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1.
Nitrate-induced polypeptides in membranes from corn seedling roots   总被引:2,自引:0,他引:2  
The polypeptide composition of the membranes from corn (Zeamays L.) seedling roots upon nitrate induction was determinedby two-dimensional gel electrophoresis and silver-staining.The synthesis of five polypeptides (49, 48, 35, 33, and 32 kDa)in the tono-plast fraction and four polypeptides (50, 49, 38,and 33 kDa) in the plasma membrane fraction was induced by both2.5 mM Ca(NO3)2 and 5 mM KNO3. Extensive washing of the membraneswith salt and NaOH demonstrated that three induced polypeptides(49, 48, and 35 kDa) in the tonoplast fraction and two inducedpolypeptides (49 and 33 kDa) in the plasma membrane fractionwere integral proteins. After incubation of seedlings in N-freemedium for 4 d, the 49 and 32 kDa polypeptides in the tonoplastfraction had disappeared. By the sixth day in N-free medium,the 35 kDa polypeptide had disappeared from the tonoplast fraction.The 50 kDa polypeptide of the plasma membrane fraction was nolonger detectable in seedlings incubated for 6 d in N-free medium.The size of the spots corresponding to the 33 kDa polypeptidesof both membrane fractions and to the 49 kDa polypeptide ofthe plasma membrane fraction was reduced following incubationof seedlings in N-free medium. The changes in nitrate-inducedpolypeptides in both membrane fractions following transfer toN-free medium correlated with a reduced capacity to take upnitrate in the treated seedlings. The results support the conclusionthat the nitrate-induced polypeptides may be involved in nitratetransport across the tonoplast and plasma membrane. Key words: Nitrate transport, induction, membrane peptides  相似文献   

2.
32P incorporation into the protein fraction of chloroplast fragmentsby short illumination was investigated under various phosphorylatingconditions. 32P incorporation was generally accompanied by cyclic and non-cyclicphotophosphorylations and also by formation of a high energyintermediate "XE". However, the addition of a DPIP-ascorbatecouple caused inhibition of 32P incorporation, while ATP formationproceeded. Effects of inhibitors and uncouplers of photophosphorylationon the formation of protein-bound 32P were generally similarto those on ATP formation. AT32P was not utilized for protein-bound 32P formation in thedark by chloroplast fragments, but its radioactivity was transferredinto the chloroplast protein fraction in the light. Oligomycininhibited ATP formation but did not inhibit protein-bound 32Pformation. m-Cl-CCP blocked both reactions. This suggests thatprotein-bound 32P is not an actual intermediate in the phosphorylativeprocess leading to formation of ATP. It is probably formed ona side pathway from an intermediate of ATP formation. Analyses of protein-bound 32P after digestion with proteaseand lipase showed that the 32P incorporated was bound to peptidesin chloroplast lamellae. The possible form of this bound 32Pis discussed. (Received November 22, 1971; )  相似文献   

3.
Early developing embryos of the toad Bufo arenarum Hensel were employed to study the content and in vivo labeling with 32P of the acid-soluble phosphates and phospholipids at the subcellular level. The radionuclide was administered to the female toad along with the pituitary extract used to induce the ovulation.Most of the total phospholipids (68%) and proteins (84%) are confined to the yolk platelet fractions. Up to the heart beat stage (130 h of development) there are no significant changes detectable in protein and phospholipid content.The total P content in trichloroacetic acid-soluble fraction was distributed mainly between postmitochondrial supernatant (58%) and yolk platelet fraction (37%) in the unfertilized oocyte. As development proceeds an increase was observed in the former and a decrease in the latter. The acid-solube phosphates in the mitochondrial fraction only amount to 4% of the total embryo throughout the examined stages.The unfertilized oocyte contains about 98% of acid-soluble phosphates labeled with 32P in the postmitochondrial supernatant and as development proceeds a striking decrease was found to occur while the radioactivity in the acid-soluble phosphates of mitochondrial and yolk platelet fractions increases significantly during the studied stages. About 11.5% of the lost radioactivity from the acid-soluble phosphates was found to be used to label the phospholipids.  相似文献   

4.
Phospholipid composition and 32P metabolism were studied in oocytes and early developing embryos of the toad, Bufo arenarum, Hensel. The content and distribution of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidic acid, sphingomyelin, phosphatidylserine, and diphosphatidylglycerol in embryos, whole oocytes, and the subcellular fractions of both were determined. Phosphatidylcholine and phosphatidylethanolamine were the major constituents of yolk platelet. Diphosphatidylglycerol was confined to the mitochondrial fraction, where it represented about 7% of the total phosphoacylglycerols. Relatively large amounts of sphingomyelin were found in microsomal and postmicrosomal supernatants. After in vivo labeling with 32P, the early development of individual phospholipids in subcellular fractions and in whole eggs was followed. The greatest uptake was found in mitochondrial and yolk platelet fractions. A steady increase in the amount of 32P present in phosphatidylethanolamine, phosphatidylcholine, and phosphatidylinositol was seen in the whole embryo from oocyte to late gastrula stage and in all subcellular fractions. Phosphatidic acid exhibited a slight decrease in specific activity, except in the yolk platelet fraction. This high 32P incorporation would indicate a rapid and uneven polar headgroup turnover determined by phospholipid class and subcellular fraction. At the same time, the phospholipid content of the subcellular fractions studied remained unchanged during early embryogenesis. Moreover, 32P was actively incorporated into the individual phospholipids in the absence of measurable net synthesis.  相似文献   

5.
Early developing embryos of the toad Bufo arenarum Hensel were employed to study the content and in vivo labeling with 32P of the acid-soluble phosphates and phospholipids at the subcellular level. The radionuclide was administered to the female toad along with the pituitary extract used to induce the ovulation.Most of the total phospholipids (68%) and proteins (84%) are confined to the yolk platelet fractions. Up to the heart beat stage (130 h of development) there are no significant changes detectable in protein and phospholipid content.The total P content in trichloroacetic acid-soluble fraction was distributed mainly between postmitochondrial supernatant (58%) and yolk platelet fraction (37%) in the unfertilized oocyte. As development proceeds an increase was observed in the former and a decrease in the latter. The acid-solube phosphates in the mitochondrial fraction only amount to 4% of the total embryo throughout the examined stages.The unfertilized oocyte contains about 98% of acid-soluble phosphates labeled with 32P in the postmitochondrial supernatant and as development proceeds a striking decrease was found to occur while the radioactivity in the acid-soluble phosphates of mitochondrial and yolk platelet fractions increases significantly during the studied stages. About 11.5% of the lost radioactivity from the acid-soluble phosphates was found to be used to label the phospholipids.  相似文献   

6.
Influence on individual types (fractions) of nucleic acids (NA) was studied in roots of wheat plants grown in various cultivation media, namely in distilled water, in sodium humate and in a nutrient solution. NA’s were prepared by means of the phenol technique. Using separation on a methylalbumine column (MAK) five fractions were obtained, namely: fraction I.— low molecular weight substances, fraction II.—soluble RNA, fraction III.—DNA-RNA, and the ribosomal RNA in two fractions, IV.—(l r-RNA) and V.—(h r-RNA). Of the NA fractions investigated, the r-RNA fraction was noticeably influenced by the kind of nutrition, its amount varying in a certain proportion to the growth intensity affected by the cultivation medium. The other NA fractions were not apparently affected. The metabolical turnover of the individual NA types (as observed from the specific32P activity) was considerably influenced by the kind of nutrition as well. The specific32P activity in all NA fractions of wheat roots cultivated in a nutrient solution was approximately double that in roots of wheat plants cultivated in distilled water and Na-humate. Changes in the specific32P activity of r-RNA were again considerably evident. With regard to root growth their relative values appeared in an inverse proportion to the changes in the r-RNA content. The specific32P activity decreased with increasing growth intensity. Besides changes in the r-RNA fraction, changes in fraction I. were apparent. An especially striking decrease in the specific32P activity was found in roots of plants grown in H2O, namely by about one order in comparison with its specific activity in fraction I. from roots of plants grown in Na-humate and in a nutrient solution.  相似文献   

7.
The rate of uptake of 32P from labelled NaH2PO4 solutions sprayedon to one leaf of swedes (Brassica napus) or French beans (Phaseolusvulgaris) was rapid during the first few hours and fell to zeroafter 4 days. 22P was detected in the root after 3 hours andcontinued to move out of the treated leaf for at least 6 daysafter application. A larger fraction of the applied 32P wasabsorbed from repeated than from a single spraying. Swedes absorbed more 32P from a single application to the lowersurface than to the upper surface of the leaf. Doubling theconcentration of the spray caused a small increase in the percentageof applied 22P that was absorbed. Absorption by French-beanleaves decreased slightly when the area sprayed with a constantamount of 32P was doubled, and decreased with increasing ageof leaf. Increasing the phosphorus supply to the roots of swedesaffected neither the initial rate nor the total amount of 32Puptake by the leaves but decreased the quantity of 32P thatwas translocated out of the treated leaf. Increasing the relative humidity of the air around the plantsalso increased32P uptake. Shading usually decreased uptake andalways decreased translocation. Rewetting the leaf to whichthe 32P had been applied, with water or sucrose solution, hadvariable effects. The significance of the results is discussed.  相似文献   

8.
The availability of six organic phosphorus substrates (OPS)to serve as sources of this nutrient for natural populationsof phytoplanktonic algae and bacteria was tested by measuringthe degree to which the organic substrates decreased uptakeof [32P]orthophosphate (32Pi). When added to samples of LakeKinneret water, six organo-phosphorus compounds usually loweredthe amount of 32Pi] taken up by microplankton retained on 0.2-µfilters.In contrast, OPS addition generally stimulated 32Pi uptake intothe mainly algal fraction (>3 µm), indicating thatthe bacteria were mostly responsible for utilising OPS. Thesparing effect of OPS addition on 32P1 uptake was very fast,suggestingthat the micro-organisms possessed constitutive or rapidly inducedenzyme systems to exploit the OPS.The results of this studyindicate that a significant flux of phosphorus may pass viaDOP into microbiota, especially bacteria, in some aquatic systems.  相似文献   

9.
1. The effect of hormones on (32)P incorporation into various RNA fractions in germinating pear embryos was studied by fractionation on methylated albumin-kieselguhr columns. Abscisic acid inhibited labelling of soluble RNA, DNA-RNA hybrid and light-ribosomal RNA fractions with (32)P and this effect was reversed by both kinetin and gibberellic acid. 2. Kinetin reversed the inhibition by abscisic acid of (32)P incorporation into total ribosomal RNA and appeared to promote labelling of heavy-ribosomal RNA. Gibberellic acid was more active than kinetin in reversing the inhibition by abscisic acid of labelling of the DNA-RNA hybrid fraction with (32)P, but in contrast with kinetin appeared to increase further the inhibition by abscisic acid of labelling of total ribosomal RNA. 3. The percentage of radioactivity in various RNA fractions showed marked variation in response to hormones. 4. The pattern of labelling of RNA in pear embryos during reversal of inhibition by abscisic acid with a combination of kinetin and gibberellic acid was similar to that after cold-treatment of dormant pear embryos. This is suggestive of hormonal interplay in dormancy release by cold-treatment in pear embryos.  相似文献   

10.
DNA-RNA hybridization-competition experiments were performedto analyze RNA heterogeneity in wheat embryos during germinationand vernalization. A significant difference was found betweenRNA populations of 24-hr and 3-day germinated embryos, whileminor differences were detected between 3- and 5-day germinatedembryo RNAs and between 20- and 40-day cold-treated embryo RNAs.RNA populations in 30-day cold-treated embryos were significantlydifferent from those in 50-day ones. The RNA species presentin 50-day cold-treated embryos were not similar to those in3- and 5-day germinated embryos. A great portion of the RNAspecies in 3-day germinated embryos was found in 30-day cold-treatedembryos. These results suggested that some new RNA species aresynthesized in wheat embryos during 30 to 50 days of cold treatment. 1 Present address: Department of Biochemistry, Faculty of PharmaceuticalSciences, Higashi Nippon Gakuen University, Ishikari-Tobetsu,Hokkaido 061–02, Japan. (Received February 21, 1977; )  相似文献   

11.
After incubation of pigeon pancreas slices with P32 and isolation of various fractions by differential centrifugation the deoxycholate extract of the microsome fraction was found to account for over half of the phospholipide P and over half of the P32 incorporated into the phospholipides. The remaining phospholipide P and P32 were fairly evenly distributed in the nuclei, zymogen granules, mitochondria, microsomal ribonucleoprotein particles, and the soluble fraction. When enzyme secretion was stimulated with acetylcholine about two-thirds of the increment in radioactivity in the total phospholipides was found in deoxycholate soluble components of the microsome fraction. The remainder of the increment was distributed in the other fractions. This indicates that the cellular component in which the increase in phospholipide turnover occurs on stimulation of secretion is a membranous structure. Evidence is presented which indicates that the increment in radioactivity in the non-microsomal fractions on stimulation of secretion is due to contamination of these fractions with fragments of the stimulated membranous structure. The distribution of P32 radioactivity in each of the chromatographically separated phospholipides in the various fractions from unstimulated tissue paralleled the distribution of radioactivity in the total phospholipide fraction, indicating that individual phospholipides are not concentrated in different fractions but are associated together in the membranous structures of the microsome fraction. The major proportion of the stimulation of the turnover of the individual phospholipides also occurred in the microsome fraction. The distribution of radioactivity from glycerol-1-C14 in the total phospholipides and in the individual phospholipides in the various fractions was similar to the distribution of P32. In the microsome fraction acetylcholine stimulated the incorporation of glycerol-1-C14 in each phospholipide which showed a stimulation of P32 incorporation. The significance of the turnover of phosphatides in microsomal membranes in relation to the mechanism of secretion is discussed.  相似文献   

12.
The metabolism of phospholipids in mouse brain slices   总被引:1,自引:1,他引:0       下载免费PDF全文
1. Slices of mouse brain grey matter were incubated with [32P]phosphate and [1-14C]acetate. Doubly labelled phospholipids were extracted from subcellular fractions prepared from the slices in a mixture of metabolic inhibitors, under conditions where there was negligible change in radioactive labelling during the preparation. Two tissue fractions were studied in detail; one contained a high proportion of mitochondria and the other was mainly microsomal. 2. In all tissue fractions the highest incorporations of both [32P]phosphate and [1-14C]acetate occurred into phosphatidylcholine. 3. After incubation for 1hr., the 32P/14C ratios for phosphatidylcholine, phosphatidylethanolamine and phosphatidic acid in the mitochondrial fraction were similar to those in the microsomal fraction. 4. The 32P/14C ratios were similar in phosphatidylcholine and phosphatidylethanolamine and much lower than those in phosphatidic acid and phosphatidylinositol.  相似文献   

13.
The fatty acid synthesizing system from developing soybean cotyledonswas fractionated intoa precipitate fraction and a supernatantfraction using carbowax-4000. Both fractions were necessaryfor fatty acid synthesis. The supernatant fraction was heatstable, non-dialyzable and lost its activity when incubatedwith pronase. Fatty acid synthesis required NADH and NADPH aswell as ATP, KHCO3 and MnCl2. The crude enzyme produced stearicand oleic acid while the recombined system produced predominadystearic acid 1Presented in part at the American Society of Plant PhysiologistMeeting in Bloomington, Indiana, August 1970 (Received November 27, 1970; )  相似文献   

14.
Contents of nicotinamide nucleotides, NAD, NADP, NADH2 and NADPH2,in wheat embryos were determined during normal germination andcold treatment. They increased markedly in the early stage ofgermination then decreased later. During cold treatment, thesenucleotides increased initially as in normal germination, butdid not decrease in the later stages. The C6/C1 ratios during germination decreased gradually withaging, but after about the fourth day increased. This suggeststhat the pentose phosphate pathway operates actively. Duringcold treatment, the C6/C1 ratio decreased slightly with aging.This suggests that the EMP pathway operates predominantly inthe glucose metabolism. Based on these results, the correlationbetween NADPH2 and glucose metabolism during cold treatmentwas discussed. Embryos which had absorbed labelled glucose were fractionatedinto several chemical components. The radioisotope accumulatedmainly in the sugar fraction, with small amounts in the organicacid, amino acid and nucleic acid fractions. Changes in thecontent of each component showed that sugar, organic acid andamino acid increased during cold treatment. 1 Present address: Department of Biochemistry, Faculty of PharmaceuticalScience, Higashi Nippon Gaguen University, Ishikari-Tobetsu,Hokkaido 061-02, Japan. 2 Present address: Hokkaido Forest Products Research Institute,Asahikawa 070, Japan. 3 Present address: Kakuto 534, Komae City, Tokyo 182, Japan. (Received November 1, 1976; )  相似文献   

15.
Summary This study concerns the development of pollen embryos as affected by various physical conditions of culture in media devoid of hormones. Freshly isolated pollen, from anthers ofDatura, failed to form embryos regardless of whether they were cultured on liquid or solid medium. In contrast, pollen isolated from anthers precultured on solid medium did form embryos and the response could be increased by prior cold treatment of anthers at 4 °C for 4 days. However, the best results were obtained when anthers were cultured from the very beginning in liquid medium and transferred serially to fresh medium. Under such conditions, the anthers dehisced, allowing spontaneous shedding of pollen grains. It was thus possible to have several fractions of shed pollen continuing their development into embryos. When serial culture was started with anthers from cold-treated buds not only were embryos formed in all the fractions of shed pollen but the frequency was also considerably higher than in any mode of culturing.  相似文献   

16.
Differential filtration was used to apportion [32p]orthophosphate(P1) uptake to predominantly bacterial (<3 µm) or algal(>3 µm) components of Lake Kinneret microplankton.Bacteria generally showed preferential 32Pi uptake in comparisonwith algae. Nevertheless, in most cases, the relative proportionof 32P counts retained on 3 µm filters was greater thanthe proportion of 14C counts from heterotrophic bacterial incorporationof [14Clglucose, indicating that algae were competing for Piwith bacteria with some measure of success. Most time courseexperiments did not show any consistent transfer of 32P frombacteria to algae. The addition of a bacterial inhibitor (garamycin)caused a relative increase in the proportion of algal to bacterial32Pi uptake. Added organic P substrates lowered the amount of32Pi uptake and appeared to be preferentially utilized by bacteria.Apparent residence times for Pi in Lake Kinneret ranged from0.4 h (prior to overturn) to 17.4 h during bomothermy. Despitelow ambient Pi concentrations, P limitation in Lake Kinneretis not as extreme as in many other aquatic environments.  相似文献   

17.
Abscission: the role of RNA synthesis   总被引:10,自引:10,他引:0       下载免费PDF全文
Holm RE  Abeles FB 《Plant physiology》1967,42(8):1094-1102
Ethylene stimulated the incorporation of 32P into RNA in the abscission zone of bean explants (Phaseolus vulgaris L. var. Red Kidney). The enhancement was observed in all fractions separated by methylated albumin kieselguhr column chromatography, although the magnitude of the increase was not the same for each fraction. Differential extraction of the nucleic acids indicated that the ethylene stimulation was confined to the fraction extracted with sodium lauryl sulfate, with the increase mainly in Fraction III (Ribosomal RNA) and Fraction IV (Messenger RNA). Actinomycin D, which blocks ethylene-stimulated abscission, inhibited 32P incorporation into all column fractions. 5-Fluorouracil, which blocked 50% of the ethylene-enhanced 32P incorporation, did not inhibit ethylene-enhanced abscission. The results indicate that ethylene may regulate abscission through control of specific RNA's.  相似文献   

18.
The sulfur-oxidizing system of a strain of Thiobacillus thiooxidanswas obtained in cell-free state. The system is resolved intothree fractions and can be reconstituted from these fractions.Both the soluble and particulate fractions are required forthe oxidation of elementary sulfur. The soluble fraction wasfurther separated into two fractions, the collodion membrane-permeable(S-P)and the impermeable(S-IP). S-P contains a low molecular weight,relatively heat stable substance(s) which is indispensable forthe reconstitution of the sulfur-oxidizing system and was identifiedas a pyridine nucleotide. The function of S-P can be replacedby NAD or NADP, but not by cysteine nor GSH. Oxidation of NADH2 and NADPH2 is catalyzed by the particulatefraction. Oxidation of the latter is much more rapid than thatof the former. Oxidation of NADPH2 as well as sulfur oxidationis inhibited by cyanide, pCMB and CO, the CO-inhibition beingphoto-irreversible. However, strong inhibitors of sulfur oxidationsuch as DDC, 8-hydroxyquinoline and salicylaldoxime have noeffect on the oxidation of NADPH2. The optimum pH values for sulfur and sulfite oxidations by thecell-free extract are shifted to the neutral side in comparisonwith pH values by intact cells. 1V = References(I). 2Partly supported by a grant from the Ministry of Education. (Received April 3, 1969; )  相似文献   

19.
天山云杉(Picea schrenkiana)是亚洲中部山地的特有物种, 在中国仅见于新疆, 主要分布在天山南北坡和昆仑山西部北坡, 对天山山地的水源涵养、水土保持和林业生产具有不可或缺的重要性。云杉天然林自然更新不良或障碍和生产力下降等问题的出现已成为长期困惑林业工作者的一大难题。该文从植物间的化感与自毒作用研究着手, 分析云杉针叶水提液的乙醚、乙酸乙酯、正丁醇萃取物对自身种子萌发和幼苗生长影响实验。以种子发芽率、发芽势和发芽指数作为种子萌发参数, 以胚根、胚芽长度和实生苗干湿重的变化作为幼苗生长参数。研究结果表明: 种子萌发实验中乙醚萃取物具有最强的抑制效应(IC50 = 5.84 mg·ml-1), 而正丁醇萃取物的抑制效应最弱(IC50> 10.00 mg·ml-1); 幼苗生长实验中, 1.25 mg·ml-1乙醚萃取物对幼苗生长具有显著抑制作用(p<0.05), 1.25 mg·ml-1正丁醇萃取物能够促进其幼苗生长, 大于2.5 mg·ml-1的正丁醇萃取物对幼苗生长具有显著抑制效应(p<0.05)。采用GC-MS-MS和NMR技术分析3种有机萃取物的化学组成, 鉴定并定量分析出了包括酚酸、长链脂肪酸、单宁酸和吲哚类物质在内的17种化合物。乙醚萃取物中存在的2-keto-4a-methyl-8-methoxy-2, 3, 4, 4a, 5, 6, 11, 12-ocahydrochrysene (在该文中, 将其命名为云杉酮)为一新的化感物质。该研究证明了天山云杉针叶中存在的化感与自毒物质是导致其种群天然更新障碍的一个最主要原因, 同时揭示了自毒作用发生的化学物质基础。  相似文献   

20.
TAYLOR  M.; FRANCIS  D. 《Annals of botany》1989,64(6):625-633
The cell cycle in Silene coeli-rosa shoot apices was measuredto test whether or not early components of the floral stimulus,produced during the 2nd and 3rd long days (LD) of an inductiveLD treatment, resulted in an increase in the duration of G2phase in constant 20–24 h cell cycles. Plants were grownat 20°C in short days (SD) of 8 h light and 16 h darknessfor 28 d (day 0). Starting on day 0, plants were given SD or3 LD each comprising an identical 8 h day and 16 h photo-extension,or 3 dark-interrupted (d.i.) non-inductive LD, interrupted at1700 h of each day with 1 h of darkness. The cell cycle (percentagelabelled mitoses method) and changes in cell number were determinedin the shoot apical meristem. During days 1–2 of the SDtreatment, the cell cycle and mean cell generation time (MCGT)was 18 and 32 h, respectively, giving a growth fraction of 56%.During days 2–3, the cell cycle and MCGT shortened to15 and 23 h, respectively (growth fraction = 65%). During days1–2 of the LD and d.i. LD treatments, cell cycles andMCGTs were 9–10 and 27–29 h, respectively, resultingin smaller growth fractions (about 33%). Thus, shortened cellcycles and altered growth fractions occurred regardless of whetheror not the treatment was inductive. The LD treatment resultedin a marked shortening of G1 and, to a lesser extent, S-phase,whilst G2 remained constant. These changes were consistent withincreases in the proportion of cells in G2 during the photoextensionof each LD which were suppressed during the comparable periodsof the d.i. LD treatment. The latter treatment resulted in eachphase occupying virtually identical proportions of the cellcycle as in the SD treatment. Thus, the unique cell cycle responsesto the initial part of the inductive LD treatment were increasesin the proportion of cells in G2 coupled with G1 and G2 beingof similar duration. Cell cycle, mean cell generation time, shoot apex, Silene coeli-rosa  相似文献   

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