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1.
Atriplex gmelini plants were regenerated via organogensis from hypocotyl explants. Callus lines were induced from the hypocotyl explants on Linsmaier and Skoog (LS) medium supplemented with 1 M benzyladenine and 5 M -naphthaleneacetic acid in the dark. Shoots were regenerated from the callus lines on LS medium supplemented with 20 M thidiazuron and 0.1 M -naphthaleneacetic acid under a high-intensity light condition (450 mol m–2 s–1). The regenerated shoots were rooted on LS medium without growth regulators to obtain fully developed plants. We succeeded in transforming Atriplex gmelini from callus lines using Agrobacterium tumefaciens.  相似文献   

2.
Shoot tip and single node explants from young shoots of 1-year old flowering plants of Rauwolfia micrantha Hook. f. were cultured on Murashige & Skoog (MS) medium variously supplemented with 6-benzyladenine (BA) and -naphthaleneacetic acid (NAA). A combination of 13.2 M BA and 2.68 M NAA induced high frequency (77%) formation of up to 3 shoots from each node in 8 weeks. The regeneration of shoot tips from the field-grown plants and in vitro shoots placed horizontally differed. Repeated subculturing of the shoot tips and single nodes at 6-week intervals for over a year in combination of 4.4 M BA and 0.27 M NAA enabled mass multiplication of shoots without any evidence of decline. Rooting of the excised shoots on medium containing 2.6 M NAA was preceded by callus formation. The rooted plants were removed off the callus, hardened off and 80% established in pots. Micropropagated plants displayed uniform morphological, growth, flowering, fruiting and seed germination characteristics.Abbreviations BA 6-benzyladenie - IAA indole-3-acetic acid - IBA indole-3-butyrie acid - 2-ip 2-isopentenyladenine - MS Murashige & Skoog (1962) - NAA -naphthaleneacetic acid  相似文献   

3.
This report describes the newly identified Sarcocystis inghami n. sp. from the skeletal muscles of opossums (Mammalia: Didelphidae) that were collected from south central Michigan (42° 43-42° 79N, 84° 18-84°mathtype="display">6'W), USA. The new species is distinguished from all species described from North and South American opossums by the distinctive morphology of the villar protrusions on the cyst wall. Sarcocysts of S. inghami are microscopic, up to 700 m long and 110 m wide. The sarcocyst wall is up to 7 m thick, with long, stalked protrusions which average 5.5 × 1.2 m. These are constricted at the base, expanded laterally, rounded off distally and occasionally bifid. The villar protrusions have numerous microtubules without electron–dense bodies that extend from the tips into the granular layer. Bradyzoites are 10.7 × 4.3 (8––12 × 4––5) m. This is the second species of Sarcocystis sarcocyst described from the Virginia opossum in North America.  相似文献   

4.
In order to uncover encoder properties of primary muscle spindle afferent fibers, time coupling (phase-locking) of action potentials on cyclic muscle stretch was studied by means of pseudo-random noise. In cats Ia action potentials were recorded from dorsal root filaments and the gastrocnemius muscles of one hind leg were stretched. The stimulus time course was a determined sequence of randomly varying muscle length which could be applied repeatedly (sequence duration 0.6 or 20 s). The noise amplitude (standard deviation of displacements) was varied between 5 and 300 m, the upper cut-off frequency of noise f c was varied between 20 and 100 Hz. The responses to the consecutive pseudo-random noise cycles were displayed as raster diagrams and cycle histograms. Phaselocking characterized the responses at all noise amplitudes outside the near threshold range (>10 m). The higher and f c , the stronger was the phase-locking of impulses on the stretch. When and f c were selected to achieve high mean stretch velocities of about 500 mm/s, phase-locking was as precise as 0.15 ms, measured as the variability of spike occurrences with respect to stretch. The rasters obtained with low noise amplitudes (<40 m) showed a loose phase-locking and this gave insight into underlying mechanisms: The elicitation of action potentials caused by dynamic stretch can be prevented by a post-spike depression of excitability. This disfacilitation was very effective in counteracting weak stretch components within the random sequence and less effective or even missing when relatively strong stretch components could force the spike elicitation. This led to the reestablishment of phase-locked patterns. The results were discussed in relation to the known encoder models.  相似文献   

5.
Plantlet regeneration in Cucumis metuliferus from several explant sources, including cotyledons, leaves, hypocotyls and petioles, was evaluated on Murashige and Skoog's medium containing various combinations of auxin (IAA, NAA, 2,4-d) and cytokinin (BA, kinetin, zeatin), Callus development was obtained within 4 to 5 weeks on all growth regulator combinations which were tested at concentrations ranging from 1.0 M to 4.0 M of each. The response was similar when the tissues were incubated under light or in continuous darkness. Differentiation of callus to form adventitious buds or shoot primordia occurred only with petiole explants on medium containing NAA/BA or 2,4-d/BA at 2.0/1.0 M; none of these calluses, however, differentiated further to form shoots. When the differentiated calluses derived from petiole explants which had been initiated on 2,4-d/BA at 2.0/1.0 M were transferred onto medium with 2.0 M zeatin, formation of shoots occurred within 2 to 3 weeks. The frequency of shoot formation was 14.6%. Subculture of these shoots onto MS medium without growth regulators gave rise to plantlets of normal appearance. Regeneration in C. metuliferus requires callus initiation on an appropriate growth regulator regime followed by transfer to a medium containing the cytokinin, zeatin, and can be achieved within 10–12 weeks.Abbreviations BA 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - NAA napthaleneacetic acid  相似文献   

6.
Endogenous and xenobiotic sulphur-containing convulsant and non-convulsant compounds containing structural moieties of, or bearing a structural resemblance to, GABA and homocysteine were tested in binding studies for their potency in displacing the GABA-mimetic [3H]muscimol from specific, high-affinity sites (K d=3.6 nM;B max=3.94 pmol/mg protein) on freeze-thawed, Triton-treated calf-brain synaptic membranes. The xenobiotic convulsants, 4-mercaptobutyric acid (MBA), 3-mercaptopropionic acid (3-MPA) and 2-mercaptopropionic acid (2-MPA) were found to be two-site competitive inhibitors exhibiting apparent inhibition affinity constants (K i app ) of 5000 M, 3750 M, and 4800 M, respectively; while homocysteic acid (K i app =4800 M) was shown to be a one-site partial competitive inhibitor. Intermediary metabolites of methionine: S-adenosyl-l-homocysteine,l-cysteine, the convulsantl-homocysteine, and its non-convulsant disulphide oxidation product, homocystine, were found to be one-site partial competitive inhibitors exhibitingK i app values of 5750 M, 8350 M, 5000 M, and 510 M, respectively. The endogenous anticonvulsant neuroeffector, taurine, and the tripeptide, reduced glutathione (GSH) were shown to be, respectively, one-site (K i=20 M) and two-site (K i app =4300 M) competitive inhibitors of [3H]muscimol binding. These findings are discussed with regard to a previously proposed mechanism for the convulsant action of homocysteine.  相似文献   

7.
Iron-efficient (WF9 corn and Coker 227 oat) and Fe-inefficient (ys1 corn and TAM 0–312 oat) cultivars were comparatively tested for their response to Fe-deficiency stress induced by the use of either ferrous or ferric chelators. Corn and oats were grown in 20 M Fe with 0, 60, and 120 M BPDS and 40 M Fe with 0, 120, and 240 M BPDS and 20 M Fe with 0 and 40 M EDDHA. All four cultivars tested, both Fe-efficient and Fe-inefficient, continuously reduced Fe3+ to Fe2+ at a low level as evidenced by the production of Fe2+ (BPDS)3 in test nutrient solutions over time. Severity of chlorosis increased as more BPDS was added to the nutrient solutions for both WF9 and ys1 corn, but unlike corn, Coker 227 and TAM 0-312 oats were both able to obtain Fe from the Fe2+ (BPDS)3 complex and were less chlorotic as a result. In short-term (4-hour) in vivo measurements, iron-stressed WF9 (Fe-efficient) corn reduced more Fe3+ to Fe2+ than similarly stressed ys1 corn, Coker 227 oat or TAM 0-312 oat. Thus, at the same time that Fe-efficient WF9 corn reduces more Fe than the other cultivars, it is also unable to compete with BPDS for that Fe in the nutrient solution. These differences coupled with the observation that only Coker 227 oat produced measureable iron solubilizing substances (phytosiderophores) suggest that these two species differ in their mechanisms for obtaining Fe during Fe-deficiency stress.  相似文献   

8.
In the field, photosynthesis of Acer saccharum seedlings was rarely light saturated, even though light saturation occurs at about 100 mol quanta m-2 s-1 photosynthetic photon flux density (PPFD). PPFD during more than 75% of the daylight period was 50 mol m-2 s-1 or less. At these low PPFD's there is a marked interaction of PPFD with the initial slope (CE) of the CO2 response. At PPFD-saturation CE was 0.018 mol m-2 s-1/(l/l). The apparent quantum efficiency (incident PPFD) at saturating CO2 was 0.05–0.08 mol/mol. and PPFD-saturated CO2 exchange was 6–8 mol m-2 s-1. The ratio of internal CO2 concentration to external (C i /C a ) was 0.7 to 0.8 except during sunflecks when it decreased to 0.5. The decrease in C i /C a during sunflecks was the result of the slow response of stomates to increased PPFD compared to the response of net photosynthesis. An empirical model, which included the above parameters was used to simulate the measured CO2 exchange rate for portions of two days. Parameter values for the model were determined in experiments separate from the daily time courses being sumulated. Analysis of the field data, partly through the use of simulations, indicate that the elimination of sunflecks would reduce net carbon gain by 5–10%.List of symbols A measured photosynthetic rate under any set of conditions (mol m-2 s-1) - A m (atm) measured photosynthetic rate at saturating PPFD, 350 l/l CO2 and 21% (v/v) O2 (mol m-2 s-1) - C constant in equation of Smith (1937, 1938) - C a CO2 concentration in the air (l/l) - C i CO2 concentration in the intercellular air space (l/l) - C i /* C i corrected for CO2 compensation point, i.e., C i -I *, (l/l) - CE initial slope of the CO2 response of photosynthesis (mol m-2 s-1/(l/l)) - CEM CE at PPFD saturation - E transpiration rate (mmol m-2 s-1) - F predicted photosynthetic rate (mol m-2 s-1) - G leaf conductance to H2O (mol m-2 s-1) - I photosynthetic photon flux density (mol m-2 s-1) - N number of data points - P m predicted photosynthetic rate at saturating CO2 and given PPFD (mol m-2 s-1) - P ml predicted photosynthetic rate at saturating CO2 and PPFD (mol m-2 s-1) - R d residual respiratory rate (mol m-2 s-1) - T a air temperature (°C) - T l leaf temperature (°C) - V reaction velocity in equation of Smith (1937, 1938) - V max saturated reaction velocity in equation of Smith (1937, 1938) - VPA vapor pressure of water in the air (mbar/bar) - VPD vapor pressure difference between leaf and air (mbar/bar) - X substrate concentration in equation of Smith (1937, 1938) - initial slope of the PPFD response of photosynthesis at saturating CO2 (mol CO2/mol quanta) - (atm) initial slope of the PPFD response of photosynthesis at 340 l/l CO2 and 21% (v/v) O2 (mol CO2/mol quanta) - I * CO2 compensation point after correction for residual respiration (l/l) - PPFD compensation point (mol m-2 s-1)  相似文献   

9.
Three glutathione-S-transferase (GST) isozymes (Q1, Q2, and Q3) from the northern quahog (Mercinaria mercinaria) were purified and separated with a combination of affinity and ion exchange chromatography. SDS-PAGE analysis of the separated quahog GSTs indicated there are four distinct subunits of the enzyme with molecular masses ranging between 23 and 27 kDa. The electrophoretic analysis in combination with GST information from literature indicates that among the quahog GST isozymes, there is a single homodimer and two heterodimers. Enzymatic kinetic analysis of the homodimeric quahog GST (Q3) using 1-chloro-2,4-dinitrobenzene and glutathione as reactants resulted in V max and K m values of 33.2 mol min–1 mg–1 and 0.40 mM, respectively. A pH profile analysis of the Q3 GST indicates that the optimum catalytic pH is 7.6. The Q3 isozyme composes about 28% of the ion exchange purified GSTs but accounts for only 9% of the total GST enzymatic activity (25 mol min–1 mg–1). An analysis investigating the dependence of the Q3 GST activity on temperature resulted in a retention of enzymatic activity (50–30% at temperature extremes from –13°C to 100°C), suggesting a unconventional role for the Q3 GST in quahog metabolism.  相似文献   

10.
Isolated embryos ofKarwinskia humboldtiana were cultured in vitro. The growth of embryos and development to plantlets on woody plant medium supplemented with indole-3-acetic acid 6.10-2 mol l–1, gibberellic acid (GA3) 3.10-2 mol l–1, and 6-benzylaminopurine (BA) 2 mol l–1 was obtained. Multiplication of shoots and rooting of excised shoots has been achieved. Callus formation on modified Murashige-Skoog medium supplemented with 1-naphthaleneacetic acid 10 mol l–1, GA3 14 mol l–1, and kinetin 5 mol l–1 on hypocotyls, or on root cultures on medium supplemented with 2.4-dichlorophenoxyacetic acid 10 mol l–1 and BA 10 mol l–1 was induced.Abbreviations BA 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - GA3 gibberellic acid - IAA indole-3-acetic acid - NAA 1-naphthaleneacetic acid - TEM transmission electron microscopy  相似文献   

11.
Catalytic properties and membrane associations of the phosphatidylglycerophosphate (PGP) and phosphatidylserine (PS) synthases of Rhodobacter sphaeroides were examined to further characterize sites of phospholipid biosynthesis. In preparations of cytoplasmic membrane (CM) enriched in these activities, apparent K m values of PGP synthase were 90 M for sn-glycerol-3-phosphate and 60 M for CDP-diacylglycerol; the apparent K m of PS synthase for l-serine was near 165 M. Both enzymes required Triton X-100 with optimal PS synthase activity at a detergent/CDP-diacylglycerol (mol/mol) ratio of 7.5:1.0, while for optimal PGP synthase, a range of 10–50:1.0 was observed. Unlike the enzyme in Escherichia coli and several other Gram-negative bacteria, the PS synthase activity had a specific requirement for magnesium and was tightly associated with membranes rather than ribosomes in crude cell extracts. Sedimentation studies suggested that the PGP synthase ws distributed uniformly over the CM in both chemoheterotrophically and photoheterotrophically grown cells, while the PS synthase was confined mainly to a vesicular CM fraction. Solubilized PGP synthase activity migrated as a single band with a pI value near 5.5 in a chromatofocusing column and 5.8 on isoelectric focusing; in the latter procedure, the pI was shifted to 5.3 in the presence of CDP-diacylglycerol. The PGP synthase activity gave rise to a single polypeptide band in lithium dodecyl sulfatepolyacrylamide gel electrophoresis at 4°C.Abbreviations CM cytoplasmic membrane - ICM intracytoplasmic photosynthetic membrane - OM outer membrane - PGP phosphatidylglycerophosphate - PS phosphatidylserine - TLC thin-layer chromatography Supported in part by a Fellowship Awards from the Charles and Johanna Busch Memorial Fund Award to the Rutgers Bureau of Biological Research  相似文献   

12.
Dihydropyrimidinase from Pseudomonas stutzeri ATCC 17588 was purified 100-fold and characterized. It was found that dihydrouracil, dihydrothymine and hydantoin could serve as substrates for the partially purified enzyme. The K m values for dihydrouracil, dihydrothymine and hydantoin were determined to be 19.6 M, 21.3 M and 36.4 M, respectively, while their respective V max values were 0.836 mol/min, 0.666 mol/min and 2.21 mol/min. Between pH 7.5 and 9.0, enzyme activity was shown to be maximal. The optimum temperature for enzyme activity was 45 °C. Using gel filtration, the molecular weight of the enzyme was calculated to be approximately 115000 Da. Metal ions were found to influence the level of enzyme activity. Dihydropyrimidinase activity was stimulated by magnesium ions and inhibited by either zinc or copper ions.  相似文献   

13.
Four recently described species of new genera of sulfate-reducing bacteria, Desulfobulbus propionicus, Desulfobacter postgatei, Desulfococcus multivorans and Desulfosarcina variabilis were examined with respect to adenylylsulfate reductase. All of the species examined contained the enzyme in sufficient concentrations to account for dissimilatory sulfate reduction.Adenylylsulfate reductase was enriched 17.1-fold from Desulfobulbus propionicus by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. The molecular weight was 175,000 and the enzyme contained 1 mol of flavin, 8 mol of non heme iron and 8 mol of labile sulfide per mol enzyme. Either ferricyanide or cytochrome c could be used as electron acceptors; the pH optimum was 7.7 with ferricyanide and 8.8 with cytochrome c. K m values for AMP and sulfite were 90 M and 1.3 M with ferricyanide and 91 M and 71 M with cytochrome c as electron acceptor. K m values for ferricyanide and cytochrome c were 89 M and 21 M, respectively. The properties of the enzyme were compared with those of purified adenylylsulfate reductases from other microorganisms.Non-common abbreviation APS adenylylsulfate  相似文献   

14.
Summary Increasing amounts of Cd, Pb, and Na salts were added to a fresh mull soil and the NO3 and NH4 concentrations measured after 2–8 weeks of incubation. The highest amounts used (CdCl2 9–18 mol/g dry weight, CdAc2 9–22 mol/g, PbAc2 121 mol/g and NaAc 250 mol/g) significantly increased NO3 accumulation. Lower concentrations had either no effect or caused a slight decrease. re]19730529  相似文献   

15.
The effect of lead on the filtration rate of the zebra musselDreissena polymorpha was investigated, together with the accumulation of Pb in the soft tissues of the mussels. The NOEC-filtration was 116 g.l–1 (0,56 mol.l–1) and the EC50-filtration was 370 g.l–1 (1.79 mol.l–1). The NOEC-accumulation was the concentration found in the control water (1.4g.l–1). These experiments show that the EC50-filtration for Pb is similar to that for Cd, higher than that for Cu and lower than that for Zn. The water quality criteria for lead allow 25 g Pb.l–1 in surface water. This will not cause short-term effects. Long-term effects may, however, occur, since an accumulation of Pb as low as 16 g.l–1 was recorded in this study.  相似文献   

16.
Studies on the growth and respiration of batch suspension cultures of rice (Oryza sativa L.) in a reference medium containing Murashige-Skoog salts, 2% (w/v) sucrose and yeast extract are reported. It was found that the yeast extract contributed 70% of the phosphate in this medium, and that the cells grew equally well in continued subculture in a defined medium which contained 6 mM phosphate and 3% (w/v) sucrose and the remaining Murashige-Skoog salts. Cell clumps (up to 1.5 mm diameter) were prevalent in the initial cultures in the reference medium. In such cultures the critical O2 pressure of cell respiration was high (125 M), and ethanol accumulated. When cell clumps were routinely removed during several weekly subcultures on the defined medium cultures were obtained in which no clumps were present, the critical O2 pressures was decreased to 40 M and no ethanol accumulated.This work was supported by grant PCM-84-03542 from the U.S. National Science Foundation.  相似文献   

17.
Cytosolic purine nucleoside phosphorylase (PNPase) is a well known, and described enzyme which exists in a variety of organisms, both procaryotic and eucaryotic. More recently this enzyme was found in bovine liver mitochondria. The mitochondrial purine nucleoside phosphorylase was purified 63 fold and has a molecular weight of 48–60 kD. From Lineweaver-Burk plots apparent Km's of 23M for inosine, 42 M for deoxyinosine, 40 M for phosphate, 2 M for hypoxanthine, and 163 M for ribose-1-phosphate were calculated. Both 8-aminoguanosine (Ki=0.5 M) and araG (Ki=381 M) are inhibitors of the enzyme. The protein's isoelectric point (pI) was calculated at a pH of 4.2. Preliminary immunological work showed no cross-reactivity between epitopes on the mitochondrial protein and those on PNPase from human erythrocytes. The apparent Km's calculated for the mitochondrial enzyme are,with the exception of that using hypoxanthine, within the range commonly associated with Km's from the cytosolic species. The mitochondrial enzyme's molecular weight and pI are less than normally described. The enzyme's isolation from mitochondria, together with several unique characteristics, suggest that it is a separate protein from that found in the cytosol.  相似文献   

18.
Summary T4-infected cells, plasmolysed 15 min after infection, incorporate low concentrations (>20 M) of deoxythymidine (TdR) into DNA at a significantly greater rate than dTMP, dTTP or thymine. At higher concentrations (>40 M), dTMP incorporation rate is high, approaching that of TdR at 200 M. TdR is selectively incorporated at all concentrations tested, and is not inhibited by the other thymine containing DNA precursors. Incorporation of low concentrations of TdR requires the T4-induced thymidine kinase (tk) and is not significantly affected by the presence or absence of T4-induced thymidylate synthetase (td). We show that, in T4-infected plasmolysed cells, exogenously added TdR is preferentially incorporated into short DNA fragments during short pulse times. To explain these and other data a model is proposed in which thymidine plays a modulatory role between leading and lagging strand precursor feeds.Preliminary accounts of these data were presented at the West Coast Phage Meetings, Evergreen State College 1980, 1981  相似文献   

19.
During recovery after a long (up to 12 h) treatment of pig embryo culture cells (PK) with nocodazole at concentrations of 0.02 g/ml and 0.2 g/ml all c-metaphase cells divide normally into two daughter cells. During recovery after a short (1–4 h) treatment with 0.6 g/ml nocodazole only multipolar mitoses (as a rule tripolar) arise. At the ultrastructural level, the increasing nocodazole concentration leads to progressive disruption of the mitotic spindle. At a nocodazole concentration of 0.2 g/ml kinetochores are not associated with microtubules. At a nocodazole concentration of 0.6 g/ml there are no microtubules around the centrosomes, and in every cell one of the two diplosomes disintegrates. In tripolar telophase centrioles are distributed among the spindle poles generally in a 2:2:0 pattern. Mother and daughter centrioles are always disoriented but not separated. The centriole-free pole contains a cloud of electron-dense material. During tripolar division two of the three daughter cells mainly fuse shortly after telophase forming one binucleate cell. Thus a multipolar mitosis arises as a result of the uncoupling of mother centrioles and spindle microtubules, but not of the duration of the c-mitotic arrest. Centriole-free poles account for the divergence of chromosomes, but mainly they are unable to ensure the normal cytokinesis of daughter cells.by M. Trendelenburg  相似文献   

20.
Data for the maximum carboxylation velocity of ribulose-1,5-biosphosphate carboxylase, Vm, and the maximum rate of whole-chain electron transport, Jm, were calculated according to a photosynthesis model from the CO2 response and the light response of CO2 uptake measured on ears of wheat (Triticum aestivum L. cv. Arkas), oat (Avena sativa L. cv. Lorenz), and barley (Hordeum vulgare L. cv. Aramir). The ratio Jm/Vm is lower in glumes of oat and awns of barley than it is in the bracts of wheat and in the lemmas and paleae of oat and barley. Light-microscopy studies revealed, in glumes and lemmas of wheat and in the lemmas of oat and barley, a second type of photosynthesizing cell which, in analogy to the Kranz anatomy of C4 plants, can be designated as a bundle-sheath cell. In wheat ears, the CO2-compensation point (in the absence of dissimilative respiration) is between those that are typical for C3 and C4 plants.A model of the CO2 uptake in C3–C4 intermediate plants proposed by Peisker (1986, Plant Cell Environ. 9, 627–635) is applied to recalculate the initial slopes of the A(pc) curves (net photosynthesis rate versus intercellular partial pressure of CO2) under the assumptions that the Jm/Vm ratio for all organs investigated equals the value found in glumes of oat and awns of barley, and that ribulose-1,5-bisphosphate carboxylase is redistributed from mesophyll to bundle-sheath cells. The results closely match the measured values. As a consequence, all bracts of wheat ears and the inner bracts of oat and barley ears are likely to represent a C3–C4 intermediate type, while glumes of oat and awns of barley represent the C3 type.Abbreviations A net photosynthesis rate (mol·m-2·s-1) - Jm maximum rate of whole-chain electron transport (mol·e-·m-2·s-1) - pc (bar) intercellular partial pressure of CO2 - PEP phosphoenolpyruvate - PPFD photosynthetic photon flux density (mol quanta·m-2·s-1) - RuBPCase ribulose bisphosphate carboxylase/oxygenase - RuBP ribulose bisphosphate - Vm maximum carboxylation velocity of RuBPCase (mol·m-2·s-1) - T* CO2 compensation point in the absence of dissimilative respiration (bar)  相似文献   

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