The Partitioning of Photosynthetically Assimilated 14C into Amino Acids in Wheat: 2. Distribution in Amino Acids of Wheat Grain Fractions

Changes in the distribution of ¹⁴C between free and bound aminoacids in wheat grains (Triticum aestivum L. cv. Arkas) at 10and 20 d post-anthesis are described. After ¹⁴CO₂, labellingof the flag leaf, ¹⁴C was initially more rapidly transferredto the grains of 20 d post-anthesis plants than for 10 d post-anthesisplants. However, after a 460 min chase period in the light theamount of ¹⁴C in the grains of the younger and older plantswere similar. In the younger, more rapidly growing grains, agreater proportion of the ¹⁴C was incorporated into structuraltissue and starch. ¹⁴C accumulation in the grains continuedduring the dark in the younger grains but not in the older grains. Although the overall ¹⁴C distribution between the free aminoacid and protein pools of the grain was similar for both treatments,the distribution within the albumin, prolamin and globulin fractionsand between the individual non-bound amino acids differed. Ofthe protein fractions, the albumins were initially the mostheavily labelled but after 460 min chase the prolamins containedmore ¹⁴C. The majority of the ¹⁴C in the albumin and globulinfractions after 280 min chase was in hydrolysable, non-aminoacid compounds. In both tissues, the free amino acid pools lostradioactivity in the dark but the solid residues and proteinscontinued to function as ¹⁴C sinks. Daily fluctuations in the radioactivity in free and bound alanineare consistent with the role of free alanine as a diurnal metabolicnitrogen pool. Wheat, Triticum aestivum¹⁴CO₂, amino acids, proteins, carbon metabolism     

Dynamics of Carbon Photosynthetically Assimilated in Nodulated Soya Bean Plants under Steady-state Conditions 2. The Incorporation of 13C into Carbohydrates, Organic Acids, Amino Acids and some Storage Compounds

Nodulated soya bean (Glycine max L.) plants at the early floweringstage were allowed to assimilate ¹³CO₂ under steady-state conditions,with a constant ¹³C abundance, for 8 h in the light. The plantswere either harvested immediately or 2 d after the end of the¹³CO₂ feeding, divided into young leaves (including flower buds),mature leaves, stems+petioles, roots and nodules; the ¹³C abundancein soluble carbohydrates, organic acids, amino acids, starchand poly-ß-hydroxybutyric acid was determined witha gas chromatography-mass spectrometry. The rapid turnover of ¹³C in the sucrose pools observed in allorgans of the plants showed that sucrose was the principal materialin the translocation stream of primary products of photosynthesis.At the end of the ¹³CO₂ exposure, sucrose in the mature leavesas the major source organs and in the stems+petioles was labelledwith currently assimilated carbon to about 75 per cent, whereasa much higher labelling of sucrose was found in the roots andin the nodules. This suggests the existence of two or more compartmentedpools of sucrose in mature leaves and also in stems+petioles. The relative labelling patterns of individual organic acidsand amino acids were similar in various plant organs. However,the rapid turnover of succinate and glycine was characteristicof nodules. Treatment with a high concentration of nitrate inthe nutrient media increased the turnover rate of amino acidcarbon in shoot organs and roots, while it markedly decreasedthe labelling of amino acids in nodules. The cyclitols, exceptfor D-pinitol, were significantly labelled with assimilated¹³C in mature leaves, but in nodules, the labelling was verymuch less. In the nodules, which were actively fixing atmospheric nitrogen,a large proportion (80–90 per cent) of currently assimilatedcarbon was found as sucrose and starch at the end of the ¹³CO₂feeding. This was also true of the roots. On the other hand,in young growing leaves, the distribution of currently assimilatedcarbon into sucrose, starch and other soluble compounds wasmuch less. This suggests that a large amount of carbon assimilatedby and translocated to young leaves was used to make up structuralmaterials, mainly protein and cell wall polymers synthesis,during the light period. Glycine max L., soya bean, ¹³CO₂ assimilation, carbon metabolism in nodules     

The Metabolism and Transport of Abscisic Acid During Grain Fill in Wheat

The metabolism and transport of (±)-l2-¹⁴Clabscisic acidand its metabolites was investigated during the period of graindevelopment in wheat. Forty-five hours after feeding the hormoneinto the flag leaf blade, or after injection into the grains,nine metabolites could be extracted with acetone. Four of thesecompounds have been identified. They are phaseic acid, dihydrophaseicacid, abscisyl-ß-D-glucopyranoside and the polar metabolite.As well as the acetone-soluble metabolites a number of othershave been found which are insoluble in acetone. These appearto be conjugated to lipids, ‘gluten-like’ proteins,and carbohydrates. ABA and its metabolites were transported to all parts of theplant above the flag leaf node when the radioactive hormonewas introduced into the blade. However, when it was injectedinto the grain the radioactivity remained there until the pointwhen dry matter accumulation ceased, after which it was foundonly in the peduncle. The results suggest that the increasing level of ABA duringthe period of dry matter accumulation is due to biosynthesiswithin the grain and that the decrease after dry matter accumulationceases is due to both metabolism and redistribution within theplant.     

Amino Acid Composition Along the Transport Pathway during Grain Filling in Wheat

The amino acid composition of endosperm cavity sap and of sieve tube saps from the flag leaf, peduncle, rachis, grain pedicel, and grain were determined for wheat plants just past the mid-half of grain filling. On a mole percent basis, glutamine accounted for almost half of the amino acids in sieve tube sap from the peduncle and ear. Other protein amino acids, plug γ-aminobutyrate, were present in varying, but mostly low (a few mole percent) proportions. The amino acid composition of phloem exudate resembled that of the mature wheat grain. The proportions of amino acids in the endosperm cavity were generally similar to those of the sieve tube sap supplying the grain. Cysteine, however, while virtually absent from sieve tube sap, comprised 1 to 2 mole percent of amino acids in the endosperm cavity, suggesting it is transported in a different form. Also, alanine and, to a lesser extent, glutamate were relatively more prominent in endosperm cavity sap than in the sieve tube sap. Thus, while most amino acids were more concentrated in the sieve tube sap than in the endosperm cavity sap, alanine and glutamate appeared to be moving from the sieve tube to the endosperm cavity in the absence of, or perhaps even against, their concentration gradients.     

Long-term Partitioning, Storage and Remobilization of 14C Assimilated by Trifolium repens (cv. Blanca)

The fourth fully expanded leaf on the main stolon of white cloverplants was exposed to ¹⁴CO₂. Thereafter, quantitative and fractionalanalysis of the partitioning, storage and remobilization afterdefoliation of the ¹⁴C labelled assimilate was sequentiallyconducted over a 2- to 3-week period. In undefoliated plants, most ¹⁴C reached its final destinationwithin 24 h of feeding. Forty percent of assimilated ¹⁴C waslost through respiration, while the rest was exported, predominantlyto meristems, but also to roots, stolons and leaves. The ¹⁴Cinitially translocated to meristems was subsequently recoveredin stolon and leaf tissue as the plants matured. Approximately 10% of assimilated ¹⁴C was invested into long-termstorage in roots and stolons. These reserves were remobilizedafter both partial and total defoliation, and a portion of theremobilized ¹⁴C was incorporated into new growth, Partly defoliatedplants regrew more rapidly than totally defoliated plants, butmore ¹⁴C reserve depletion took place in the totally defoliatedtreatment. Reserve depletion took place from both stolons androots, but stolon reserves were preferentially utilized. Bothhigh and low molecular weight storage compounds were involved. Trifolium repens, white clover, assimilate partitioning, storage, remobilization, defoliation     

Long-term Partitioning, Storage and Re-mobilization of 14C Assimilated by Lolium perenne (cv. Melle)

The youngest fully expanded leaves of single tillers of vegetativeperennial ryegrass plants were exposed to ¹⁴CO₂. Thereafter,quantitative and fractional analysis of the partitioning, storageand re-mobilization after defoliation of the ¹⁴C-labelled assimilatewas sequentially conducted over a 22 d period. In undefoliated plants, most ¹⁴C reached its final destinationwithin 5–6 of feeding. Forty per cent of assimilated ¹⁴Cwas subsequently lost through respiration, while 13.5, 8.5 and34 per cent remained in roots, stem bases and tops respectively.At least some ¹⁴C was distributed to tillers throughout theplant, but secondary tillers subtended by the fed tiller madethe greatest demand on ¹⁴C translocated from the fed tiller. A small, but significant portion of ¹⁴C was invested into longterm storage in undefoliated plants, four per cent of the totalassimilated still being present in a labile chemical form inroots and stem bases 22 d after feeding. In plants that wereseverely defoliated 4 d after feeding, depletion of reserve¹⁴C was observed relative to undefoliated plants. The depletiontook place from stem bases, not roots, and both low and highmolecular weight storage compounds were involved. A portionof the depleted ¹⁴C was incorporated into new growth after defoliation. Lolium perenne, perennial ryegrass, assimilate partitioning, storage, re-mobilization, defoliation     

The Distribution Pattern of 14Carbon Assimilated by the Third Leaf of Wheat

The third leaf of wheat, variety Jufy I, was allowed to assimilate¹⁴CO₂ for 2 hrs.; after a further hour the distribution patternof the assimilates was determined. Uptake of ¹⁴CO₂ and assimilatesleaving the leaf increased until the leaf was fully expanded,then slowly decreased. High proportions of labelled translocates were recorded in boththe stem and the the root system, that in the roots increasinggreatly as movement of translocates to the leaves decreased.The two fully grown leaves, L₁ and L₂, imported only slightamounts of labelled translocates. Movement of labelled translocateto each of the younger leaves in turn occurred in a strikingpattern, such that import into a given leaf reached a maximumwhich coincided with its maximum rate of growth, subsequentlyfalling rapidly and reaching a very low level by the time theleaf is fully grown. The results are discussed in relation to what is known of thegeneral pattern of growth and translocation in the wheat plant.     

Re-interpretation of an Experiment on the Role of Assimilated Transport Resistance in Partitioning in Tomato

The importance of transport resistance (distance between sourceand sink) on assimilate partitioning in tomato is questioned.Slack and Calvert ( Journal of Horticultural Science 52 : 309–315,1977) concluded that, in tomato, excising of fruit trusses showeda direct influence of distance from source on assimilate partitioning.A dry matter distribution model for tomato, based on the hypothesisthat distribution is regulated by the sink strengths of theplant organs and that no influence of transport resistance onpartitioning exists, has been described and validated by Heuvelink( Annals of Botany 77 : 71–80, 1996). Using this model,it is shown that the results of Slack and Calvert (1977) canbe explained more simply on the basis of the succession of trusseswith growth shifted with respect to time. Therefore, their resultsdo not prove that transport resistance plays a role in assimilatepartitioning. Allocation; distance; dry matter distribution; model; assimilate pool; partitioning; simulation; transport resistance; tomato     

Carrier Protein-mediated Transport of Neutral Amino Acids into Mung Bean Mitochondria

The transport of neutral amino acids into mitochondria isolated from the hypocotyl of mung bean (Roxb.) was studied by the swelling technique. Isolated mitochondria swelled when added to an isosmotic solution of proline, serine, methionine, threonine, alanine, and glycine. The swelling was stereospecific in that it was faster in the l-amino acid than in the corresponding d-amino acid. Preincubation of the mitochondria with the sulfhydryl modifying reagents, p-mercuribenzoate and mersalyl, resulted in an inhibition of the swelling caused by proline, serine, threonine, and glycine. The swelling induced by alanine was inhibited only by mersalyl, whereas that by methionine was inhibited only by p-mercuribenzoate. In all cases, the inhibition caused by the sulfhydryl modifying reagents was readily reversible by the subsequent treatment of the mitochondria with dithiothreitol. N-Ethylmaleimide, another sulfhydryl-modifying reagent, did not cause any inhibition of the swelling. The findings indicate the existence of a protein mediated mechanism for the transport of neutral amino acids into plant mitochondria.     

Effects of Ear Removal on Photosynthesis, Carbohydrate Accumulation and on the Distribution of Assimilated 14C in Wheat

Removal of an ear from a tiller of a wheat plant growing inthe field did not result in any marked change in the net photosyntheticrate of the subtending flag leaf, even during the period whenthe ear would normally have received large amounts of assimilatefrom the flag leaf. Following ear removal, there was an increasein the amount of ethanol-soluble and ethanol-insoluble carbohydratesin the remaining organs of the tiller. ¹⁴C labelling studiesshowed that a new pattern of translocation was established within2–3 days of ear removal, and the tiller exported assimilateto other tillers on the plant, and possibly to the roots.     

Secondary Transport of Amino Acids in Prokaryotes

Amino acid transport is a ubiquitous phenomenon and serves a variety of functions in prokaryotes, including supply of carbon and nitrogen for catabolic and anabolic processes, pH homeostasis, osmoprotection, virulence, detoxification, signal transduction and generation of electrochemical ion gradients. Many of the participating proteins have eukaryotic relatives and are successfully used as model systems for exploration of transporter structure and function. Distribution, physiological roles, functional properties, and structure-function relationships of prokaryotic α-amino acid transporters are discussed.     

Effect of Source-to-sink Ratio on Partitioning of Dry Matter and14C-photoassimilates in Wheat during Grain Filling

Increasingly, wheat (Triticum aestivumL.) is being grown intropical environments, but there is inadequate information aboutthe physiological processes limiting yield. In this investigation,the source:sink ratio was manipulated to examine the performanceof source-sink interactions after anthesis and the factor(s)limiting grain filling in tropical conditions. Plants of threewheat cultivars, Cuba C-204, Candeias and IAC-60, were artificiallymodified to give different source:sink ratios. The treatmentswere: I, Control; II, all spikelets on one side of the spikeremoved; III, all spikelets removed except the four centralspikelets of the spike; and IV, flag leaf blade removed. Thedistribution of dry matter between kernels and stem internodeswas analysed at harvest in all three cultivars. Partitioningof¹⁴C-photoassimilates was measured on three occasions afteranthesis in the cultivar Cuba C-204. Modifications of source:sinkratio led to different patterns of allocation of dry matterbetween cultivars and sowing dates. The reduction in sink sizein treatment II produced no significant change in the mass pergrain in the January sowing, but this was enhanced in two cultivarsin the November sowing. In treatment III, both mass per grainand translocation of¹⁴C-photoassimilates declined, apparentlydue to feedback inhibition of photosynthesis. The participationof stem reserves in grain filling and the existence of genotypicdifferences in response to availability of photoassimilateswere corroborated. The pattern of partitioning of dry matterobserved in plants in this investigation suggests a source limitation,particularly during the November sowing. This pattern differedmarkedly from that in other studies, most of which have beenmade in temperate areas.Copyright 1999 Annals of Botany Company Photoassimilates, sink, source, partitioning, grain filling, wheat.     

Incorporation of 5-Diazouracil-2-14C into Ribonucleic Acids of Escherichia coli During Division Inhibition

Radioactive diazouracil (DZU-2-(14)C) was rapidly incorporated into acid-insoluble material of Escherichia coli B. Isolated ribonucleic acid contained essentially all of the incorporated label, and this was solubilized by ribonucleases but not by deoxyribonuclease. A maximum of 45 to 50% of added label was incorporated at division-inhibitory and subinhibitory concentrations. Incorporation levels and filament-forming capacity were concomitantly depressed by preincubation of DZU in various medium components. The lower levels of incorporation brought about by preincubation were apparently related to an inherent instability or high reactivity of the DZU. The limit on incorporation of fresh DZU suggests the presence of geometrical isomers in the original DZU. The nature of these isomers and the reactivity of DZU are discussed in relation to its use in future cell division studies.     

Transport of Recently Assimilated15N Nitrogen to Individual Spikelets in Spring Wheat Grown in Culture Solution

Two cultivars of spring wheat ( Triticum aestivum L.), Sport(high protein) and WL4 (low protein), were grown to maturityin culture solution. Nitrogen in the form of nitrate was addedin daily doses at stepwise-decreasing relative rates to ensurenormal development, and both cultivars received the same totalamount of N during development. At weekly intervals from anthesisto maturity the daily nitrate dose was, for selected groupsof plants, labelled with¹⁵N. After the labelling period theselected plants were harvested and analysed. The cultivar WL4produced more biomass than Sport, as well as more spikeletsand more grains per ear, with a higher mean grain weight, suchthat grain yield of WL4 was 57% greater than Sport. The earsof both cultivars were heterogenous: mean grain weight was highestin middle spikelets, which also contained more grains; the Ncontent followed the pattern of dry weight with more N in themiddle spikelets; but the N concentration was practically thesame in all spikelets (2.15% of d. wt in WL4 and 3.33% in Sport).The distribution of¹⁵N showed that the main stem ear maturedmuch earlier than tiller ears. The results of this nitrogen-labellingexperiment show that, late in development, substantial amountsof recently-absorbed N were immediately assimilated and transportedto the ears. Transport of¹⁵N decreased earlier to the top spikeletsthan to the bottom spikelets. As both cultivars were grown underidentical conditions and both received the same amount of Nit was concluded that the difference in grain N concentrationwas not caused by differences in the capacity of N assimilationand translocation but rather by different rates of accumulationof non-nitrogenous dry matter in the grains. Ear; grain; nitrate; nitrogen transport; Triticum aestivum L.; yield     

Allocation and Turnover of Photosynthetically Assimilated CO(2) in Leaves of Glycine max L. Clark

The allocation and turnover of photosynthetically assimilated ¹⁴CO₂ in lipid and protein fractions of soybean (Glycine max L. Clark) leaves and stem materials was measured. In whole plant labeling experiments, allocation of photosynthate from a pulse of ¹⁴CO₂ into polymeric compounds was: 25% to proteins in 4 days, 20% to metabolically inert cell wall products in 1 to 2 days, 10% to lipids in 4 days, and 4% to starch in 1 day. The amount of ¹⁴C labeled photosynthate that an actively growing leaf (leaf 4) used for its own lipid synthesis immediately following pulse labeling was about 25%. The ¹⁴C of labeled proteins turned over with half-lives of 3.8, 3.3, and 4.1 days in leaves 1, 2, and 3, respectively; and turnover of ¹⁴C in total shoot protein proceeded with a half-life of 5.2 days. Three kinetic ¹⁴C turnover patterns were observed in lipids: a rapid turnover fraction (within a day), an intermediate fraction (half-life about 5 days), and a slow turnover fraction. These results are discussed in terms of previously published accounts of translocation, carbon budgets, carbon use, and turnover in starch, lipid, protein, and cell wall materials of various plants including soybeans.     

Phloem Transport of Amino Acids in Relation to their Cytosolic Levels in Barley Leaves

A comparison of barley (Hordeum vulgare L.) leaves was made between the cytosolic content of amino acids and sucrose as determined by subcellular fractionation and the corresponding concentration in phloem sap, which was collected continuously for up to 6 days from severed aphid stylets. Because amino acids were found to be almost absent from the vacuoles, and because the amino acid patterns in the stroma and cytosol are similar, whole leaf contents could be taken as a measure of cytosolic amino acid levels for a comparison of data during a diurnal cycle. The results show that the pattern of amino acids in the phloem sap was very similar to the pattern in the cytosol. Therefore, we concluded that the overall process of transfer of amino acids from the cytosol of the source cells into the sieve tubes, although carrier mediated, may be a passive process and that the translocation of amino acids via the sieve tubes requires the mass flow of sucrose driven by the active sucrose transport involved by the phloem loading.