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1.
Phytochrome, activated by continuous red light, increases the amount of total polyadenylated RNA during photomorphogenesis of mustard (Sinapis alba L.) cotyledons. In-vitro translation of total polyadenylated RNA in a reticulocyte translation system has shown that the activity of translatable -amylase mRNA is increased by phytochrome about threefold in the 3-d-old cotyledons, based on equal amounts of polyadenylated RNA, and about eightfold on a per-cotyledon basis. Cordycepin prevents the accumulation of translatable -amylase mRNA. It is concluded that the phytochrome-mediated control of -amylase synthesis is exerted on the level of mRNA synthesis. During seedling development in continuous red light, a phytochrome-dependent increase of -amylase mRNA can be observed at least 6 h before the onset of -amylase synthesis. If, after a period of enzyme synthesis, phytochrome action is interrupted by long-wavelength far-red light followed by darkness, -amylase mRNA as well as -amylase synthesis remain at a high level for 8–10 h and then decline sharply. It is concluded that -amylase mRNA, having an apparent lifetime of the order of 8–10 h, can be formed under the influence of phytochrome during early seedling development but it activates -amylase synthesis only after a lag-phase of about 8 h, when the cotyledons acquire competence to synthesize the enzyme. The consequences of these findings for the signal-transduction chain of phytochrome are discussed.Abbreviations EDTA
Na2-ethylenediaminotetraacetic acid
- PAGE
polyacrylamide gel electrophoresis
- poly(A)+RNA
polyadenylated mRNA
- Pr, Pfr
red- and far-red-absorbing forms of phytochrome
- SDS
sodium dodecyl sulfate
- Tris
2-amino-2-(hydroxymethyl)-1,3-propanediol 相似文献
2.
The intracellular localisation of phytochrome and ubiquitin in irradiated oat coleoptiles was analysed by electron microscopy. We applied indirect immunolabeling with polyclonal antibodies against phytochrome from etiolated oat seedlings or polyclonal antibodies against ubiquitin from rabbit reticulocytes, together with a goldcoupled second antibody, on serial ultrathin sections of resin-embedded material. Immediately after a 5-min pulse of red light-converting phytochrome from the red-absorbing (Pr) to the far-redabsorbing (Pfr) form-the label for phytochrome was found to be sequestered in electron-dense areas. For up to 2 h after irradiation, the size of these areas increased with increasing dark periods. The ubiquitin label was found in the same electrondense areas only after a dark period of 30 min. A 5 min pulse of far-red light, which reverts Pfr to Pr, given immediately after the red light did not cause the electron-dense structures to disappear; moreover, they contained the phytochrome label immediately after the far-red pulse. In contrast, after the reverting far-red light pulse, ubiquitin could only be visualised in the electron-dense areas after prolonged dark periods (i.e. 60 min). The relevance of these data to light-induced phytochrome pelletability and to the destruction of both Pr and Pfr is discussed.Abbreviations FR
far-red light; Pfr
- Pr
far-red-absorbing and red-absorbing forms of phytochrome, respectively
- R
red light 相似文献
3.
The amount of in-vitro translatable mRNA of the light-harvesting chlorophyll a/b-binding protein (LHCP) of photosystem II strongly increases in darkness (D) after a 5-min red-light pulse while continuous illumination of mustard seedlings with far-red (FR), red or white light leads only to a slight increase in the amount of translatable LHCP-mRNA. No increase can be observed after a long-wavelength FR (RG9-light) pulse. However, a FR pretreatment prior to the RG9-light pulse strongly increase LHCP-mRNA accumulation in subsequent D. This is not observed in the case of the mRNA for the small subunit of ribulose-1.5-bisphosphate carboxylase. The increase of LHCP-mRNA in D after a FR pretreatment can be inhibited by a reillumination of the seedlings with FR. The inhibition of LHCP-mRNA accumulation during continuous illumination with FR and the strong increase in D following a FR illumination was found to be independent of chlorophyll biosynthesis since no correlation between chlorophyll biosynthesis and translatable LHCP-mRNA levels could be detected. Even strong changes in the amount of intermediates of chlorophyll biosynthesis caused by application of levulinic acid or 5-aminolevulinic acid did not affect LHCP-mRNA levels. Therefore, we conclude that the appearance of LHCP-mRNA is inhibited during continuous illumination, even though illumination leads to a storage of a light singal which promotes accumulation of translatable LHCP-mRNA in D.Abbreviations c
continuous
- Chl
chlorophyll
- D
darkness
- FR
far-red light (3.5 W·m-2)
- LHCP
light-harvesting chlorophyll a/b-binding protein of photosystem II
- NF
Norfluration
- PChl
protochlorophyll(ide)
- Pfr
far-red absorbing form of phytochrome
- Ptot
total phytochrome
- R
red light (6.8 W·m-2)
- RG9-light
long-wavelength FR (10 W·m-2)
- SSU
small subunit of ribulose-1.5-bisphosphate carboxylase
- WL
white light
- ()
Pfr/Ptot=wavelength-dependent photoequilibrium of the phytochrome system 相似文献
4.
Characterisation of a new monoclonal antibody (mAb), designated LAS 41, directed against 124-kilodalton (kDa) etiolated-oat (Avena sativa L.) phytochrome, indicates that it recognises an epitope unique to the red-light-absorbing form, Pr. In a solid-phase enzyme-linked immunosorbent assay (ELISA), LAS 41 exhibits a seven- to eight-fold higher affinity for Pr than for the far-red-light-absorbing form of phytochrome, Pfr. In addition, in immunoprecipitation assays LAS 41 effectively precipitates 100% of phytochrome presented as Pr but only precipitates a maximum of 24.5% of phytochrome presented as Pfr. These values are indicative of binding exclusively to Pr. Peptide-mapping studies show that LAS 41 recognises and epitope located within a region 6–10 kDa from the aminoterminus of the phytochrome molecule. Since binding of LAS 41 to Pr induces alterations in the spectral properties of Pr, this indicates that at least part of the 4 kDa domain to which the antibody binds is essential for protein-chromophore interaction. Subsequent photoconversion of LAS 41-Pr complexes produces native Pfr spectra, with concomitant production of free antibody and antigen, as shown by a modified ELISA. The specificity of LAS 41 for Pr has facilitated the purification of Pfr which is free of contaminating Pr. This has enabled direct determination of the mole fraction of Pfr established by red light to be 0.874.Abbreviations ELISA
enzyme-linked immunsorbent assay
- kDa
kilodalton
- mAb
monoclonal antibody
- Pfr
far-red-absorbing form of phytochrome
- Pr
red-absorbing form of phytochrome
- SDS-PAGE
sodium dodecyl sulphate polyacrylamide gel electrophoresis
- (A)
difference in absorbance (A
665
Pr
–A
730
Pr
)-(A
665
Pfr
–A
730
Pfr
)
- Ar/Afr
spectral change ratio (SCR)
- max
mole fraction of Pfr following saturating red light 相似文献
5.
6.
Phytochrome-mediated germination of fern spores of Dryopteris paleacea Sw. was initiated by a saturating red-light (R) irradiation after 20 h of imbibition. For its realization external Ca2+ was required, with a threshold at a submicromolar concentration, and an optimum was reached around 10-4 M. At concentrations 10-1 M only a reduced response was obtained, based probably on an unspecific osmotic or ionic effect. The germination response was inhibited by La3+, an antagonist of Ca2+. From these results it is concluded that Ca2+ influx from the medium into the spores may be an important event in phytochrome-mediated germination. In the absence of Ca2+ the R-stimulated system remained capable of responding to Ca2+, added as late as 40 h after R. Moreover, Ca2+ was effective even if added after the active form of phytochrome, Pfr, had been abolished by far-red (FR) 24 h after R. Thus, the primary effect of Pfr, that initiates the transduction chain, does not require calcium. Coupling of Pfr to subsequent dark reactions has been investigated by R-FR irradiations with various dark intervals. The resulting escape kinetics were characterized by a lag phase (6 h) and half-maximal escape from FR reversibility (19 h). These kinetics were not significantly changed by the presence or absence of calcium. Thus, direct interaction of Pfr and calcium is not a step in the transduction chain initiated by the active form of photochrome.Abbreviations EGTA
ethyleneglycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid
- FR
far-red light
- Pr
red-light-absorbing form of phytochrome
- Pfr
far red-light-absorbing form of phytochrome
- Pipes
piperazine-1,4-bis(2-ethanesulfonic acid)
- R
red light
A preliminary report of this work was presented at the XIV Int. Bot. Congr., Berlin (West), Germany, Book of Abstracts, 2-116a-5 (1987) 相似文献
7.
Nils G. A. Ekelund 《Archives of microbiology》1989,151(3):187-190
The effects of the calcium channel blockers, verapamil, diltiazem and lanthanum ions and the Ca2+ dependency on motility as well as the photophobic response (stop-response) of Gyrodinium dorsum were studied. At Ca2+ concentrations below 10-3 M, motility was inhibited. La3+ inhibits the stop-response, in contrast to verapamil and diltiazem. The only calcium channel blocker that increased the amount of non-motile cells was verapamil. The results indicate that motility are Ca2+ dependent and that the stop-responses of G. dorsum could be affected by extracellular Ca2+. Effects of the photosythesis inhibitor (DCMU) on the stop-response was also determined. With background light of different wavelength (614, 658 and 686 nm) the stop-response increased. DCMU inhibited this effect of background light. Negative results with the monoclonal antibody Pea-25 directed to phytochrome and the results with DCMU, indicate that the stop-response of G. dorsum is coupled to photosynthesis rather than to a phytochrome-like pigment. Oxygen evolution, but not cell movement, was completely inhibited by 10-6 M DCMU.Abbreviations DCMU
3-(3,4-dichlorophenyl)-1,1-methylurea
- DILT
diltiazem
- DMSO
dimethylsulfoxide
- SDS-PAGE
sodium dodecyl sulphate-polyacrylamide gel electrophoresis
- VER
verapamil 相似文献
8.
The red light-stimulated component of unrolling in sections from 7-d-old dark-grown barley (Hordeum vulgare L.) leaves is inhibited by ethyleneglycol-bis-(-aminoethyl ether)-N,N,N,N-tetracetic acid (EGTA). A free-Ca2+ activity of less than 40 M restores the ability to respond to red light, but only if supplied within 1 h of red light. Magnesium ions are an ineffective substitute. At least two processes in unrolling appear to be Ca2+-sensitive.Fluence-response measurements indicate that the levels of the far-red-absorbing from of phytochrome (Pfr) still present 4 h after red-light treatment should be above saturation for the unrolling response; consequently, loss of Pfr does not explain the loss in effectiveness of Ca2+ during prolonged EGTA treatment. However, if a further red-light treatment is given simultaneously with Ca2+ addition 4 h after the initial light stimulus, then full unrolling occurs in EGTA-treated sections. These data indicate that, under normal circumstances, a functional change in the properties of Pfr must occur, uncoupling it from the transduction chain.Abbreviations EGTA
ethyleneglycol-bis-(-aminoethylether)-N,N,N,N,-tetracetic acid
- FR
far-red light
- Mes
2-(N-morpholino)ethanesulphonic, acid
- Pfr
far-red absorbing form of phytochrome
- Pr
red-absorbing form of phytochrome
- R
red light 相似文献
9.
Under continuous white light (WL), extension growth of the first internode in Sinapis alba L. was promoted by low red (R): far-red (FR) ratios reaching the stem and-or the leaves. Conversely, the growth promotion by end-of-day light treatments was only triggered by FR perceived by the leaves and cotyledons, while FR given to the growning internode alone was tatally ineffective. Continuous WL+FR given to the internode was also in-effective if the rest of the shoot remained in darkness. Both the background stem growth, and the growth promotion caused by either an end-of-day FR pulse or continuous WL+FR given to the internode, increased with increasing fluence rates of WL given to the rest of the shoot. The increase by WL of the growth-stimulatory effect of low phytochrome photoequilibria in the internode appears to be mediated by a specific blue-light-absorbing photoreceptor, as blue-deficient light from sodium-discharge lamps, or from filtered fluorescent tubes, promoted background stem growth similarly to WL but did not amplify the response to the R:FR ratio in the internode. Supplementing the blue-deficient light (94 mol·m-2·s-1) with low fluence rates of blue (<9 mol·m-2·s-1) restored the promotive effect of low R:FR reaching the internode.Abbreviations BL
blue light
- FR
far-red light
- PAR
photosynthetically active radiation
- Pfr/P
ratio between the FR-absorbing form and total phytochrome
- R
red light
- SOX
low-pressure sodium lamp
- WL
white light
Supported by the Consejo Nacional de Investigaciones Cientificas y Técnicas (República Argentina) and the ORS scheme (UK) 相似文献
10.
On the opportunity cost of the photosynthate invested in stem elongation reactions mediated by phytochrome 总被引:5,自引:0,他引:5
Summary Seedlings of shade-intolerant species react to alterations of the light climate caused by their neighbors with morphological changes that may influence the pattern of resource acquisition and utilization at the whole-canopy level. One such change, the increased stem elongation rate that is triggered by low red (R, 660 nm) to far-red (FR, 730 nm) ratios (R:FR) in dense canopies, might reduce the amount of assimilates available for leaf area expansion or root growth, and in that way affect resource capture by the canopy. We have tested this hypothesis by comparing the growth of both isolated individuals and canopies of the weed Amaranthus quitensis under conditions differing only in the spectral distribution of the incident light. When canopies received the full spectrum of sunlight, the stems were a large proportion (40–57%) of total biomass. Filtering the FR waveband (and hence raising the R:FR ratio to eliminate the neighbors' proximity-signal) resulted in shorter canopies with lighter stems. However, the growth of leaves and roots was not promoted by this treatment, indicating that the opportunity cost of the assimilates invested in the stems was nil or very small. Filtering the FR had no effect on biomass accumulation when plants were grown as isolated individuals. The higher growth of the canopics under full spectrum could be due to a higher light interception or to a higher efficiency of light conversion into biomass. The first possibility is weakened by the observation that filtering the FR had no effect on the dynamics of soil covering by the crops. The second is indirectly strengthened by results of an experiment with isolated plants showing that stem elongation, stem growth, and total plant biomass can be increased by reducing the flux of R light received by the stems without affecting the light climate of the leaves. Further work is needed to distinguish between these two possibilities; whatever the cause, our results show that the elongation responses to decreased R:FR may lead to a net increase in canopy productivity, and do not necessarily have a negative impact on the growth of resource-harvesting organs. 相似文献