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1.
Using the min-LD method, light requirements of the L1- and L2-phasesof L. gibba G3 were found to be satisfied by only 5 min illuminationgiven respectively from CT 0:00 to 0:05 and from CT 11:55 to12:00. This rigorous time sense was displayed without any alterationeven in the presence of iron reagents, e.g., 10–5 M o-phenanthroline,10–5M,'-dipyridyl and 10–6 M kinetin, which completely eliminatedcircadian rhythmicity in reproductive (flower production) aswell as vegetative (frond production) response to a light pulsescanning a continuous dark period. Circadian rhythms of metabolicactivities, e.g., active K+ ion uptake and respiratory CO2 output,were not changed at all by the iron reagents. These and relevantresults suggested that in this long-day duckweed, the circadianoscillator, probably located in the meristem and sensitive toiron deficiency, only modulates the frond and flower productionin the meristem and is not related to the critical daylengthmeasurement. (Received December 18, 1978; )  相似文献   

2.
DL-Epinephrine (10–8–10–7 M), DL-norepinephrine(106 M) and DL-isoproterenol (10–8–10–6 M)alleviated floral inhibition due to 1% of sucrose, in Lemnagibba G3. The induction period extended by sucrose was curtailedby epinephrine, frond multiplication enhanced by the sugar beingleft unaltered. The pattern of action of catecholamines appearedto be very similar to that of cAMP. DL-Epinephrine, however,was ineffective in the presence of 10–7 M DL-propranololwhich affected neither flower nor frond production by itself.Quabain and nicotinic acid also nullified the epinephrine actionon duckweed flowering. These and other relevant findings supportthe hypothesis that the cAMP-adenyl cyclase system participatesin the processes of flower induction in this long-day plant. (Received August 21, 1973; )  相似文献   

3.
Inhibition of flower initiation in light culture of a long-dayduckweed, Lemna gibba G3 by an inserted period of brief darknesschanges its magnitude with the application time of the dark-break.‘Response vs. time of the dark-break’ curve consistsof superimposed thermo-insensitive oscillatory and thermo-sensitivehourglass components. Oscillation is diurnal and damps in afew days. The situation is very similar to what has previouslybeen revealed concerning the non-photosynthetic light-requirementof the same strain of duckweed for frond multiplication. Increasingphotophily for flower production is likely accompanied by decreasingphotophily for frond production, and vice versa. The presenceof iron chelators such as o-phenanthroline and ,'-dipyridylor reduced iron concentration in culture medium abolishes theoscillatory change, without modifying the hourglass change,in either vegetative or reproductive photophily. Ferrous, butnot ferric, ions likely mediate between the basic clock andthe photophily. Critical daylength for flower induction is notaffected by exogenous chelators. (Received November 17, 1970; )  相似文献   

4.
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   

5.
Action patterns of IAA, KIN and GA on the growth of Lemna gibbaG3 are similar to those reported for agents chelating both cupricand ferric ions. Relatively high doses of growth substances,e.g.10–6 M IAA or KIN and –4 M GA, inhibit developmentof photoperiodically induced flower buds and antagonisticallypromote frond multiplication; whereas, at relatively low doses,e.g. 10–9 M IAA or KIN and –5 M GA, they acceleratethe process occurring in the latter half of the induction periodto enhance flower induction. Complex forming abilities of IAA,KIN and GA with cupric and ferric ions are demonstrated spectrophotometrically.Moreover, the ferrous ion-dependent oscillatory change in reproductiveand vegetative photophilies of duckweed is eliminated by KINbut not by IAA and GA. Of the three growth substances tried,KIN alone shows an affinity for ferrous ions. (Received April 5, 1971; )  相似文献   

6.
Under short-day conditions the growth or the production of fronds in Lemna gibba G3 was stimulated by KIN (10–5 M);the longer the nyctoperiod, the greater the stimulation was.Under long-day conditions KIN was slightly inhibitory for thefirst 2 days, but promotive thereafter. IAA (10–5 M) reversedthe growth inhibition by KIN in long-days, and DCA (10–4to 10–6 M) and also CA (10–4 to 10–6 M), althoughless effectively, cancelled the growth promotion by KIN in short-days.DCA (10–5 M) little altered the KIN inhibition in thefirst 2 days of light culture, nevertheless it alleviated strikinglythe KIN promotion in the subsequent illuminated days. These and relevant findings explained the above-described promotionand inhibition by KIN of the duckweed growth in terms of invivo level of a cofactor (probably auxin) which may be underthe regulative influence of photoperiodic conditions given.Moreover, some bearing of the above conclusion on floweringmechanism in this long-day duck-weed was suggested. (Received September 12, 1966; )  相似文献   

7.
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   

8.
The luciferin-luciferase method was used to determine ATP extractedfrom darkmaintained and light-exposed samples of the green algaChlorella pyrenoidosa and of the blue-green alga Anacystis nidulans.A few measurements on Synechococcus lividus (a bluegreen thermophile,clone 65?C) are also reported.
  1. The light-minus-dark ATP levels (ATP) from aerobic cells ofChlorella and Anacystis were negative; however, ATP from Synechococcuswas positive. Large positive ATP was obtained in regularly grown(RG: moderate light) Chlorella treated with oligomycin; darklevels were reduced, light levels remained essentially unaffected.In high-light exposed (HLE) Chlorella, oligomycin reduced bothlight and dark ATP levels, but positive ATP was still obtained.However, in Anacystis, which has a different organization ofthylakoid membrane, oligomycin severely reduced both the lightand the dark ATP levels and the ATP remained negative.
  2. Theoligomycin (12 µM) treated Chlorella and the untreatedAnacystis and Synechococcus show the presence of cyclic photophosphorylationunder conditions in which the non-cyclic electron flow fromphotosystem II to photosystem I is blocked by 10 µM 3-(3,4-dichlorophenyl)-l,l-dimethylurea(DCMU), or not allowed to operate by the absence of CO2. Cyclicphotophosphorylation ranged from 10–30% of the maximumATP in RG, to 40–50% in HLE Chlorella. In RG Chlorella,cyclic and non-cyclic (in the absence of DCMU) photophosphorylation(ATP) saturate at about 103 ergs cm–2 sec–1 and104 ergs cm–2 sec–1 and 104 ergs cm–2 sec–1red (>640 nm) light, respectively; a lag was observed inthe light curve.
  3. In Chlorella, the addition of the photosystemI electron acceptormethyl viologen (MV; 1 mM) increased ATPby twofold. Furtheraddition of DCMU (25 µm) reduced thisto the level observedwith DCMU alone. If 1 mM reduced dichlorophenolindophenol orphenazine methosulphate (DCPIPH2 or PMSH2, respectively)wasadded along with DCMU, the ATP level was 30–40% ofthecontrol. Further addition of MV increased the JATP to be70–80%of that of the control. These and other resultsconfirm thepresence of both non-cyclic and cyclic photophosphorylationin vivo, the former predominating in Chlorella, and the latterin Anacystis and Synechococcus.
(Received May 1, 1973; )  相似文献   

9.
In Vigna mungo cotyledons, the -amylase activity increased markedlyduring germination at 27°C in the dark, while the activityof other amylases was very low. The -amylase was purified from4-day-old cotyledons by affinity chromatography on epoxyactivatedSepharose 6B substituted with rß-cyclodextrin andby column chromatography on Bio-Gel P-200. Gel filtration andpolyacrylamide gel electrophoresis showed that the enzyme existsmostly as a monomer (43,000 daltons), but partially aggregatesto form dimer, trimer and further multimers. Ca2+ protectedthe -amylase against heat inactivation. Incubation of the enzymewith 5 mM EDTA or dialysis against 10 mM EDTA resulted in a50–90% loss of activity. The inactivation was partiallyreversed by the addition of Ca2+. Other properties, such asthe amino acid composition, Km value, pH optimum and activationenergy were similar to those of other plant -amylases. (Received May 6, 1981; Accepted June 22, 1981)  相似文献   

10.
A gamete recognition mechanism in Antithamnion sparsum Tokidais proposed based on experiments using various lectins and carbohydrates.Spermatial binding to trichogynes is inhibited by pre-incubationof spermatia with concanavalin A (ConA) and/or L-fucose, whiletrichogyne receptors are blocked by the complementary carbohydrate-methyl D-mannose and/or the lectin Ulex europaeus agglutinin(UeA1). Binding inhibition (40–50%) was observed with10–50 mM carbohydrates and 25–50 µg ml-1 lectins.The inhibitory effects of ConA and UeA1 is partially reversed(to 80–90% of controls) by addition of -methyl D-mannoseand L-fucose, respectively. Lectin binding to spermatial surfaceswas visualized by Fluorescein isothiocyanate (FITC) conjugatedConA, whereas carbohydrate receptors along the trichogyne andspermatium were localized with -mannosylated-FITC-albumin andL-fucosylated-FITC-albumin, respectively. These results suggestthat gamete recognition in Antithamnion sparsum is mediatedby a double-docking recognition system consisting of spermatiapossessing surface L-fucose receptors and -methyl D-mannosemoiety, and trichogynes possessing the complementary receptors. (Received December 5, 1995; Accepted April 22, 1996)  相似文献   

11.
Nitrite reductase was extracted from the red alga Porphyra yezoensisUeda and purified through precipitation with ammonium sulfate,column chromatographies, and polyacrylamide gel disk electrophoresis.The enzyme preparation thus obtained showed a single band ondisk electrophoresis. The absorption spectrum had three maxima at 385 nm (Soret band),580 nm (-band), and 278 nm; the ratio of absorbance of the Soretband to the -band was 4.3. The molecular weight and the numberof amino acid residues were estimated to be 63,000 and 601,respectively. The enzyme activity was optimal at around pH 7.5, and its activitywas heat labile as indicated by reduction of activity by about70% when heated at 37°C for 10 min. The enzyme used ferredoxin and methyl viologen, but not NADP+or NAD+, as the electron carriers. Moreover, reduced forms ofthe latter two showed no effect on its activity. Km values ofthis enzyme for NO2, Fd, and MV were 8.1 x 10–4M, 4.3 x 10–8 M, and 3.7 x 10–4 M, respectively.Almost half of its activity was lost when potassium cyanidewas added at a concentration as low as 10–5 M, and theKi value was 1.8 x 10–5 M. Thus, the nitrite reductaseof Porphyra must be systematically grouped in EC 1.7.7.1 [EC] . Itresembled closely that of Chlorella, except for the amountsof some amino acids. 1 Present address: Department of Biological Sciences, Universityof Tsukuba, Sakura-Mura, Ibaraki, 300-31 Japan. 2 Present address: Department of Fisheries, College of Agricultureand Veterinary Medicine, Nihon University, Shimouma, Setagaya-ku,Tokyo, 154 Japan. (Received June 10, 1975; )  相似文献   

12.
-Amanitin and cordycepin at various concentrations were testedfor their inhibitory effect on the fresh weight increase ofVigna unguiculata embryonic axes after the onset of imbibitionand on the incorporation rate of 3H-labeled leucine into proteinin axes of the 36–38 h stage. -Amanitin at 0.5–5µ/Kg/ml clearly exerted an inhibitory effect on both thefresh weight increase and the protein synthesis. This drug at1 µg/ml, however, showed no significant effect on theprotein synthesis at an early stage of imbibition (4 h), whereascycloheximide was a very potent inhibitor. By experiments inwhich ‘dry’ axes were allowed to imbibe 3H-lebeledadenosine solution for 4 and 12 h in the presence of -amanitin,it was found that poly A+RNA was newly synthesized to some extentin axes as early as 4 h after the onset of imbibition and thatthe drug effectively inhibited the poly A+RNA synthesis. Theresults may indicate the occurrence of stored mRNA in embryonicaxes of V. unguiculata seeds. (Received June 11, 1983; Accepted August 16, 1983)  相似文献   

13.
Oryzains, cysteine proteinases of rice seeds, are induced byGA3 in germinating rice seeds [Abe et al. (1987) Agric. Biol.Chem. 51: 1509]. The effects of GA1, GA3, GA4, GA9, and GA20on the production of oryzain and -amylase were investigatedin embryoless half- and whole-seeds of rice (cv. Nipponbare).When gibberellins (GAs) were incubated with embryoless half-seeds,GA1, GA3 and GA4 induced oryzain and -amylase, but GA9, andGA20 did not. GA9 and GAM induced oryzain and -amylase productionin whole seeds, but this production was inhibited by the simultaneousapplication of prohexadione, an inhibitor of 2ß- and3ß-hydroxylation of GAs. Prohexadione did not inhibitthe activities of oryzain and -amylase induced by GA1. Theseresults suggest that GAs possessing the 3ß-hydroxylgroup induce activities of oryzain and -amylase in rice seedsand that GA9 and GA20 have activity only after they are convertedmetabolically to active GAs, probably GA4 and GA1, respectively.GA1, was more active than GA4 in both half seeds and wholeseeds incubation. Oryzain and -amylase activities induced byGA4 were significantly inhibited in the presence of 10–4M prohexadione. This suggests that the conversion of GA1, toGA4 (13-hydroxylation) might be inhibited at a high dose ofprohexadione in whole seeds. 4Present address: Institute of Food Development, Kyung Hee University,Suwon 449-701, Korea  相似文献   

14.
-Aminolevulinic acid dehydratase (5-aminolevulinate hydro-lyase,EC 4.2.1.24 [EC] ) was purified from greening radish cotyledons. Thefinal product was homogeneous on polyacrylamide disc gel electrophoresisand had a molecular weight, estimated by gel filtration, of282,000 daltons. The enzyme seems to require magnesium ion aswell as sulfhydryl compounds for maximum activity. EDTA anda low concentration of zinc ion markedly inhibited the activity.The optimum pH was 8.0; the Km value for -aminolevulinic acidwas 3.85?10–4M. Levulinic acid was a competitive inhibitorof the enzyme, with a Ki of 2.14?10–4M. These propertieswere compared with those of microorganism and animal -aminolevulinicacid dehydratases. (Received November 22, 1976; )  相似文献   

15.
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   

16.
-Glucans (average mol wt, 1.3 ? 104) extracted with perchloricacid from 8-day-old suspension-cultured nonglutinous (var. Sasanishiki)and glutinous rice (var. Miyakogane) cells were compared. Theresults of hydrolysis by alpha;-, ß- and iso-amylasesand methylation analysis of the -glucans suggested that theirbasic structures are almost the same. These -glucans are highly-branchedpolysaccharides with an average chain length of about 9–10,with exterior and interior chain lengths of about 6–7and 2–3, respectively. 1Current address: Laboratory of Food Science, Faculty of Education,Hirosaki University, Hirosaki, Aomori 036, Japan. (Received April 27, 1987; Accepted March 2, 1988)  相似文献   

17.
Properties of the nucleotide binding sites on chloroplast couplingfactor 1 (CF1) were studied by equilibrium dialysis and UV spectroscopy.From our direct binding studies, we identified at least fourkinds of ADP binding sites on CF1; a barely dissociable ADPbinding site (site A), a slowly exchangeable high affinity sitewith dissociation constant (Kd) 0.021 µM (site B), anotherslowly exchangeable high affinity site with Kd 1.6 µM(site C) and several low affinity (Kd {small tilde}30 µM)sites. The Kd values for sites B and C of the other nucleotidestested were 0.5 µM and 16 µM (GDP), 8 µM and34 µM (CDP), 17 µM and 20 µM (UDP) and 1.4µM and 1.4 µM (PP1). From a comparison of the observed UV spectral change and theamount of nucleotide bound to these sites, as calculated fromthe above Kd values, we concluded that the nucleotide bindingto site B or G induces UV spectral changes that are almost thesame in shape and magnitude. The estimated difference molarabsorption coefficient () was 3.4?103M–1ADP cm–1for ADP at 278 nm. Our conclusions were strengthened by thegood agreement between the observed spectra and the calculatedspectra (derived from the Kd and values of ADP and GDP) whenADP and GDP were added together to CF1. The cause of the unusual behavior of GDP in the UV differencespectrum which was unexplained in our previous report was shownto be competition between the GDP added and previously boundADP at sites B and C; this distorted the real spectrum inducedby GDP. (Received October 3, 1983; Accepted February 13, 1984)  相似文献   

18.
The inhibitory effects of the pyrone derivatives, 6-()-decenyl-2,3-dimethyl--pyrone(DDP) and 6-farnesyl-2,3-dimethyI--pyrone (FDP), on the photosyntheticelectron flow system was investigated using the blue-green algaAnabaena variabilis and the green alga Chlorella pyrenoidosa. Both reagents inhibited photosynthesis in intact cells; 50%inhibition occurred at 2.7 x 10–5 M with DDP and at 4.3 lO–6 M with FDP in Anabaena photosynthesis. The reagentssuppressed the photosystem II reaction [water to 2,6-dichlorophenolindophenol (DCIP)] of Anabaena membrane fragments, but werefar less inhibitory on the photo-system I reaction (DCIPH2 tomethyl viologen). The kinetics of the fluorescence inductionindicated that the reagents do not block Q-reduction, but dosuppress the oxidation of reduced Q indirectly. Oxygen evolutionunder repetitive flashes at a low repetition rate (5 Hz) wasinsensitive to the reagents even at concentrations which inducedmore than 50% inhibition. These results are evidence that DDPand FDP inhibit the plastoquinone reaction by slowing down itsturnover rate. The advantages of pyrone derivatives are that they are inactivein the oxidation-reduction reaction and do not quench the fluorescenceof chlorophyll in vivo. (Received April 14, 1980; )  相似文献   

19.
Uptake of the toxic heavy-metal, thallium, was studied in thecyanobacterium Synechococcus R-2 (PCC 7942) using clinicallyavailable 201Tl +. Thallium was found to distribute across theplasmalemma passively, and so the accumulation ratio of theion ([Tl+]i/[Tl+]o) could be used to calculate the apparentmembrane potential (­i,o) of the cells (ETI+i,o = ­i,o).The permeability of the plasmalemma to TI+ (PTI+ 1 to 5 nms–1)is higher than that of K+. Valinomycin does not increase thepermeability of TI+. Transient changes in the ­i,o of cells,because of electrogenic transport of ions, could be detectedfrom its effects upon the uptake rate of TI+. HCO3 hyperpolarizedSynechococcus cells, whereas NH+4, CH3NH+, and K+ led to depolarization.The use of TI+ as a reporter of ­i,o has some inherent limitations.Tl+ is toxic at very low concentrations (inhibitory effectsare apparent after about 6 h at concentrations as low as 1 mmolm–3). The rate of equilibration is slow (t1/25 to 20 min).Equilibration of TI+ takes about 2 h, which limits its valueas a membrane potential probe. Large amounts of TI+ bind tothe surface of the cells making the method impracticable formeasuring accumulation ratios of less than about 10 (­i,o)values smaller than about –60 mV). Cultures continuouslyexposed to Tl+ (10 mmol m–3) eventually become TI+ resistantby actively extruding TI+ (µTI+i,o= –3±0.2kJ mol–1) and so thallium cannot be used as a ­i,oprobe in such cells. (Received October 28, 1997; Accepted August 31, 1998)  相似文献   

20.
To a certain extent the flowering of Lemna paucicostata 6746is evoked in continuous light by application of abscisic acid(ABA) and CCC. Moreover, the action of the combined substancesappears in two separate concentration ranges. In the range ofABA 2?10–9 M/CCC 10–7–10–6 M floweringis initiated without inhibition of vegetative growth and proceedsonly in the presence of high intensity light and sucrose. Acombination of ABA 2?10–5 M/CCC 10–3M simultaneouslycauses a strong inhibition of frond multiplication. Here theeffect can be observed also under low intensity light conditionsand without sucrose in the medium. A range with flower inhibitingactivity lies between the two flower promoting concentrationranges. (Received November 16, 1972; )  相似文献   

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