The Organic Acid Metabolism of Cox's Orange Pippin Apples: I. SOME EFFECTS OF THE ADDITION OF ORGANIC ACIDS TO THE PEEL OF THE FRUIT

1. The basic respirations (CO₂-output and O₂-uptake) of Cox'sOrangePippin apples and of the peel tissue prepared from themwerecompared in fruit in various stages of development, bothinitiallyand after storage at 12°C. Both show the samegeneral trend,although as the apples become mature the peakvalue of the respirationclimacteric tends to rise in the wholefruit and fall in thepeel.
2. The effect of adding malate orcitrate on the respiration ofthe same samples of peel was studied.
3. Three broad stages of development were observed. During thefirst stage (petal fall to 60 days after) the metabolic patternappears to be different from the two later stages. Here O₂-uptakeas well as C₂-output are influenced by the addition of bothmalate and, to a considerably less extent, citrate. In stage2 (60–125 days from petal fall), the malate effect (CO₂-output)is small until after detachment from the tree, when it risessharply. In stage 3 (125 days to full maturity) the malate effectfollows the course expected for earlier work, namely, it developsat the same time as the climacteric rise in respiration. Thepossible reasons for the different behaviour of the peel atthe three stages is discussed.
4. Results were similar in generaltrend for Cox's Orange Pippinapples grown on different rootstocksand under different culturalconditions.
5. It is suggested thatthe malate effect is most active in theepidermal and hypodermaltissues of the fruit.

     

FLAVOPROTEINS IN THE LEAVES OF HIGHER PLANTS CATALYZING THE OXIDATION OF L-GLUTAMATE

1. Two forms of enzyme capable of catalyzing the oxidation of L-glutamate(and L-aspartate) were isolated from the leaves of spinach andseparated from each other by column-chromatographic purificationon calcium phosphate and anion exchangers. They were distinguishedas GD₁ (L-glutamate dehydrogenase 1) and GD₂ (L-glutamate dehydrogenase2). The purification procedures and some fundamental propertiesof the partially purified enzymes were investigated.
2. It wasdiscovered that the enzymes did not require any cofactor,ie., neither dialysis nor precipitation with ammonium sulfatecaused a fall in enzyme activities and the addition of DPN andTPN to the reaction mixture did not accelerate the reactionrate
3. From the results of spectroscopic investigation GD₁ andGD₂were shown to be flavoproteins, although their prostheticgrouphas not yet been identified The activity of GD₁ was enhancedby the addition of FAD or FMN, while GD₂ was not acceleratedby these factors.
4. The characteristics of the two enzymes includingsubstrate specificity,MICHAELIS constant, optimum pH of thereaction and specificityfor electron acceptors were compared.
5. From the stoichiometric study of the oxidation of L-glutamatewith these enzymes, it was confirmed that the reaction is representedby the following equation: L-glutamate+oxidized dye+h₂o
6. Among various inhibitors tested,molecular oxygen which couldfunction as electron acceptor ofL-glutamate oxidation in thepresence of GD₁ was found to causea strong inhibition uponthe same reaction with TTC as el acceptor.The inhibition wasconfirmed to be due to hydrogen peroxideproduced as a resultof the aerobic oxidation of L-glutamate.

(Received July 25, 1962; )     

Organic Acid Metabolism of Sedum praealtum

1. The organic acids present are citric, isocitric, and l-malic,with a small residue of unidentified acids.
2. The diurnal variationin acidity is due chiefly to changes,in malic acid, with aparallel fluctuation shown by citric acid.Under these conditionsisocitric acid shows little change.
3. The importance of carbondioxide during acidification is confirmed,and it is shown thatat room temperatures or higher the CO₂produced in respirationis sufficient to produce maximum acidification.At lower temperaturesthe supply of CO₂ limits acid production.
4. In the absence ofoxygen no acidification occurs, but even smallquantities (approx.1 per cent.) are sufficient to cause someacid production.
5. Completebalance-sheets are presented for acids, carbohydrates,CO₂ andoxygen for leaves maintained in the dark at high andlow temperatures.As acids are produced there is a correspondingloss of carbohydrate(chiefly starch). A scheme of reactionsis suggested to explainthe experimental results.

     

Studies in Stomatal Behaviour: IV. THE WATER-RELATIONS OF THE EPIDERMIS

1. It is shown that a dry external atmosphere exerts the followingeffects on stomatal movement:

1. A striking accelaration ofclosure in darkness.
2. A slight acceleration of opening in light.
3. If the water-supply to the leaf is impaired, an inabilitytomaintain full opening in the light.

Conversely, a saturatedexternal atmosphere induces sluggishness of movement and a tendencyto incomplete closure in darkness.

1. These results are consideredto support La Rue's contentionthat the epidermal water-supplyis drawn solely by lateral movementfrom the main veins, andnot from the underlying mesophyll.The stomatal phenomena themselvesdo not appear capable of anysimple explanation based on currentknowledge of guard-cellphysiology.
2. The biological significanceof these results is discussed, withparticular reference tothe problem of xeromorphic structures,for which a new interpretationis suggested.

     

Physiological Studies in Plant Nutrition: XVI. The Mineral Nutrition of Bracken

1. The nutritional requirements of the bracken sporophyte wereexamined in a factorial combination of 3 potassium levels x2phosphorus levels x3 solution types, in which the cations weremainly Na, Ca, or NH₄.
2. The effects of nitrogen and phosphorusdeficiency and of shadingunder conditions of high and low potassiumsupply were alsoexamined.
3. Leaf area, total dry weight, andnet assimilation rates aremuch depressed by lack of eitherK or P.
4. Water contents of leaves and rhizomes are generallyincreasedby lack of K when Na is present in the culture mediumbut notwhen Ca is in excess.Phosphorus causes diminished succulence.
5. Starch content increases as phosphorus supply is lowered.Withincreasing doses of K, starch content falls in the highcalciumsolution, while increasing in the other two solutiontypes.
6. Reduction of light intensity is shown to have a beneficialeffectunder conditions of K deficiency.
7. Analysis revealscomplex interactions between the various factors,and possibletoxic effect of Na, NH₄, and excess P. An estimateof the relativeimportance of net assimilation rate, leaf numberand area perleaf in determining total plant size has been calculatedforthe different nutrient treatments. Comparisons are madewithprevious results for barley and flax.

     

Further Studies on the Acid Metabolism of Aspergillus niger: THE FORMATION AND UTILIZATION OF OXALIC ACID

1. Some recent works on the formation of oxalic acid by variousfungi are critically considered.
2. The present work deals withthe role of oxalic acid in the metabolismof Aspergillus niger.
3. When glucose solutions were supplied to preformed mats ofthefungus oxalic acid accumulated, attaining an equilibriumlevelwhich was not exceeded despite the presence of a considerableconcentration of glucose.
4. When the glucose supplies were depletedthe oxalic acid concentrationfell steeply to a low level.
5. Theconcentration of oxalic acid was dependent on the glucoseconcentration.In three separate series of experiments it wasshown that theoxalic acid concentration diminished with increasingglucoseconcentration.
6. Similar results were obtained when the cultureswere rearedfrom spores on culture solutions with the normalamounts ofnutrient salts but different glucose concentrations.
7. In all cases the CO₂ output increased with the glucose concentration.
8. When cultures were supplied with glucose+oxalic acid, theconcentrationof the latter fell steeply to the equilibriumlevel attainedon glucose only. In a culture receiving glucose+oxalicacid,with the oxalic acid concentration somewhat below thenormalequilibrium concentration, the formation of oxalic acidfromthe glucose ceased as soon as the equilibrium level hadbeenattained.
9. When 1 per cent. oxalic acid only was suppliedto the fungusthe concentration gradually diminished to a lowlevel. When3 per cent. oxalic acid was supplied the rate ofacid utilizationsoon fell to low value.
10. In several experimentsit was shown that the rate of CO₂ outputwas higher from culturessupplied with glucose+excess oxalicacid than from culturessupplied with glucose only.
11. The rate of oxalic acid carbonloss was always below that ofthe CO₂ carbon output both incultures supplied with oxalicacid only and in cultures receivingglucose+oxalic acid.
12. The cultures were incapable of utilizingneutral sodium oxalateand the presence of this substance hadno effecft on the ofCO₂ output.
13. The results indicate thatthe utilization of oxalic acid isassociated with the liberationof at least an equivalent amountof CO₂.
14. It is suggested thatthe utilization of oxalic acid is promotedby the presence ofglucose, thus accounting for the lower oxalicacid concentrationsand higher rates of CO₂ output of cultureswith higher glucoseconcentrations.

     

The Organic Acid Metabolism of Bramley's Seedling Apple Peel1

1. The effect of various Krebs cycle acids on the respirationofdisks of apple peel at various stages of maturity was measuredin a Warburg respirometer.
2. Peel tissue from apples at thepre-climacteric and early post-climactericstages apparentlycontain sufficient of the Krebs cycle acidsused, with the exceptionof succinate, to maintain oxidativeprocesses at a maximum.
3. The addition of malate causes a large increase in the CO₂-outputof peel from post-climacteric and senescent fruit but not frompre-climacteric fruit, and a close correlation exists betweenthe climacteric and this decarboxylation of malate. The decarboxylationof malate does not affect the rate of O₂-uptake of peel tissue.The possible part played by the decarboxylation of malate inthe increased CO₂-output at the climacteric is discussed.
4. Addedpyruvate is decarboxylated by the tissue at all stagesof storagelife.
5. The decarboxylation of added malate is an aerobic fermentation,resulting in the quantitative production of acetaldehyde. Althoughthe presence of oxygen is necessary, the rate of O₂-uptake isnot affected by the reaction. Pyruvate decar boxylation doesnot require the presence of oxygen.
6. The O₂-uptake of peelfrom senescent apples can be stimulatedby addition of malate,succinate, and a-ketoglutarate. No evidencewas obtained, however,of oxidation of fumarate, citrate, orpyruvate. The additionof malate to senescent tissue restoresthe lower endogenousrate of O₂-uptake to that of early postclimacteric tissue.
7. Succinate and fumarate are toxic to peel tissue at concentrationabove 0.02M.

     

METABOLISM OF GLUCOSE IN THE PROCESS OF "GLUCOSE-BLEACHING" OF CHLORELLA PROTOTHECOIDES

1. As previously demonstrated, normal cells of Chlorella protothecoidesare bleached with degeneration of chloroplasts when they areincubated, under aerobic conditions—either in the lightor in darkness—, in a glucose-containing medium withoutadded nitrogen source ("glucose-bleaching"). It was found inthe present study that under the atmosphere of N₂, neither bleachingnor growth of algal cells occurs in the dark, while in the lighta significant growth of cells takes place with formation ofa certain amount of chlorophyll.
2. Studies on the effects ofvarious inhibitors (ammonium ion,DNP, CMU, -hydroxysulphonates,arsenate, cyanide, azide, andantimycin A) under different conditionsshowed that oxidativephosphorylation is a necessary processfor the occurrence ofthe glucosebleaching as well as the assimilationof glucose(cellular growth). Under light-anaerobic conditionsin the presenceof glucose, assimilation of glucose (cellulargrowth) takesplace being supported by photophosphorylation,but no bleachingoccurs.
3. When the algal cells in the courseof bleaching were transferredto the glucose-free mineral medium,the cell growth ceased immediatelybut the cell bleaching proceededfor several hours before itscessation. The respiratory activity,which was high in the glucose-containingmedium, became loweron transferring the algal cells into theglucose-free medium.The lowered level of respiration was maintained,for more than8 hr after the transfer of cells to the glucose-freemedium.
4. When the cells in the course of bleaching were placed underthe atmosphere of N₂, the cell bleaching ceased almost instantaneously.
5. Based on these observations and other inhibition experiments,it was inferred that a certain intermediate(s) produced by theaerobic respiration of glucose is closely associated with theoccurrence of cell bleaching, and that an O₂-requiring stepmay be involved in the process of chlorophyll degradation.

(Received September 9, 1965; )     

The Time Factor in Studies of Growth Inhibition in Excised Organ Sections

1. Apprehension over the adequncy of current techniques stimulateda detailed study of the time factor in the arsenate inhibitionof growth and respiration in excised stem and root sectionsof Pisum sativum.
2. Growth inhibition by arsenate sets in veryslowly, its rateof onset being related to the molar concentration(C) of arsenateate by the relation where T₅₀ is the time taken in hours to reduce the growthrateto 50 per cent of the control and K is a constant. An explanationof the physiological basis of this relationship is attempted.
3. Estimates were made of the final steady growth rate (relativeto control) in various arsenate concentrations. The inhibitionscalculated from this rate are held to approximate to the truearsenate effect and are shown to be very different from thosecalculated from ‘total growth’ measures.
4. Respirationof growing stem sections is not inhibited by thelow arsenateconcentrations that inhibit growth. Some inhibitionis indicatedat high concentrations (3 ? 10^–4M. and over)but onlyafter 15-20 hours of exposure.
5. Two per cent sucrose has noeffect on the arsenate inhibiitionof stem growth. Sucrose,however, markedly stimulates respirationin stem sections, butthis stimulation is prevented by arsenate.
6. The misinterpretationswhich may arise as a result of ignoringthe time factor in inhibitionstudies in excised organ sectionsare discussed and the desirabilityof constructing completegrowth curves in all such studies isstressed.

     

Photosynthetic and light-enhanced dark fixation of 14CO2 from the ambient atmosphere and 14C-bicarbonate infiltrated through vascular bundles in maize leaves

1. The capacity of light-enhanced dark fixation of ¹⁴CO₂ from theambient atmosphere decayed following time-course characteristicsof a first-order reaction (half-life, 1–2 min). The levelof phosphoenolpyruvate in maize leaves under CO₂-free air didnot decrease in the dark subsequent to preillumination. Theseresults indicate that phosphoenolpyruvate carboxylase is activatedin light and quickly inactivated in the following darkness.
2. Removal of oxygen from the atmosphere did not exert any effecton the products of light-enhanced dark fixation of ¹⁴CO₂ providedfrom the atmosphere, the major labeled compounds being malateand aspartate. This confirms that the transfer of carboxyl carbonof C₄-acids to form 3-phosphoglycerate is light-dependent.
3. WhenNaH¹⁴CO₃ solution was vacuum-infiltrated through vasculartissuesof maize leaves, the main initial photosynthetic ¹⁴CO₂fixationproducts were phosphate esters. This indicates thatby thistechnique, ¹⁴CO₂ could be directly provided to the bundlesheathcells, and was fixed via the reductive pentose phosphatecycle.On the other hand, the main initial ¹⁴CO₂-fixation productswere malate and aspartate even when ¹⁴CO₂ was provided throughvascular tissues in the dark immediately following preillumination.The possible regulatory mechanisms underlying the above findingsare discussed.

¹ This work was reported at the 4th International Congress onPhotosynthesis, Reading, September 1977. Request for reprintsshould be addressed to S. Miyachi, Institute of Applied Microbiology,University of Tokyo, Bunkyo-ku, Tokyo 113, Japan ² Present address: Okinawa Branch of Tropical Agriculture ResearchCenter, Ishigaki-shi, Okinawa 907, Japan. (Received October 28, 1977; )     

CHANGES IN LEVELS OF PYRIDINE NUCLEOTIDES IN CHLORELLA CELLS DURING THE COURSE OF PHOTOSYNTHESIS

1. Using intact cells of Chlorella, the effects of CO₂ on thelevelsof oxidized and reduced forms of DPN and TPN in the lightandin the dark were investigated.
2. It was found that the light-inducedchanges of the DPNH-levelwere not affected by the presenceor absence of CO₂. On theother hand, the light-induced increaseof TPNH was suppressedin the presence of CO₂ and the levelof TPNH which was raisedon illumination in the absence of CO₂was lowered by the provisionof CO₂.
3. On the basis of thesefindings, it was concluded that TPNH,but not DPNH, is participating,in some way, in the mechanismof photosynthesis.
4. Discussionswere made on the difference in the sites of participationofTPNH and of the photogenic reducing agent (R) in the pathofcarbon in photosynthesis.

(Received February 28, 1960; )     

PHOTOCHEMICAL INTERCONVERSION BETWEEN PRECURSORS OF PHYCOBILIN CHROMOPROTEIDS IN TOLYPOTHRIX TENUIS

1. The photochemical conversion between the precursors of phycocyaninand phycoerythrin in Tolypothrix tenuis was investigated.
2. Itwas found that the conversion of phycocyanin-precursor intophycoerythrin-precursor was induced by green light, and thereverse reaction by red light. These reactions proceeded exponentially, indicating that the photochemical process was acceleratedautocatalytically by the reaction-product.
3. The rates of thesephotochemical reactions were found to beunaltered by varyingthe incubation temperature (0? to 35?)and the composition ofthe gas atmosphere (presence or absenceof CO₂ and of O₂ orby an inhibitor of photosynthesis, p-chlorophenyldimethylurea.
4. The action spectra of the photochemical interconversions betweenprecursors of phycobilin chromoproteids were found to be distinctlydifferent from the absorption spectra of chlorophyll and carotenoids.The most effective wavelength for inducing the conversion ofphycocyanin- into phycoerythrin-precursor (541 mµ) isnear the absorption maximum of phycoerythrin (565 mµ),and that of the reverse reaction (641 mµ) is near theabsorption maximum of phycocyanin (620 mµ). Additionaldata, indicating that the phycobilin chromoproteids themselvesdo not participate in these processes as light absorber, werealso presented.
5. On the basis of these results, a possiblemechanism of the photochemicalinterconversion between the precursorsof phycobilin chromoproteidsis proposed.

(Received March 13, 1962; )     

Studies on Plant Growth Hormones: II. FURTHER BIOLOGICAL PROPERTIES OF 3-INDOLYLACETONITRILE

1. 3-Indolylacetonitrile is more active than 3-indolylacetic acidin the Avena straight-growth test, but less active in the Avenacurvature test at comparable concentrations. Reasons for thisare discussed, and results of previous work on plant extractsusing the curvature test as a means of assay are considered.
2. Transport of both the acid and the nitrile is polar, fromapexto base of the coleoptile. The nitrile can reach the growingcells as easily, and possibly more easily, than the acid. Thesignificance of these findings for a theory on the mechanismof action of the nitrile is discussed.
3. The nitrile is inactivein the pea curvature test and straight-growthof pea stem sectionsexcept at high concentrations. It is alsoinactive or only slightlyactive in lateral bud inhibition,root initiation, and petioleabscission at the concentrationstested.
4. It is less activethan the acid in root inhibition in cress,but approximatelyas active in Avena. It is approximately asactive as the acidin parthenocarpic fruit development, andinitiation of cambialactivity.
5. The significance of these results is discussed.

     

COMPONENTS OF THE ELECTRON-TRANSFERRING SYSTEM IN ACETOBACTER SUBOXYDANS AND RECONSTRUCTION OF THE LACTATE OXIDATION SYSTEM

1. Cytochromes a₁⁵⁹⁰, b⁵⁶⁰, c₁⁵⁵⁴ and c₁⁵⁵² were isolated andpurifiedfrom a strain of Acetobacter suboxydans. The proceduresusedwere described in detail.
2. The main cytochrome band at550-560 mµ in intact cellssplitted at liquid air temperatureinto two bands, 551 mµ(strong) and 559 mµ (weak).
3. Optical and physiological properties of the four cytochromeswere investigated. Lactic dehydrogenase activity was found tobe associated with cytochrome c₁⁵⁵⁴. The two c₁-type cytochromes,especially cytochrome c₁⁵⁵⁴, persisted in their reduced formafter the purification through many steps.
4. By some combinationsof isolated components reconstruction ofthe oxygen uptake systemcould be realized.
5. The oxygen-consuming activity of purifiedoxidase preparationswas accelerated by a-tocopherol but notby Emasoll 4130 andTween 80.
6. Some discussions were made onthe nature of terminal oxidase,the role of cytochrome c₁⁵⁵²in the electron-transport system,and persistence of reducedstate of c₁-type cytochromes.
7. A possible scheme of the electron-transferringsystem of Acetobactersuboxydans was presented.

(Received May 16, 1960; )     

EFFECT OF CHROMATIC LIGHTS ON PHYCOBILIN FORMATION IN A BLUE-GREEN ALGA, TOLYPOTHRIX TENUIS

1. The formation of phycobilin pigments in a blue-green alga Tolypothrixtenuis was investigated with special reference to the effectsof preillumination with colored lights.
2. It was discoveredthat the algal cells are capable of formingphycobilin pigmentsin the dark, if they have been previouslyilluminated for severalhours in the presence of CO₂.
3. The color of light applied inthe later period of preillumination(chromatic illumination)was found to affect the ratio of phycoerythrinto phycocyaninformed in the subsequent dark period. A greenlight acceleratesthe dark-formation of phycoerythrin, a redlight that of phycocyanin,and the two lights counteractingwith each other in their effects.
4. These directive effects of the "chromatic illumination" canbe accomplished within a very short period, for instance, in3 minutes if it is preceded by sufficient "preillumination"with an incandescent or day light fluorescent light. The reactionsoccurring during the period of chromatic illumination does notrequire the presence of CO₂ and the aerobic condition.
5. Thealga can be grown heterotrophically when supplied with casaminoacids and glucose. Under such a condition the alga forms phycocyanintogether with chlorophyll and carotenoids, but not phycoerythrin.
6. On the basis of the results obtained, a tentative scheme forthe biosynthesis of phycobilin pigments in the alga was proposed,assuming the light-induced formation of unknown precursors whichare converted into phycocyanin and phycoerythrin in the subsequentdark period.

(Received July 4, 1960; )     

STUDIES ON THE METABOLISM OF A SULFUR-OXIDIZING BACTERIUM III. POSTOXIDATIVE FIXATION OF CARBON DIOXIDE

1. The cells of Thiobacillus thiooxidans, which had been in contactwith sulfur or sulfide in air (or CO₂-free air), could fix addedCOa very rapidly after replacing air with nitrogen. This fixationis designated as the postoxidative fixation.
2. "Preoxidation"of the sulfur compounds is mandatory for theoccurrence of thepostoxidative fixation.
3. The cells which had preliminarilyoxidized sulfide could notshow the CO₂-fixation, when theywere placed under an anaerobiccondition in the absence of thesulfur compound.
4. These results indicate that sulfur compoundsmay have an importantrole as the electron donor for the reductionof CO₂, besidestheir role as the substrate of respiration tosecure energyfor the fixation of CO₂

(Received March 6, 1962; )     

Mineral nutrition of cultured chlorophyllous cells of tobacco IV. Kinetic studies of K and Mg uptake by the cells

1. When applying the adsorption theory to the selectivity coefficient,K and Mg uptakes by cells in a K-Mg replacement series of mediaare not regulated only by a common mechanism, under the assumptionthat b values, which indicate the affinities between the ionand the adsorptive surface, do not change.
2. Regulation of Kand Mg uptakes by a common multiphasic mechanismin the cellsis possible when the selectivity coefficient b_K/b_Mgvaries inverselywith the M_K/M_Mg ratio in the external medium.
3. K and Mg uptakesare not regulated only by their respectivesingle specific mechanisms.
4. Another possibility is regulation of K and Mg uptakes bothbycommon and specific mechanisms. The common mechanism maybemultiphasic.

(Received December 2, 1975; )     

EFFECT OF LIGHT ON THE CHLOROPHYLL FORMATION IN THE "GLUCOSE-BLEACHED" CELLS OF CHLORELLA PROTOTHECOIDES

1. Previous studies have shown that when Chlorella protothecoidesis grown in a medium rich in glucose and poor in nitrogen source(urea), apparently chlorophyll-less cells with profoundly degeneratedplastids—referred to as "glucose-bleached cells—areproduced either in the light or in darkness. When the glucose-bleachedcells are incubated in a medium enriched with the nitrogen sourcebut without added glucose, an active formation of chlorophylloccurs after a certain lag period under illumination, whilein darkness a very small amount of chlorophyll is formed atabout the same time as in the light. The stimulating effectof light on the chlorophyll formation is not appreciably affectedwhen the photosynthetic CO₂-fixation of greening algal cellsis blocked by the addition of CMU. In the present study, itwas further found that the light-enhanced chlorophyll formationproceeds, although at a somewhat lower rate, under aerationof CO₂-free air. All the experiments in this work were doneunder these non-photosynthetic conditions to exclude any influenceof photosynthates.
2. The effect of light (from daylight fluorescentlamps) on thechlorophyll formation in the glucose-bleachedalgal cells wassaturating at about 1,000 lux. Blue light wasfound to be mosteffective; yellow, green and red light followingin the orderof decreasing effectiveness.
3. When the bleachedalgal cells were illuminated for a short periodin the lag phaseof chlorophyll formation and subsequently incubatedin darkness,there occurred an appreciable enhancement of chlorophyllformationin the dark. When the short illumination was appliedat differenttimes of the lag phase, the enhancement was inducedto almostthe same extent. But the longer the duration of theilluminationduring the lag phase, the greater was the enhancementof chlorophyllformation in the subsequent dark incubation.In such experimentsblue light was most effective and red lightleast, as it wasthe case in the experiments of continuous illumination.An intervenientillumination of the bleached cells at lowertemperatures orunder the atmosphere of N2 produced little orno enhancementof the chlorophyll formation in the subsequentdark incubation.
4. Based on these results, it was concluded that the light enhancementof chlorophyll formation in the glucose-bleached algal cellsis mediated by a non-chlorophyllous photoreceptor(s), absorbingmaximally blue and yellow light, and that a light-induced changeof the photoreceptor is immediately followed by a certain dark(temperaturedependent and aerobic) process(es) which is connected,directly or indirectly, to the chlorophyll synthesis.

(Received August 10, 1967; )     

Absorption and Assimilation of Inorganic Nitrogen from Different Sources by Storage Root-Tissue

1. The rate of ammonium and nitrate absorption and assimilationby storage tissues of sweet potato tubers, radish, and carrotroots cultured in aerated dilute solutions (5 mg. eq./1.) ofNH₄Cl, KNO₃ or NH₄NO₂ at 25° C. and the effect of the sourceof nitrogen on the nitrogenous composition of the tissues werestudied.
2. Both ions were absorbed at more or less equal ratesunder theprevailing experimental conditions by cells of thedifferentplant materials used.
3. The rate of nitrate assimilationwas lower than absorption,leading to the accumulation of nitrate-Nin the cells of alltissues examined. Ammonium-N, on the otherhand, was assimilatedalmost as soon as it entered the cellsof sweet potato tubersand carrot roots, and there was onlya small increase in thelevel of arnmonium-N; but in radishroot cells ammonium assimilationlagged very much behind absorption,resulting in a great increasein the level of arnmonium-N.
4. Sweetpotato tubers and carrot tissues were shown to be adaptedtothe utilization of more ammonium than nitrate nitrogen, butradish root tissues are adapted to the utilization of more nitratenitrogen.
5. Ammonium or nitrate-N caused similar increases inthe complexorganic-N level (protein-+rest-N) in radish root,but the rateof synthesis was higher with ammonium in sweetpotato tubersand with nitrate in carrot roots.

Furthermore, with both sources changes in the content of amideand amino-N were negligible in radish tissues of both series.In contrast to radish, carrot tissues showed marked increasesin the two fractions. The increase of these fractions in sweetpotato tuber tissues was much lower. The marked contrast in nitrogenous composition in the threedifferent plant materials fed with ammonium and nitrate saltsseparately indicated that the actual course of protein metabolismis most probably dependent on the source of nitrogen utilized.     

The Absorption of Ions by Excised Root Systems: I. APPARATUS AND PRELIMINARY EXPERIMENTS

1. An apparatus is described by means of which the absorptionofions from a complete nutrient solution of constant compositionby excised root systems of plants, grown under known nutrientdeficiencies, may be measured in standard conditions of aerationand temperature. Results of some prelimi nary experiments aredescribed.
2. It was found that the roots readily absorbed theelement inwhich they were deficient, but tended to lose thoseelementswhich were already present in normal amounts.
3. Therewas almost invariably a loss in fresh weight of the rootsafterthe absorption period and also a loss in dry weight. Thislossappears to be complex and is partly attributable to lossofrespiratory material.
4. The addition of 2 per cent. sucroseto the solution from whichthe root systems of phosphorus-deficientbarley plants wereabsorbing increased the nitrogen and phosphoruscontents ofthe roots and maintained the potassium content,while in absenceof sucrose only the phosphorus content increased,but this increasewas significantly less than in the presenceof sucrose.
5. It was shown that roots excised from plants growingin soilwere capable of absorbing phosphorus or nitrogen—elementsin which they were apparently deficient.
6. The interpretationof data obtained from excised roots is discussed,and it isconcluded that excised roots from plants grown incomplete nutrientare not likely to behave in the same way,as regards absorption,as corresponding roots of intact plants,but that roots grownunder conditions of deficiency will behaverather similarlywhether excised or intact. This fact providesa potential methodfor diagnosing and evaluating nutrient deficiencies.
7. The low-saltcondition of roots postulated by Hoagland and Broyeris notnecessarily the primary requisite for rapid absorptionof aparticular ion. It is rather that the roots should be deficientin that ion. The roots could be high in other salts.