A Population Study to Aid the Selection of Improved Dried Pea (Pisum sativum) Crop Plants

The effect of genotype and plant density, over the range from100 to 277 plants m^–2, on plant to plant variation inprecision sown microplots has been assessed for three ‘leafless’(afafstst) pea (Pisum sativum) lines. This range of plantingdensities did not significantly affect the total above groundbiological yield per unit area of two of the genotypes (BS5and BS4) whereas the biological yield of the third (BS151) declinedat densities above 156 plants m^–2. The differences weredue to changes in seed yield. The effect of planting densityon the variation between plants for biological yield withinthe microplots differed between the genotypes. The distributionpattern of BS4 and BS5 changed from normal to skewed with increaseddensity, while the distribution for BS151 remained skewed atall planting densities. The differences between the three genotypes in the proportionof biological yield partitioned into seed yield (harvest index)on a unit area basis was due almost entirely to the differencesin structure of the plant populations. The maximum level ofpartitioning by individual plants was similar for all threelines. The difference between this maximum for an individualand the crop harvest index therefore represents the area forimprovement of crop harvest index through breeding. It is suggestedthat improvements in dried pea yields will come, therefore,by selecting plants which form more uniform populations withregard to plant size and to the proportion of plant biomasspartitioned into seed (plant harvest index). Pisum sativum, leafless pea, population, genetic variation, distribution patterns, harvest index     

Turnover of Storage Protein in Seeds of Soya Bean and Pea

We were interested in determining whether the low protein contentof pea seeds (Pisum sativum L.) as compared to soya bean seeds(Glycine max L. Merrill) might be due to faster degradationof the pea storage proteins during development of the seed.Pea and soya bean cotyledons were subjected to a ‘pulse-chase’experiment using [³H]glycine in in-vitro cultures. In peas,legumin had a half-life of 146 days, while vicilin had a half-lifeof 39 days. There was no measureable degradation of soya beanstorage proteins. Even with the pea storage proteins, the half-liveswere so much longer than the maturation time of seeds that degradationof storage proteins could not account for the lower proteincontent of peas as compared to soya beans. The validity of theseresults was indicated by the finding that non-storage proteinshad much shorter half-lives and that omission of a carbon ora nitrogen source greatly accelerated degradation. Labelledglycine was found to be a good probe for protein turnover studiesbecause it was very rapidly metabolized. Glycine max L. Merrill, soya bean, Pisum sativum, L. pea, protein turnover, storage proteins, legumin, vicilin     

The Effect of Paclobutrazol on Physiological and Biochemical Changes in the Primary Roots of Pea

Primary roots of pea (Pisum sativum L. cv. Taichung No. 11)were treated with 0, 10 and 50 mg dm^–3 paclobutrazol [(2RS,3RS)-1-(4-chlorophenyl)-4, 4-dimethyl-2-(l,2,4-triazol-l-yl)pentan-3-ol]for 1 h at 48 h after germination. Paclobutrazol treatment inhibitedroot extension, promoted swelling (cell expansion was radialrather than longitudinal), and increased cell volume and theactivity of catalase and peroxidase enzymes. Paclobutrazol alsodecreased root respiration and ethylene production. However,under non-stressed conditions, paclobutrazol treatment did notaffect soluble carbohydrate content, water potential, osmoticpotential or water loss. Under osmotic stress with polyethyleneglycol (PEG), paclobutrazol diminished the increase of waterpotential and decreased the rate of water loss caused by theimposed stress, but had no effect on osmotic potential. Catalaseand peroxidase activity were increased in osmotically-stressedroots of treated plants. Key words: Root growth, paclobutrazol, pea, Pisum sativum, water shortage     

Comparison of [14C]oryzalin Uptake in Root Segments of a Sensitive and a Resistant Species

Uptake of the dinitroaniline herbicide oryzalin (3,5-dinitro-N⁴,N⁴-dipropylsulfamlamide) and its effect on root growth werestudied using 5 mm corn (Zea mays L.) and pea (Pisum sativum)root apices. Pea root growth was much less susceptible to oryzalinthan corn root growth. Uptake studies showed that pea root apicesalso accumulated much less [¹⁴C]oryzalin and had a lower bindingaffinity for this herbicide. [¹⁴C]oryzalin was not metabolizedin root apices from either species. Thus, the differential susceptibilityto oryzalin in the case of corn versus pea can be explained,at least in part, by differences in oryzalin uptake and accumulationby roots. Oryzalin, dinitroaniline herbicides, Zea mays, Pisum sativum     

The Effect of pH on the Binding of Calcium to Pea Epicotyl Cell Walls and its Implications for the Control of Cell Extension

Cell walls were prepared from the epicotyls of dark-grown pea(Pisum sativum L.) seedlings. The walls were found to bind externally-added⁴⁵Ca²⁺, with a binding constant of 4 ? 10^–4 mol dm^–3and a maximum capacity of 1.5 ? 10^–8 g-ions of Ca²⁺ perg fresh weight of epicotyl. The binding capacity decreased asthe pH of the medium was decreased below 6.0, suggesting thatthe calcium was bound by an anionic group with an apparent pKof 4.7. More than half the calcium binding was due to polygalacturonicacid in the wall, since up to 60% of the calcium binding capacitywas removed by pre-incubation of the cell walls with polygalacturonase(E.C.3.2.1.15). Only small decreases in calcium binding wereseen following pre-incubation with protease, nucleases, phospholipaseand hemicellulase. These results indicate that calcium willbe displaced from the cell wall at hydrogen ion concentrationswhich are known to occur in the wall during wall extension.They are consistent with a mechanism by which calcium inhibitswall extension by forming ionic bridges between polygalacturonicacid molecules, and also with the hypothesis that calcium andhydrogen ions exert opposing influences on cell wall extensionby competing for the same binding sites on the polygalacturonicacid. Key words: Pea epicotyl, Cell wall, Calcium, pH     

Requirements for the light-stimulated degradation of stromal proteins in isolated pea (Pisum sativum L.) chloroplasts

Chloroplasts from 17-d-old pea leaves (Pisum sativum L.) wereisolated to elucidate the requirements for the light-induceddegradation of stromal proteins. The influence of electron transportthrough the thylakoids and the influence of ATP on protein degradationwere investigated. When chloroplasts were incubated in the light(45 µmol m^–2s^–1), glutamine synthetase, thelarge subunit of ribulose-1,5-bisphosphate carboxylase and glutamatesynthase were degraded, whereas phosphoribulokinase, ferredoxin-NADP⁺reductase and the 33 kDa protein of photosystem II remainedmore stable. Major protein degradation was not observed over240 mm in darkness. The electron transport inhibitor dichlorophenyldimethylureareduced protein degradation in the light over several hours,whereas dibromothymoquinone was less effective. Inhibiting theproduction of ATP with tentoxin or by destroying the     

Buckminsterfullerene (C-60 Carbon Allotrope) Inhibits Ethylene Evolution from 1-Aminocyclopropane-1-carboxylic acid (ACC)-treated Shoots of Pea (Pisum sativum), Broadbean (Vicia faba) and Flowers of Carnation (Dianthus caryophyllus)

When applied either in the form of a colloidal solution or inliposomes, buckyballs (C-60—buckminsterfullerene) markedlyreduced ethylene evolution from cut carnation (Dianthus caryophyllus)flowers, as well as pea (Pisum sativum) and broadbean (Viciafaba) foliage treated with ethylene precursor l-aminocyclopropane-l-carboxylicacid (ACC). The liposome preparation was approximately twiceas effective as colloidal solutions. Moreover, upon being incubatedin a closed atmosphere with ethylene, buckyballs induced a significantdepletion of ambient ethylene which was temperature and C-60—concentrationdependent. This mode of C-60 action is attributed to ethyleneadsorption stemming from the vast C-60 surface area, calculatedto be 1317 m² g_-1, and the affinity of its carbon atoms forthe component in the ethylene double bond.Copyright 1993, 1999Academic Press Dainthus caryophyllus, Pisum sativum, Vicia faba, adsorption, ethylene, fullerene     

Ion Absorption and Allocation of Carbon Resources in Excised Pea Roots Grown in Liquid Medium in Absence or Presence of NaCl

Freshly excised pea roots (Pisum sativum cv. Alaska) when transferredto growth medium (130 mOsm) or growth medium containing salt( 370 mOsm) suffer an initial osmotic shock and lose water.Contol roots tended to accumulate potassium, particularly inthe apical zone, while those exposed to NaCl accumulated mainlysodium, potassium accumulation being depressed. Exposure tosalinity for 6 d caused increases in root protein, cellulose,uronic acid and lignin content per cell. In roots supplied with¹⁴C-glucose for 24 h immediately after excision there was littledifference in uptake of glucose and in its use in respirationbetween control and salt treated roots. However, there werenoticeable differences in incorporation of labelled carbon intoseveral cell fractions, and particularly into the cellulosefraction in the upper parts of the root. When roots were grownfor six days in culture before being supplied with [¹⁴C]glucose,uptake per root was greater in the 120 mM NaCl treatment, andthe fraction diverted to respiration was decreased by salinity.On a per cell basis incorporation into soluble starch, uronicacid and cellulose fractions was increased in the salt treatedroots. The data obtained are in accord with the previous findingsand are suggestive of increased synthesis of cell wall materials.No conclusion could be drawn as to whether the changes describedare of adaptive value. Pisum sativum L. cv. Alaska, root culture, salinity, osmoregulation, cell wall     

Effect of Potassium on Sucrose and Amino Acid Release from the Seed Coat of Developing Seeds of Pisum sativum

After removal of the embryo from developing seeds of Pisum sativum,the ‘empty’ ovules (seed coats without enclosedembryo) were filled with a solution (pH 5.5) containing mannitol(usually 400 mM) to which various salts were added. A solutioncontaining two isotopes ((a) [²H]-sucrose/[–¹⁴C]aminoisobutyricacid (AIB) or (b) [³H]valine/[₁₄C]asparagine mixture) was administeredto the plant via the petiole subtending the fruiting node, and[²H]solute and [¹⁴C]solute unloading from the seed coat wasmeasured, in pulse-labelling experiments of about 5 h. The presenceof 25 or 50 mM K⁺ in the ‘empty’ ovule enhancedthe release of sucrose from the seed coat particularly duringthe first hours of the experiment, but the stimulating effectof K⁺ on the release of labelled solutes derived from aminoacids was much smaller. The presence of 25 mM CaCl₂ did notaffect the release of sucrose or amino acids from the seed coat.The effect of K⁺ on sucrose and amino acid release is explainedas an inhibition of sucrose and amino acid resorption from theseed coat apoplast into seed coat cells, after unloading fromthe seed coat unloading sites. It is suggested that amino acidrelease is much less affected by K⁺ than sucrose release, becausefar less resorption of amino acids by seed coat parenchyma cellstakes place during amino acid transport into the seed coat cavity. Pisum sativum, pea, assimilate transport, assimilate unloading, seed-coat exudate, seed development, sucrose resorption, surgical treatment     

Red and blue light-stimulated proton efflux by epidermal leaf cells of the Argenteum mutant of Pisum sativum

Light stimulates leaf expansion in dicotyledons by increasingapoplastic acidification, cell wall loosening and solute accumulationfor turgor maintenance. Red and blue light enhance growth viadifferent photo-systems, but the cellular location and modesof action of these systems is not known. Here, the effect of red and blue light was studied on transportprocesses in epidermal cells of expanding leaves of the Argenteummutant of Pisum satlvum. Both red and blue light caused extraceiiuiaracidification by isolated epidermal tissue, which was stimulatedby extracellular K⁺ and inhibited by DCCD at 0.1 mol m^–3.Acidification induced by red compared with blue light showeddifferent saturating kinetics in fluence rate-response curves.Under near saturating light conditions the effects of red andblue light were additive. The red light-induced acidificationwas inhibited by far-red light while the blue light-inducedacidification was not. Light caused a hyperpoianzation of themembrane potential in epidermal strips, and stimulated ⁸⁶Rb⁺uptake by epidermal protoplasts. These results show that phytochromeand an additional blue light-photoreceptor function in isolatedepidermal cells to promote proton efflux, hyperpolarization,and cation uptake. Key words: Pisum sativum, light-induced acidification, ion transport, epidermis, photoreceptor     

Measurement of the Water Potential of Intact Plant Tissues1

A modified design of a thermocouple psychrometer is describedwhich utilizes the Peltier effect for producing an exceedinglysmall wet-bulb thermometer. The thermocouple assembly has beenreduced to miniature dimensions and the thermocouple chamberconsists of a silver cylinder (volume 0.06 cm³) whose base isformed by the material under observation. This makes it possibleto measure water potentials over limited surfaces of intactplant tissues, e.g. germinating pea seeds (Pisum sativum L.).The apparatus is capable of measuring water potentials downto a magnitude of about –6,000 joules/kg.³     

Time of Flowering of Pea (Pisum sativum L.) as a Function of Leaf Appearance Rate and Node of First Flower

In order to assess the influence of environmental conditionson time of flowering of pea (Pisum sativum L.), a serial sowingtrial was conducted over 2 years at Dijon, France, on two wintercultivars Frisson and Frilene. Time of flowering was analysedaccording to two variables: the leaf appearance rate R_L andthe node of first flower N_I. R_L was linearly related to temperature (r² = 0·94). Thebase temperature was 2°C for both varieties. Growth rateaccounted for the residual variability of R_L . Photoperiod andtemperature acted on N_I in an additive way. Frilene, the latergenotype, was more responsive than Frisson. A model for predicting time of flowering based upon these resultsis proposed. Deviations from this model were related to N nutritionin interaction with the plant water relations. Steps for improvingthe model are then discussed.Copyright 1993, 1999 Academic Press Pisum sativum L., pea, flowering, temperature, photoperiod, phyllochron, model     

Theoretical Basis of Protocols for Seed Storage III. Optimum Moisture Contents for Pea Seeds Stored at Different Temperatures

In previous work, we demonstrated that there was an optimummoisture level for seed storage at a given temperature (Vertucciand Roos, 1990), and suggested, using thermodynamic considerations,that the optimum moisture content increased as the storage temperaturedecreased (Vertucci and Roos, 1993b). In this paper, we presentdata from a two year study of aging rates in pea (Pisum sativum)seeds supporting the hypothesis that the optimum moisture contentfor storage varies with temperature. Seed viability and vigourwere monitored during storage under dark or lighted conditionsat relative humidities between 1 and 90%, and temperatures between-5 and 65°C. The optimum moisture content varied from 0·015g H₂O g^-1 d.wt at 65°C to 0·101 g H₂O g^-1 d.wt at15°C under dark conditions and from 0·057 at 35°Cto 0·092 g H₂O g^-1 d.wt at -5°C under lighted conditions.Our results suggest that optimum moisture contents cannot beconsidered independently of temperature. This conclusion hasimportant implications for 'ultra-dry' and cryopreservationtechnologies.Copyright 1994, 1999 Academic Press Seed storage, seed aging, seed longevity, water content, temperature, glass, desiccation damage, ultradry, Pisum sativum L., pea, cryopreservation     

Brefeldin A: a specific inhibitor of cell wall polysaccharide biosynthesis in oat coleoptile segments

The effect of brefeldin A (BFA) on the synthesis and incorporation of polysaccharides, proteins and glycoproteins into the cell wall of subapical coleoptile segments isolated from etiolated oat seedlings (Avena sativa L. cv. Angelica) has been investigated. In the presence of D-[U-¹⁴C]-glucose, the incorporation of radioactive glycosyl residues into buffer-soluble, membrane (matrix polysaccharides) and cell wall polysaccharides was drastically inhibited by increasing concentrations of BFA up to 10 μ·mL⁻¹. BFA also altered the pattern of these polysaccharides suggesting a different sensitivity of glycosyltransferases toward the action of the drug. The incorporation of [U-¹⁴C]-glycine or L-[U-¹⁴C]-leucine into non-covalently- and covalently-bound cell wall proteins as well as the incorporation of radioactive N-acetylglucosamine residues into the newly synthesised oligosaccharidic chains of cytosolic, membrane and cell wall glycoproteins remained unchanged in the presence of 10 μg·mL⁻¹ BFA. The data demonstrate that, in oat coleoptile segments, BFA specifically inhibits the synthesis of cellulose and matrix polysaccharides without altering the synthesis and incorporation of proteins and glycoproteins into the cell wall. In addition, it is demonstrated that BFA does not affect the in vivo activity of glycosyltransferases involved in the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to the oligosaccharidic chains of glycoproteins.     

Immunofluorescence Microscopy of Microtubule Arrangement in Root Cells of Pisum sativum L. var Alaska

The arrangement of wall microtubules (MTs) in Pisum sativumroots was viewed immunofluorescently using cryosectioning. Mostcells in the tip region of pea roots (0–2 mm from tip)had wall MTs arranged transversely to the root axis. In theregion elongating at a higher rate (2–4 mm), wall MTsof epidermal, cortical and stelar cells were all transverselyarranged. In the region of about 5 mm from the tip, in whichcell elongation had already ceased, wall MTs in cortical cellschanged from a transverse to an oblique arrangement in relationto the root axis. Some cells had a crossed arrangement of wallMTs, which was interpreted as representing two sets of unidirectional,oblique wall MTs in opposite cell cortices of a single cell.This change was completed within a region of 1-mm width. Sinceroots elongated at a rate of 0.6 mm h^–1, it means thatthe arrangement of wall MTs changed within 2 h. An oblique arrangementof wall MTs was also observed in stelar cells. As the cellsaged, the oblique arrangement tended to change to a steeperor even a longitudinal one. (Received January 24, 1986; Accepted May 15, 1986)     

Effects of fluorinated pyrimidines on the growth of excised pea embryos

The effects of some fluorinated pyrimidines on the growth ofexcised pea embryos {Pisum sativum var. Alaska) in sterile culturewere studied. Even the lowest concentrations of the compoundstested inhibited growth in length of the embryos. In order ofdecreasing activity, the compounds tested were : 5-fluorodeoxyuridine,5-fluorodeoxycytidine, 5-fluorouridine, 5-fluoroorotic acidand 5-fluorouracil. Inhibition of growth in length of the rootprimordia was found to be mainly due to inhibition of cell divisionwith no effect on cell elongation. Reversal of fluoropyrimidineinduced inhibition of growth by pyrimidine bases and their relatedmetabolites indicated that the analogues primarily inhibitedDNA synthesis. ¹Part of a thesis submitted by the senior author for the degreeof M.Sc. of the University of Malaya ²Present address: Department of Botany, University College ofWales, Aberystwyth, U.K. (Received October 27, 1969; )     

Inhibition of Gibberellic Acid-Induced Elongation-Growth of Pea Epicotyls by Xyloglucan Oligosaccharides

The biological activity of a cell wall-derived xyloglucan nonasaccharide(XG9) was investigated using a bioassay with entire pea epicotyls(Pisum sativum cv. Progress). The xyloglucan fragment was foundto inhibit gibberellic acid-induced elongation of etiolatedpea epicotyls with maximum inhibition at concentrations rangingfrom 10^–11 to 10^–9M. Growth of etiolated epicotylsin the absence of exogenously applied GA₃ was also inhibitedby XG9 in the same concentration range. A cell wall-derivedheptasaccharide (XG7) lacking the fucosyl-galactosyl-side chainshowed no inhibitory effect in the pea epicotyl bioassay withand without exogenous GA₃. Furthermore, the biological activityof a synthetic pentasaccharide (XG5), containing the fucosylgalactosyl-sidechain which is necessary for the biological activity was investigatedin the same bioassay. Compared to XG9 the pentasaccharide hada similar inhibitory activity on GA₃-promoted elongation aswell as on the endogenous growth in the absence of exogenouslyapplied GA₃, but did not exhibit a distinct concentration optimum. Key words: Elongation-growth, gibberellic acid (GA₃), oligosaccharides, pea, XG9     

Heat Shock Induced Change in Protein Ubiquitination in Chlamydomonas

Ubiquitin was purified from pea (Pisum sativum L.) and its antibodywas produced. Western blot analysis showed that the antibodycross-reacted with ubiquitins from a green alga Chlamydomonasreinhardtii, a brown alga Laminaria angustata and a red algaPorphyridium cruentum but not with ubiquitin from a blue-greenalga Synechococcus sp. In Chlamydomonas, the antibody also reactedwith some ubiquitinated proteins including 28- and 31-kDa polypeptides.The isoelectric points of Chlamydomonas ubiquitin and the 28-and 31-kDa ubiquitinated proteins were 8.0, 8.9 and 10.3, respectively.The ubiquitinated proteins, including the 28- and 31-kDa polypeptideswere detected after in vitro ATP-dependent ubiquitination ofChlamydomonas cell extract with ^l25I-labeled bovine ubiquitin.Heat treatment of Chlamydomonas cells (>40°C) causeddrastic increase of ubiquitinated proteins with high mol wt(>60kDa), and coordinated redistribution or decrease of otherubiquitinated proteins and free ubiquitin. Quantitative analysisrevealed that the 28- and 31-kDa ubiquitinated proteins showeddifferent responses against heat stress, i.e. the former beingmore sensitive than the latter. (Received July 10, 1988; Accepted October 4, 1988)     

Some Properties of an Acid Phosphatase Isolated from the Seed Coats of Developing Pea Seeds

An acid phosphatase (EC 3 1 3 2)isolated from the seed-coatsof developing pea seeds was estimated to have MW 30000 by gelfiltration on Sephadex G-75 It was shown to act on a broad spectrumof physiological substrates, the most preferred being ß-glycerophosphate,3-phosphoglycerate and ADP, wich all showed rates of about halfthe maximum rate shown with p-nitrophenyl phosphate (p-NPP)Another model substrate frequently used in enzyme localizationstudies, -naphthyl acid phosphate, was hydrolysed at about 30% of the rate shown with p-NPP This acid phosphatase was enhancedor stabilized by the chelators EDTA and 1, 10-phenanthrolme,unaffected by Mg²⁺ and N-ethyl maleimide, but strongly inhibitedby Zn²⁺ and F^– Both oxidized and reduced glutathione werewithout effect at low concentration and slightly inhibitoryat high concentration (15 mm) Thiol groups are clearly not involvedin regulating the activity of this acid phosphatase, a featurewhich distinguishes it from acid phosphatases from several otherplant species. Pisum sativum L, pea, acid phosphatase, seed-coats, seed development     

Growth and Biomass Allocation, with Varying Nitrogen Availability, of Near-isogenic Pea Lines with Differing Foliage Structure

The single-gene mutation afila in pea (Pisum sativum L.) resultsin the replacement of proximal leaflets with branched tendrils,thereby reducing leaf area. This study investigated whethertheafila line could adjust biomass partitioning when exposedto varying nutrient regimes, to compensate for reduced leafarea, compared with wild-type plants. Wild-type and afila near-isogeniclines were grown in solution culture with nitrate-N added toinitially N-starved seedlings at relative addition rates (R_N)of 0.06, 0.12, 0.15 and 0.50 d^-1. The relative growth rate (R_W)of the whole plants closely matched R_Nat 0.06 and 0.12 d^-1,but higher R_Nresulted in a slightly higher growth rate. At agiven R_N, the wild-type line had lower plant nitrogen statusthan the afila line. R_Wof the roots of the afila line was lessthan R_Wof the roots of the wild-type at the three higher ratesof N supply despite a greater accumulation of N in the rootsof the afila plants. Consequently, plant nitrogen productivity(growth rate per unit nitrogen) was lower for afila. Dry matterallocation was strongly influenced by nitrogen status, but nodifferences in shoot–root dry matter allocation were foundbetween wild-type and afila with the same plant N status. Theseresults imply that decreased leaf area as a result of the single-genemutation afila affects dry matter allocation, but only accordingto its effect on the nitrogen status. Copyright 2000 Annalsof Botany Company Pisum sativum, pea, nitrogen limitation, growth, shoot–root allocation, relative growth rate, nitrogen productivity, isolines