Preparation of Radioactive Starch, Glucose and Fructose from C14O2

Radioactive starch, glucose and fructose have been preparedfrom tobacco leaves after assimilation of C¹⁴O₂. The apparatusused for photosynthesis consisted of a shallow Perspex leafchamber connected to a closed gas system, in which C¹⁴O₂ wasgenerated from BaC¹⁴O₂. Six leaves, area 14 to 18 sq. dm. whenexposed to bright sunlight with an initial CO₂ concentrationof 8 to 10 per cent., assimilated 3.35 g. of C¹⁴O₂ in 8 to 10hours. At least 80 per cent. of the C¹⁴O₂ supplied appearedin the leaves as starch and sugar and over 80 per cent. of theradioactivity was accounted for in these carbohydrates. Thespecific activity per m. atom of carbon of the isolated productswas 85 to 90 per cent. of that of the C¹⁴O₂. Small amounts ofradioactive carbon were also incorporated in the leaf proteinand in the celluose, hemicellulose and polyuronides.     

Photosynthesis of Lamina and Sheath of Barley Leaves

Apparent photosynthesis, in mg. CO₂ absorbed per dm.² per hour,of the sheath and enclosed stem of a barley leaf was about 50per cent. of that of the lamina of the same leaf, when the photosynthesizingarea was measured as one side of the lamina and the outer exposedsurface of the sheath. Apparent photosynthesis of a particularlamina or sheath was about 70 per cent. of that of the one aboveon the same stem. Respiration per dm.², though not per g. dry weight, of sheathwith enclosed stem was greater than of lamina in one experimentdone with low-intensity illumination so that true rates of photosynthesisof lamina and sheath were similar. Differences in respirationrates per unit area of laminae and sheaths probably accountedfor most of the greater apparent photosynthesis of the formerin other experiments done with higher intensity illumination. It is suggested that for growth-analysis studies the size ofthe photosynthetic system of cereals should be measured as thatof one side of the leaf laminae plus the outer surface of thecombined leaf sheaths. In the later stages of growth the surfacearea of exposed stem and peduncle should also be included.     

Photosynthesis of Ears and Flag Leaves of Wheat and Barley

Immediately after anthesis ears of spring wheat absorbed lessthan 0.5 mg CO₂, per hour in daylight and later evolved CO₂,in the light and in the dark. The rate of apparent photosynthesisof the combined flag-leaf lamina and sheath and peduncle (collectivelycalled flag leaf) of two spring wheat varieties, Atle and JufyI, was 3–4 mg per hour; the rates of the flag leaf andthe ear of two spring barleys, Plumage Archer and Proctor, wereeach about 1 mg per hour. The gas exchange of ears and flag leaves between ear emergenceand maturity accounted for most of the final grain dry weight.The CO₂, fixed by the wheat ear was equivalent to between 17and 30 per cent of the grain weight, but more than this waslost by respiration, so assimilation in the flag leaf was equivalentto 110–20 per cent of the final grain weight. In barley,photosynthesis in the flag leaf and the net CO₂ uptake by theear each provided about half of the carbohydrate in the grain. Barley ears photosynthesized more than wheat ears because oftheir greater surface, and flag leaves of wheat photosynthesizedmore than those of barley because they had more surface anda slightly greater rate of photosynthesis per dm².     

Varietal Differences in Photosynthesis of Ears and Leaves of Barley

Rates of apparent photosynthesis of ears and of the combinedflag leaf and sheath and peduncle of Proctor barley grown inpots or in the field were similar to those of Plumage Archer,or slightly smaller when the dimensions of the ear and leafarea of Proctor were less than those of Plumage Archer. Thephotosynthesis rate of the ear—about 1.0 mg. CO₂ per earper hour—was similar or slightly less than the rate ofthe flag leaf and sheath and peduncle. These rates of photosynthesisindicated that 40-50 per cent, of the carbohydrate in the grainwas provided by photosynthesis in the shoot and about 40 percent, by photosynthesis in the ear. The total CO₂ fixed by theear was equivalent to about 60 per cent, of the grain weight,20 per cent, being lost by respiration. Shading the ear underestimatedthe total amount of CO₂ fixed by the ear and decreased dry weightof grain per ear of both Proctor and Plumage Archer by 26 percent., as in pots. The contribution of ear photosynthesis toyield of grain per acre was greater for Proctor than for PlumageArcher because Proctor had more ears. The rate of apparent photosynthesis per dm.² of leaves of Proctorwas similar to that of Plumage Archer both before and afterear emergence. Before ear emergence, the photosynthesis rateof a particular leaf decreased linearly with time and was slowerfor lower than for higher leaves on the shoot. Respiration ratesper g. dry weight of ears of Proctor and Plumage Archer weresimilar; in one experiment the leaves of Proctor respired slightlyfaster than those of Plumage Archer.     

Effects of Nitrogen Fertilizer on the Distribution of Photosynthate during Grain Growth of Spring Wheat

The distribution of photosynthate labelled with ¹⁴C was studiedin spring wheat grown with different amounts of nitrogen fertilizerin the three years 1972–4, after exposing the flag leafor the leaf below the flag leaf to ¹⁴CO₂ at 6–10 or 19–26days after anthesis. The movement of ¹⁴C to ears was unaffectedby nitrogen fertilizer except after early exposure in 1973,when nitrogen increased the retention of ¹⁴C in stems at maturity The concentration of sugar in the top part of the shoot at theend of the day was unaffected by nitrogen in 1973, but at 22days after anthesis in 1974 the concentration of sucrose inthe glumes and rachis, and in the flag leaf lamina was increasedby nitrogen. Loss of sugar by translocation and respirationduring the night may explain why this increase in concentrationwas not reflected in the ¹⁴C distribution 24 h after supplying¹⁴C. The proportion of the total ¹⁴C content of the shoot that wasin the ear at maturity ranged from 68 to 95 per cent dependingon when and to which leaf the ¹⁴CO₂ was supplied. Less than5 per cent remained in the leaf exposed to ¹⁴CO₂. The proportionof the final ear weight contributed by the leaf below the flagleaf was about half that contributed by the flag leaf. In 1974 about 24 per cent of the ¹⁴C absorbed by the flag leaf,and 56 per cent of that absorbed by the second leaf, was lostby maturity, presumably by respiration. Most loss occurred inthe first 24 h.     

Dark Respiration of Several Varieties of Winter Wheat Given Different Amounts of Nitrogen Fertilizer

Carbon dioxide production in the dark by ears and by the restof the shoot of winter wheat grown in the field was measuredin 2 years during grain growth. The respiration rate per g d.wt of the ears was increased by nitrogen fertilizer. Ears ofthe semi-dwarf varieties Maris Fundin and Hobbit respired moreslowly than ears of Maris Huntsman and Cappelle-Desprez. Respirationrates of the rest of the shoot were unaffected by nitrogen orvariety. The amount of carbohydrate required to provide the CO₂ respiredduring the whole period of grain growth varied from 163 to 443g m^–2, or 42 to 76 per cent of the dry weight of the grain.More than half the CO₂ lost was respired by the ear. The additionof 180 kg N ha^–1, which increased grain yield by 78 percent in 1975, almost trebled the amount of CO₂ lost by the ears.The semi-dwarf varieties lost less CO₂ from ears and shootsthan did the taller ones, and had larger yields of grain. Respiration was also estimated from the difference between the¹⁴C contents of shoots sampled immediately after a 30 s exposureto ¹⁴CO₂ and at maturity. When 14C was supplied 10 days afteranthesis, the loss by maturity amounted to 16–28 per centof that initially absorbed by flag leaves and 40 per cent ofthat absorbed by the leaf below the flag leaf. Most of the lossoccurred in the first day. The loss of 14C by maturity was significantlyincreased by nitrogen fertilizer in 1975. Triticum aestivum L., wheat, respiration, nitrogen supply, fertilizer treatment     

Nitrogen Deficiency in Small Closed Communities of S24 Ryegrass. I. Photosynthesis, Respiration, Dry Matter Production and Partition

Small communities of S24 ryegrass were grown under supplementarylights in a glasshouse at 20°C, and abundantly suppliedwith a complete nutrient solution containing 300 p.p.m. of nitrogen,until they had a leaf area index of 5 and fully interceptedthe light. Half were then given a solution containing only 3p.p.m. of nitrogen (LN) while the rest were kept at 300 p.p.m.(HN). The LN plants had a rate of single leaf photosynthesis lowerthan that of the HN plants at all but the lowest light intensities(33 per cent lower at the saturating irradiance of 170 W m^–2).Similarly, the LN communities had rates of canopy gross photosynthesis(P_sc) markedly lower than those of the HN communities. A comparisonof the observed rates of P_sc with those predicted by a mathematicalmodel of canopy photosynthesis indicated that it was the effectof nitrogen on single leaf photosynthesis, rather than differencesbetween the communities in leaf area, which led to the observeddifferences in P_sc. The superiority of the HN communities in terms of P_sc was partlyoffset by a higher rate of respiration so that they only exceededthe LN communities in terms of canopy net photosynthesis atirradiances in excess of 180 W m_–2, and produced only15 per cent more total dry matter. Nevertheless, the HN plantsdirected less of that dry matter into root and more into topsso that they came to possess twice the weight of live laminae,and the HN communities twice the leaf area, of their nitrogendeficient counterparts. Lolium perenne, S24 ryegrass, photosynthesis, respiration, dry matter production and partition, nitrogen dekieacy     

Effect of Temperature on Photosynthesis and Photorespiration of Wheat Leaves

Wheat plants were grown in a controlled environment with daytemperatures of 18 ?C and with 500 µ Einsteins m^–28^–1 of photosynthetically active radiation for 16 h. Beforeanthesis and 2 to 3 weeks after, rates of net photosynthesiswere measured for leaves in 2 or 21% O₂ containing 350 vpm CO₂at 13, 18, 23, and 28 ?C and with 500 µEinsteins m^–2s^–1 of photosynthetically active radiation. Also, underthe same conditions of light intensity and temperature, therates of efflux of CO₂ into CO₂-free air were measured and,for mature flag leaves 3 to 4 weeks after anthesis, gross andnet photosynthesis from air containing 320 vpm ¹⁴CO₂ of specificactivity 39?7 nCi µmol^–1. When the O₂ concentration was decreased from 21 to 2% (v/v)the rate of net photosynthesis increased by 32 per cent at thelowest temperature and 54 per cent at the highest temperature.Efflux of CO₂ into CO₂-free air ranged from 38 per cent of netphotosynthesis at 13 ?C to 86 per cent at 28 ?C. Gross photosynthesis,measured by the ¹⁴C assimilated during 40 s, was greater thannet photosynthesis by some 10 per cent at 13 ?C and 17 per centat 28 ?C. These data indicate that photorespiration was relativelygreater at higher temperatures.     

Respiratory Loss of Recently Assimilated Carbon in Wheat

A series of experiments was undertaken to assess the amountof respiration associated with the growth of wheat at differentstages. Plants (or in some cases just the flag leaf) were labelledwith ¹⁴CO₂ and the amount of ¹⁴CO₂ respired during the subsequent48 or 72 h was measured. The evolution of ¹⁴C, expressed asa percentage of the amount initially assimilated (referred toas the R/A value) was used as a measure of the overall efficiencyof dry matter production. Respiratory ¹⁴CO₂ evolution from labelledplants was most rapid in the first 12 h after labelling, thereafterdeclining rapidly. Evolution was also more rapid following labellingsat the end of the light period (dusk) than at the beginningof it (dawn). The R/A values were greatest (42 and 50 per centrespectively for dawn- and dusk-labelled plants) for young plantsand least (13 and 28 per cent respectively) for plants duringmid grain filling. When flag leaves, as distinct from wholeplants, were labelled, R/A values were lower still (9 and 21per cent respectively), indicating that flag leaf assimilatewas used efficiently in grain production. The calculated minimum R/A for the formation of grain material(10 per cent protein, 90 per cent starch) was 6.2 per cent.That the experimentally determined values were greater thanthis is attributed to the turnover of carbon in enzymes, toother maintenance processes, and possibly to the operation ofthe pentose phosphate pathway of glucose oxidation. R/A valueswere lower in those plants labelled at the beginning than thoseat the end of the photoperiod. This was considered to be a consequenceof refixation of respiratory ¹⁴CO₂ during the light. The higherR/A values found for young plants were considered to be a consequenceof the greater percentage of ¹⁴C translocated to the roots (rootsbeing unable to refix respired CO₂) and of greater turnoverof enzymes associated with more active metabolism. Triticum, wheat, respiration, carbon assimilation, carbon loss, grain-filling     

Effects of Certain Inhibitors on Photorespiration by Wheat Leaf Segments

The effect on the carbon metabolism of wheat leaf segments ofcertain inhibitors of photorespiration was studied. Sodium 2-hydroxy-3-butynoatesupplied for 40 min resulted in accumulation of ¹⁴C in glycolicacid with only a 7% inhibition of photosynthesis; when suppliedfor 90 min, photosynthesis was inhibited by 47%. When ¹⁴CO₂was replaced by 1000 vpm ¹²CO₂, radioactivity in glycine decreasedbut increased more rapidly in sucrose with less release of ¹⁴CO₂.Isonicotinyl hydrazide (INH) inhibited photosynthesis from ¹⁴CO₂by 50% and glycine replaced sucrose as the main product. When,after 15 min, ¹⁴CO₂ was replaced by 150 vpm ¹²CO₂, in the presenceof INH less ¹⁴CO₂ was released, ¹⁴CO in glycine decreased moreslowly, and less [¹⁴CO]sucrose accumulated. Glycidate (potassium2,3-epoxypropionate) at 2 mM had no effect on photosyntheticrate and little effect on carbon metabolism; 20 mM glycidateinhibited photosynthesis by 64% and resulted in less radioactivityin glycine, more in phosphate esters, and less ¹⁴CO₂ released.When photosynthesis was measured in 1000 vpm CO₂ the inhibitorsgave smaller effects on metabolism than during photosynthesisfrom 150 vpm ¹⁴CO₂ but 20 mM glycidate still resulted in a 42%inhibition of photosynthesis. When U- [¹⁴CO]glycerate was appliedto leaf segments in air with 320 vpm ¹⁴CO₂ the total uptakeof glycerate was not changed by the inhibitors. INH and glycidateboth decreased the amount of glycerate metabolised. More ¹⁴COaccumulated in glycine in the presence of INH and in phosphateesters and serine in the presence of glycidate. Hydroxybutynoateincreased the production of glycolate from glycerate but didnot affect the total amount of glycerate metabolised. Although all three inhibitors affected photorespiratory metabolismnone stimulated photosynthesis. The results are consistent withthe main release of CO₂ in photorespiration arising from theconversion of glycine to serine.     

The Production, Storage, and Translocation of Carbohydrates in Developing Potato Plants

¹⁴C-tracer experiments showed that the export of assimilateslabelled with a short pulse of ¹⁴CO₂ continued for 5 weeks.There appeared to be an approximately exponential loss of ethanol-insoluble¹⁴C from the haulm with a half-time of about 3.5 weeks. Initiallythere was a poor correlation between the fresh weight of thetubers and their ¹⁴C content but the correlation became goodafter 5 weeks. Field data showed that, in many instances, the rate of tubergrowth was greater than the rate of growth of the whole plant.This suggests the transfer of large amounts of dry matter fromthe haulm to the tubers. Dry-matter loss from the haulm couldaccount for the discrepancy and the amount transferred in thisway was about 10 per cent of the final dry weight of the tuber. There was an increase in net assimilation rate when the leafarea of the plants started to decline. It is suggested thatthis increase was due to an increase in the rate of photosynthesisbrought about by the influence of the rapidly growing tubers.     

Metabolism of Radioactive Sugars by Tobacco leaf Disks

Destarched tobacco-leaf disks were floated on per cent. (w/v)solutions of sucrose uniformly labelled with ¹⁴C in either theglucose or fructose moiety, and on invert sugar in which onehexose only was so labelled. The experiments were carried outin an atmosphere of oxygen at 25° C. Seventy-five per cent,of the sugar lost from the external solutions was recoveredas starch, sucrose, fructose, glucose, and CO₂. With sucroseas the substrate, 30 per cent, of the material was recoveredas CO₂ and 17 per cent. each as starch, sucrose, fructose, andglucose. With invert sugar as the substrate, 30 per cent, wasagain recovered as CO₂ only 20 per cent. as the three sugarstogether, and 50 per cent. as starch. Whichever hexose was initiallylabelled and whether the sugar was supplied as sucrose or hexose,the relative specific activities of starch and sucrose in theleaf disks and of the CO₂ evolved were equal or nearly equalto that of the sugar supplied. With sucrose as the substratethe sucrose in the disks retained its asymmetry of label, andfree hexoses produced were similarly asymmetrically labelled.When invert sugar was the substrate the sucrose synthesizedwas strongly labelled in both moieties, as also were the freehexosea. It is concluded that fructose and glucose free or combinedin sucrose were equally available for starch synthesis and CO₂,formation, and that there can be no question of preferentialutilization of one or other hexose. Starch and CO₂ must arisefrom a common source in which readily formed derivatives ofthe hexoses are rapidly equilibrated. Free hexose cannot participatedirectly in either sucrose or starch synthesis. Accumulationof sugar not immediately metabolized and inversion of sucrosetake place at a site remote from the common pool. A scheme toaccommodate the results is discussed.     

Carbon Translocation in the Tomato: Carbon Import and Fruit Growth

The rates of carbon import by fruits were measured over 48 has the sum of the change in the total organic carbon contentof the fruit and the respiratory loss of carbon. Over a rangeof fruit sizes from 20–90 per cent of the maximum volumethe smaller fruits imported carbon at an absolute rate (mgCfruit^–1 h^–1) nearly twice that of the larger fruits.The imported leaf assimilates, identified as the ¹⁴C-compoundsalong the pathway between a ¹⁴CO₂-fed leaf and a young fruit,comprised 90 per cent sucrose and 10 per cent glutamic acid,aspartic acid and malic acid. Within the fruit the imported¹⁴C-sucrose was hydrolysed into hexoses. The changes in thelevels of starch and insoluble residue in the fruit were positivelycorrelated with the carbon import rates. In the largest fruitswith the lowest import rates, there was breakdown of insolubleresidue and less accumulation of starch, but a significant increasein the level of sucrose. The sink strength of a tomato fruitis dependent more on sink activity than on sink size.     

Cell Wall Metabolism in Developing Strawberry Fruits

Cell wall metabolism was studied in strawberry receptacles (Fragariaananassa, Duchesne) of known age in relation to petal fall (PF).Polysaccharide and protein composition, incorporation of [¹⁴C]glucoseand [¹⁴C]proline by excised tissue, and the fate of ¹⁴CO₂ fixedby young, attached fruits were followed in relation to celldivision, cell expansion, fine structure, and ethylene synthesis. Cell division continued for about 7 d after PF although vacuolationof cells was already beginning at PF and the subsequent cellexpansion was logarithmic. There was an associated logarithmicincrease in sugar content per cell and a decreasing rate ofethylene production per unit fresh weight. During cell expansion radioactivity from [¹⁴C]glucose was incorporatedinto fractions identified as starch and soluble polyuronideand into glucose and galactose residues in the cell wall. Radioactivityfrom [¹⁴C]proline was also incorporated into the cell wall,but only 10 per cent of this activity was found in hydroxyproline.Correspondingly wall protein contained a low proportion of hydroxyprolineresidues. The proportion of radioactivity from ¹⁴CO₂ fixed byfruitlets remained constant in most sugar residues in the cellwall. The proportion of radioactivity in galactose fell, indicatingturnover of these residues. Between 21 and 28 d after PF receptacles became red and softenedbut there was no change in the rate of ethylene production.Cell expansion continued for at least 28 d. Tubular proliferationof the tonoplast and hydration of middle lamella and wall matrixmaterial had begun 7–14 d after PF but became extremeduring ripening. Associated with the hydration of the wall,over 70 per cent of the polyuronide in the wall became freelysoluble, and arabinose and galactose residues lost from thewall appeared in soluble fractions. There was no increase intotal polysaccharide during ripening and incorporation of [¹⁴C]glucoseinto polysaccharides ceased, although protein increased andincorporation of [¹⁴C]proline into wall protein continued.     

Effect of Temperature on Starch Degradation in Chlorella vulgaris 11h Cells

Analysis of products formed in Chlorella vulgaris 11 h cellsduring photosynthesis in air containing 3,000 ppm ¹⁴CO₂ at varioustemperatures revealed that the level of ¹⁴C-starch was maximumaround 20–24?C and decreased with further rise in temperatureuntil 40?C, while ¹⁴C-sucrose greatly increased at temperaturesabove about 28?C. Elevating the temperature from 20 to 38?Cduring photosynthetic ¹⁴CO₂ fixation resulted in a remarkabledecrease in ¹⁴C in starch and a concomitant increase in ¹⁴Cin sucrose. This conversion of starch to sucrose when shiftingthe temperature from 20 to 38?C proceeded even in the dark.Hydrolysis of sucrose by rß-fructosidase showed that,irrespective of the experimental conditions, the radioactivitiesin sucrose were equally distributed between glucose and fructose.The enhancement of starch degradation with temperature risewas more remarkable than that of the activity of ribulose bisphosphatecarboxylase from the same cells. When Chlorella cells whichhad been preloaded with ¹⁴C-starch after photosynthesis for30 min at 20?C were incubated in the dark for an additional30 min at 20?C, ¹⁴C-starch was degraded by only about 4%. However,the values after 30-min dark incubation at 28, 32, 36 and 40?Cwere increased by about 10, 19, 36 and 50%, respectively. Duringthe temperature-dependent conversion of starch to sucrose, nosignificant amount of radioactivity accumulated in free glucoseand maltose. (Received October 27, 1981; Accepted January 9, 1982)     

Effects of CO2 Enrichment at Different Irradiances on Growth and Yield of Wheat: II. EFFECTS ON KLEIBER SPRING WHEAT TREATED FROM ANTHESIS IN CONTROLLED ENVIRONMENTS IN RELATION TO EFFECTS ON PHOTOSYNTHESIS AND PHOTORESPI RATION

Spring wheat plants were grown in a cage with a glass roof untilthree days after anthesis and then subjected to treatments inconstant environment rooms with any one of all combinationsof four irradiances and two concentrations of carbon dioxide.The photoperiod was 16 h and day/night temperatures 19?C/14?C.Growth and yield of grain were saturated at the two brightestirradiances. Carbon dioxide enrichment from 350 to 1200 mm³dm^–3 increased shoot dry weight and grain yield at finalharvest at all irradiances, by averages of 10.5 (not significant)and 23.5 (significant) percent respectively. However, increasingthe irradiance from 150 to 613 µE m^–2 s^–1caused much larger yield increases (approximately 3-fold). Increasedgrain production by increased light was caused by both increasesin dry weight per grain and by increases in grain number perspikelet. The increase caused by CO₂ enrichment was mainly becauseof increased dry weight per grain. Increase in ear dry weightcaused by CO₂ enrichment took place between 30 and 60 d afteranthesis. The increase in shoot dry weight took place immediatelyafter exposure to increased CO₂ from 3 to 15 d after anthesis.Net photosynthesis by flag leaves on the main shoots was almostdoubled 16 d after anthesis by the CO₂ enrichment even thoughstomatal resistance was also doubled. However, this increasewas not reflected by a proportional increase in yield, probablybecause increased mutual shading by bigger stems and late tillersreduced total assimilation and because of increased respirationby the shoots. The increase in photosynthesis was not due toa decrease in photorespiration but to an increase in gross photosynthesis. Key words: CO₂enrichment, Photosynthesis, Photorespiration     

Effect of CO2 Concentration on Growth of Sugar-beet, Barley, Kale, and Maize

Increasing the concentration of CO₂ in the air from the usual300 ppm to 1, 000 ppm in growth rooms with temperatures of 20°C during the 16-h light period and 15° C during the 8-hdark period increased the total dry weight of sugar-beet, barley,and kale by about 5o per cent. A further increase in CO, concentrationto 3, 300 ppm increased dry weight slightly more. These effectsoccurred with light intensities ranging from 3.7 to II.6 caldm–² min–¹ of visible radiation supplied by a mixtureof fluorescent and tungsten lamps, and were only slightly greaterwith the brighter light. Extra CO₂ also increased leaf area,though relatively less than dry weight, and the number of barleyshoots but not of sugar-beet or kale leaves; it decreased leaf-arearatio, specific leaf area, and the ratio of tops to roots. Maizewas taller with extra CO₂. Net assimilation rates in 1, 000 and 3, 300 ppm CO₂ were about20 and 30 per cent respectively greater than in 300 ppm. Uptakeof CO₂ in the light by complete tops and single leaves alsoincreased with increase in CO₂ concentration. Photosynthesisof leaves of plants recently transferred to a new CO₂ concentrationdepended only on that concentration and not on the originalone. Doubling the light intensity from 3.7 to 7.7 cal dm–²min–¹ affected dry weight, leaf area, net assimilationrate, etc., similarly to a tenfold increase in CO₂ concentration.     

Effects of Nitrogen Fertilizer on Growth and Yield of Spring Wheat

Nine amounts of nitrogen fertilizer, ranging from 0 to 200 kgN ha^–1, were applied to spring wheat cv. Kleiber in the3 years 1972-1974. In 1972 grain dry weight with 125 kg N ha^–1or more was 100 g m^–2 (23 per cent) greater than withoutnitrogen. Grain yield was unaffected by nitrogen in the otheryears. Leaf area at and after anthesis was increased throughoutthe range of nitrogen tested, most in 1972 and least in 1973.Consequently, the addition of 200 kg N ha^–1 decreasedthe amount of grain produced per unit of leaf area by approximately25 per cent in all years. The dry weight of leaves and stems at anthesis and maturitywas increased by nitrogen in all years, similarly to leaf area.However, the change in stem dry weight between anthesis andmaturity was not affected by nitrogen; stems increased in dryweight for about 20 days after anthesis and then decreased tovalues similar to those at anthesis. The uptake of CO₂ per unit area of flag leaf or second leaf(leaf below the flag leaf) was slightly decreased by nitrogenwhen the increase in leaf area caused by nitrogen appreciablydecreased the light intensity at the surface of these leaves.In spite of such decreases the CO₂ absorbed by flag and secondleaves per unit area of land was always increased by nitrogen,and relatively more than was grain yield. It is suggested that increases in respiratory loss of CO₂ withincreasing nitrogen fertilizer may explain why nitrogen increasedvegetative growth and leaf area relatively more than grain yield.     

Effect of Contrasting Patterns of Nitrate Application on the Nitrate Uptake, N2-Fixation, Nodulation and Growth of White Clover

White clover (Trifolium repens L.) plants were grown from seedin perlite, inoculated with effective rhizobia and exposed tothe same ‘concentration x days’ of ¹⁵N-labellednitrate in four contrasting patterns of doses. Acetylene reductionwas measured at intervals using an open, continuous-flow sytem.Mean dry weight per nodule and rates of acetylene reductionfell rapidly (2–3 d) during periods of exposure to highnitrate concentrations (> 7 mM N) and rose again, equallyrapidly, when nitrate was withdrawn or substantially reduced.The fall in mean dry weight per nodule (50–66 per cent)was almost certainly too large to be accounted for by loss ofsoluble or storage carbohydrate only. No new nodules were formedduring periods of high nitrate availability. When nitrate wassupplied continuously at a moderate concentration (5.7 mM N)nodule numbers stabilised although existing nodules increasedin dry weight by almost four-fold over the 30 d measurementperiod. Treatment had no effect on the percentage nitrogen in planttissues although there were large differences in the proportionsderived from nitrate and N₂-fixation. Plants exposed continuouslyor frequently to small doses of nitrate took up more nitrate,and hence relied less heavily on N₂-fixation, than those exposedto larger doses less often. Increased reliance on nitrate broughtwith it increased total dry weight and shoot: root ratios. Possiblemechanisms involved in bringing about these differences in nitrogennutrition and growth are discussed. White clover, Trifolium repens, nitrate, N₂-fixation, nodule, acetylene reduction, ¹⁵N     

Nitrogen Fixation in the Phyllosphere of Tropical Plants: Occurrence of Phyllosphere Nitrogen-Fixing Micro-organisms in Eastern India and their Utility for the Growth and Nitrogen Nutrition of Host Plants

Of the 560 leaf samples belonging to 259 species of green plantsexamined more than 50 per cent of the Angiosperms and 25 percent of the Pteridophytes and Gymnosperms revealed the presenceof N₂-fixing micro-organisms in their phyllosphere. Plants particularlyremarkable in this respect are orchids and several other epiphytes,Scindapsus officinalis, Ficus and cucurbits. Most of the isolatesappear to be biotypes of Klebsiella pneumoniae. The more activestrains fixed more than 5 mg N g^–1 glucose utilized andreduced more than 100 nmol C₂H₂ mg^–1 cell d. w h^–1. The efficacy of the phyllosphere N₂-fixing isolates for N-nutritionof host plants was studied by spraying suspensions of the culturesgrown on N-free media on rice and wheat seedlings. In IR-26rice or Sonalika and Janak wheat grown on soil in wooden flatsor earthenware pots, 22 per cent of the 161 cultures studiedcaused increased height and about three-quarters of the culturesenhanced dry weight by more than 50 per cent; chlorophyll andN-contents were enhanced more than 50 per cent by about halfand two-thirds of the cultures respectively. In N-free sandculture 26 of the 50 promising strains doubled N-content, and30 doubled dry weight of the tested plants. In some cases dryweight, number of grains per panicle, and 1000 grain weightwere increased by 300, 70–83 and 126–158 per centrespectively; N-content of straw and seed was increased three-or fourfold. In several cases the beneficial effects were foundto match closely the performance of plants receiving ammoniumsulphate. Nitrogen-fixing micro-organisms, nitrogen nutrition, phyllosphere, rice, tropical plants, wheat