Darkness and the activity of diquat and paraquat on tomato, broad bean and sugar beet

A period of darkness after treatment increased the activity of diquat and paraquat on tomato, broad bean and sugar beet. The extent of the increase was dependent on the time of day (and year) of treatment, on the light quality and intensity before spraying and on the duration of the darkness. Damage from morning treatments, unlike those in the afternoon, increased rhythmically with increasing darkness. The rhythm showed a maximum about every 24 hr and continued through several days of darkness. The results underline not only the complexity of diquat and paraquat action but also the importance of strictly controlling experimental procedure when studying their mode of action.     

The influence of darkness on the uptake and movement of diquat and paraquat in tomatoes, sugar beet and potatoes

Uptake of diquat and paraquat was doubled and occasionally quadrupled when tomatoes were darkened after treatment. The increase was not directly related to the duration of darkness because, after a time, uptake decreased. Three possible explanations for this decrease are considered, namely: diquat exudation from the leaves, downward movement into the roots and the complexing of diquat in plant tissue. Evidence was against exudation from leaves and downward movement into the roots. Diquat activity also depends on its degree of movement in the plant but the possible influence of other factors is discussed when treatments are made in the afternoon or in the morning after a period of reduced light intensity. In a CO₂-free atmosphere, uptake and activity increased in light and were comparable to those in darkness. Uptake is therefore not impeded by phyto-toxic effects of diquat but possibly by the presence of a high concentration of solutes or by photosynthetic intermediates.     

Sucrose uptake by sugar beet tap root tissue

Sucrose uptake by discs of mature sugar beet root tissue incubated in [¹⁴C]-sucrose exhibited nonsaturating kinetics over the concentration range of 1 to 500 millimolar. Uptake was inhibited by dinitrophenol, sodium cyanide, low O₂, and penetrating sulfhydryl inhibitors. ATPase inhibitors, sodium fluoride, and oligomycin reduced uptake by 20 and 40%, respectively. Uptake as asymmetrically labeled sucrose ([¹⁴C]glucose) occurred with approximately 80% retention of asymmetry, indicating a nonhydrolytic pathway. Uptake was against a concentration gradient and required metabolic energy.     

Inhibition of water uptake in sugar beet roots by ammonia

Ammonium sulfate, ammonium carbonate or ammonia gas inhibited water uptake in sugar beet roots whenever the pH was sufficiently high to cause the production of ammonia. When ammonia was removed by aeration, inhibition of the water uptake by roots was rapidly reversed. ATP at 0.2 mm appeared to either wholly or partially prevent the ammonia-induced inhibition of water uptake by roots. ATP may be involved in maintaining the structure of water pathways through the root. In roots lacking epidermis, ammonia did not inhibit water uptake by the roots. This may indicate that the site of the inhibition lies within the root epidermis.     

Atomic force microscopy of tomato and sugar beet pectin molecules

Atomic force microscopy has been used to image the structure of pectin molecules isolated from unripe tomato and sugar beet tissue. The tomato pectin molecules were found to be extended stiff chains with a weight average contour length of L_W = 174 nm and a number average contour length of L_N = 132 nm (L_W/L_N = 1.32). A proportion of the pectin molecules (30%) were found to be branched structures. Chemical analysis of the sugar beet pectin extracts showed that the samples contained protein (8.6%). This protein proved difficult to remove and is believed to be covalently attached to the polysaccharide. Imaging of the extracted pectin revealed largely un-aggregated chains: a small fraction (33%) of which were extended stiff polysaccharide chains and a major fraction (67%) of which were of polysaccharide–protein complexes containing a single protein molecule attached to one end of the polysaccharide chains (‘tadpoles’). In addition the sample contained a small number of aggregated structures. The un-aggregated pectin molecules were found to be predominately linear structures with a small fraction (17%) of branched structures. The branched structures were all in the free polysaccharide fraction and no branched pectin chains were observed in the protein–polysaccharide complexes. Alkali treatment was found to remove the protein. For the alkali-treated, un-aggregated structures the average contour lengths were found to be L_W = 137 nm, L_N = 108 nm with L_W/L_N = 1.27. It is proposed that the ‘tadpole’ structures contribute to the unusual emulsifying properties of sugar beet pectin.     

Stereoselective degradation of benalaxyl in tomato, tobacco, sugar beet, capsicum, and soil

The stereoselective degradation of the racemic benalaxyl in vegetables such as tomato, tobacco, sugar beet, capsicum, and the soil has been investigated. The two enantiomers of benalaxyl in the matrix were extracted by organic solvent and determined by validated chiral high-performance liquid chromatography with a cellulose-tris-(3, 5-dimethylphenylcarbamate)-based chiral column. Rac-benalaxyl was fortified into the soil and foliar applied to vegetables. The assay method was linear over a range of concentrations (0.5-50 microg ml(-1)) and the mean recoveries in all the samples were more than 70% for the two enantiomers. The limit of detection for both enantiomers was 0.05 microg g(-1). The results in soil showed that R-(-)-enantiomer dissipated faster than S-(+)-enantiomer and the stereoselectivity might be caused by microorganisms. In tomato, tobacco, sugar, beet, and capsicum plants, there was significantly stereoselective metabolism. The preferential absorption and degradation of S-(+)-enantiomer resulted an enrichment of the R-(-)-enantiomer residue in all the vegetables.     

Natural infection of sugar beet with tomato bushy stunt virus

A virus transmissible toChenopodium quinoa was isolated from leaves of sugar beet showing large chlorotic ring spots and line pattern. The virus was serologically unrelated to tobacco necrosis virus and tomato black ring virus or to its beet ringspot strain either. A positive result was obtained with antiserum against tomato bushy stunt virus. Reactions of herbaceous indicators and properties of the virus in crude sap were in accordance with the serological diagnosis. A survey of natural hosts of tomato bushy stunt virus demonstrated recently by the authors is given.     

Relationship between EUF-K contents,K uptake and sugar yield of sugar beet in deep loess soils

Summary In field experiments with varying K fertilization (1981 and 1982) changes in EUF-K contents were studied in deep loess soils of Southern Lower Saxony under sugar beet. A significant positive linear relationship was found between EUF-K contents at 20°C and 200 V (15 mA) of the topsoils and quantities of K absorbed by sugar beet in both years. The corresponding regression lines for 1981 and 1982 are almost parallel, the only difference being the yield level which was higher in 1982.The relationship between EUF-K contents at 20°C of topsoils and sugar yields showed the same parallelism for the two years. Not much increase in sugar yield was found at EUF-K contents over 12 mg/100 g soil at EUF-K 80°C/EUF-K 20°C ratios between 0.5 and 0.7. To attain a sugar yield of 10 t/ha an EUF-K 20°C value of at least 12 mg/100 g soil is required for these deep soils at the beginning of the K uptake period. This finding confirms experiences gained over an 8-year period at the Tulln Sugar Factory (Austria) with fertilizer recommendations based on EUF.     

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     

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.     

Effect of plant hormones on sucrose uptake by sugar beet root tissue discs

Abscisic acid (ABA), auxins, cytokinins, gibberellic acid, alone or in combination were tested for their effects on short-term sucrose uptake in sugar beet (Beta vulgaris cv USH-20) roots. The effect of ABA on active sucrose uptake varied from no effect to the more generally observed 1.4-to 3.0-fold stimulation. A racemic mixture of ABA and its trans isomer were more stimulatory than ABA alone. Pretreating and/or simultaneously treating the tissue with K⁺ or IAA prevented the ABA response while cytokinins and gibberellic acid did not. While the variable sensitivities of beet root to ABA may somehow be related to the auxin and alkali cation status of the tissue, tissue sensitivity to ABA was not correlated with ABA uptake, accumulation, or metabolic patterns. In contrast to ABA, indoleacetic acid (IAA) and other auxins strongly inhibited active sucrose uptake in beet roots. Cytokinins enhanced the auxin-induced inhibition of sucrose uptake but ABA and gibberellic acid did not modify or counteract the auxin effect. Trans-zeatin, benzyladenine, kinetin, and gibberellins had no effect on active sucrose uptake. None of the hormones or hormone mixtures tested had any significant effect on passive sucrose uptake. The effects of IAA and ABA on sucrose uptake were detectable within 1 h suggesting a rather close relationship between the physiological activities of IAA and ABA and the operation of the active transport system.     

Effect of irrigation frequency on root water uptake in sugar beet

A 2-year trial was performed on autumn-sown sugar beet grown in pots in order to study the influence of irrigation frequency on the water used by plants along the soil profile. The outdoor pots, containing one plant each, were 1.3 m high and had circular openings, through which Time Domain Reflectometry (TDR) apparatus wave guides could be inserted. Three irrigation intervals were compared and plants were watered whenever the soil layer explored by roots had lost 30% (SWD₁), 50% (SWD₂) and 70% (SWD₃) of the total available water (TAW). During the irrigation season, the water extracted by the plants from each layer along the soil profile (RWU) was determined by monitoring volumetric soil moisture content (), by TDR. At harvest time, root length density (RLD) along the soil profile was assessed using the Tennant method. The applied irrigation frequencies significantly affected the RWU. With the SWD₃ protocol, irrigation was at longer irrigation intervals (9 days) and watering volumes were as high as 84 mm. In this treatment, the plants lost almost 60% of total water from the lower soil layer (0.6–1.0 m). In treatment SWD₁, the irrigation interval was very short (3 days), and water extraction from 0.0–0.6 m soil depth was 92.0%. In the intermediate treatment, the irrigation interval was 5.5 days and a more uniform water depletion was observed along the root zone, approximately equal between the 0–0.6 and 0.6–1.0 m soil layer. Water extraction of sugar beet plants at the deeper soil layers in response to long irrigation intervals was related to an increase in water uptake efficiency of the deeper younger roots and not to an increase in root length density, which, on the contrary, decreased. This morpho-physiological acclimatization to progressive soil water deficit was coupled with an increase of the root/shoot ratio.     

Specific uptake of 6-deoxy-d-glucose and of 2-aminoisobutyric acid by sugar beet leaves and roots

Sugar beet was grown in pots and the transport of 6-deoxy-d-glucose (6-dg) and of 2-aminoisobutyric acid (AIB) into leaf and root tissue segments was examined. The uptake of 6-dg into both tissues displayed two kinetic components, one apparently of diffusional nature, the other saturable (half-saturation constant 0.22 mM in leaves, 0.10 mM in roots; limiting rate 0.37 and 0.10 nmol mg-1 (fresh mass) h-1 for leaves and roots, respectively). It was suppressed by d-glucose and inhibited by diethyl-stilbestrol (DES), suloctidil (SUL) and 2,4-dinitrophenol (DNP), but little by D2O and not at all by sodium vanadate. It showed a temperature optimum at 30 °C and a pH optimum at 6.0 - 6.5 in both tissues. The uptake of AIB was also two-componental but even the specific component had an immeasurably high half-saturation constant although it was inhibited by high concentrations of AIB as well as by DES, SUL, DNP and D2O. However, vanadate stimulated the uptake by as much as 40% in both leaves and roots. The temperature optimum in leaves lay at 30 °C while in the roots no maximum was present; its pH optimum was at 5.5 in leaves and again no maximum was observed in roots. Both transport systems were unaffected by 0.1M NaCl or 0.1 M KCl and by the presence of cytokinin in the incubation vessel. However, spraying with the cytokinin N6-(m-hydroxybenzyl) adenosine decreased both the half-saturation constant and the limiting velocity of 6-dg uptake slightly in the leaves and substantially in the roots. Spraying had an insignificant effect on the uptake of AIB. This revised version was published online in July 2006 with corrections to the Cover Date.     

Mutational studies with diquat and paraquat in vitro.

Diquat and paraquat were assayed in the following tests. (1) Ames test in Salmonella typhimurium (strains TA1535, TA1537, TA1538, TA98 and TA100) with and without rat-liver microsomal fractions. (2) Resistance to 8-azaguanine in Salmonella typhimurium (strain hisG46, TA92 and TA1535. (3) Repair test in Salmonella typhimurium (strains TA1538 and TA1978). (4) Gene mutations in Aspergillus nidulans: 8-AG resistance and methionine suppression (meth A1 locus). (5) Lethal recessive damage in Aspergillus nidulans. (6) Unscheduled DNA synthesis (UDS) in human epithelial-like cells (EUE). Diquat and paraquat were positive in S. typhimurium (in the repair test and the 8-AG resistance system), in A. nidulans (for gene mutations and lethal recessive damage induction) and in EUE cells (UDS induction).     

The action of paraquat and diquat on the respiration of liver cell fractions

1. Paraquat and diquat produce only a slight increase in the oxygen uptake of rat liver mitochondria, and it is likely that they do not penetrate the mitochondrial membrane. 2. In mitochondrial fragments inhibited by antimycin A or by Amytal, both substances stimulate oxygen uptake with NADH or beta-hydroxybutyrate as substrate but not with succinate. The NADH dehydrogenase of the respiratory chain appears to be involved, at a site only partially inhibited by Amytal. 3. An NADPH oxidase activity is stimulated in rat liver microsomes by diquat, and to a smaller extent by paraquat; diquat also causes an NADH oxidase activity to develop. The effect is not inhibited by carbon monoxide or p-chloromercuribenzoate, and it is probable that a flavoprotein is involved by a mechanism not requiring thiol groups. 4. One molecule of oxygen can oxidize two molecules of NADPH in the stimulated microsomal system, the hydrogen peroxide produced being broken down by a catalase activity in the microsomes. 5. Diquat can stimulate NADH oxidase and NADPH oxidase activity in the postmicrosomal soluble fraction; the enzyme involved may be DT-diaphorase. 6. The mechanism of these reactions and their significance in relation to the toxicity of the dipyridilium compounds are discussed.     

Invertase inhibitors from red beet, sugar beet, and sweet potato roots

Invertase inhibitors have been isolated and partially purified from red beets, sugar beets, and sweet potatoes. These inhibitors are thermolabile proteins with molecular weights of 18,000 to 23,000. They do not inhibit yeast and Neurospora invertases, but they are reactive with potato tuber invertase and other plant invertases with pH optima near 4.5. There are differences in reactivity of the inhibitors with some of the plant invertases, however. For most invertases, red beet and sugar beet inhibitors are most effective at pH 4.5 while sweet potato inhibitor is most effective at pH 5.     

Foliar uptake of Cd by pea (Pisum sativum) and sugar beet (Beta vulgaris)

Pea ( Pisum sativum L. cv. Fenomen) and sugar beet ( Beta vulgaris L. cv. Monohill) were cultivated in nutrient media without or with 10 μM CdCl₂. Leaves of the same size and stage of development, detached or still attached to the intact plants, were submerged into redistilled water containing 1 to 250 μM CdCl₂. The uptake experiments were run for 1 to 8 h at pH 3.6 and 5.1. Cuticular transpiration rate, density of leaf and density of stomata were also measured. Percentage of open stomata was studied at different pH.
Foliar uptake of Cd into the leaf is evident since Cd is transported from the exposed part of the pea leaves, through the petioles and into the stipules, and since the Cd concentration of the leaves increases with time and external Cd concentration. The foliar uptake depends on the permeability of the cuticular membrane, which is increased by a high intrinsic Cd level, which in turn enhances the foliar uptake of Cd in sugar beet. Higher cuticular permeability in pea than in sugar beet is shown by a 2.5 times higher cuticular transpiration rate and a 4 times lower density of leaf for pea, which causes a 7 times higher foliar uptake in pea than in sugar beet. Low pH decreases the net uptake of Cd, probably by an exchange reaction in the cutin and pectin of the cuticular membrane. Stomata are not directly involved in the Cd uptake, and the differences in the sum total of stomatal aperture area per unit leaf area is not related to differences in foliar uptake of Cd. Percentage of open stomata, calculated as average of both sides of the leaves, was not affected by changes in pH: but especially at high pH. proportionally more stomata were open on the adaxial than on the abaxial side.