absorption.The effect of boron levels on the plant and root sugar concentration has also been studied.     

Probing by aphids in the leaf tissues of resistant and susceptible sugar beet

The probing of Aphis fabae and Myzus persicae in the leaves of sugar beet with inherited resistance or susceptibility to aphids was studied by microscopic examination of samples of whole leaves, prepared after 48 h exposure to adult aphids at approximately three aphids cm^-2.The density of saliva stylet-sheaths left by the aphids (cm^-2) and the proportion reaching phloem differed between sugar beet stocks and were inversely associated. Differences in resistance between stocks could not, however, be related directly to either. All beet stocks examined were probed freely. Seasonal differences in sugar beet grown in the glasshouse affected the proportion of sheaths reaching the phloem, but the differences between beet stocks were similar at all times.The densities of sheaths left by different clones of M. persicae corresponded with the aphids' response to sugar beet as a host plant. Among aphid clones which readily colonize sugar beet, the densities of stylet sheaths which reached phloem suggested that the adults of both A. fabae and M. persicae gained sufficient access to sieve tubes to satisfy their nutritional needs. The phloem of sugar beet from the glasshouse was always within the estimated maximum depth to which the aphids probe; but, in leaves from the field, it appeared that the phloem might be inaccessible to young M. persicae in the sugar beet crop during late summer.

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Zusammenfassung Das Proben von Aphis fabae und Myzus persicae in Blättern von Zuckerrüben mit erblicher Blattlausresistenz bzw.-anfälligkeit wurde untersucht durch mikroskopische Durchmusterung von Speichelscheiden in Proben von ganzen Blatt. Rübenblätter wurden mit genähert drei adulten Läusen cm^-2 besetzt und nach 48 Stunden quergeschnittene Streifen der Blätter in Alkohol fixiert, gefärbt und mit der Unterseite nach oben auf Objektträgern eingeschlossen.23890 Speichelscheiden wurden registriert. Die Dichte der Scheiden von M. persicae (cm^-2) und der Anteil der das Phloem erreichenden Scheiden (SRP) unterschieden sich signifikant zwischen den Rübenstämmen. Bei A. fabae ergaben sich entsprechende, aber nicht gesicherte Unterschiede. Scheidendichte und Prozentsatz SRP waren gegenläufig, zwei Rübenstämme zeigten eine hohe Scheidendichte, zwei andere hatten weniger Scheiden, aber einen höheren Prozentsatz SRP. Diese Gruppierung der Stämme korrespondierte aber nicht mit ihrer Blattlausresistenz. Aus der Scheidendichte ergab sich, dass M. persicae und A. fabae auf allen geprüften Rübenstämmen, resistenten und anfälligen, unbehindert probten, so dass jede Laus das Phloem durchschnittlich etwa viermal am Tag erreichte. Ein Klon von M. persicae, der sich an Rüben nicht entwickelt, hinterliess weniger Scheiden in den Blättern aller Stämme.Der Anteil von SRP war bei Prüfungen im März grösser als im November. Dieser Unterschied war besonders deutlich bei Scheiden von Larven, die im übrigen zu allen Zeiten das Phloem weniger oft erreichten als ihre Eltern. Messungen des Abstandes von der unteren Blattfläche zum Phloem ergaben, dass das Phloem den Läusen in Gewächshaus-Zuckerrüben immer zugänglich war. M. persicae-Larven konnten jedoch in Blättern von Freilandrüben das Phloem nicht erreichen.

     

Characterization of sterol uptake in leaf tissues of sugar beet

The uptake of cholesterol has been characterized in leaf discs from mature leaves of sugar beet (Beta vulgaris L.). This transport system exhibited a simple saturable phase with an apparent Michaelis constant ranging from 30 to 190 M depending on the sample. When present at 10 M excess, other sterols were able to inhibit cholesterol uptake. Moreover, binding assays demonstrated the presence of high-affinity binding sites for cholesterol in purified plasma membrane vesicles. In the range 1–60 M, cholesterol uptake showed an active component evidenced by action of the protonophore carbonyl cyanide m-chlorophenylhydrazone. Energy was required as shown by the inhibition of uptake induced by respiration inhibitors (NaN₃), darkness and photosynthesis inhibitors [3-(3,4-dichlorophenyl)-1,1-dimethylurea, methyl viologen]. Moreover, the process was strongly dependent on the experimental temperature. Uptake was optimal at acidic pH (4.0), sensitive to ATPase modulators, inhibited by thiol reagents (N-ethylmaleimide, p-chloromercuribenzenesulfonic acid, Mersalyl) and by the histidyl-group reagent diethyl pyrocarbonate. The addition of cholesterol did not modify H⁺ flux from tissues, indicating that H⁺-co-transport was unlikely to be involved. MgATP did not increase the uptake, arguing against involvement of an ABC cassette-type transporter. By contrast, cryptogein, a sterol carrier protein from the Oomycete Phytophtora cryptogea, greatly increased absorption. Taken together, the results reported in this work suggest that plant cells contain a specific plasma membrane transport system for sterols.Abbreviations CCCP carbonyl cyanide m-chlorophenylhydrazone - PCMBS p-chloromercuribenzenesulfonic acid - PMV plasma membrane vesicle - TLC thin-layer chromatography     

Sources of sucrose translocated from illuminated sugar beet source leaves

A search for source leaf sucrose pools that differed in their relation to export was carried out in photosynthesizing leaves of Beta vulgaris L. The time course of depletion of [¹⁴C]sucrose in a leaf in unlabeled CO₂ following steady state labeling provided evidence for two distinct sucrose pools. After the start of the light period, leaf blade sucrose remained constant although it exchanged between the two pools. Newly synthesized sucrose destined for export passed through one pool more rapidly than through the other. All of the leaf blade sucrose appeared to exchange with export sucrose. Modeling and regression analysis of [¹⁴C]sucrose data provided a means for estimating the size of the two pools. From 20 to 40% of the sucrose was calculated to be present in the pool that provided the less direct path to export; this was likely vacuolar sucrose. The remainder of the sucrose in the blade was probably in the cytoplasm and veins. Added amounts of leaf blade sucrose, produced in response to elevated CO₂, appeared to be stored mainly in the vacuolar compartment.     

Chromatin- and nuclei-directed ribonucleic Acid synthesis in sugar beet root

The synthesis of RNA by chromatin-bound RNA polymerase prepared from sugar beet (Beta vulgaris) root tissue is completely dependent on the presence of a divalent metal (Mg²⁺ or Mn²⁺) and the presence of four ribonucleoside triphosphates. Accumulation of labeled acid-insoluble product is inhibited by the addition of RNase and actinomycin D to the reaction. When beet root slices are washed for 25 hours, chromatin-associated RNA polymerase activity increases 7-fold over that of unwashed tissue. This enzyme activity declines with further washing. DNA template availability, as measured by saturating levels of added Escherichia coli RNA polymerase, was also found to follow a pattern similar to that for RNA polymerase. Nearest neighbor frequencies of the RNA synthesized by chromatin isolated from unwashed and washed tissue are different.     

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.     

Abscisic Acid localization and metabolism in barley aleurone layers

Aleurone layers of Hordeum vulgare, cv. `Himalaya' took up [¹⁴C]-abscisic acid (ABA) when incubated for various times. Radioactivity accumulated with time in a low speed, DNA-containing pellet accounting for 1.6 to 2.3% of the radioactivity recovered in subcellular fractions at 18 hours. Thin layer chromatography of ethanolic or methanolic extracts of the cytosol, which contained greater than 95% of the radioactivity taken up by layers, revealed that labeled ABA was metabolized to phaseic acid (PA) and 4′-dihydrophaseic acid (DPA) and three polar metabolites Mx₁, Mx₂, and Mx₃. ABA was not metabolized by endosperm, incubated under conditions used for layers, indicating that metabolism was tissue-specific. Layers metabolized [³H]DPA to Mx₁ and Mx₂. ABA, PA, and DPA-methyl ester and epi-DPA-methyl ester inhibited synthesis of α-amylase by layers incubated for either 37 or 48 hours. These layers converted the methyl DPA and epi-methyl-DPA esters to their respective acids. DPA did not inhibit Lactuca sativa germination or root and coleoptile elongation of germinating Hordeum vulgare seeds, or coleoptile elongation of germinating Zea mays seeds.     

Carboxylate metabolism in sugar beet plants grown with excess Zn

The effects of Zn excess on carboxylate metabolism were investigated in sugar beet (Beta vulgaris L.) plants grown hydroponically in a growth chamber. Root extracts of plants grown with 50 or 100 μM Zn in the nutrient solution showed increases in several enzymatic activities related to organic acid metabolism, including citrate synthase and phosphoenolpyruvate carboxylase, when compared to activities in control root extracts. Root citric and malic acid concentrations increased in plants grown with 100 μM Zn, but not in plants grown with 50 μM Zn. In the xylem sap, plants grown with 50 and 100 μM Zn showed increases in the concentrations of citrate and malate compared to the controls. Leaves of plants grown with 50 or 100 μM Zn showed increases in the concentrations of citric and malic acid and in the activities of citrate synthase and fumarase. Leaf isocitrate dehydrogenase increased only in plants grown with 50 μM Zn when compared to the controls. In plants grown with 300 μM Zn, the only enzyme showing activity increases in root extracts was citrate synthase, whereas the activities of other enzymes decreased compared to the controls, and root citrate concentrations increased. In the 300 μM Zn-grown plants, the xylem concentrations of citric and malic acids were higher than those of controls, whereas in leaf extracts the activity of fumarase increased markedly, and the leaf citric acid concentration was higher than in the controls. Based on our data, a metabolic model of the carboxylate metabolism in sugar beet plants grown under Zn excess is proposed.     

Effect of heavy metals on nitrate and protein metabolism in sugar beet

Nitrate content, activities of nitrate reductase and glutamine synthetase, soluble protein content, and proportion of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) protein in total proteins were measured in leaves of Beta vulgaris L. plants affected by nickel, cadmium, and molybdenum in concentrations of 10^-4, 10^-2, and 1 mM. The most harmful effect on the above mentioned parameters had Cd, less harmful Ni, whereas Mo stimulated the investigated parameters. The proportion of RuBPCO protein showed a high tolerance to heavy metals. This revised version was published online in July 2006 with corrections to the Cover Date.     

Polyamine metabolism and ethylene biosynthesis in normal and habituated sugar beet callus

The optimal assay conditions and the trend with time in culture (28 days) of arginine decarboxylase (ADE; EC 4.1.1.19), omithine decarboxylase (ODC; EC 4.1.1.17) and diamine oxidase (DAO; EC 1.4.3.6) activities in habituated (H) and normal (N) auxin- and cytokinin-requiring sugar beet callus were compared. Although the response to variations in buffer pH and EDTA and pyridoxal phosphate (PLP) concentrations varied for ADC and ODC activities between the two callus types, pH 8.3, 50 μ M PLP and 5 m M EDTA were generally optimal or near-optimal for both H and N callus. In most cases the addition of ornithine or arginine in the ADC and ODC assays, respectively, given to block the interconversion between the two substrates, resulted in lower ¹⁴CO₂ recovery. DAO activity was very differently affected in H and N callus by the presence of polyvinylpyrrolidone in the extration buffer. However, in both cases, this activity increased with time in culure. ADC activity was always predominant in both cell lines and always higher in N callus. In the latter, ADC activity rose sharply between days 14 and 21 and then leveled off while in H callus it incresed steadily from day 14 onwards. ODC activity was also higher in N callus and peaked sharply on day 21 while in H callus it was not detectable in the second half of the culture period. In both cell lines this activity was low or nil on day 28. 3,4-[¹⁴C]-methionine incorporation into ethylene and polyamines was also compared in N and H callus. In the latter, ethylene synthesis was lower and [¹⁴C]-spermidine formation higher than in N callus. This is in accord with the significantly higher spermidine titres found in H callus.     

Effect of altered sink: source ratio on photosynthetic metabolism of source leaves

When seven crop species were grown under identical environmental conditions, decreased sink:source ratio led to a decreased photosynthetic rate within 1 to 3 days in Cucumis sativus L., Gossypium hirsutum L., and Raphanus sativus L., but not in Capsicum annuum L., Solanum melongena L., Phaseolus vulgaris L., or Ricinus communis L. The decrease was not associated with stomatal closure. In cotton and cucumber, sink removal led to an increase in starch and sugar content, in glucose 6-phosphate and fructose 6-phosphate pools, and in the proportion of ¹⁴C detected in sugar phosphates and UDPglucose following ¹⁴CO₂ supply. When mannose was supplied to leaf discs to sequester cytoplasmic inorganic phosphate, promotion of starch synthesis, and inhibition of CO₂ fixation, were observed in control discs, but not in discs from treated plants. Phosphate buffer reduced starch synthesis in the latter, but not the former discs. The findings suggest that sink removal led to a decreased ratio inorganic phosphate:phosphorylated compounds. In beans ¹⁴C in sugar phosphates increased following sink removal, but without sucrose accumulation, suggesting tighter feedback control of sugar level. Starch accumulated to higher levels than in the other plants, but CO₂ fixation rate was constant for several days.     

Phenotypic polymorphism and allele differentiation of isozymes in fodder beet,multigerm sugar beet and monogerm sugar beet

Summary Thirteen enzymes (MDH, SDH, LAP, PGM, PX, IDH, GPI, 6PGD, APH, GOT, GDH, ME and SOD) of 3 cultivated beet (B. vulgaris L.) gene pools, comprising 12 accessions of fodder beet, 11 of old multigerm sugar beet and 10 of modern monogerm sugar beet, were investigated using horizontal starch gel electrophoresis. Eleven accessions of primitive or wild B. vulgaris were also included for the comparison of isozymes. Variation in isozyme phenotypes was investigated to detect diversity in the three cultivated forms of beet. Phenotypic variation was observed in all except ME and SOD, which were monomorphic. A high degree of phenotypic polymorphism (Pj) was found in GDH, PGM, IDH, APH and MDH. Differences in phenotypic polymorphism in MDH, GPI and PX were recognized between fodder beet and both sugar beet groups. Average polymorphism for 13 enzymes in both sugar beets was significantly higher than that in fodder beet. For 13 enzymes, the existence of high isozyme diversity in both sugar beet gene pools was revealed. Allele frequencies in 13 alleles of five enzyme-coding loci, Lap, Px-1, Aph-1, Got-2 and Gdh-2, were investigated. New alleles, Px-1 ¹ and Got-2 ¹, were found in fodder beet accessions. No significant differences of average allele frequencies of five loci between fodder beet and both sugar beets were recognized. Several unique alleles and different isozyme phenotypes were observed in the accessions of B. vulgaris ssp. macrocarpa and ssp. adanensis. Future utilization of cultivated beet gene pools for sugar beet breeding is discussed from the viewpoint of genetic resources.     

Chemistry of Abscisic Acid, Abscisic Acid Catabolites and Analogs

Recently there have been breakthroughs on a number of fronts in abscisic acid (ABA) biology research that have advanced the field significantly, including discovery of genes involved in ABA metabolism, along with progress in understanding of ABA signaling (Finkelstein and others 2002; Kushiro and others 2004; Lim and others 2005; Saito and others 2004). At the same time, the chemistry of ABA has advanced. New analytical methods have been developed for profiling ABA and catabolites (Ross and others 2004; Zaharia and others 2005). Novel bioactive catabolites have been discovered from feeding studies with deuterated ABA and catabolites (Zaharia and others 2004; Zhou and others 2004). This review covers recent advances and prospects in natural products chemistry, analysis of ABA catabolism, and applications of ABA analogs for biochemical studies and horticultural uses.     

Sodium absorption by intact sugar beet plants

Sodium absorption by intact sugar beet plants (Beta vulgaris) was found to be mediated by at least two distinct mechanisms when uptake was studied over a wide range of Na and K concentrations. The first mechanism operates at low Na concentrations (<1 milliequivalent per liter); presence of K completely blocks this mechanism for Na. The second mechanism operates at high Na concentrations (>1 milliequivalent per liter), transporting Na as well as K; but apparently this mechanism is not active for Na absorption in young sugar beet plants up to the 10-leaf stage.     

Effect of Azotobacter strains on sugar beet callus proliferation and nitrogen metabolism enzymes

The effect of five Azotobacter chroococcum strains and nitrogen content in nutrient media on callus growth of two Beta vulgaris L. cultivars were investigated, as well as the activity of nitrate reductase (NR), glutamine synthetase (GS) and glutamate dehydrogenase (GDH) in inoculated callus tissue. On medium with full nitrogen content (1 N) the inoculation with A. chroococcum strain A2 resulted in the highest calli mass, while strains A8 and A14 maximally increased NR activity. On media with 1/8 N the highest effect on calli growth, GS and GDH activity had the strain A8. The strain A2/1 significantly increased callus proliferation on medium without N. Asymbiotic association between sugar beet calli and Azotobacter depended on genotype/strain interaction and was realised in presence of different nitrogen levels. This revised version was published online in July 2006 with corrections to the Cover Date.     

Treatment of beet sugar wastewater by UAFB bioprocess

The aim of this work was to study the treatment of strong beet sugar wastewater by an upflow anaerobic fixed bed (UAFB) at pilot plant scale. Three fixed bed bioreactors (each 60 L) were filled with standard industrial packing, inoculated with anaerobic culture (chicken manure, cow manure, anaerobic sludge digested from domestic wastewater) and operated at 32-34 degrees C with 20 h hydraulic retention time (HRT) and influent COD ranging between 2000-8000 mg/L. Under these conditions the maximum efficiency of organic content reduction in the reactor ranged from 75% to 93%. The reactor filled with standard pall rings made of polypropylene with an effective surface area of 206 m(2)/m(3) performed best in comparison to the reactor filled with cut polyethylene pipe 134 m(2)/m(3) and reactor filled with PVC packing (50 m(2)/m(3)). There was 2-7% decrease in efficiency with PE while it was 10-16% in case of PVC when compared to standard pall rings. The study provided a very good basis for comparing the effect of packing in reduction efficiency of the system.