Some further studies on metabolism of acetate-l-14C in potassium deficient rice leaves

Acetate-1-¹⁴C was fed to excised leaves of normal and potassiumdeficient rice plants. The rate of respiratory evolution of¹⁴CO₂ was increased by a potassium deficiency. Malonate, glyoxylateor malate supplied enhanced the metabolism of acetate-l-¹⁴C.These results suggest an accelerated turn of the TCA cycle inpotassium deficient leaves. Malate synthetase activity was notrecognized in either normal or deficient leaves. However, condensingenzyme activity was higher in deficient leaves than in normalones. An addition of DNP to the leaves increased ¹⁴CO₂ productionfrom acetate-l-¹⁴C though its effect was smaller in deficientleaves than it was in normal ones. This result may suggest anincrease in the turnover rate of ATP or loose coupling of electrontransfer with oxidative phosphorylation in deficient leaves.Chromatographic separation of cold acidsoluble nucleotides hasshown that the ATP level was lowered by a potassium deficiency,though the ADP level was not affected. ¹Present address: Sericultural Experiment Station, Suginami-Ku,Tokyo     

Distribution of radioactivities in sugars from acetate-2-14C incubation and elemental content of some vascular plants in two diverse habitats of Glacier Bay, Alaska

Essentially six sugars became labeled following acetate-2-¹⁴Cmetabolism in plants distributed in two different communities—pioneerand forest. Of these the bulk of radioactivity resided in glucose,fructose and sucrose. Except for willows, labeled glucose andlabeled fructose dominated the plants of the pioneer habitat,while labeled sucrose in addition to labeled glucose and fructosecharacterized those of the forest. For willows, the radioactivitieswere present in both monosaccharides and disaccharides. Thissugar labeling pattern occurred in the alder which for the twohabitats tested, was consistent with the general observation.The relative level of the content of magnesium and phosphorusfor various plants in the pioneer habitat were no differentfrom those in the forest habitat. Levels of sulfur, chlorine,potassium and calcium were generally higher in plants of theforest than in those of the pioneer community. Plants with alow level of K contained a higher level of labeled hexoses;while plants with a high level of K contained both labeled hexosesand disaccharide, indicating that potassium may be involvedin the incorporation of the ¹⁴C into sugar during acetate-2-¹⁴C.     

Effect of agmatine on putrescine formation in tobacco plants

Incorporation of L-arginine-U-₁₄C, fed to leaf disks of tobaccoplants, into putrescine decreased about 70% in the presenceof agmatine, but that of L-ornithine-U-₁₄C decreased only slightly.These results indicate that putrescine is synthesized from argininevia agmatine, but is also synthesized from ornithine withoutpassing through arginine and agmatine. (Received April 23, 1969; )     

Metabolism of Glycollate by Lemna minor L. Grown on Nitrate or Ammonium as Nitrogen Source

Marques, I. A., Oberholzer, M. J. and Erismann, K. H. 1985.Metabolism of glycollate by Lemna minor L. grown on nitrateor ammonium as nitrogen source.—J. exp. Bot. 36: 1685–1697. Duckweed, Lemna minor L., grown on inorganic nutrient solutionscontaining either NH₄⁺ or NO₃^– as nitrogen source wasallowed to assimilate [1-¹⁴C]- or [2-¹⁴C]glycollate during a20 min period in darkness or in light. The incorporation ofradioactivity into water-soluble metabolites, the insolublefraction, and into the CO₂ released was measured. In additionthe extractable activity of phosphoenolpyruvate carboxylasewas determined. During the metabolism of [2-¹⁴C]glycollate in darkness, as wellas in the light, NH₄⁺ grown plants evolved more ¹⁴CO₂ than NO₃^–grown plants. Formate was labelled only from [2-¹⁴C]glycollateand in NH₄⁺ grown plants it was significantly less labelledin light than in darkness. In NO₃^– grown plants formateshowed similar radioactivity after dark and light labelling.The radioactivity in glycine was little influenced by the nitrogensource. Amounts of radioactivity in serine implied that thefurther metabolism of serine was reduced in darkness comparedwith its metabolism in the light under both nitrogen regimes.In illuminated NH₄⁺ plants, serine was labelled through a pathwaystarting from phosphoglycerate. After [1-¹⁴C]glycollate feedingNH₄⁺ grown plants contained markedly more radioactive aspartateand malate than NO₃^– plants indicating a stimulated phosphoenolpyruvatecarboxylation in plants grown on NH₄⁺. Key words: Photorespiration, glycollate, nitrogen, Lemna     

木荷属和柃木属植物的嘌呤代谢及嘌呤碱合成研究

以［8-¹⁴C］标记的腺嘌呤和黄嘌呤为底物,对两种可以合成少量咖啡碱和茶叶碱的木荷属和柃木属植物（Schima mertensiana,Eurya japonica）叶片的嘌呤代谢进行了检测研究。发现木荷属和柃木属植物中嘌呤代谢相似,¹⁴C标记的腺嘌呤可以整合到嘌呤核苷酸、RNA、酰脲（包括尿囊素和尿囊酸）、二氧化碳中。经过24 h培养,在叶片吸收的放射能中,仅有6%～7%用于甲基黄嘌呤类化合物的合成（3-甲基黄嘌呤、7-甲基黄嘌呤核苷、7-甲基黄嘌呤、茶叶碱）。和其他植物一样,绝大多数¹⁴C标记的黄嘌呤整合到嘌呤的分解代谢物中（二氧化碳和酰脲）,少量的放射能分布在3-甲基黄嘌呤及茶叶碱中。根据结果可以推断木荷属和柃木属植物具有N-甲基转移酶活性,可以用来合成咖啡碱和茶叶碱,相对于茶树而言,活性不高。综上,本文对木荷属和柃木属植物的嘌呤代谢以及嘌呤碱合成进行了研究。     

PHOTOSYNTHESIS OF GLYCINE AND SERINE IN GREEN PLANTS

1. The effects of "carbonyl" reagents on the photosyntheticin-corporation of ¹⁴CO₂ into the assimilation products of tobaccoand spinach leaves were studied. The presence of "carbonyl"reagents causes an increase in the ratio of ¹⁴CO₂ incorporatedin glycine and a decrease in serine. The incorporation of ¹⁴Cfrom glycolate-1-¹⁴C and glycolaldehyde-2-¹⁴C into glycine andserine was also affected by "carbonyl" reagents, as in the caseof ¹⁴CO₂-experiment. 2. The feeding experiments of glycine-1-¹⁴C and serine-1-¹⁴Cin the presence and in the absence of "carbonyl" reagents revealedthat these reagents inhibit the conversion of glycine to serine. 3. The results obtained above, together with the effects ofthiols on ¹⁴CO₂ incorporation presented in this paper, supportthe assumption that glycine and serine are formed via glycolateand glyoxylate during photosynthesis in green plants. 4. Comparison of ¹⁴C incorporation in malate from ¹⁴CO₂, glycolate-1-¹⁴C,glycine-1-¹⁴C and serine-1-¹⁴C in the presence and in the absenceof "carbonyl" reagents suggested the occurrence of the pathwayof the malate formation via glycolate and glyoxylate, not passingthrough glycine and serine, during photosynthesis. ¹ A part of this paper was presented at the Symposium on "Nitrogenand Plant" by the Japanese Society of Plant Physiologists, inOctober, 1963 ² Present address: Radiation Center of Osaka Prefecture, Sakai,Osaka     

Inhibition of Sterol Biosynthesis and Stem Elongation of Tobacco Seedlings Induced by Some Hypocholesterolemic Agents

Incorporation of DL-[2-¹⁴C]mevalonic acid ([2-¹⁴C]MVA) into4-desmethylsterols in Nicotiana tabacum cv. Turkish Samson seedlingswas inhibited by SK&F 7997-A₃,¹ SK&F 7732-A₃, AY 9944,and the plant growth retardant, Amo 1618. Reductions in 4-desmethylsterol levels resulted from treatmentwith AY 9944 and Amo 1618, but not the SK&F compounds. Amo1618 and SK&F 7997-A₃ both significantly reduced the specificactivity of each of the four major 4-desmethylsterols examined.Although SK&F 7732-A₃ reduced the specific activity of campesterol,and AY 9944 reduced the specific activity of stigmasterol, neitherhad an effect on the specific activity of ß-sitosterol. Stem elongation of tobacco seedlings was retarded by SK&F7997-A₃, AY 9944, and SK&F 7732-A₃, particularly the former,and the retarded plants thus produced were morphologically indistinguishablefrom the Amo 1618-treated plants. Application of exogenous stigmasterol,or GA₃, to the chemically-retarded plants resulted in a reversalof stem growth retardation.     

Time sequence of the effect of ethylene on transport, uptake and decarboxylation of auxin

The effect of ethylene on the uptake, decarboxylation and basipetaltransport of IAA-1-¹⁴C, IAA-2-¹⁴C and NAA-1-¹⁴C in cotton stemsections (Gossypium hirsutum L., var. Stoneville 213) was studied.A reduction in the capacity of cotton stem sections to transportauxin basipetally appears in sections excised from plants exposedto ethylene for only 3 hr and increases with fumigation time. In addition to reducing transport, increasing ethylene pretreatmentperiods from 3 to 15 hr also progressively reduced the uptakeof ¹⁴C and increased the release of ¹⁴C as ¹⁴CO₂ from IAA-1-¹⁴C.The effect of ethylene on the decarboxylation of IAA-1-¹⁴C wassignificant when expressed as either the cpm of ¹⁴C releasedper hr per mg dry weight or the cpm released per hr per mm²in contact with the IAA donor. Comparative experiments usingIAA-1-¹⁴C and IAA-2-¹⁴C demonstrated that the effect of ethyleneon the decarboxylation of IAA was primarily a cut surface effectwhich apparently contributes to the reduction of IAA uptakeby ethylene. Although ethylene significantly reduced the transport of NAA-1-¹⁴C,uptake was significantly increased rather than decreased aswith IAA-1-¹⁴C while decarboxylation was unaffected. Ethylene pretreatment caused no significant changes in the dryweight or the cross-sectional area of the absorbing surfaceof the transport tissue. ¹A contribution of the Texas Agricultural Experiment Station.Supported in part by Grant GB-5640, National Science Foundationand grants from the Cotton Producers Institute and the NationalCotton Council of America. ²Present address: Central Research Department, E. I. Du PontDe Nemours and Company, Wilmington, Delaware 19898, U. S. A. (Received May 29, 1969; )     

The translocation of photosynthetic products from mesophyll into midrib in tobacco plant III. The transformation of translocated 14C-sugars in the veins

¹⁴C-U-sugars were introduced into tobacco plants through themesophyll, the veins of the first order of branching, and themidrib, and ¹⁴C-compounds in the veins and the midrib whichtranslocated towards the base of the midrib were traced duringthe period of 120 min after the ¹⁴C-sugar introductions. 1) When ¹⁴C-U-sucrose was introduced into the leaf, no matterwhat the means of feeding was, most of the ¹⁴C which translocatedbasipetally in the veins and the midrib was found in the formof sucrose. 2) When ¹⁴C-U-glucose or ¹⁴C-U-fructose was administered tothe leaf dirough the cut vein of the first order of branching,most of the ¹⁴C which translocated basipetally in the veinsand the midrib was found in the form of sucrose. 3) ¹⁴C-U-glucose or ¹⁴C-U-fructose injected into the vascularbundles of the midrib was translocated basipetally, as such,10 and 30 min after injection; and at 30 min, the amount ofthe ¹⁴C-sucrose in the midrib attained 9–22% of the 80%ethanol-soluble ¹⁴C in the midrib. 4) When ¹⁴C-U-glucose or ¹⁴C-U-fructose was supplied to themesophyll, the radioactivities of these hexoses were predominantin the first and second veins soon after application, then decreasingwith a concomitant increase in the radioactivity of the ¹⁴C-sucrose. From these results, it was inferred that in the veins of thefirst and second order of branching, glucose and fructose whichmoved from the mesophyll did not translocate as such, but wereutilized for the synthesis of sucrose available for translocationvia the midrib to the stem. ¹A part of this paper was presented at the Crop Science Societyof Japan, in April, 1969 (Received December 8, 1969; )     

Phytochemical studies on tobacco alkaloids XIV. The occurrence and properties of putrescine N-methyltransferase in tobacco roots

Putrescine N-methyltransferase, a new enzyme catalyzing theformation of N-methylputrescine from putrescine and S-adenosyl-L-methioninewas found in roots of tobacco plants. The enzyme was purified30-fold from crude extracts of tobacco roots. NMethylputrescinewas identified as the reaction product by comparison with theauthentic compound. The enzyme had a pH optimum between pH 8and 9, and a molecular weight of about 60,000, as determinedby gel filtration. Km values for putrescine and 5-adenosyl-L-methioninewere 4.0 x 10^–4 M and 1.1 x 10^–4 M, respectively.Enzyme activity was inhibited by N-chloromercuribenzoate andAg⁺. No cofactors were required. Of the various substrates tested,only putrescine served as a methyl acceptor. The enzyme waslocalized exclusively in the roots and its activity was greadyenhanced by decapitation. The presence of putrescine N-methyltransferase in tobacco rootsstrongly suggests that N-methylputrescine participates as anintermediate in nicotine biosynthesis. (Received March 2, 1971; )     

Uptake, transport and stability of [2-14C] abscisic acid in cucumber following foliar application

The uptake, transport and stability of ABA after foliar applicationto cucumber plants was studied using [2-¹⁴C] ABA. ABA uptakethrough uninjured epidermis of cucumber leaves appeared to bevery slow compared with the uptake through abraded leaves. Transportof [2-¹⁴C] ABA in the same system was also found to be veryslow. No metabolic changes in the absorbed [2-¹⁴C] ABA weredetected 24 hr after its application as checked by TLC and GLCtechniques. These results are compared and discussed in relationto previous reports. ¹The Irene and David Schwartz Professor of Plant Genetics (Received April 26, 1976; )     

Effect of mineral deficiency on amine formation in higher plants

Potassium deficiency caused putrescine accumulation in the leaves of barley, radish, pea, bean and spinach plants. Magnesium deficiency caused putrescine accumulation in barley, pea and bean leaves, and also in the leaves of older radish plants. In young radish plants less putrescine was found in magnesium deficiency, and in spinach magnesium deficiency was without effect on putrescine levels. Putrescine content may be a useful guide to the mineral status of legumes, since accumulation of this amine may be detected before deficiency symptoms appear. Radioactivity from l-arginine-[U-¹⁴C] fed to barley seedlings was detected in agmatine within 2 hr, and probably also in the hordatines after 24 hr, feeding. After 2 hr the label in the agmatine was greatest in the potassium-deficient plants, but after 24 hr the level declined to that found in the agmatine of the leaves of the magnesium-deficient and control seedlings. The rate of putrescine formation was high in both potassium and magnesium deficiency. Incorporation of radioactivity in spermidine and spermine on feeding putrescine-[1,4-¹⁴C] to barley seedlings was estimated in the dansylated amines after separation by TLC. Activity was higher in spermidine and lower in spermine in the potassium-deficient plants than in the controls. The spermidine/spermine ratio declined on excision of barley leaves.     

BIOGENESIS OF ETHYLENE IN APPLE TISSUE: I. FORMATION OF ETHYLENE FROM GLUCOSE, ACETATE, PYRUVATE, AND ACETALDEHYDE IN APPLE TISSUE

1. With the aim of elucidating the path of carbon in the formationof ethylene in plants, studies were made on the incorporationof ¹⁴C into ethylene evolved from apple slices, using several¹⁴C- labeled compounds as substrates. The effects of inhibitorswere also investigated. 2. The formation of ethylene-¹⁴C from glucose-¹⁴C was inhibitedby fluoride, but unaffected by arsenite, thus suggesting thatglucose is converted to ethylene via pyruvate. 3. Acetate is converted to ethylene after cleavage of C-l andC-2. Only a small portion of the latter (C-2) enters the moleculeof ethylene, the former (C-l) is detected in carbon dioxide.On the other hand, 2, and 3-carbons of pyruvate are converted,without splitting, to ethylene. 4. On removal of air, the incorporation of ¹⁴C into ethylenefrom acetate-2-¹⁴C was depressed, while that from pyruvate-¹⁴Cwas unaffected. 5. Acetaldehyde-l,2-¹⁴C is converted to ethylene without conversioninto ethanol. 6. These results are interpreted to suggest the occurrence ofthe pathway in which pyruvate and acetaldehyde may serve asprecursors of ethylene. ¹ A part of this paper was read at the regular Meeting of KansaiBranch of the Agricultural Chemical Society of Japan in Kyoto,October, 1964, and at the Annual Meeting of the Japanese Societyof Plant Physiologists in Tokyo, April, 1965 and presented ina preliminary form elsewhere (10).     

Long chain fatty acid synthesis in developing castor bean seeds IV. The synthetic system in proplastids

The proplastid fraction containing no cytosol and mitochondrionwas isolated from developing castor bean endosperm by stepwisesucrose density centrifugation. This fraction possesses thecapacity to synthesize LFAs from [u-14C]sucrose, [u-¹⁴C]-glucose,[u-¹⁴C]G-1-P, [u-¹⁴C]G-6-P, [2-¹⁴C]pyruvate and [1-¹⁴C]acetate.Little was incorporated from [1-¹⁴C]pyruvate into LFAs, butmuch into ¹⁴CO_a. Addition of cytosol to the proplastid fractiondid not enhance the LFA synthesis. From these data, the wholepath from sucrose to LFAs through glycolytic path and pyruvatedecarboxylation seems to be located within the proplastid indeveloping castor bean endosperm. The difference in utilizationof substrates indicates that the rate of LFA synthesis in castorbean proplastids is limited at a step between sucrose and hexosephosphate. In addition, experiments with CO₂ output and LFAsynthesis from [1-¹⁴C]glucose, [6-¹⁴C]glucose and [u-¹⁴C]G-6-Pstrongly suggest that the path flow branches actively throughG-6-P to the pentose phosphate path and little through acetylCoAto the TCA cycle. (Received May 12, 1975; )     

New evidence for the metabolic conversion of aspartate-4-14C to glutamine-l-14C during the initial period of ammonia assimilation in the roots of barley plants

Metabolic conversion of aspartate was investigated in rootsof barley plants fed a 30-min pulse of aspartate-4-¹⁴C, followedby transfer to an ammonia or nitrate medium for 1 hr. Glutamate was the predominant labeled amino acid after the pulse;whereas, glutamine was after ammonia assimilation. Each labelappeared almost entirely at the C-1 position. The organic acidfraction was also labeled with¹⁴C. Negligible labeling of asparagineshowed that it was not a primary product of ammonia assimilation. The data demonstrated that aspartate-4-¹⁴C was transformed intoglutamate-1-¹⁴C by the tricarboxylic acid cycle mechanism, withglutamate being converted to glutamine-1-¹⁴C during ammoniaassimilation. The physiological significance of this metabolicconversion in plant metabolism is discussed. This conversionplays an important role in preventing a drop in the oxalacetatelevel when -ketoglutarate has been drained off by rapid synthesisof glutamate and glutamine during the initial period of ammoniaassimilation. This paper also presents, for die first time, evidence for theexistence of a new pathway, named the "aspartate-glutamine pathway",in higher plants. (Received August 5, 1972; )     

In vivo citrate studies with developing soybean cotyledons

The in vivo data presented here are strong evidence for theinvolvement of citrate cleavage enzyme in lipid synthesis indeveloping soybean cotyledons. The incorporation of ¹⁴C fromcitrate into crude lipid fraction in vivo had a pH optimum of4.5; was linear with time; had a temperature optimum of 35?C;and was inhibited by (–)-hydroxycitrate. The point ofcitrate cleavage was between carbons 3 and 4 of the citratemolecule and therefore ¹⁴C was incorporated into crude lipidfraction from citrate-5-¹⁴C but not citrate-1-¹⁴C or citrate-6-¹⁴C. ¹ Cooperative investigations of the Agricultural Research Service,U.S. Department of Agriculture, and the Illinois AgriculturalExperiment Station. ² This research represents partial fulfillment of the Ph.D.requirements of Daniel R. Nelson. Presently at Monsanto AgriculturalProducts Co., St. Louis, MO 63141, U.S.A. (Received September 20, 1976; )     

Fate of ($ ) S-methyl-L-cysteine sulfoxide in Chinese cabbage, Brassica pekinensis RUPR.

($) S-methyl-L-cysteine sulfoxide (MCS) was scarcely found inseeds of Chinese cabbage, but was present in relatively largeamounts in all plant parts after germination. Changes in MCScontent paralleled those for soluble sulfur content of tissue. When Na₂³⁵SO₄ was fed to plants, the ³⁵S was predominantly incorporatedinto MCS in the free amino acid fraction in both sulfur-sufficientand deficient plants, but it was incorporated to a greater extentin the former. ³⁵S-MCS was metabolized more readily in deficientthan in sufficient plants, and its sulfur was found not onlyin various soluble compounds but in the insoluble fractionsfrom plants as well. These results indicate that MCS is a conspicuousconstituent in the free amino acid pool of Chinese cabbage andmay play an important role in sulfur metabolism by acting asa soluble pool for organic sulfur. (Received June 22, 1970; )     

Participation of farnesol in the biosynthesis of ipomeamarone

Farnesol-2-¹⁴C was readily incorporated into ipomeamarone, oneof the furanoterpenoids produced in sweet potato infected withthe black rot fungus, Ceratocystis fumbriata. This was demonstratedby isolating labeled ipomeamarone and analyzing its radioauto-gramby silica gel thin layer chromatography of the extracts solublein chloroformmethanol (1: 1 , v/v), after farnesol-2-¹⁴C feedingto the tissue. Further proof for farnesol-2-¹⁴C incorporationinto ipomeamarone comes from the fact that the specific radioactivityof ipomeamarone semicarbazone was constant throughout the crystallizations.Fractionation of the label of farnesol-2-¹⁴C showed that radioactivitywas little distributed in the methanol-water fraction and wasmainly incorporated into ipomeamarone. Accordingly, it is notlikely that farnesol is incorporated into ipomeamarone afterits degradation to a small molecule(s) such as acetate. An additionalexperiment indicatedthat the incorporation of farnesol-2-¹⁴Cinto ipomeamarone markedly decreased under strict anaerobicconditions. This shows that some oxidative reactions are involvedin ipomeamarone biosynthesis from farnesol. ¹ This paper constitutes Part 91 of the Phytopathological Chemistryof Sweet Potato with Black Rot and Injury. (Received February 3, 1971; )     

Distribution of labeled auxin in geotropically stimulated steins of cucumber and pea

The distribution of IAA-2-¹⁴C or IAA-5-³H applied to the apexin the upper and lower (with respect to gravity) halves of geotropicallystimulated stems of cucumber and pea was examined and the resultsobtained are as follows: 1. A larger amount of IAA-2-¹⁴C or IAA-5-³H was detected inthe lower than upper half of cucumber hypocotyls with 3-hr geostimulation. 2. A larger amount of IAA-2-¹⁴C was distributed in the lowerthan upper epidermis of pea epicotyls with 1-hr geostimulation. 3. Freezing autoradiography revealed that IAA-2-¹⁴C was concentratedin the vascular bundles and epidermis of cucumber hypocotyls,the distribution being affected by geostimulation only in thelatter. 4. Application of 1% TIBA in lanolin inhibited the distributionof IAA-2-¹⁴C in the lower epidermis of pea epicotyls, causingsuppression of geotropic curvature. 5. From these results, we concluded that IAA which accumulatedin the lower epidermis of the stem upon geostimulation causedthe negative geotropic curvature of the stem. (Received October 13, 1975; )     

The Oxidation of 14C-labelled Glucose by Chlorella vulgaris: 1. The Time Course of 14CO2 Production

Glucose, either uniformly labelled with¹⁴C, or specificallylabelled in the I, 2, or 6 position, was added to C. vulgaris.Radio-active carbon dioxide was produced initially ten timesfaster from glucose-I-¹⁴C than from glucose-6-¹⁴C. This differencewas found with carbohydrate-starved cultures, exponentiallygrowing cultures, and cultures assimilating ammonia or nitraterapidly. A similar difference was also found with C. pyrenoidosaand Ankistrodesmus. 37 per cent. of the ¹⁴C added as glucose-¹-¹⁴Cto exponentially growing cells was recovered as carbon dioxidebut generally the recovery was less than this. Only 5 per cent.of ¹⁴C added as glucose-6-¹⁴C was recovered as carbon dioxide.The specific activity of the carbon dioxide produced was considerablylower than that of the carbon in the added glucose.