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1.
Various Cucurbita seed globulins showed patterns similar toone another on SDS-gel electrophoresis, and ß bandsfor unreduced globulins and , ', and ' bands for reduced ones.On gel electrophoresis in 6 M urea, reduced globulin gave twoacidic and two basic bands. These corresponded to and ' chainsand 1 and 2 chains, respectively, identified by two-dimensionalurea-SDS gel electrophoresis. The compositions of the and ßsubunits were proposed. (Received September 8, 1977; )  相似文献   

2.
Subunits (, ß, ) and mixtures of subunits ( ß, , ß , ß ) were isolated without denaturationfrom a chloroform extract of chloroplast coupling factor 1 (CF1)from maize (Zea mays var. Ushiku 5-4) and from spinach by fastprotein liquid chromatography (FPLC), on an anion-exchange columnof Mono-Q in the presence of n-octylglucoside (OG) and on achromatofocusing column of Mono-P. The ß -subunitcomplex (CF1 ß ) was the minimum unit required forATPase activity, as was confirmed by the reconstituted complexof ß and subunits. An subunit isolated from maizeinhibited the ATPase activity of CF1 ß from bothmaize and spinach. CF1 ß was found to contain anOG-dependent Mg2+-ATPase. The ATPase activity of CF1 ß required divalent cations, such as Mg2+ or Mn2+, for its expressionin the presence of OG; its optimum pH was 8.0 and it was markedlyinhibited by NaN3. The enzyme hydrolyzed ATP in prefernece toGTP but not CTP, UTP, ADP, AMP or pNPP. Lineweaver-Burk plotsof its activity were curvilinear in the range of 0.6–0.7mM ATP.Mg2+. 1Present address: Department of Biology, School of Education,Waseda University, Shinjuku-ku, Tokyo, 160 Japan. (Received February 15, 1989; Accepted April 20, 1989)  相似文献   

3.
The subunit of plant heterotrimeric G proteins (G) plays pivotalroles in multiple aspects of development and responses to planthormones. Recently, several lines of evidence have shown thatG participates in brassinosteroid (BR) responses in Arabidopsisand rice plants. In this study, we conducted a comprehensiveanalysis of the roles of the rice G in the responses to BR usinga defective mutant of the G gene, T65d1. Decreased sensitivityto 24-epi-brassinolide (24-epiBL) in the T65d1 mutant was observedin many processes examined, e.g. in the inhibition of root growthand the promotion of coleoptile elongation. The T65d1 mutantalso showed similar phenotypes to those of BR-deficient mutants,such as the specifically shortened second internode and theconstitutive photomorphogenic growth phenotype under dark conditions.However, a negative feedback effect by 24-epiBL on the expressionof BR biosynthetic genes was observed in the T65d1 mutant, andthe levels of BR intermediates did not fluctuate in this mutant.To determine the epistatic relationship between the T65d1 mutantand d61-7, a weak allele of a rice BR receptor mutant, the twomutants were crossed. The T65d1/d61-7 double mutant showed noepistasis in the elongation inhibition of the internodes, theinternode elongation pattern, the leaf angle and the morphologicalabnormality of leaf, except for the vertical length of seedand the seed weight. Our results suggest that the rice G affectsthe BR signaling cascade but the G may not be a signaling moleculein BRI1-meditated perception/transduction.  相似文献   

4.
The angiospermous plant parasite Cuscuta derives reduced carbonand nitrogen compounds primarily from its host. Free amino acidsalong Cuscuta vines in three zones, viz., 0 to 5 cm, 5 to 15cm, and 15 to 30 cm, which in a broad sense represent the regionof cell division, cell elongation and differentiation and vasculartissue differentiation respectively, were quantitatively estimated.The free amino acid content was the highest in the 0 to 5 cmregion and progressively decreased along the posterior regionsof the vine. The haustorial region showed the lowest contentof free amino acids. In general, the free amino acid contentin samples collected at 7 p.m. was found to be higher than thatin the samples collected at 7 a.m. Three basic amino acids,histidine, the uncommon amino acid -hydroxyarginine, and arginineconstituted more than 50% of the total free amino acids in allthe zones studied except the haustorial region. Aspartic acidand glutamic acid constituted the major portion in the acidicand neutral fraction of amino acids. Glutamine, asparagine,threonine, and serine were eluted together and occurred in substantialamounts. -Hydroxyarginine constituted the largest fraction inthe cut end exudate of Cuscuta and presumably appeared to bethe major form of transport amino acid. -Hydroxyarginine wasalso a major constituent of the basic amino acids in Cuscutavines parasitizing host plants from widely separated families,suggesting that this amino acid is a biosynthetic product ofthe parasite rather than that of the hosts. Also, U-14C argininewas converted to -hydroxyarginine by cut Cuscuta vines, suggestingthat -hydroxyarginine is synthesized de novo from arginine byCuscuta. (Received March 30, 1987; Accepted June 7, 1988)  相似文献   

5.
Pumpkin seed globulin is composed of heterogeneous polypeptidechains, acidic and chains and basic 1 and 2 chains (12). This study showed that the basicchains had similar N-terminal sequences, Gly-Leu-Asp-Glu-Thr-Ile-for the 1 chain and Gly-Leu-Glu-Glu-Thr-Ile- for 2. On the contrary,the N-terminal sequences of the acidic and chains were dissimilar, Ile-Gln-Gly-Tyr- for the chain and no N-terminal residue for the chain, according to routine terminal analysis. Pyrrolidonylpeptidase digestion of the chain and its thermolysin digestion followed by Edman degradationsrevealed that the N-terminal sequence of the chain was < Glu-Ile-Glu-Gln-Gln-Glu-Pro(Trp,Ser)-. The N-terminal sequences and the C-terminal residuesindicated that the acidic and chains were more heterogeneous than the basic 1 and 2 chains.A preliminary study on the degradation of storage globulin isalso presented. (Received November 9, 1979; )  相似文献   

6.
A Ras-related NTP-binding protein was partially purified froma membrane fraction derived from the mycelia of Neurospora crassa.[-32P]ATP and [-32P]GTP were incubated with mem brane and solublefractions which were then irradiated with UV light to inducecrosslinking of tightly bound nucleotides. After SDS-polyacrylamidegel electrophoresis, blotting onto a nitrocellulose filter andautoradiography it was apparent that most of the proteins thatbound [-32P]-GTP also bound [-32P]ATP. Pretreatment of the membranefraction with Ras-specific antibody effectively blocked thebinding of [-32P]ATP and [-32P]GTP to several ATP-GTP-bindingproteins. The band of a protein with a molecular weight of 26kDa on the SDS-polyacrylamide gel cross-reacted strongly withthe Ras-specific antibody. The protein was extracted from thegel and further purified by repeated gel electrophoresis. Thepurified protein bound [-32P]ATP, [-32P]-GTP, [-32P]CTP and[-32P]UTP at 1.6x10 M and was autophosphorylated in thepresence of [-32P]ATP and [-32P]GTP at 1.7x10 M. Pretreatmentof the protein with Ras-specific antibody partially blockedthe autophosphorylation in the presence of these nucleotides.The binding of [-32P]ATP to the NTP-binding protein was blockedby addition of ATP at 10–4–10–3 M. ATP ata concentration of 10–4 M prevented the binding of [-32P]to a greater extent than did GTP at the same concentration.Binding of [-32P]CTP and [-32P]UTP to the protein was also observed. (Received October 7, 1991; Accepted July 14, 1992)  相似文献   

7.
Nucleoside triphosphate(NTP)-binding proteins were detectedin the crude extract of mycelia of Neurospora crassa, whichwas treated with 1% Lubrol PX and fractionated by gel filtration.Protein fractions showing the capacity to bind [35S]ATPS or[35S]GTPS were designated as AGN1 to 6. The binding of [35S]ATPSor [35S]GTPS was prevented in the presence of 0.1 mM ATP orGTP except that in fractions AGN1 and 2, the presence of GTPstimulated the binding of [35S] ATPS to ATP(NTP)-binding proteins.ATP or GTP was 1 to 2 orders of magnitude more effective thanCTP or UTP in preventing the binding of [35S]GTPS in AGN1, 2and 5. Among these fractions AGN1, 2, 5 and 6 showed activityto hydrolyze 1 nM [–32P]ATP or [–32P]GTP. NTP-bindingproteins bound with [35S]ATPS or [35S]GTPS had lower apparentmolecular weights than the same proteins without bound nucleotide.Proteins bound with [35S]ATPS or [35S]GTPS and those [32P]ADP-ribosylatedby endogenous ADP-ribosyl transferase in each fraction wereanalyzed by SDS-PAGE. About 20 species of ATP or ATP-GTP-bindingproteins were detected, several of which were ADP-ribosylated.The binding of [35S]ATPS or [35S]GTPS to NTP-binding proteinswas confirmed by the comparison of non-boiled and boiled samplesimmediately before loading to SDS-PAGE. ATP, GTP, CTP or UTPat the concentration of 0.1 mM effectively removed [33S]ATPSor [35S]GTPS bound to NTP-binding proteins. (Received December 10, 1990; Accepted April 18, 1991)  相似文献   

8.
Eleven different types of bacteria were isolated which werecapable of growing on -caprolactam, the monomeric material fornylon 6 polyamide, as the sole source of both carbon and nitrogen. The optimal concentration of -caprolactam for the bacterialgrowth was about 0.6% in a synthetic liquid medium enrichedwith a small amount of yeast extract. The bacterial strains grew also on -butyrolactam, -valerolactamand the -amino acids corresponding to these lactams and -caprolactam.Ammonium adipate was a good substrate for the growth of allthe strains. One strain of Corynebacterium aurantiacum was found to be capableof utilizing cyclic and linear oligomers of 6-aminocaproic acidwith an exception of cyclic dimer. The strains of corynebacteria required vitamin B1 for growth. Metabolism of -caprolactam and related compounds is discussedbriefly. (Received September 9, 1965; )  相似文献   

9.
Aquaporin is a water channel that increases water permeabilitythrough membranous structures. In plants, vacuoles are essentialorganelles that undergo dynamic volume changes during cell growth.To understand the contribution of aquaporins to plant cell growth,we developed a transgenic tobacco BY-2 cell line overexpressingthe tonoplast intrinsic protein (TIP), TIP. Vacuolar membranesof isolated vacuoles from TIP-overexpressing cells showed higherwater permeation activities than those from wild-type cells.We then examined the role of TIP in vacuolar regeneration ofevacuolated tobacco BY-2 protoplasts (miniprotoplasts). Vacuolarregeneration from thin to thick tube-network vacuoles and subsequentdevelopment of large vacuoles was accelerated in miniprotoplastsof this cell line. A parallel increase in the rate of cell expansionindicated a tight relationship between vacuolar developmentand cellular volume increases. Interestingly, overexpressionof tobacco TIP also enhanced cell division. Thus, increasedvacuolar aquaporin activity may accelerate both cell expansionand cell division by increasing water permeability through thevacuolar membrane.  相似文献   

10.
A b-type cytochrome having an -band at 560 nm was isolated fromspinach leaves (Spinacia oleracea). A method is described forpreparing this cytochrome, cytochrome b-560 (spinach), in apurified state. The cytochrome has, in its reduced state, absorption bands at560 nm (), 530 nm (ß) and 427 nm (); and in the oxidizedstate at 562 nm (), 529 nm (ß) and 417 nm (). Thepyridine ferro-haemochrome prepared from cytochrome b-560 hadan -band at 556.5 nm, indicating the protohaem-nature of theprosthetic group. The cytochrome has an oxidation-reduction potential (E'0) of+0.13V at pH 7.0, as measured using the ferri-ferro oxalate system. The cytochrome is rapidly reduced on illumination with red orfar-red light in the presence of spinach chloroplasts and isoxidized at a slower rate in the dark. This photoreduction isinhibited by 1x10–6 M 3-(3,4-dichlorophenyl)-1,1-dimethylurea(DCMU). The molecular weight of the cytochrome is 30,000 asestimated by the dextran gel filtration method. (Received December 3, 1971; )  相似文献   

11.
Tea seedlings were treated with 14C-methylamine to cause synthesisof 14C--glutamylmethylamide (N-methyl-14C). The metabolic conversionof -glutamylmethylamide was studied by tracing 14C. 14C--Glutamylmethylamide (N-methyl-14C) translocated from rootsand cotyledons to shoots of tea seedlings, was converted almostentirely into caffeine. Conversion was greater in light-exposedsamples. For those grown in the dark, the converted amount didnot correspond to the total caffeine produced. More 14C--glutamylmethylamidewas present in stems than in leaves, but with 14C-caffeine,the opposite was found. When 14C--glutamylmethylamide or 14C-methylamine was appliedto leaf disks, 14C-caffeine was biosynthesized from both substances. 1 Present address: Department of Agricultural Chemistry, ShizuokaUniversity, Iwata, Shizuoka 438, Japan. (Received September 25, 1971; )  相似文献   

12.
-Caprolactam-utilizing bacteria split -caprolactam, -valerolactamand -butyrolactam, and produce the -amino acids correspondingto them. This activity is lost when cells are grown on 6-amino-caproicacid or ammonium adipate, and reappears when cells are incubatedwith either -caprolactam or -valerolactam as the sole majororganic nutrient. Chloramphenicol inhibits this adaptation.The enzyme splitting those lactams is one and the same. It maybe called "lactam-splitting enzyme". But attempts to demonstratethe enzymic activity in a cell-free system has not yet beensuccessful. (Received September 9, 1965; )  相似文献   

13.
1. Localization of carbon in caffeine molecule biosynthesizedfrom the N-methyl carbon of -glutamylmethylamide in tea plantswas observed. 14C-Caffeine produced from 14C--glutamylmethylamidewas isolated and degraded. Approximately 26–55% of the14C was observed in the three methyl carbons in caffeine, withonly 2–3% at the C-2 carbon, 3–7% at the C-8 carbonposition. The amount of 14C at the C-4, C-5 and C-6 positionswas calculated from the results obtained. 2. The role of the N-methyl carbon of -glutamylmethylamide inthe formation of RNA in tea plants was examined. Incorporationof the N-methyl-14C of 14C--glutamylmethylamide into AMP andGMP in RNA was found. These facts indicate that in tea plants, -glutamylmethylamideis metabolized and most of its N-methyl carbon is utilized asa precursor for caffeine formation and little, if any, as aprecursor for nucleic acid formation. 1 Present address: Department of Agricultural Chemistry, ShizuokaUniversity, Iwata, Shizuoka 438, Japan. (Received February 2, 1972; )  相似文献   

14.
The sexual agglutinability of haploid cells of heterothallicSaccharomyces cerevisiae was repressed when they were culturedin the absence of easily fermentable sugars, such as glucoseand mannose. The repression was reversed by the action of hormone-likesubstances of the opposite mating types. The substance producedby mating type cells was identical to subtsance-I which isknown to induce sexual agglutinability of inducible matingtype cells. The mating type cells produce a new hormone-likesubstance which induces or enhances sexual agglutinability of mating type cells. A crude fraction of the mating type-specific substance ( substance-I)was obtained by passing the culture filtrate of mating typecells through Amberlite CG-50 (H+ form), followed by elutionwith 1.5 M ammonia. 2 On leave from Osaka City University. (Received December 25, 1975; )  相似文献   

15.
Enzymatic Degradation of Chlorophyll in Chenopodium album   总被引:3,自引:0,他引:3  
The breakdown of chlorophyll (Chi) in crude extracts of Chenopodiumalbum (white goose foot) in the dark was examined. Derivativesof pheophorbide were formed when Chi or chlorophyllide wasincubated with depigmented crude extracts. The formation ofpheophorbide was completely prevented by heat treatment of extracts,indicating that the reaction was enzymatic, and the presenceof a Mg-releasing enzyme, the so called Mg-dechelatase, waspostulated. This hypothesis was strongly supported by the observationthat the formation of pheophorbide was inhibited by 51% by 10mM MgCl2. Analysis by high-performance thin-layer chromatography(HPTLC) and liquid chromatography (HPLC) showed that the appearanceof chlorophyllide , pheophorbide 132-hydroxychlorophyllide and pyropheophorbide was accompanied by a concomitant decreasein levels of Chi The formation of 132-hydroxychloro-phyllide was not clearly an enzymatic reaction and requires furtherexamination. It appears that Chl is degraded in a crude extractof C. album via the following enzymatically catalyzed reactions (Received September 10, 1990; Accepted November 15, 1990)  相似文献   

16.
Polyamines and Anaerobic Elongation of Rice Coleoptile   总被引:2,自引:0,他引:2  
The role of polyamines in the anaerobic elongation of rice (Oryzasativa L.) coleoptiles was studied. The reduced growth of ricecoleoptiles under anoxic conditions was accompanied by a massiveaccumulation of free putrescine. Putrescine was synthesizedfrom arginine in a reaction catalyzed by arginine decarboxylase(ADC). The anoxic titer of putrescine was closely correlatedwith elongation of coleoptiles. In experiments in which putrescineand inhibitors [-difluoromethylarginine (DFMA) and -difluoromethylornithine(DFMO)] of the synthesis of polyamines were exogenously supplied,we demonstrated an absolute requirement for putrescine, synthesizedby ADC, for anaerobic elongation of coleoptiles. The presenceof exogenous putrescine (alone or in combination with DFMA)increased the rate of anaerobic elongation of coleoptile by30–40%. (Received December 1, 1988; Accepted June 19, 1989)  相似文献   

17.
A cell-free ethylene-forming system of Pseudomonas syringaepv.phaseolicola (Kudzu strain) was characterized by its psychrophilictrait. Ethylene was most effectively produced from -ketoglutaricacid (-KG) at 0.5 mM followed by glutamate and then istidineat 5 to 10 mM. The presence of FeSO4 was essential to the cell-freesystem. DTT and histidine greatly stimulated ethylene production;the latter could be substituted to some extent by its analogues.The optimum pH value and temperature for the ethylene-formingreactions were pH 7.0 and 25?C, respectively. Ethylene formationfrom -KG was inhibited in the presence of carbonates or organicacids of the TCA cycle, whereas that from glutamate was inhibitedin the presence of ammonium salts. Ethylene production from-keto--methylthiobutyric acid in the cell-free system was largelydependent on non-enzymical processes in the presence of DTTand FeSO4. The ethylene-forming reactions were inhibited completelyby 1 mM n-propyl gallate and 1 mM p-chloromercuribenzoic acidand partly by coenzymes such as pyridoxal-1-phosphate, folicacid, and flavin mononucleotide at 5mM. The complete systemfor the highest ethylene production consisted of: 0.5 mM -KG,50 mM HEPES (pH 7.0), 5 mM DTT, 0.5 mM FeSO4, and 10 mM histidine.The amount of ethylene produced in this system was equivalentto 40 to 50% of that produced by the living cells. (Received October 22, 1986; Accepted January 19, 1987)  相似文献   

18.
Hypocotyl growth of seedlings of dark-grown squash (Cucurbitamaxima Duch.) was greatly reduced by the addition of 60mM polyethyleneglycol (PEG) to hydroponic solution (water stress). Apoplastic solution (A) and cell sap (C) were separately collectedfrom the hypocotyl segments by a centrifugation method. Theosmotic potentials of A (A) and C (c), and (=cA) ofstressed hypocotyls were always lower than those of unstressedhypocotyls. Suction force was measured by immersing the segments into solutionsof different concentrations of mannitol. Suction force was significantlycorrelated with C (r= –0.99). The mechanical properties of the cell wall of hypocotyl segmentswere measured by stressrelaxation technique. Minimum stressrelaxation time (To), relaxation rate (R) and residual stressof unstressed hypocotyls were low during the growth period andincreased when the growth ceased. To and R of stressed hypocotylsdecreased one day after the stress treatment, but the residualstress was not decreased by the water stress throughout theexperiment. These results suggest that the suppressed growth of dark-grownsquash hypocotyls under water stress was due neither to thereduction of the osmotic potential difference between innerand outer space of the cell, nor to the decrease in suctionforce, but was partly due to the unchanged mechanical propertiesof the cell wall, as represented by one stress-relaxation parameter,residual stress. (Received February 5, 1988; Accepted September 8, 1988)  相似文献   

19.
Aeroponically grown sunflower seedlings (Helianthus annuus L.cv. Russian Giant) were droughted or treated with abscisic acid(ABA) for 7 d. Drought stress prompted a three-phase growthresponse in sunflower roots: an initial phase of increased rootelongation was followed by a period of almost complete inhibitionbetween about 6 h and 72 h; this was followed, in turn, by aphase of partial recovery in the rate of root elongation. Droughtdecreased the size of the apical meristem as cells in the proximalregion of the meristem vacuolated and elongated. Root-to-shootbiomass ratios (R:S) increased initially but declined after72 h. Drought stress decreased water potential () and osmoticpotential ( and increased turgor pressure p in the apical 30mm of the roots. These initial changes were transitory, lastingabout 3 h. Thereafter, and began to rise; p fell back to controllevels. In the later stages of treatment, fell as the stressgrew more severe, but fp was maintained by osmotic adjustment.Desiccation for 1 h increased turgor pressures in excised 30mm apical segments. The transitory increase in root elongationwas contemporary with the initial rise in p in the root apices,while the periods of greatest inhibition and partial recoveryin root elongation were contemporary with the periods of declineand partial recovery in the length of the apical meristem respectively.The inhibition of root elongation and the anatomical changesin the root apices were not determined by loss of turgor orlack of photosynthate, but rather appeared to be an active responseby the meristem to a drop in external . Treatment with ABA triggeredmany of the same changes as drought stress: ABA promoted a three-phasegrowth response, increased R:S, triggered the same initial changesin , , and p, increased p in excised 3.0 mm apical segments,and induced the same pattern of anatomical changes in the rootapices as drought stress. It is proposed that ABA mediates drought-inducedchanges in the primary development of sunflower roots. Key words: Abscisic acid, apical meristem, drought, osmotic adjustmen  相似文献   

20.
The effects of the inhibitors of polyamine biosynthesis, canavanineand -methyl ornithine on growth, the activities of argininedecarboxylase (EC 4.1.1.19 [EC] ) and ornithine decarboxylase (EC4.1.1.17 [EC] ) and on polyamine content were examined in two differentgrowth regions of Phaseolus vulgaris L. cv. Taylor's Horticulturalroots. Separately, in the same manner, in the same bean rootsystem exogenous putrescine effect and the interaction of canavaninewith putrescine were determined. The arginine and ornithine decarboxylase activities found inroot apex were high where cell division activity was highest.Polyamine (putrescine and spermine) content did not correlatewith these activities, but polyamine level was high in the rootbase where cell elongation is the main process. The arginineanalogue, canavanine, inhibited arginine decayboxylase activityand polymine liters. Putrescine partially reversed the canavanineinhibition of root growth as well as arginine decarboxylaseactivity and polyamine content. Similarly -methyl ornithineslightly inhibited the root length and ornithine decarboxylaseactivity in the root apex. Besides, exogenous putrescine didnot effect significantly the endogenous polyamine titers. Theseresults reinforce the growing connection between polyaminesand the rates of cell devision in the roots of bean plants.Separately, arginine decarboxylase is the main enzyme in thebean roots. (Received November 10, 1986; Accepted March 3, 1987)  相似文献   

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