ERRATA

Effects of coupled solute and water flow in plant roots withspecial reference to Brouwer's experiment. Edwin L. Fiscus. p. 71 Abstract: Line 3 delete ‘interval’ insert‘internal’. p. 73 Materials and Methods: line 6: delete ‘diversion’ insert ‘division’ line 9 equation should read J_v=L_p P–RT(C⁰–C¹). 74 Last line of figure legend: 10^–1 should read 10^–11. 75 Line 11: delete ‘seems’ insert ‘seem’. le 1 column heading—10⁶ should read 10¹¹. 77 delete ‘...membrane in series of...’ insert ‘membranein series or...’ Delete final paragraph.     

Pyrimidine Nucleotide Biosynthesis in Vinca rosea Cells: Changes in the Activity of the de novo and Salvage Pathways during Growth in a Suspension Culture

Marked changes in the activity of the ‘de novo’and ‘salvage’ pathways of pyrimidine biosynthesisduring growth of Vinca rosea cells in a batch suspension culturewere observed. The activity of these pathways was investigated by determiningthe contribution of ¹⁴C of [2-¹⁴Cluracil, 12-¹⁴Cluridine. and[6-¹⁴Clorotate to the cell constituents and by measuring theactivity of the several enzymes of these pathways. During the lag phase of the culture, ‘uracil-’ and‘uridine-salvage’ pathways made the predominantcontribution to nucleotide biosynthesis, but, following theinitiation of cell division, the ‘de novo’ pathwayfor nucleotide biosynthesis operated appreciably. These results suggest that nucleotide synthesis during cellgrowth in a suspension culture can be divided into two stages:a ‘turnover stage’, during the lag phase of cellgrowth, and a ‘true biosynthetic stage’, which isinitiated in the cell division phase.     

Effect of Inorganic Phosphate on the Biosynthesis of Purine and Pyrimidine Nucleotides in Suspension-Cultured Cells of Catharanthus roseus

The levels of purine and pyrimidine nucleotides in suspensioncultures of Catharanthus roseus were determined 24 h after stationary-phasecells were transferred to fresh complete (‘+Pi’)or phosphate-deficient (‘–Pi’) Murashige-Skoogmedium. The levels of ATP, GTP, UTP and CTP were from approx.3 to 5-fold greater in the cells grown in ‘+Pi’medium than in the cells grown in ‘–Pi’ medium.The levels of almost all other nucleotides were slightly higherin the cells in ‘+Pi’ medium. The rates of de novoand salvage biosynthesis of purine and pyrimidine nucleotideswere estimated from the rates of incorporation of radioactivityfrom [¹⁴C]formate, [2–¹⁴C]glycine, NaH¹⁴CO₃, [6–¹⁴C]orotate,[8–¹⁴C]adenine, [8–¹⁴C]adenosine, [2–¹⁴C]uraciland [2–¹⁴C]uridine. The results indicated that the activityof both the de novo and the salvage pathway was higher in thecells in ‘+Pi’ medium than in the cells in ‘–Pi’medium. The rate of degradation estimated from the rate of releaseof ¹⁴CO₂ from labelled purines and pyrimidines indicated thatdegradation of uridine was significantly reduced in the cellsin ‘+Pi’ medium, but no significant difference wasfound in the degradation of adenine, adenosine and uracil. Thepossible role of Pi in the control of the biosynthesis of nucleotidesand in the degradation of uridine is discussed. Catharanthus roseus, Madagascar periwinkle, suspension culture, inorganic phosphate, nucleotides, purines, pyrimidines, biosynthesis, degradation     

Behavioral responses of the brittle star Ophiura ophiura to amino acids, acetylcholine and related low-molecular-weight compounds

Feeding behavior of the brittle star Ophiura ophiura includesorienting posture, orienting movements, arm ‘walking’,changing the direction of ‘walking’ arm coilingand ingestion. All sequential behavior patterns were releasedor enhanced by single low-molecular-weight compounds. Stimuliwhich released ‘walking’ behavior at high concentrations(10^–4 M) in all the test animals are listed in decreasingorder of sensitivity: sarcosine, glycine, urea, L-valine, L-leucine,L-methionine, L-homocysteine, L-norvaline, L-norleucine, L-threonine,L-serine, S-methyl-L-cysteine, L-proline. Threshold values forsingle amino acids were as much as 100 times different in differentindividuals and ranged from 3 x 10^–9 to 3 x 10^–7M for the most effective stimulus, sarcosine, and from 10^–6to 10^–4 M for proline. Above 10^–5 M, only L-prolineregularly released a second behavior pattern, the arm coilingresponse, which temporarily inhibited the ‘walking’behavior. Behavioral thresholds for the ‘walking’behavior for L(+)-lactate and L-alanine were higher than thosefor the orienting movements. Thyoglycolic acid and ß-alaninereleased tube feet walking, which is not part of the feedingbehavior. Structure—activity comparisons were studied at estimated10^–5 M concentrations. Gycline, sarcosine, L-valine, L-norvaline,L-leucine, L-isoleucine, DL-norleucine and DL-homocysteine releasedarm ‘walking’ behavior in more than 75% of all thetests. With the exceptions of S-methyl-and S-ethyl-cysteine,and glycine methylester, derivatives of amino acids were noteffective behavioral stimuli in Ophiura ophiura. L-Isomers ofvaline and leucine regularly stimulated the ‘walking’behavior while their D-isomers were effective in some testsand ineffective in others. Acetylcholine iodide, acetyl-ß-methylcholine chloride and choline phosphate chloride regularly released‘walking’ behavior at concentrations above 10^–5M.     

Free Proline and Prolamin Protein in the Grain of Three Barley Varieties Subjected to a Gradient of Water Supply

Three barley varieties (Atem, Arabic Abiad and Beecher) weregrown on a line source sprinkler to establish a gradient ofwater supply from rain-fed (218 mm) to fully irrigated (330mm). Developing ‘seeds’ were analysed for free grainproline at three stages up to maturity. At the milk stage allthree varieties showed a strong inverse correlation (r>0–95)between free proline and irrigation treatment. A pronouncedinter-varietal difference was evident with the relatively drought-intolerantAtem having 213 nmol seed^–1 free proline when rain-fedas compared with Arabic Abiad or Beecher which had about 120nmol seed^–1 free proline at this stage. By maturity thiscorrelation was lost in all varieties, with most of the freeproline being incorporated into protein. Prolamin protein production was not impaired by drought in anyof the varieties, when measured on a per seed basis. There wassome indication that it may even rise in concentration in severelydrought-stressed plants. The results are discussed in relation to the decline in finalgrain yield with increased water deficit. Each of the threevarieties tested showed different yield responses along theirrigation gradient. These may reflect the physiological tolerancesto drought of each variety. At the milk stage the measurementof free ‘grain’ proline as a diagnostic for droughtstress tolerance may be of some value. Key words: Drought stress, seed physiology, proline     

Determinate Floral Buds of Plantain (Musa AAB) as a Site of Adventitious Shoot Formation

Floral buds of the ‘False Horn’ plantain clonesMusa (AAB) ‘Harton Verde’, ‘Harton Negra’,and ‘Currare’ terminate in a large single floralstructure. The apices of these floral buds are here designatedas determinate since they have lost the ability to produce additionalfloral initials or buds. Terminal peduncle segments can be culturedin a modified Murashige and Skoog (1962) medium supplementedwith N⁶-benzyl-aminopurine (5 mg I^–1). Under these conditions,this apparent inability to yield buds can be overcome as vegetativeshoot clusters form in the axils of the bracts. Rooted plantletsare obtainable by treating shoots with naphthaleneacetic acid(1 mg I^–1) and activated charcoal (0.025%). The adventitiousorigin of the shoots has been established. Musa cultivars, plantains, floral bud, adventitious buds, tissue culture     

Particle-bound phytochrome: Differential pigment release by surfactants, ribonuclease and phospholipase C

Surfactants and hydrolytic enzymes were used to probe the natureof the constituents) to which phytochrome binds in paniculatefractions from red-irradiated Cucurbita. [¹⁴C]-choline and [³H]-uridinepre-labelled tissue was used to monitor the release of phospholipidsand RNA by these agents. Ribonuclease (RNase) digestion of 20,000xgpellets eliminates both the phytochrome and ribonucleoprotein(RNP) which cosediment at 31S. Little [¹⁴C]-choline occurs inthe 31S fraction and the amount is not changed by RNase digestion.This is further evidence that phytochrome binds directly tothe RNP in the 31S fraction rather than to any membranous materialpresent. The distribution profile of the RNA in a second ( =‘heavy’)phytochrome fraction does not correlate with that of the pigment.This suggests that the phytochrome in this fraction is not boundto RNP. The RNA is of ribosomal origin but much less degradedthan that of the 31S RNP and is resistant to RNase digestion.Phospholipase C releases>80% of the [¹⁴C]-choline from the‘heavy’ fraction without freeing phytochrome. Thisindicates that the pigment does not bind to the polar head groupsof the membrane phospholipids present. Low concentrations ofdeoxycholate dissociate phytochrome from this fraction withoutreleasing substantial quantities of integral membrane proteinsor phospholipids. Some RNP is dislodged by the surfactant butthe phytochrome and RNP are not released as a complex. The datasuggest that the pigment in the ‘heavy’ fractionmay be loosely bound to a protein constituent rather than toRNP or polar phospholipids. ¹This work was done while on sabbatical leave from the WeizmannInstitute of Science, Rehovot, Israel. (Received April 1, 1976; )     

Developmental control of xylem hydraulic resistances and vulnerability to embolism in Fraxinus excelsior L.: impacts on water relations

The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 10^–4M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar ‘Alaska’, an ABA-deficientmutant ‘wil’, and its ‘wildtype’. Freezinginjury was determined graphically as the temperature that caused50% injury (T₅₀) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T₅₀ for root tissuewas 1C for all genotypes, regardless of treatment. For ‘Alaska’shoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by –1.5 to –4.0C, while coldtreatment increased the freezing tolerance by –7.5 to–14.8C. Cold treatment increased the freezing toleranceof shoot tips by –9 and –15C for ‘wil’and ‘wild-type’, respectively. Cold acclimationincreased endogenous ABA concentrations in ‘Alaska’shoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of ‘Alaska’, respectively.In ‘Alaska’ shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in ‘wil’and ‘wild-type’ shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization     

Freezing tolerance, protein composition, and abscisic acid localization and content of pea epicotyl, shoot, and root tissue 3

The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 10^–4M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar ‘Alaska’, an ABA-deficientmutant ‘wil’, and its ‘wildtype’. Freezinginjury was determined graphically as the temperature that caused50% injury (T₅₀) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T₅₀ for root tissuewas 1C for all genotypes, regardless of treatment. For ‘Alaska’shoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by –1.5 to –4.0C, while coldtreatment increased the freezing tolerance by –7.5 to–14.8C. Cold treatment increased the freezing toleranceof shoot tips by –9 and –15C for ‘wil’and ‘wild-type’, respectively. Cold acclimationincreased endogenous ABA concentrations in ‘Alaska’shoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of ‘Alaska’, respectively.In ‘Alaska’ shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in ‘wil’and ‘wild-type’ shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization     

Effects of Monochromatic Radiations on Growth of Pelargonium Callus Tissue

Pith callus tissues were grown under continuous blue (450 mµ),green (545 mµ), red (650 mµ), and ‘white’(full-spectrum) light, and in the dark for 22 days at 27±2°C at energy levels of 15,000 ergs cm^–2 sec^–1. Mean increases in fresh weight of tissues grown under ‘white’and blue light were significantly greater than those of tissuesgrown in green and red light and in the dark. Tissues grownin the dark yielded mean fresh weight increases significantlylower than tissues grown under blue, red, and ‘white’light. No significant differences were shown between blue and‘white’, red and green, and green and dark treatmentsrespectively. Cell differentiation occurred in all treatmentsonly to the extent of vessel element formation. There were nodifferences in degree of differentiation between treatments. It was proposed that the high-energy reaction of photomorphogenesiswas in operation in the Pelargonium callus tissue. The resultsindicated the presence in the tissue of high-energy photoreceptor(s).The use of high-intensity, incandescent illumination for experimentalprocedures approximating natural conditions of irradiation wasindicated as desirable for pith callus tissues of Pelargoniumzonale var. Enchantress Fiat.     

Polyamine Content in Relation to Embryo Growth and Dedifferentiation in Barley (Hordeum vulgare L.)

Putrescine, spermidine, and spermine content were analysed inzygotic embryos of barley (Hordeum vulgare L.). Changes in polyaminecontent were observed during zygotic embryo growth. In two cultivars,‘Bomi’ and ‘Golden Promise’, the totalpolyamine content in the embryos was 2.6–2.9 nmol mg^–1fresh weight 10 d after anthesis, the highest content observed.It dropped to 1.3 nmol mg^–1 fresh weight 14 d after anthesis.This drop was caused by decreases in all three polyamine concentrations.From 14 to 35 d after anthesis the putrescine content continuedto decrease while the spermidine and spermine content increased,thus the total polyamine content remained constant until 35d after anthesis. The mutant ‘Ris? 1508’ showeda constant polyamine content around 1.3 nmol mg^–1 freshweight from 14 to 35 d after anthesis. The polyamine patternwas conserved in all three lines throughout the period of investigationshowing a spermidine content higher than putrescine contentwhich was, in turn, higher or equal to the spermine content.The polyamine content measured as nmol µg^–1 proteindecreased from 14 to 21 d post anthesis in all three lines,because the protein content (µg mg^–1 fresh weight)increased during the period. In dedifferentiating zygotic embryoscultured in vitro the putrescine content (nmol mg^–1 freshweight) rose by a factor of nine and the spermidine contentdoubled within the first week of cultivation, whereas sperminecontent did not change. For embryoderived calli a repeated patternof change in polyamine content was observed throughout the subculturingperiod. Key words: Polyamines, Hordeum vulgare L., embryo development     

A Cytochrome P450 Mediated Naringenin 3'-Hydroxylase from Sweet Orange Cell Cultures

A microsomal flavonoid 3'-hydroxylase (F3'H) catalyzing themetabolism of naringenin to eriodictyol in Citrus sinensis (L.)Osbeck cv. ‘Hamlin’ cell suspension cultures wasshown to be a cytochrome P450 enzyme. This reaction requiredO₂ and NADPH and was inhibited by CO, with partial reversalof CO-inhibition by light at 450 nm. Cytochrome P450 contentranged from 10–20 pmol (mg microsomal protein)^–1.The F3'H reaction was shown to be linear in regard to proteinconcentration between 2.5 and 25 µg of microsomal protein.The optimum pH for the reaction was 7.4–7.6 and the temperatureoptimum was between 30 and 37°C. The apparent K_m and V_maxfor naringenin were 24 µM±3.2 and 81.4±7.9pmol eriodictyol min^–1 (mg protein)^–1, respectively.The microsomal F3'H was also capable of forming dihydroquercetinfrom dihydrokaempferol (40 pmol min^–1 (mg protein)^–1)and of quercetin from kaempferol (3.25 pmol min^–1 (mgprotein^–1). Cytochrome c and ketoconazole were the bestinhibitors of WH activity followed by piperonyl butoxide anda-naphthoflavone. Light was shown to be an inducer of the F3'Halmost doubling the specific activity and increasing the microsomalcytochrome P450 content by 30% over that of dark grown cells.F3'H activity was also confirmed in microsomal preparationsof young (new flush) leaves from ‘Hamlin’ treesand flavedo of ‘Hamlin’ oranges, ‘Marsh’grapefruit, and ‘Lisbon’ lemon. No activity wasobserved in older, hardened leaves and albedo of all the fruittested. Initiation of embryogenesis in the ‘Hamlin’cell suspension cultures by switching from a sucrose mediumto a glycerol-based medium resulted in the down-regulation ofF3'H. ¹Mention of a trademark, warranty, proprietary product, or vendordoes not constitute a guarantee by the U.S. Department of Agricultureand does not imply its approval to the exclusion of other productsor vendors that may also be suitable.     

The Sensitivity of Net Photosynthesis in Several Plant Species to Short-term Fumigation with Sulphur Dioxide

The effects of exposure of up to 2 h with sulphur dioxide ona range of plant species was observed by measuring changes inthe rate of net photosynthesis under closely controlled environmentalconditions. Ryegrass, Lolium perenne ‘S23’ was thespecies most sensitive to SO₂; significant inhibition was detectedat 200 nl l^–1. Fumigations at 300 nl l^–1 also inhibitedphotosynthesis in field bean (Vicia faba cv. ‘Three FoldWhite’ and ‘Blaze’) and in barley (Hordeumvulgare cv. ‘Sonja’). No effect was detected inwheat (Triticum aestivum cv. ‘Virtue’) at concentrationsup to 600 nl l^–1 SO₂, or in oil-seed rape (Brassica napuscv. ‘Rafal’) except at 800 nl l^–1 SO₂). Recoverycommenced immediately after the fumigation was terminated andwas complete within 2 h when inhibition had not exceeded 20%during the SO₂ treatment. Key words: Sulphur dioxide, short-term fumigation, photosynthesis     

Azetidine-2-carboxylic Acid and Lily Pollen Tube Elongation

Azetidine-2-carboxylic acid (AZC), which occurs naturally inLiliaceous plants, is reported to be a proline (pro) analoguePlant cell walls contain ‘extensin’, which is richin hydroxyproline (hyp). Peptidyl hyp arises through hydroxylationof peptidyl pro followed by glycosylation (arabinose attachment)of hyp Because AZC replaces peptidyl prolyl residues, it maybe a useful tool for evaluating the significance of hyp-o-arabinoselinkages in cell elongation. Therefore, we determined the effectof AZC on [¹⁴C]pro uptake, incorporation and conversion to wall-bound[¹⁴C]hyp in relation to elongation of lily pollen tubes whosewalls consist, in part, of hyp-containing glycopeptides TheAZC suppressed pollen germination 9–42 per cent (1–10mM) and subsequent tube elongation 40–54 per cent (0·1–1mM without affecting respiration In contrast, similar hyp concentrationswere without effect on tube elongation Whereas uptake of [¹⁴C]prowas 16·5–6·2 per cent of the control at0·1–1 mM AZC, [¹⁴C]leucine uptake was 85–25per cent of the control. Light microscope radioautography revealedfewer silver grains over tubes elongated in 0·1–1mM AZC than in its absence. Incorporation of [¹⁴C]pro into tnchloroaceticacid (TCA)-precipitable cytoplasm was reduced by only 10 percent at 0·01–1 mM but 43 per cent at 10 mM AZCGel filtration of cytoplasm from pollen germinated without AZCbut with [¹⁴C]pro resulted in labelled void volume (V) and threeretarded peaks (RI–III) Incorporation into V and RI wasinhibited at both 0·01 and 1 mM AZC These AZC concentrationsreduced conversion of [¹⁴C]pro to wall-bound hyp by 20 percent However, total incorporation of [¹⁴C]pro into salt-water-purifiedwall fractions was suppressed 47–53 per cent (0·1–1mM AZC). Lilium longiflorum, lily, hydroxyproline, proline, azetidine-2-carboxylic acid, pollen, pollen tube elongation     

Guard Cell Metabolism in Epidermis of Commelina communis L. During Stomatal Opening and Closing

The rates of CO² incorporation into the epidermis of C. communiswere linear and were similar during the completion of opening(2 h) and closing (1 h) movements of stomata. The kinetics of¹⁴C turnover between metabolites and the rates of ‘leakage’of metabolites were determined for opening and closing movements.When stomata were opening there was a slow turnover of ¹⁴C frommalate chiefly into sugars. Upon stomatal closure ¹⁴C was initiallymainly in sugars, malate, and sugar phosphates. Thereafter,there was a slight loss of label from sugar phosphates witha corresponding increase in malate. Starch became labelled duringopening and closing movements. Rates of incorporation of CO₂found in the ‘leakage’ fraction were greatest whenstomata were opening. Of the labelled compounds Most‘from the tissue, malate was the most highly labelled whetherstomata were opening or closing. Although interpretation of the turnover patterns is difficultwithout knowledge of pool sizes for the metabolites it is suggestedthat a pool of sugars exists within the guard cells, which havefairly direct and reversible access to carbon from starch andmalate. The implications of loss of malate from guard cellsduring stomatal opening and closing are discussed.     

Relative Sensitivity and Tolerance of some Gladiolus Cultivars to Sulphur Dioxide

Five Gladiolus cultivars, namely ‘Aldebaran’, ‘BrightEye’, ‘Illusion’, ‘Manisha’ and‘Manmohan’, were exposed to 1 and 2 µg l^–1sulphur dioxide to test their relative-sensitivity toleranceto the pollutant Plants were fumigated experimentally for 2h daily Foliar injury symptoms were observed first in ‘Manisha’followed by ‘Aldebaran’ and ‘Illusion’at the higher dose Photosynthetic pigments and leaf extractpH were significantly decreased, particularly in ‘Manisha’and ‘Illusion’ Overall disturbances in the plantmetabolism due to SO₂ treatment led to retarded growth of plants,as evident from decreased shoot length and phytomass valuesThe order of sensitivity of the five Gladiolus cultivars toSO₂ was as follows, with the greatest first Manisha, Illusion,Aldebaran, Bright Eye, Manmohan Cultivars, Gladiolus, sensitivity, sulphur dioxide, tolerance     

Nitrogen Fixation, Assimilation and Transport During the Initial Growth Stage of Phaseolus vulgaris L

In nodulated common bean (Phaseolus vulgaris L.), there is typicallya period of N stress between 15 and 20 d after emergence (DAE),due to a lack of synchronization between the depletion of Nin the cotyledons and the beginning of N² fixation and transport.Screening trials identified some Rhizobium leguminosarum bv.phaseoli strains with which symptoms of N deficiency were notvisible (‘precocious’ strains). Cultivar Negro Argelwas then inoculated with two ‘traditional’ strains(C-05 and CIAT 727) and two ‘precocious’ strains(CNPAF 146 and CNPAF 512), and plants were harvested from 8to 30 DAE. There were no differences between the two groupsof strains in nodule dry weight or in the acetylene reductionrates between 8 and 16 DAE. However, nodules induced by the‘precocious’ strains showed earlier onset of glutaminesynthetase (GS) (EC 6.3.1.2 [EC] ) and glutamate synthase (GOGAT)(EC 1.4.1.14 [EC] ) activities, and ureide synthesis. The N concentrationin the nodules formed by ‘precocious’ strains variedfrom 4.2 to 4.5%, whereas with the ‘traditional’strains, it increased from 3.2% at 8 DAE to 65% at 18 DAE, atwhich time plants exhibited N-deficiency symptoms. By 21 DAE,GS and GOGAT activities in ‘traditional’ noduleswere increased, as well as the ureide-N-concentration in thexylem sap, nodule N content declined to 4.5% and the leavesbecame green. These results suggest that the N stress with ‘traditional’strains is not a limitation in early N² fixation activity butrather in the rates of expression of the processes of N assimilationand transport. Key words: Glutamate synthase, glutamine synthetase, nitrogen fixation, Phaseolus vulgaris, Rhizobium     

Fruit Softening II. Precursor Incorporation into Pectin by Pear Tissue 6

The incorporation of radioactivity from precursors of methylester and galacturonosyl residues into pectin was investigatedusing tissue slices cut from ripening pear fruits. Incorporationfrom ¹⁴CH₃ methionine into methyl ester of water soluble pectinincreased 10 fold in 4 d at 18 °C and declined in laterstages of ripening. Activity from [³H]inositol could not bedetected in gaJacturonic acid released enzymically from solublepolysaccharides. When l³H]glucose was used as a precursor, activitycould be detected in galacturonic acid released from both thesoluble and insoluble polysaccharide fractions. Methionine wasa more efficient precursor of methyl ester groups than S-adenosylmethionine or S-methyl methionine; incorporation from all threeprecursors was inhibited under nitrogen. Radioactively labelledmethyl ester did not decline during a 225 min ‘chase’following a 15 min ‘pulse’ of [¹⁴CH₃]methionine;the total pectin content of slices increased by 20% during this4 h incubation.     

Uptake of 32P by Young Plants of Banksia hookeriana Meissner When Healthy and Infected with Phytophthora cinnamomi Rands

Young plants of Banksia hookeriana were grown in acid-washedsand with adequate phosphate and water supply, and a proportionwere inoculated with Phytophthora cinnamomi. There were no majordifferences in growth between uninoculated and infected plants,but there was a large increase in uptake of ³²P with increasingroot disease. In healthy plants ³²P uptake was greatest in youngleaf tissue, but in diseased plants labelled phosphate was directedmore towards older leaves where the activity was almost twicethat of young leaves. Enhanced uptake with disease was ascribed to possible blockageof the ‘message’ or ‘signal’ of phosphatetranslocation from shoot to root, such that the diseased rootincorrectly treated the shoot as P deficient and increased Puptake. Key words: Banksia hookeriana, Proteaceae, ³²P uptake, Phytophthora cinnamomi     

Effects of Inorganic Phosphate on the Utilization of Sucrose by Suspension-cultured Catharanthus roseus Cells

The metabolic fate of [U-¹⁴C]sucrose in suspension culturesof Catharanthus roseus cells was monitored for 96 h after thecells were transferred to fresh complete (‘+Pi’)or to phosphate-deficient Murashige and Skoog (‘–Pi’)medium. Sucrose was hydrolysed extracellularly to glucose andfructose. The rate of uptake of sugars by the cells was 1.5–3times higher in ‘+Pi’ culture than in ‘–Pi’culture. Little difference in the rate of incorporation of radioactivityinto the ethanol-soluble fraction was found between the ‘+Pi’and ‘– Pi’ cultures during the initial 24h of culture, but after 48 h the rate in ‘ +Pi’cultures was higher than that in ‘–Pi’ cultures.Incorporation of radioactivity into ethanol-insoluble macromoleculeswas always significantly higher in the cells in ‘+Pi’cultures than in those in ‘–Pi’ cultures.The results suggest that Pi strongly affects the utilizationof sugars by cultured plant cells through the stimulation oftransport of sugars as well as through the activation of metabolism. Catharanthus roseus, Madagascar periwinkle, suspension culture, inorganic phosphate, sucrose, transport, metabolism