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1.
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   

2.
Effects of l, N6-ethenoadenylates (e-adenylates) were testedon phosphorylation, and electron transport under phosphorylation,arsenylation and quasi-arsenylation (stimulation of electrontransport in the presence of ATP, AMP and arsenate) conditionsin isolated spinach chloroplasts. -ATP as well as ATP partially inhibited ferricyanide reductionthrough binding to the chloroplast coupling factor 1 with anapparent dissociation constant (KDapp) of around 5µM,which was remarkably larger than that for ATP (ca. 2µM).e-ATP at below 500 µM had no effect on phosphorylationbut inhibited quasi-arsenylation in competition with ATP withan apparent inhibition constant (K1app) of around 60 µM. -ADP as well as ADP partially inhibited ferricyanide reductionwith a KDapp value close to that for -ATP. -ADP was phosphorylated(the apparent Michaelis constant, Kmapp=80µM) accompanyingstimulation of ferricyanide reduction to the magnitude predicted(P/e=l). -ADP-arsenylation was also detected by stimulationof ferricyanide reduction. -AMP alone caused little inhibition of ferricyanide reductionas AMP, but competitively depressed the electron transport inhibitionby ADP and ATP with a K1app value of around 200 µM. -AMPwas not effective for ADP phosphorylation but inhibited stimulationdue to quasi-arsenylation coupling in competition with AMP K1app=150µM Among the possible combinations of adenylates and -adenylatesfor quasiarsenylation, only [ATP+AMP] could couple with theenergy transduction mechanism. Based on the specificity of binding sites to adenylates and-adenylates, an attempt was made to distinguish at least four(two pairs) kinds of binding sites (at least six sites in toto)on the chloroplast coupling factor 1 for photosynthetic energytransduction. When one pair of sites is occupied by the designatedadenylates or -adenylates (allosteric effectors), the couplingfactor is thought to be in a conformation for coupling withthe energy transduction mechanism in the presence of phosphateor arsenate. 1Presented to the 1st Symposium of Japan Bioenergetics Group,December 19, 1975, Osaka. (Received February 17, 1976; )  相似文献   

3.
Sixteen legumes were grown in N-free media so that N was suppliedentirely by symbiotic N2 fixation. The plant tissues were analyzedfor natural 15N abundance (expressed as 15N per mil relativeto air N2) with a ratio mass spectrometer. The nodules of desmodium,centro, siratro, soybean and winged bean showed high enrichmentin 15N (+9), while red clover showed slight enrichment (+2).The nodules of 9 other forage legumes (Townsville stylo, whiteclover, alsike clover, common vetch, Chinese milk vetch, senna,alfalfa, ladino clover, and hairy vetch) showed little enrichmentin 15N. In all the legumes investigated, particularly in the ureide-transportingplants such as desmodium, centro, siratro, soybean, winged beanand field bean, the 15N value of the shoots was negative (–3.2).The 15N value of the shoots in winged bean and field bean variedby about 1 depending on the Rhizobium strains used. The isotopicmass balance of 13 legumes indicated that isotopic fractionationoccurs during N2 fixation by the legume-rhizobia symbiosis witha preference for 14N over 15N, resulting in a 15N value of –0.2to –2 in the whole plant. The results indicate that 15N/14N isotopic discrimination witha preference for the lighter atom may occur in both N2 fixationand export of fixed N from nodules. 1Present address: Department of Soils and Fertilizers, NationalAgriculture Research Center, Kannondai, Tsukuba, Ibaraki 305,Japan. (Received October 8, 1985; Accepted April 7, 1986)  相似文献   

4.
This work aimed to study the impacts of acquisition and assimilationof various nitrogen sources, i.e. NO3, NH4+ or NH4NO3,in combination with gaseous NH3 on plant growth and acid-basebalance in higher plants. Plants of C3 Triticum aestivum L.and C4 Zea mays L. grown with shoots in ambient air in hydroponicculture solutions with 2 mol m–3 of nitrogen source asNO3, NH4+ or NH4NO3 for 21 d and 18 d, respectively,had their shoots exposed either to 320 µg m–3 NH3or to ambient air for 7 d. Variations in plant growth (leaves,stubble and roots), and OH and H+ extrusions as wellas the relative increases in nitrogen, carbon and carboxylatewere determined. These data were computed as H+/N, H+/C, (C-A)/N,and (C-A)/C to analyse influences of different nitrogen sourceson acid-base balance in C3 Triticum aestivum and C4 Zea maysplants. Root growth in dry weight gain was significantly reduced bytreatment with 320 µg m–3 NH3 in Triticum aestivumand Zea mays growing with different N-forms, whereas leaf growthwas not significantly affected by NH3. In comparison with C3Triticum aestivum, non-fumigated C4 Zea mays had low ratiosof OH/N in NO3–3-grown plants and of H+/N in NH4+- and NH 4NO3-grown plants. Utilization of NH3 from the atmospherereduced both the OHN ratios in NO3 -grown plantsand the H+/N ratio in NH4+ - and NH4NO3 -grown plants of bothspecies. Furthermore, Zea mays had higher ratios of (C-A)/Nin NH4+ - and NH4NO3-grown plants than Triticum aestivum. Thismeans that C4 Zea mays had synthesized more organic anion perunit increase in organic N than C3 Triticum aestivum plants.Within both species, different nitrogen sources altered theratios of (C-A)/N in the order: NH4NO3>NH4+>NO3.Fumigation with NH3 increased organic acid synthesis in NO3- and NH4+ - grown plants of Triticum aestivum, whereas it decreasedorganic acid synthesis in Zea mays plants under the same conditions.Furthermore, these differences in acid-base regulation betweenC3 Triticum aestivum and C4 Zea mays plants growing with differentnitrogen sources are discussed. Key words: Acid-base balance, ammonia, ammonium, nitrate, ammonium nitrate, C3 Triticum aestivum L., C4 Zea mays L.  相似文献   

5.
An -glucan was isolated from 11-day-old suspension-culturedrice cells by extraction with hot Na-phosphate buffer (pH 6.8).The -glucan had []D=+234? (C = 0.14, in water) and its averagemolecular weight was estimated to be about 1.4 ? 104, basedon elution characteristics on acalibrated Sepharose CL-6B column.Upon partial acid hydrolysis, the -glucan gave mainly malto-oligosaccharides.The maximum absorption of the iodine complex of the -glucanin the presence of Na2SO4 was at 470 nm. The results of hydrolysisby , ß- and iso-amylases and methylation analysisindicated that the isolated -glucan is a highly branched polysaccharidewith an average chain length of 9. The exterior and interiorchain lengths of the -glucan were calculated to be 5 and 3,respectively. (Received July 23, 1986; Accepted February 7, 1987)  相似文献   

6.
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   

7.
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   

8.
Two proteolytic activities I and II involved in the globulindegradation were detected in pumpkin seeds. Activity I, hydrolyzing and ß subunits of the globulin to form Fß,was found in both dry seeds and cycloheximide-treated cotyledons,and decreased during germination. Activity II, hydrolyzing Fßto produce small peptides and amino acids, was not observedin dry seeds but found in cycloheximide-treated cotyledons,increased up to 4 days, and gradually decreased during germination. Activity I gave limited hydrolytic products from the globulinand the chain, but not from Fß, the chain and some animal proteins. It was inhibitedby EDTA. On the other hand, activity II hydrolyzed Fßand the chain faster than the globulin, the chain and some animal proteins. It was inhibitedby EDTA and p-chloromer-curibenzoate, and activated by ß-mercaptoethanol,dithiothreitol and CoCl2. Optimum pH's were at about 6.8 andat 6.0 to 6.8 for activities I and II, respectively. The degradation process of the globulin can be divided intotwo steps: the first step is the conversion of globulin to Fßand the second step, Fß to small peptides and aminoacids. (Received November 9, 1979; )  相似文献   

9.
A non-linear Arrhenius plot of diaphorase activity of the membrane-boundferredoxin-NADP+ oxidoreductase of thylakoids, showed a breakat 25–30°C, whereas a straight-line was obtained forthe soluble reductase. The Ea values of the thylakoid-boundenzyme was 4–7 and 15–18 kj/mol above and belowthe transition temperature, respectively. In contrast, the Eaobtained for the soluble enzyme was in the range 25–30kj/mol. An Arrhenius plot with similar transition to that ofthylakoids, but with higher Ea values, was obtained when thereductase was isolated as a complex closely associated withthe 17.5 kDa-intrinsic polypeptide. The differences in H* and in S* values below and above the transitiontemperature for thylakoids and the complex were almost the samesuggesting that lipids were not important for the transition. Comparing the H* and S* values for the diaphorase activity ofsoluble and membrane-bound enzyme suggest that the conformationof the latter is more favorable for catalysis. 1Present address: Depto. de Bioquímica, Fac. de Biologiay C.S.I.C., Univ. de Sevilla, Apdo. 1095, 41080-Sevilla, Spain (Received May 1, 1986; Accepted June 18, 1986)  相似文献   

10.
The effect of a pure preparation of substance-IA (S-IA) whoseamino acid sequence is identical to that of one of the factorpeptides (2), on sexual agglutinability and DNA synthesis wascomparatively studied. The optimum concentration of S-IA forthe induction of sexual agglutinability of cells of an inducible strain was 1 ng/ml. The inducing action of S-IA was detectedin 20 min and reached a maximum in 60 min. Only 8.7% inhibitionof DNA synthesis by S-IA in the same strain was detected in1 hr and 40.4% inhibition in 2 hr at a concentration of 1 µg/ml.These results suggest that the primary action of the peptidyl sex fractor on a mating-type cells is the induction of sexualagglutinabiity. (Received October 25, 1977; )  相似文献   

11.
The possibility that gibberellin-induced -amylase synthesisin barley endosperm might be mediated by cyclic-3',5'-adenosinemonophosphate (3',5'-AMP) was examined. Promotion of -amylasesynthesis by 3',5'-AMP (5 mM) was observed in the absence ofgibberellic acid (GA3) and in combination with GA3 at concentrationsbelow 2 mµM. When combined with gibberellin at concentrationsabove 2 mµM, however, 3',5'-AMP reduced the amount of-amylase obtained. The cyclic nucleotide showed slight activityat concentrations as low as 0.05 mM. These promotions were shownto be due to increased synthesis of -amylase rather than toan increased secretion of the enzyme. Of a variety of adeninecompounds and nucleoside diphosphates tested only 3',5'-AMPand adenosine diphosphate (ADP) induced -amylase synthesis.Longer incubation times were necessary to obtain maximal -amylaseinduction with the nucleotides than with GA3. ADP and 3',5'-AMPwere about one third and one fifth as active, respectively,as GA3 in promoting -amylase synthesis, although GA3 was morethan 107 times more effective. AMO-1618 did not inhibit theaction of the nucleotides and methanolic extracts of the nucleotidesshowed no gibberellin-like activity. Both nucleotides were synergisticwith GA3 in overcoming the inhibitory effects of acetate andcitrate buffers on -amylase synthesis. (Received February 24, 1969; )  相似文献   

12.
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   

13.
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   

14.
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   

15.
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   

16.
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   

17.
The effects of transpiration rate on the vertical gradientsof leaf and stem xylem water potential ( and ) were examinedusing hydroponic sunflower plants. Transpiration was variedby stepwise alterations of environmental conditions. The gradientsof and were relatively small (2.3 and 0.8 x 105 Pa m–1)when transpiration rates approached zero, but increased sharplyto 5.4 and 2.3 x 105 Pa m–1 as transpiration increased.However, the gradients were independent of transpiration ratesabove 0.4 g dm–2 h–1 owing to variability of theplant resistance. The gradients of I were usually less thanhalf those of I. 1 in individual leaves remained constant over a wide range oftranspiration rates (0.4—2.4 g dm–2 h–1) andeach leaf possessed a characteristic plateau value related toits elevation. I responded similarly but was approximately 2.0x 105 Pa higher than I at the same elevation. Identical resultswere obtained regardless of the procedure employed to vary transpiration. The drop in water potential between stem and leaf implies thatthe leaf resistance is appreciable. This was confirmed usingrapidly transpiring excised leaves freely supplied with water.I increased by 2.0–2.5 x 105 Pa following removal of theroot resistance but remained 2 x 105 Pa lower than similar excisedleaves in darkness. Furthermore, I in excised leaves remainedconstant over a wide range of transporting rates, demonstratingthat the leaf resistance is also variable. The results are discussed in relation to previous reports.  相似文献   

18.
19.
SYNOPSIS. Studies are described on the adenosine triphosphatase(ATPase) properties of myosin isolated from skeletal muscleof Coryphaenoides, a benthic fish captured at 2,200 meters depth.Ca2+-ATPase and EDTA-ATPase of Coryphaenoides myosin show thesame pH dependence as ATPase of mammalian myosin; however, ratesof ATP hydrolysis by Coryphaenoides myosin are only 5–10%of rates of ATP hydrolysis by rabbit skeletal myosin. Coryphaenoidesmyosin ATPase shows a decrease from Q10 of 2.0 at 25°C toQ10 of 1.4 a t 2°C, and undergoes irreversible denaturationat temperatures above 25°C. At pH 6.8 to pH 8.5, Coryphaenoidesmyosin ATPase undergoes activation by pressure at 25°C,but at 2°C shows negligible effect of pressure at valuesbelow 3,000 psi. The kinetic data on Ca2+-ATPase indicate valuesof 11 kcal/mole for H, –7.5 kcal/mole for TS, and –5.7cc/mole for V at 25°C, pH 7.6. Comparable data at 2°Cindicate values of 5 kcal/mole for H. –13 kcal/mole forTS, and negligible V. According to the results of 25°C,Ca2+-activatkm of myosin-ATP may involve disruption of fouror five hydrophobic or polar groups, presumably due to an "opening-up"of the myosin molecule at or near the site for ATP binding.It would also appear that Coryphaenoides myosin has undergonean adaptive change in the enzyme mechanism for ATPase such thatthe rate of ATP hydrolysis is relatively insensitive to pressureand temperature under conditions encountered by the living fish.  相似文献   

20.
The euryhaline charophyte Lamprothamnium papulosum (Wallr.)J. Gr. was adapted to media with decreasing salinities rangingfrom 550 to 0 mosmol kg–1. Vegetative plants grown inmedia with osmotic pressures (0) in the range of 550 to 130mosmol kg–1 maintained a constant turgor pressure () at309 + 7 mosmol kg–1. The ions K+, Na+ and Cl–, werethe predominant solutes in the vacuole. Changes in their concentrationsaccount for the variation in internal osmotic pressure (1) with,0. The divalent ions Mg2+, Ca2+ and were also present in significant amounts, but their concentrationsdid not alter with changes in, 0. In cells subjected to hypo-osmotic shock the regulation of was incomplete. The turgor pressure increased from 302 to 383mosmol kg–1. The first rapid response to the sudden decreasein 0 was a loss of K+ and Cl. In contrast to the decreasein ionic concentrations an accumulation of sucrose occurredwhich could account for the increase of . The increase in sucroseconcentration started 24 to 48 h after the downshock and reachedits highest value after 3 to 4 weeks. The sucrose concentrationin the vacuole was up to 320 mol m–3. During this timethe ionic content continued to decrease but did not counterbalancethe sucrose concentration sufficiently to regain the original. High sucrose levels accompanied by an enhanced were also observedduring the period of fructification (sexual reproduction: formationof antheridia and oogonia) in Lamprothamnium kept under conditionsof constant salinity. It is concluded that high sucrose content and elevated arecharacteristic of sexual reproduction in this charophyte. Lamprothamniumis able to tolerate different during various developmentalstages (e.g. vegetative and reproductive phases). Key words: Lamprothamnium papulosum, sucrose, turgor pressure  相似文献   

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