3-Dehydroshikimate hydro-lyase ofPhaseolus mungo seedlings

3-Dehydroshikimate hydro-lyase was extracted and partially purified by ammonium sulfate fractionation from the hypocotyls of etiolatedPhaseolus mungo seedings. The enzyme was most active at pH 7.0 and the Km for DHS was 1.4 mM. Enzyme activity was inhibited byp-chloromercuribenzoate and arsenite, the inhibition being reversed by reduced glutathione. Enzyme activity was found in extracts from young hypocotyls and cotyledons, especially from 5–6 day-old hypocotyls and 3–4 day-old cotyledons. Changes in enzyme activity in both cotyledons and hypocotyls were measured during the early stage of germination. Participation of this enzyme in the catabolism of alicyclic acids is suggested.     

Identification of cytokinins present in root nodules ofPhaseolus mungo L

Root nodules ofPhaseolus mungo were extracted for their cytokinin content. Four cytokininsviz. zeatin, zeatin riboside, N⁶(‡²-isopentenyl) aminopurine and its riboside were identified on the basis of Rf values obtained by co-chromatography with authentic samples of cytokinins on thin layer of a mixture of polyvinylpyrrolidone/CaSO₄.     

Studies on pachytene and somatic chromosomes ofPhaseolus mungo L.

The pachytene and somatic chromosomes ofPhaseolus mungo L. (2n=22), were identified and classified on the basis of their relative length, arm ratio, chromomere pattern and nucleolar association. A comparison of the karyotypes during the somatic and pachytene stages, revealed that some chromosomes are of different relative lengths during the two phases. Thus, the two nucleolar organizer chromosomes are the 2nd and 3rd longest at somatic metaphase, but only 7th and 9th longest during pachytene. The pachytene chromosomes belong to the differentiated group.     

Alicyclic acid metabolism in plants 4. Effect of external supplies of shikimate and quinate to excised hypocotyls of Phaseolus mungo seedlings

When excised hypocotyls of etiolated Phaseolus mungo seedlingswere allowed to take up shikimate solutions of various concentrationsfor 4 to 32 hr at 25°C in the dark, a marked rise in endogenousquinic acid level appeared in the organ. A similar effect wasobserved, to a lesser extent, in other organs, i.e. roots andcotyledons. When quinate solution was supplied to hypocotyls,the effect on the increase of endogenous shikimic acid concentrationwas discernible, although it was not as remarkable as the profoundeffect of shikimate on quinic acid level. A supply of eitherof the acids to excised hypocotyls in a culture solution resultedin a significant increase in dehydroquinate dehydratase activityin the organ, whereas shikimate dehydrogenase activity was slightly,if at all, affected by this treatment. On the basis of theseexperimental results, the metabolic role of quinic acid in aromaticbiosynthesis in this plant is discussed. (Received February 26, 1972; )     

Glucose catabolism during alkaloid production in a strain ofClaviceps purpurea

Badiorespirometric experiments with glucose labelled in positions 1; 2; 3,4 and 6 were carried out inClaviceps purpurea strain pepty 695. The pattern of¹⁴CO₂ evolution from 5 – 50 mM glucose indicated the operation of the pentose phosphate cycle during alkaloid production. The authors thank Mrs. H. Pechfelder for her skilled technical assistance and Prof. Dr. H. Reinbothe and Dr. C. Wasternack for helpful discussions.     

Glucose catabolism during germination of Bacillus megaterium spores.

When heat-activated spores of Bacillus megaterium germinated in glucose-containing medium, 10 to 30% of the glucose was found to be oxidized to gluconate.     

Variations in the catabolism of the acetylcholine receptor during aging

The time and modalities of degradation of the muscles acetylcholine receptor have been studied by several investigator, both in vivo and in vitro and under normal and pathological condition. The metod employed in this study for assessment of possible changes in the degradation to age involves the valutation of several parameters. These are explained and discussed. The preliminary results show the turnover of the receptor for junction acetylcholine, which is known to be considerably accellerated in conditions such denervation and myastenia, is slowed down during senescence.     

Effect of maleic hydrazide on peroxidase isoenzymes in relation to rooting hypocotyl cuttings ofPhaseolus mungo

A time course study of changes in the pattern of peroxidase isoenzymes shows that two new isoenzymesa andb appeared in hypocotyl cuttings cultured either in indolyl-3 acetic acid (IAA) + sucrose or in water but not in maleic hydrazide (MH). Roots were also initiated in the former two cases but not in the last case. That these isoenzymes may be associated with root initiation is also evident from the fact that these appeared when hypocotyl cuttings were transferred from MH to IAA + sucrose. The time of appearance of these isoenzymes in different cultures also synchronized with the time of microscopic root initiation. While the isoenzymea disappeared,b persisted in cuttings transferred from IAA + sucrose to MH. The number of roots produced on cuttings transferred to MH was very low, though their development was normal. It is suggested that isoenzymesa andb may be associated with root initiation andc andd with root development.     

Pattern of glucose-14C catabolism in barley seedlings during vernalization

The pattern of glucose catabolism in barley shoots during vernalizationwas examined using glucose-1-¹⁴C and glucose-6-¹⁴C. The pentosephosphate pathway became progressively predominant over theEmbden-Meyerhof-Parnas (EMP) pathway after germination in unvernalizedseedlings, while the EMP pathway maintained a superior positionduring the corresponding developmental stages in seedlings undervernalization. (Received August 20, 1973; )     

NaCl and wounding induced changes in NAD reductase in hypocotyls and root tips ofPhaseolus vulgaris L.

The effect of 300 mM NaCl and wounding on the nicotinamide adenine dinucleotide (NAD) kinase, revealed by the guanosine triphosphate-dependent NAD reductase activity, was studied in two differently resistant bean cultivars using densitometric analysis of electrophoretic gels. In the presence of NaCl the total activity of NAD reductase was increased, in hypocotyls and root tips of resistant cultivar. The contribution of each of NAD reductase isoforms to the total activity was not significantly different between cultivars. Conversely, after wounding the hypocotyl, an increase could be observed in both cultivars and differences were demonstrated in the contribution of the different isoforms.     

b-Type cytochromes in plasma membranes ofPhaseolus vulgaris hypocotyls,Arabidopsis thaliana leaves, andZea mays roots

Summary The plasma membrane of higher plants contains more than one kind ofb-type cytochromes. One of these has a high redox potential and can be fully reduced by ascorbate. This component, the cytochromeb ₅₆₁ (cytb ₅₆₁), has its characteristic -band absorbance close to 561 nm wavelength at room temperature. Cytb ₅₆₁ was first isolated from etiolated bean hook plasma membranes by two consecutive anion exchange chromatography steps. During the first step performed at pH 8, cytb ₅₆₁ did not bind to the anion exchange column, but otherb-type cytochromes did. In the second step performed at pH 9.9, cytb ₅₆₁ was bound to the column and was eluted from the column at an ionic strength of about 100 mM KCl. However, when the same protocol was applied to the solubilized plasma membrane proteins fromArabidopsis thaliana leaves and maize roots, the ascorbate-reducible cytb ₅₆₁ bound already to the first anion exchange column at pH 8 and was eluted also at an ionic strength of about 100 mM KCl. Otherb-type cytochromes than the ascorbate-reducible cytb ₅₆₁ from the plasma membranes of Arabidopsis leaves and maize roots showed similar Chromatographic characteristics to that of bean hypocotyls. These results demonstrate particular differences in the Chromatographic behavior of cytb ₅₆₁ from different sources.Abbreviations cyt b ₅₆₁ cytochromeb ₅₆₁ - PM plasma membrane - PAGE polyacrylamide gel electrophoresis     

3-Dehydroquinate synthase in germinating Phaseolus mungo seedlings.

Dehydroquinate synthase, an enzyme catalyzing the conversion of 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) to 3-dehydroquinate, was detected in cell-free extracts of etiolated Phaseolus mungo seedlings. The reaction product, 3-dehydroquinate, formed from [1-14C]DAHP was identified by paper-radiochromatography. The enzyme required NAD+ and Co2+ for activity.     

Changing cell-wall compositions in hypocotyls of dark-grown bean seedlings

Hypocotyls of dark-grown 6-day-old seedlings of Phaseolus vulgaris L. proved to be sufficiently homogeneous to permit studies relating the rate of cell elongation to the composition of the primary cell walls. Whereas the levels of cellulose and uronic acids remained practically constant during and after cell extension, all other components showed major or minor changes. Cell-wall protein, as such, decreased by more than 50%, but indications are that hydroxyproline-rich glycoprotein increased with a decreasing rate of cell elongation, concomitant with a rise in the degree of arabinosylation of wall-bound hydroxyproline. As cell elongation slowed down, non-cellulosic glucose accumulated, presumably in the form of a -(1–4)glucan closely associated with cellulose. These findings confirm the notion that the primary cell wall is a highly dynamic structure.     

Polyamine catabolism is involved in response to salt stress in soybean hypocotyls

The possible relationship between polyamine catabolism mediated by copper-containing amine oxidase and the elongation of soybean hypocotyls from plants exposed to NaCl has been studied. Salt treatment reduced values of all hypocotyl growth parameters. In vitro, copper-containing amine oxidase activity was up to 77-fold higher than that of polyamine oxidase. This enzyme preferred cadaverine over putrescine and it was active even under the saline condition. On the other hand, saline stress increased spermine and cadaverine levels, and the in vivo copper-containing amine oxidase activity in the elongation zone of hypocotyls. The last effect was negatively modulated by the addition of the copper-containing amine oxidase inhibitor N,N'-diaminoguanidine. In turn, plants treated with the inhibitor showed a significant reduction of reactive oxygen species in the elongation zone, even in the saline situation. In addition, plants grown in cadaverine-amended culture medium showed increased hypocotyl length either in saline or control conditions and this effect was also abolished by N,N'-diaminoguanidine. Taken together, our results suggest that the activity of the copper-containing amine oxidase may be partially contributing to hypocotyl growth under saline stress, through the production of hydrogen peroxide by polyamine catabolism and reinforce the importance of polyamine catabolism and hydrogen peroxide production in the induction of salt tolerance in plants.     

Glucose and pyruvate metabolism during mouse gonadal sex differentiation

In the mouse, gonadal sex differentiation starts around E12 and meiosis begins in the ovary shortly after E13. In the search for metabolic changes that might be correlated to gonadal sex differentiation and/or possibly the onset of meiosis, we investigated the metabolism of glucose and pyruvate in the developing mouse ovary before (E11.5-E12.5), during (E14.5-16.5), and after meiosis (E18.5), and in fetal testes without meiosis. Gonads were cultured with 14C-labeled glucose (0.02 and 5.58 mM) and 14C-pyruvate (0.17 mM). The oxidation expressed as 14CO2 production and the organification expressed as retention of 14C in the tissues were measured and correlated to the protein content of the gonads. Using 0.02 mM glucose, a decline in oxidation and organification was found in ovaries as well as in testes, which is probably related to starvation. Using 5.58 mM glucose, a continuous decline in oxidation was seen only in the testis. Organification of 0.17 mM pyruvate increased at E12.5 and E14.5 in the ovary but not in the testis. This was in despite of an exponential increase of protein content in the testes compared to only a moderate increase in the ovary. The CO2 production from 5.58 mM glucose was equal to that from 0.17 mM pyruvate in gonads of both sexes. In conclusion, an increased metabolism of 5.58 mM glucose and 0.17 mM pyruvate in the ovaries as compared to the testes is related to sex differences during gonadal formation and onset of meiosis in the ovaries. J. Exp. Zool. 288:130-138, 2001.     

Investigation on the role of nodule cytokinins in regulation of nitrate reductase activity ofPhaseolus mungo (L.)

The effects of some nodular cytokinis, zeatin (Z), zeatin riboside (ZR), N⁶ (Δ²-isopentenyl) adenine (IPA), and N⁶ (Δ²-isopentenyI) adenosine (IPAS) on nitrate reductase (E.C 1.9.6.1) activity of root nodules ofPhaseolus mungo were investigated. The cytokinis were also tested for their effect on nitrate uptake by nodules. The results show that IPAS is the most effective of all the four cytokinins tested. Z and IPA, which caused higherin vivo activity than ZR and IPAS, stimulated uptake of nitrate by nodules. The other two (ZR and IPAS) while inhibiting uptake showed greaterin vitro activity than Z and IPA. It may be concluded that some cytokinins, in addition to their direct effects on the enzyme, may increase the substrate availability to it, whereas others may have only an direct effect on the enzyme activation or degradation.     

Gravitropism of hypocotyls of wild-type and starch-deficient Arabidopsis seedlings in spaceflight studies

The major purpose of this spaceflight project was to investigate the starch-statolith hypothesis for gravity perception, and a secondary goal was to study plant growth and development under spaceflight conditions. This research was based on our ground studies of gravity perception in the wild type and three starch-deficient (one starchless and two reduced starch) mutants of Arabidopsis thaliana (L.) Heynh. Dark-grown seedlings that developed in microgravity were given one of several (30 min, 60 min, or 90 min) 1-g stimuli by an on-board centrifuge, and additional controls for seedling development also were performed. These latter control experiments included a morphological study of plants that developed in space in microgravity (F μg), in space on a centrifuge (F 1g), on the ground (G 1g), and on a rotating clinostat on the ground. Since elevated levels of ethylene were reported in the spacecraft atmosphere, additional controls for morphology and gravitropism with added ethylene also were performed. While exogenous ethylene reduced the absolute magnitude of the response in all four strains of Arabidopsis, this gas did not appear to change the relative graviresponsiveness among the strains. The relative response of hypocotyls of microgravity-grown seedlings to the stimuli provided by the in-flight centrifuge was: wild type > starch-deficient mutants. Although the protoplast pressure model for gravity perception cannot be excluded, these results are consistent with a statolith-based model for perception in plants. Received: 12 February 1999 / Accepted: 9 March 1999     

Organization of enzymes in the shikimate pathway of Phaseolus mungo seedlings

DEAE-cellulose and Sephadex G-100 column chromatography showedthe separability of 3-dehydroquinate synthase, 3-dehydroquinatehydro-lyase-shikimate: NADP oxidoreductase complex, shikimatekinase and 5-enolpyruvylshikimate-3-phosphate synthase of Phaseolusmungo seedlings. The approximate molecular weights of theseenzymes were estimated using a gel filtration column. (Received October 30, 1978; )