Fermentative products and dark CO2 fixation during germination of seeds of Cicer arietinum

During the germination of Cicer arietinum L. the amounts of ethanol, lactate and malate reached their highest values at 24 hr, the concentration of ethanol being about 4 times that of lactate and twice that of malate. The activities of phosphoenolpyruvate carboxylase and malic enzyme seem to be correlated with the ability of cotyledons to fix CO₂ from NaH¹⁴CO₃ into malate and with the further decrease in this metabolise from 36 hr onwards.     

Control of glutamate dehydrogenase from Pisum sativum roots

-Glutamate dehydrogenase (GDH) was found in soluble and particulate (mitochondrial) fractions of pea roots. The activity of NADH-dependent GDH in fresh mitochondrial extract was increased about 10-fold by addition of zinc, manganese or calcium, but high concentrations of zinc were inhibitory. During storage, GDH activity of the mitochondrial extract slowly increased. The NADH activity was inhibited by citrate and other chelating agents. NADH-dependent reductive amination was also inhibited by glutamate, the product of the reaction; by contrast NADPH dependent activity was relatively unaffected by zinc, chelating agents or glutamate. Sensitivity (of NADH-GDH) to glutamate was lost on purification, but was restored when the enzyme was immobilized by binding to an insoluble support (AE cellulose). Glutamate appears to change the affinity of the enzyme for 2-oxoglutarate.     

Interconversion of gibberellin A20 to gibberellin A29 by etiolated seedlings and germinating seeds of dwarf Pisum sativum

Tritium labelled gibberellin A₂₀ ([³H]-GA₂₀) applied to etiolated shoots and germinating seeds of dwarf pea (Pisum sativum L. cv. Meteor) was converted to gibberellin A₂₉. Identifications were made by GLRC and GC-MS.     

De novo synthesis of acetylcholinesterase in roots of Pisum sativum

Pisum sativum seeds contain a conserved acetylcholinesterase (AChE) which is active during the early stages of germination. The enzyme activity soon disappears and reappears after 72 hr of germination. A protein devoid of catalytic ability, but exhibiting similar chromatographic and electrophoretic properties as the active AChE, could be detected after 24 hr of germination. The pattern of incorporation of labelled amino acids into AChE and the influence of cycloheximide revealed that the AChE found in the roots from 72 hr onwards was entirely new. During this period of growth, the AChE protein accounts for 4–10% of the total proteins in the root tissue.     

Isolation and properties of a lectin from the roots of Pisum sativum (Garden pea)

Abstract The roots of pea (Pisum sativum L. ev. Feltham First) seedlings contained haemagglutinating activity and a protein which reacted with antibodies directed against pea seed lectin. This protein was shown to be present on the surface of root hairs and in the root cortical cells by immunofluorescence. Lectin (haemagglutinin) was purified from pea seedling roots by both immunoaffinity chromatography and affinity chromatography on Sephadex G-100. The pea root lectin was similar to the seed lectin when analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, and was antigenically identical: however, the isoelectric focussing band patterns of the proteins differed. The sugar specificity of the root lectin differed from that of the seed lectin, and the haemagglutinating activity of the root lectin was less than the seed lectin. These results are discussed with reference to the hypothesis that lectins mediate in the symbiotic association of legume and Rhizobium through their carbohydrate-binding properties.     

Effects of anaerobiosis on carbohydrate oxidation by roots of Pisum sativum

The aim of this work was to discover the effects of anaerobiosis on the breakdown of sugars by the apical 6 mm of the roots of 5-day-old seedlings of Pisum sativum. Estimates of the maximum catalytic activities of alcohol dehydrogenase, lactate dehydrogenase, phosphoenolpyruvate carboxylase and NADP-specific malic enzyme showed them to be comparable to that of phosphofructokinase. Metabolism of sucrose-[U-¹⁴C] by excised apices was restricted by anoxia mainly to conversion to ethanol, CO₂ alanine and glycolytic intermediates. Measurements of metabolites over a period of 240 min after transfer of excised apices to nitrogen showed a marked and continual accumulation of ethanol, a smaller continual accumulation of alanine, a small initial rise in lactate and no detectable accumulation of malate or pyruvate. The rates of CO₂ production, of accumulation of ethanol and alanine, and of the labelling of these compounds by sucrose-[¹⁴C] declined markedly during the first 240 min of anaerobiosis. The conclusion is that under anaerobic conditions carbohydrate metabolism in the pea root apex is largely restricted to alcoholic fermentation, and, to a lesser degree, to alanine production.     

Sucrose metabolism by roots of Pisum sativum

The aim of this work was to relate [¹⁴C]-sucrose metabolism to the activities of sucrose synthetase and acid and alkaline invertases in roots of Pisum sativum. We fed [U-¹⁴C]-sucrose to 5-day-old plants and then excised the apical 6 mm of the roots and dissected the regions 6–24 mm from the root apices into stele and cortex. The detailed distribution of ¹⁴C in these parts of the roots was determined at the end of the feeding period and after a chase. The data indicate that sucrose arriving in the stele is divided between storage, conversion to polysaccharide, and consumption in respiration, whereas sucrose arriving in the rest of the root is used in respiration or converted to polysaccharide or hexose so rapidly that little is stored. Fractionation of carefully prepared extracts of pea roots, tubers of Solanum tuberosum, and spadices of Arum maculatum showed that sucrose synthetase was recovered in the soluble fraction. The results are discussed in relation to the roles of the aforementioned enzymes.     

Conversion of gibberellin A14 to other gibberellins in seedlings of dwarf Pisum sativum

Gibberellin A₁₄-[17-³H] applied to seedlings of dark grown dwarf pea (Pisum sativum L. cy. Meteor) was converted to GA₁, GA₈, GA₁₈, GA₂₃, GA₂₈, and GA₃₈. The sequence of interconversion of GA₁₄→ GA₁₈ → GA₃₈ → GA₂₃ → GA₁ → GA₈ is indicated. Identifications were made by gas-liquid radiochromatography using three liquid stationary phases.     

DNA polymerase activity and DNA synthesis in roots of pea (Pisum sativum) seedlings

Soluble DNA polymerase (DNA polymerase-α) and chromatin-bound DNA polymerase (DNA polymerase-β) have been assayed in serial sections cut from the roots of 5-day-old pea seedlings. The activity of DNA polymerase-α is high in regions of the root which exhibit high rates of DNA replication, and declines during cell differentiation and maturation. The activity of DNA polyrnerase-β is low in cells which show high rates of DNA replication, and increases during differentiation and maturation.     

CO2 dark fixation in the halophytic species mesembryanthemum crystallinum

The time course of ¹⁴CO₂ dark fixation was studied in leaves of the facultatively halophytic plant species Mesembryanthemum crystallinum cultivated with and without 400 mM NaCl in the nutrient medium. It is generally known from the literature that plants grown under saline conditions incorporate ¹⁴C predominately into amino acids. By contrast in leaves of M. crystallinum grown on NaCl and exposed to ¹⁴CO₂ in the dark, relatively more radioactivity is incorporated in the organic acids (especially malate) than in amino acids. The data obtained are discussed in relation to the NaCl induced Crassulacean acid metabolism in M. crystallinum reported earlier.     

Purification and specificity of RNase A3 from Vicia faba root cells

Vicia faba root ribonucleases are bound to Cibacron blue F3GA. A Blue dextran-Sepharose column was used to purify RNase A₃, the more abundant enzyme from V. faba root. Using dinucleoside monophosphate as substrates, it appears that this enzyme behaves as a cyclizing phosphotransferase. With high enzyme/substrate ratios on prolonged digestion a partial release of a nucleoside 3′ phosphate occurs. The specificity is relatively high since only the purine-purine phosphodiester linkages out of 16 types of possible links are easily cleaved. When a pyrimidine is involved in the phosphodiester bond, a much slower rate of attack (Py in 5′) or no attack (Py in 3′) was detected.     

Effects of anoxia and flooding on alcohol dehydrogenase in roots of Glyceria maxima and Pisum sativum

Flood tolerant Glyceria maxima and intolerant Pisum sativum were compared in respect of the effects of anoxia and flooding on the maximum catalytic activities of alcohol dehydrogenase in their roots. Small (<73%) increases in enzyme activity occurred when excised roots of both species were incubated in nitrogen for up to 2 days. Further incubation in nitrogen rapidly and permanently damaged the roots of both species. Enzyme activity in flooded roots of Glyceria was about double that in corresponding non-flooded roots. A marginally greater difference was found for roots of Pisum. It was concluded that the two species respond so similarly to the above treatments that variation in the extent of induction of alcohol dehydrogenase is unlikely to be a significant factor in determining their ability to tolerate flooding.     

Effect of number and configuration of fruits, photon flux and age on the growth and dry matter distribution of fruits of Pisum sativum L.

Abstract. In experiments with Pisum sativum cv. Sleaford Orbiter in a controlled environment, the effect of fruit number and position, photon flux density and developmental stage on fruit growth was studied. During early development (up to 22 d from anthesis) growth of the first fruit was unaffected by the presence of one or two additional fruits irrespective of their position. When grown to maturity in competition with fruits at the same node a small decrease in weight of this fruit was observed. Where plants retained a full complement (20-30) of fruits the growth of the first fruit was markedly decreased at all stages of development (6-40 d). In all instances where competition was observed, the pericarp was more affected than the seeds. This was particularly so when photon flux was decreased 18-22 d from anthesis compared with a decrease at an earlier stage. Partition of dry matter between fruits showed a progressively increasing allocation to the later-formed fruits with time for all treatments. The actual proportions allocated to different fruits were not changed by the number of competing fruits. Decreasing photon flux by more than two-thirds decreased fruit growth rates but had little effect on dry matter partitioning in most cases, although where all fruits were retained, there was a tendency for fruits at the lower reproductive nodes to be less affected. These findings are discussed in relation to known sources of assimilate for fruits, assimilate transport and sink demand. It is suggested that partition of dry matter between fruits can be estimated on the basis of fruit size and the developmental trend in relative growth rate of fruits grown in the absence of competition for assimilate from other fruits.     

Xylogliicae- and cello-oligosaccharides: Antagonists of the growth-promoting effect of H+

Xyloglucan-oligosaccharides and cello-oligosaccharides, both of which are potential products of the action of cellulase on plant cell wail polysaccharides, inhibited acid-induced elongation in pea ( Pisum sativum L. cv. Alaska) stem segments. Xyloglucan-derived nonasaccharide (XG9; Glc₄-Xyl₃)Gal-Fuc) and decasaccharide (XG10; Glc₄-Xyl₃-Gal₂-Fue) inhibited acid-induced growth at 1.0 and 0.1 n M , respectively, whereas the heptasaccharide (XG7; Glc₄-Xyl₃) and octasaccharide (XG8; Glc₄-Xyl₃-Gal)₂ which lack L-fucose, did not. XG9 at 1 n M inhibited acid-induced growth as effectively as it inhibits auxin-induced elongation. This suggests that XG9's effect as an inhibitor of auxin action is not mediated by a suppresion of H⁺-efflux, but rather that XG9 blocks some step that is common to the action of both auxin and H⁺ on growth. Cello-oligosaccharides (degree of polymerisation 4–7) also inhibited acid-induced growth at 10 n M ; these are therefore a new class of possible oligosaccha-rin. The inhibitory effect of xyloglucan- and cellooligosaccharides on acid-induced growth was rapidly reversed by washing.     

Carbon and nitrogen partitioning in young nodulated pea (wild type and nitrate reductase-deficient mutant) plants exposed to NH4NO3

Carbon and nitrogen partitioning was examined in a wild-type and a nitrate reductase-deficient mutant (A317) of Pisum sativum L. (ev. Juneau), effectively inoculated with two strains of Rhizobium leguminosarum (128C23 and 128C54) and grown hydroponically in medium without nitrogen for 21 days, followed by a further 7 days in medium without and with 5 mM NH₄NO₃. In wild-type symbioses the application of NH₄NO₃ significantly reduced nodule growth, nitrogenase (EC 1.7.99.2) activity, nodule carbohydrates (soluble sugars and starch) and allocation of [¹⁴C]-labelled (NO₃⁻, NH₄⁺, amino acids) in roots. In nodules, there was a decline in amino acids together with an increase in inorganic nitrogen concentration. In contrast, symbioses involving A317 exhibited no change in nitrogenase activity or nodule carbohydrates, and the concentrations of all nitrogenous solutes measured (including asparagine) in roots and nodules were enhanced. Photosynthate allocation to the nodule was reduced in the 128C23 symbiosis. Nitrite accumulation was not detected in any case. These data cannot be wholly explained by either the carbohydrate deprivation hypothesis or the nitrite hypothesis for the inhibition of symbiotic nitrogen fixation by combined nitrogen. Our result with A317 also provided evidence against the hypothesis that NO₃⁻ and NH₄⁺ or its assimilation products exert a direct effect on nitrogenase activity. It is concluded that more than one legume host and Rhizobium strain must be studied before generalizations about Rhizobium /legume interactions are made.     

Complete mechanical impedance increases the turgor of cells in the apex of pea roots

In this paper we describe an experimental approach which allows turgor (p) in an impeded root to be measured without the need to remove the root from the impeding environment. The maximum axial growth pressure (σ_max) generated by completely impeded pea (Pisum sativum L.) roots was measured using a novel apparatus incorporating a force transducer. The apparatus was designed so that it was possible to gain access to the impeded root with the microcapillary of a pressure probe and so obtain in situ measurements of P. Turgor in cells in the apical region of impeded roots was 0.78 MPa, compared with 0.55 MPa in unimpeded roots. In impeded roots, σ_max was 0.52 MPa, showing that the pressure component resulting from cell wall tension (W, where W=P–σ) decreased from 0.55 to 0.26 MPa as the roots became impeded. When impeded roots were removed from the apparatus, there was no decrease in P over the following 90 min. Impedance did not cause P to change in the non-elongating part of the roots further from the apex.     

Interconversion of gibberellin A5 to gibberellin 3 in seedlings of dwarf Pisum sativum

[³H]-Gibberellin A₅ ([³H]-GA₅) applied to seedlings of dark-grown dwarf pea (Pisum sativum L. cv. Meteor), was converted to two acidic compounds, GA₃ and a chromatographically similar unknown. Identification of GA₃ was made by gas-liquid radiochromatography using three stationary phases.     

Low temperature spectrophotometry of the phototransformation of Pfr to Pr in pelletable pea phytochrome

Manabe, K. 1987. Low temperature spectrophotometry of the phototransformation of P_fr to P_r, in pelletable pea phytochrome.
Low temperature spectrophotometry was used to study the phototransformation of P_fr to P_r in 1000–7000 g pelletable fractions extracted from dark grown pea ( Pisum sativum L. cv. Alaska) epicotyls which had been irradiated with red and then far-red light. At -170°C, far-red irradiation of the pelletable phytochrome which had been pre-irradiated with saturating fluence of red light before freezing caused formation of an intermediate (named I₆₆₀), the difference spectrum of which showed a marked ab-sorbance decrease at 740 nm and a concomitant small increase at about 660 nm. The inermediate I₆₆₀ was converted to another intermediate (I₆₆₀) when it was warmed above -80°C. The difference spectrum of this intermediate showed a positive peak at 670 nm. This intermediate was photoconverted to P_fr by red irradiation and also underwent dark reversion to P_fr at -60°C. I₆₆₀ formed P_r if the temperature was above -10°C. The basic features of the phytochrome intermediates resemble those obtained in vivo and in degraded purified phytochrome.     

Homoserine dehydrogenase in Pisum sativum and Ricinus communis

Homoserine dehydrogenase was extracted from Ricinus communis and Pisum sativum. The kinetic parameters of the forward and reverse reactions were determined. In the forward reaction only the enzyme from Ricinus is inhibited by threonine. The response to K⁺ is different for the enzyme from the two sources.     

14CO2 dark fixation in the halophytic species mesembryanthemum crystallinum

The time course of ¹⁴CO₂ dark fixation was studied in leaves of the facultatively halophytic plant species Mesembryanthemum crystallinum cultivated with and without 400 mM NaCl in the nutrient medium. It is generally known from the literature that plants grown under saline conditions incorporate ¹⁴C predominately into amino acids. By contrast in leaves of M. crystallinum grown on NaCl and exposed to ¹⁴CO₂ in the dark, relatively more radioactivity is incorporated in the organic acids (especially malate) than in amino acids. The data obtained are discussed in relation to the NaCl induced Crassulacean acid metabolism in M. crystallinum reported earlier.