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1.
Aphototropic mutants of the moss Ceratodon purpureus with spectrally normal and with spectrally dysfunctional phytochrome 总被引:3,自引:1,他引:2
Following UV mutagenesis of protonemal tissue of the moss Ceratodon purpureus we have isolated different aphototropic mutant lines that can be divided into two distinct classes. One class, represented by the line ptr1, shows characteristic features of phytochrome chromophore deficiency. ptrl shows negligible photoreversibility (<5% of wild type), whereas immunoblots show normal apoprotein levels. The aphototropic phenotype could be partially restored with biliverdin, a precursor of the phytochrome chromophore. It was found that, whereas in wild type formation of Pfr leads to suppression of gravitropism, there is no such suppression ptrl. In addition, ptr1 shows lower chlorophyll levels than the wild type. These findings indicate that, as expected for a chromophore-deficient mutant, multiple phytochrome effects are lost. The other class of mutants, represented by the line ptr103, shows more specific effects. In this mutant, only phototropism is affected. Suppression of gravitropism, the content of chlorophyll and photoreversibility of phytochrome were similar to those of the wild type. 相似文献
2.
The mobility of phytochrome within protonemal tip cells of the moss Ceratodon purpureus, monitored by fluorescence correlation spectroscopy
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Fluorescence correlation spectroscopy (FCS) is a versatile tool for investigating the mobilities of fluorescent molecules in cells. In this article, we show that it is possible to distinguish between freely diffusing and membrane-bound forms of biomolecules involved in signal transduction in living cells. Fluorescence correlation spectroscopy was used to measure the mobility of phytochrome, which plays a role in phototropism and polarotropism in protonemal tip cells of the moss Ceratodon purpureus. The phytochrome was loaded with phycoerythrobilin, which is fluorescent only in the phytochrome-bound state. Confocal laser scanning microscopy was used for imaging and selecting the xy measuring position in the apical zone of the tip cell. Fluorescence correlation was measured at ancient z-positions in the cell. Analysis of the diffusion coefficients by nonlinear least-square fits showed a subcellular fraction of phytochrome at the cell periphery with a sixfold higher diffusion coefficient than in the core fraction. This phytochrome is apparently bound to the membrane and probably controls the phototropic and polarotropic response. 相似文献
3.
In darkness, protonemal filaments of Ceratodon purpureus (Brid.) grow negatively gravitropically (upwards). Red light induces a positive phototropic response mediated by the photoreceptor
phytochrome. A red light treatment also has an inhibitory effect on the gravitropic response, an effect also mediated by phytochrome.
In this study the effects of blue light on phototropism and on gravitropism were analysed. Unilateral blue light resulted
in only a weak phototropic response, but markedly randomised growth direction. Blue light given together with a gravitropic
stimulus reversed the gravitropism, changing it from negative to positive (filaments grow downward). The effect of blue light
was also analysed with the mutant ptr116, which is defective in the biosynthesis of the phytochrome chromophore, and in a newly isolated mutant wwr2, which is positively gravitropic in darkness. Blue light induced the same reversal of gravitropism in ptr116 as in the wild type, indicating that phytochrome is not involved in this process. In wwr2 the direction of gravitropism was unaltered by the blue light treatment. Light also affects chlorophyll content and the size
of plastids, potential statoliths for gravitropism. Red light induced an increase in plastid size and chlorophyll content
in the wild type but not in ptr116. Blue light induced a similar change in wild type plastids. It seems as though light-induced alterations of gravitropism
are not simply mediated by alterations in plastid properties, and that red light and blue light evoke fundamentally different
responses.
Received: 11 July 1997 / Accepted: 30 January 1998 相似文献
4.
The moss Ceratodon purpureus has long been used as a model system in plant development and physiology. However, the molecular population genetics of the species remains virtually unexplored. In this study, we used population genetic analyses of DNA sequence data from three unlinked loci (atpB-rbcL spacer, adk, and phy2) to examine biogeographical patterns in a global sample of this species. The three loci differed significantly in mutation frequency spectra and implied population structure. Pairs of haplotypes from single populations were frequently more divergent than haplotypes sampled from widely disjunct populations. In the atpB-rbcL spacer and adk samples, Australasian haplotypes were more closely related to Northern Hemisphere haplotypes than to haplotypes found in the equatorial regions. In contrast, the phy2 sample showed that the north and south temperate regions were genetically divergent, with the equatorial regions intermediate. Maximum-likelihood estimates (MLE) of the rates of migration between the two hemispheres were significantly different for the two nuclear genes. The frequency spectra of mutations indicated that differences in implied population structure among the three loci resulted from directional selection on the chloroplast genome and on the chromosomal segment containing adk. Collectively, these data suggest that long-distance migration within the Northern Hemisphere and Australasian regions is common (relative to the mutation rate) and that migration between these two regions, potentially via equatorial populations, is more frequent than migration among equatorial populations. 相似文献
5.
The aphototropic mutant line ptr116 of the moss Ceratodon purpureus shows characteristics of a deficiency in the phytochrome chromophore. Photoreversibility measurements indicate an approximately 20 time lower concentration of spectrally active phytochrome compared to wild-type, whereas normal phytochrome apoprotein levels are found on immunoblots. Feeding with the tetrapyrroles biliverdin, the proposed precursor of the phytochrome chromophore, or phycocyanobilin, which may replace the phytochrome chromophore, resulted in the rescue of ptr116 phototropism. The ptr116 mutant and the phenotypically-related mutant ptr1 contain lower chlorophyll levels than the wild-type. Chlorophyll content of wildtype and mutant tissue grown under different light conditions was estimated using conventional spectrophotometry of extracts and fluorimetrically, on single apical cells. Dark-grown tissue contained about 100 times less chlorophyll than tissue grown under standard white light conditions. Red light given for 24 h to dark adapted filaments induced an increase in the chlorophyll content in the wildtype, but not in ptr116. Blue light induced an increase in chlorophyll both in wildtype and in ptr116. The red light effect on the wildtype was partially reversible with far-red. If ptr116 was grown on phycocyanobilin, an increase in chlorophyll was also found when cells were irradiated with red light. The results indicate that phytochrome as well as a blue light photoreceptor regulate chlorophyll accumulation in C. purpureus protonemata. It can be assumed that in ptr116, the synthesis of the phytochrome chromophore is blocked specifically beyond the synthesis common to chlorophyll and the phytochrome chromophore and affects an enzymatic step between protoporphyrin and biliverdin. 相似文献
6.
Fritz Thümmler Monika Dufner Peter Kreisl Peter Dittrich 《Plant molecular biology》1992,20(6):1003-1017
The phytochrome gene (phyCer) of the moss Ceratodon purpureus was isolated and characterized. phyCer is composed of three coding exons: exon I of 2035 bp, exon II of 300 bp and exon III of 1574 bp. The deduced polypeptide encoded by exon I and II exhibits substantial sequence homology to the conserved NH2-terminal chromophore domain of known phytochromes. In contrast, the COOH-terminal polypeptide encoded by exon III shows no sequence homology to any phytochrome molecule. phyCer most likely represents a single-copy gene and is expressed in a light-independent manner. From the DNA sequence analysis it can be deduced that the PhyCer polypeptide is composed of 1303 amino acids (including the starting Met) which predicts a molecular mass for PhyCer of 145 kDa. The polypeptide encoded in exon III exhibits striking homology within the 300 carboxy-terminal amino acids to the catalytic domain of protein kinases. The carboxy terminus of PhyCer was found to be most homologous to protein-tyrosine kinases of Dictyostelium discoideum and to the products of retroviral oncogenes which belong to the Raf-Mos serine/threonine kinase family. From the hydropathy profile PhyCer appears to be a soluble protein. The predicted structure suggests that PhyCer represents a soluble light-sensor protein kinase which is linked with a cellular phosphorylating cascade. 相似文献
7.
A phytochrome-regulated phototropic response of the moss Ceratodon purpureus was investigated. Chlorotetracycline (CTC) was used to visualize membrane-associated calcium gradients in the tip cell of moss caulonemal filaments. A tip-to-base Ca2+ gradient was observed. The ionophore monensin rapidly inhibited the growth of the tip cell and abolished the CTC fluorescence. Six hours after transferring to inhibitor-free medium, protonemal growth resumed and reached the normal growth rate within 12 h. The growth was accompanied by a reappearance of the CTC-fluorescence gradient. Unilateral irradiation given during the monensin treatment or after the treatment during the period when growth inhibition persisted led, with the re-initiation of growth, to a typical positive phototropic bending in complete darkness. Far-red light applied just before the growth response started, or during growth inhibition, abolished the phototropic response. The phytochrome-mediated signal was qualitatively (position) and quantitatively (degree of bending) memorized. Signal perception and response could be separated temporally. This result indicates that at least under some circumstances, e.g. under the influence of monensin, the phytochrome-mediated signal can be stored for several hours in darkness. Calcium seems to be essential for the processing of polar growth only. A specific function (second messenger) in phytochrome-dependent signal transduction could not be confirmed.Abbreviation CTC
chlorotetracycline 相似文献
8.
The action spectrum for polarotropism was determined, using the Okazaki large spectrograph, by brief irradiation with light between 260 nm and 850 nm in single-celled protonemata of the fern Adiantum capillus-veneris L., which had been cultured for 6 days in red light and then in the dark for 15 h. The action spectrum had a peak at around 680 nm. This effect was nullified by subsequent irradiaton with far-red light, and typical red/far-red reversibility was observed, indicating the involvement of phytochrome. Polarized ultraviolet or blue light had no effect on the direction of apical growth. The action spectrum for phototropism was also determined in the red light region by means of brief microbeam irradiation of a flank of the subapical region of the protonema. This spectrum showed a peak at 662 nm which was consistent with the absorption peak of phytochrome, but not with the peak of the action spectrum for polarotropism. 相似文献
9.
Summary The F-actin distribution in caulonemal tip cells of the mossCeratodon purpureus was examined by rhodamine-phalloidin staining. Gravitropically-growing caulonemal tip cells of the moss possess a distinct alignment of microfilaments (MFs) in their apices. Axially oriented actin bundles run from subapical regions to the apex where they converge towards a central area of the tip, although bundles are absent from the central area itself thus forming a collar-like structure. During a unilateral red light irradiation the actin strands of the apical dome become reoriented towards the irradiated apical flank and still surround an area free of MFs, the point of prospective outgrowth. This process is closely correlated with the morphological effect of bulging and precedes the light-directed outgrowth. The collar structure is essential for the tubular growth form. In darkness, under the influence of antimicrotubule agents the structure is decomposed, the actin strands drift along the cell flanks and finally accumulate in randomly distributed areas where further growth takes place. The microtubules (MTs) are not involved in the phytochromemediated reorientation of the microfilaments. Unilateral red light suppresses the distorting effect of antimicrotubule drugs and restores the collar structure with a pronounced light-directed orientation. Instead, the MTs seem to be responsible for restricting the reorientation to the cell tip. This notion is based on the observation that the small area in the apical dome, which is normally the exclusive location of the light-regulated MF rearrangement, extends towards the cell base when MT inhibitors are applied before the unilateral red light irradiation. This in turn leads to a non-tubular expansion of the light-directed cell flank.Abbreviations DIG
differential interference contrast
- DMSO
dimethyl sulfoxide
- EGTA
ethyleneglycol-bis-(beta-aminoethylether) N,N,N,N-tetraacetic acid
- MF
microfilament
- MT
microtubule
- MTSB
microtubule stabilizing buffer
- MBS
3-maleimidobenzoic 相似文献
10.
Apical cells of protonemata of the moss Ceratodon purpureus (Hedw.) Brid. are negatively gravitropic in the dark and positively phototropic in red light. Various fluence rates of unilateral
red light were tested to determine whether both tropisms operate simultaneously. At irradiances ≥140 nmol m−2 s−1 no gravitropism could be detected and phototropism predominated, despite the presence of amyloplast sedimentation. Gravitropism
occurred at irradiances lower than 140 nmol m−2 s−1 with most cells oriented above the horizontal but not upright. At these low fluence rates, phototropism was indistinct at
1 g but apparent in microgravity, indicating that gravitropism and phototropism compete at 1 g. The frequency of protonemata that were negatively phototropic varied with the fluence rate and the duration of illumination,
as well as with the position of the apical cell before illumination. These data show that the fluence rate of red light regulates
whether gravitropism is allowed or completely repressed, and that it influences the polarity of phototropism and the extent
to which apical cells are aligned in the light path.
Received: 19 January 1999 / Accepted: 19 March 1999 相似文献
11.
Protonemal cells ofAdiantum capillus-veneris were grown under red light conditions over 6 days and exposed to blue light for 8 hr (and then dim green light for 1 hr for
technical reasons), before they were centrifuged acropetally over at least 1 hr at 2,000×g. After this treatment, an arrangement
of endoplasmic microtubules (MTs) that resembled the shape of a tadpole could be detected some distance below the nucleus
in about 40% of the cells. The percentage of protonemata bearing this Mtstructure was dependent on centrifugation time as
well as the time of blue light irradiation. The size of the structure was constant at any time of its existence. Additionally,
a wide belt of transversally oriented cortical MTs in the upper part of the protonemata was detected in many cells after blue
light irradiation and acropetal centrifugation. Its formation rate seemed to be also dependent on the period of blue light
irradiation and centrifugation time. None of the endoplasmic and few of the cortical transverse MT patterns could be seen
without blue light irradiation. A strict coincidence in the formation of both MT patterns was not seen. Further, a few tadpole-shaped
MT arrays remained during mitosis, whereas the cortical transverse MT pattern was found in stages other than metaphase and
anaphase. 相似文献
12.
Wild-type (WT) protonemata of the moss Ceratodon purpureus grow upwards in darkness (negative gravitropism), whereas protonemata of the mutant, wrong-way response (wwr-1) grow down. Since Ceratodon protoplasts regenerate to form new protonemata, we analyzed whether the direction of filament emergence was influenced by
gravity (gravimorphism) and determined the cytological events that correlated with the onset of gravitropism in WT and wwr-1 filaments formed de novo. In the WT the direction of filament emergence appeared to be gravimorphic as more than 66% of the
new filaments emerged above the horizontal. In contrast, the direction of filament emergence was random in wwr-1. Tip-growing cells of both genotypes became gravitropic within a total of one to two cell divisions. Gravitropic curvature
in wwr-1 was opposite in direction to that of WT, and the timing of curvature was comparable, indicating that the wwr-1 mutation acts during the onset of gravitropic competence. In time-lapse studies of both genotypes, neither a plastid-free
zone nor obvious and extensive plastid sedimentation characteristic of mature dark-grown protonemata was observed in the new
filaments prior to gravitropic curvature. Thus, it appears that these latter two features are not required for gravitropism
in new protonemal filaments from protoplasts.
Received: 24 October 1997 / Accepted: 18 November 1997 相似文献
13.
The growth of cotyledons and primary leaves of I-day-old Sinapis alba L. plants were studied under various light conditions and action spectra produced. For both responses blue and red light are most effective and a strong fluence rate dependency can be observed. The red light effect appears to be mediated through phytochrome, that of blue light being due to a separate blue light receptor, although this receptor requires the presence of far-red absorbing phytochrome (Pfr ) in order to be effective. 相似文献
14.
Both phytochrome A and phytochrome B are required for the normal expression of phototropism in Arabidopsis thaliana seedlings 总被引:3,自引:0,他引:3
Abdul-kader Janoudi Radomir Konjevic Garry Whitelam William Gordon Kenneth L. Poff 《Physiologia plantarum》1997,101(2):278-282
The role of phytochrome A (phyA) and phytochrome B (phyB) in phototropism was investigated by using the phytochrome-deficient mutants phyA-101 , phyB-1 and a phyA/phyB double mutant. The red-light-induced enhancement of phototropism, which is normally observed in wild-type seedlings, could not be detected in the phyA/phyB mutant at fluences of red light between 0.1 and 19 000 μmol m−2 . The loss of phyB has been shown to have no apparent effect on enhancement, while the loss of phyA resulted in a loss of enhancement only in the low fluence range (Janoudi et al. 1997). The conclusions of the aforementioned study can now be modified based on the current results which indicate that phototropic enhancement in the high fluence range is mediated by either phyA or phyB, and that other phytochromes have no role in enhancement. First positive phototropism was unaffected in phyA-101 and phyB-1 However, the magnitude of first positive phototropism in the phyA/phyB mutant was significantly lower than that of the wild-type Landsberg parent. Thus, the presence of either phyA or phyB is required for normal expression of first positive phototropism. The time threshold for second positive phototropism is unaltered in the phyA-101 and phyB mutants. However, the time threshold in the phyA/phyB mutant is about 2 h, approximately six times that of the wild type. Finally, the magnitude of second positive phototropism in both phyA-101 and phyB-1 is diminished in comparison with the wild-type response. Thus, phyA and phyB, acting independently or in combination, regulate the magnitude of phototropic curvature and the time threshold for second positive phototropism. We conclude that the presence of phyA and phyB is required, but not sufficient, for the expression of normal phototropism. 相似文献
15.
Summary Two dynamic changes are associated with the phytochrome-regulated phototropic response in tip cells of the mossCeratodon purpureus: a tip-located gradient shift of chlortetracy-cline (CTC)-stained calcium and a structural reorganization of apical microfilaments (MFs). We examined the interdependence of these processes. Cells were treated with the antimicrotubule drug oryzalin, the antimicrofilament drug cytochalasin-D, and the calcium channel blocker nifedipine. respectively. The effects on phototropic growth, on the structural alignment of the cytoskeleton (microtubules, MTs; microfilaments) and on the distribution of CTC-stained calcium were studied under each of these conditions. In gravitropically growing tip cells the apical MFs form a cortical collar-like structure, consisting of actin bundles with a parallel axial alignment. These MFs point towards the presumptive growing point, a weakly stained region in the tip of the cell from which bundles are absent. MTs are present in the cortex and in the endoplasm of the tip, predominantly oriented longitudinally. The MTs converge within the central apex. The cells show a steep tip-to-base CTC-calcium gradient with its highest signal in the central apex. Destruction of MTs by 1 M oryzalin induces several translocational effects: (i) the growing zone and phototropic outgrowth shift from the apex to subapical parts of the cell; (ii) the structural integrity of the apical MFs and the tip-to-base alignment of the CTC-calcium gradient are disturbed; and (iii) the red light induced gradient shift and the reorientation of MFs proceed in an expanded area spanning from the tip to subapical parts of the cell. Cytochalasin-D (10 g/ml) destroys the MFs. Under these conditions tip growth stops and the phototropic outgrowth is suppressed. The apical MT-structure and the CTC-calcium gradient are not influenced by the agent. Unilateral red light still induces the light-directed translocation of the gradient. Tip cells memorize a unilateral irradiation applied during growth inhibition with cytochalasin-D. After recovery in darkness the cells start to grow in the former light direction. The restoration of the MFs precedes the outgrowth. The structural alignment of the rebuilt actin bundles indicates the future growth direction. The calcium channel blocker nifedipine (10 M) also inhibits tip growth and concurrently phototropic outgrowth. Nifedipine destroys the CTC-calcium gradient and apical MFs; MTs are not influenced by the channel blocker.Abbreviations CTC
chlortetracycline
- DIC
differential interference contrast
- DMSO
dimethyl sulfoxide
- EGTA
ethyleneglycol-bis-(-aminoethylether) N,N,N-N-tetraacetic acid
- MBS
3-maleimidobenzoic acid N-hydroxysuccimide ester
- MF
microfilament
- MT
microtubule
- MTSB
microtubule stabilizing buffer
- PIPES
piperazine-N,N-bis(2-ethane-sulfonic acid)
- RP
rhodamine labeled phalloidin 相似文献
16.
Photogeneration of a bioelectric field by red and far-red irradiation in soybean hypocotyls 总被引:1,自引:0,他引:1
TAKLJMA TANADA 《Plant, cell & environment》1983,6(1):69-72
Abstract. Changes in the bioelectric field potential at the apical end of dark-grown soybean ( Glycine max) hypocotyls after exposure to several wavelengths of light have been monitored with an electrometer. A small dose of 660 nm radiation brings about a rapid positive rise in the potential. A following small dose at 760 nm prevents the increase, suggesting a phytochrome action. Pre-irradiation with 660 nm enables a subsequent larger dose of 760 nm, which by itself is without effect, to elicit a delayed rise in the potential.
A large dose at 710 nm induces a delayed increase in the field potential. The increase is prevented by a following dose at 550 nm, but not at 660 nm. 相似文献
A large dose at 710 nm induces a delayed increase in the field potential. The increase is prevented by a following dose at 550 nm, but not at 660 nm. 相似文献
17.
Chloroplast-avoidance response induced by high-fluence blue light in prothallial cells of the fern adiantum capillus-veneris as analyzed by microbeam irradiation 总被引:3,自引:0,他引:3
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Chloroplast movement was induced by partial cell illumination using a high-fluence blue microbeam in light-grown and dark-adapted prothallial cells of the fern Adiantum capillus-veneris. Chloroplasts inside the illuminated area moved out (high-fluence response [HFR]), whereas those outside moved toward the irradiated area (low-fluence response [LFR]), although they stopped moving when they reached the border. These results indicate that both HFR and LFR signals are generated by high-fluence blue light of the same area, and that an LFR signal can be transferred long-distance from the beam spot, although an HFR signal cannot. The lifetime of the HFR signal was calculated from the traces of chloroplast movement induced by a brief pulse from a high-fluence blue microbeam to be about 6 min. This is very short compared with that of the LFR (30–40 min; T. Kagawa, M. Wada [1994] J Plant Res 107: 389–398). These data indicate that the signal transduction pathways of the HFR and the LFR must be distinct. 相似文献
18.
In the caulonema tip cells of Funaria hygrometrica, chloroplasts, mitochondria, and dictyosomes have differences in structure which are determined by cell polarity. In contrast to the slowly growing chloronema tip cells the apical cell of the caulonema contains a tip body. Colchicine stops tip growth; it causes the formation of subapical cell protrusions, redistribution of the plastids, and a loss of their polar differentiation. Cytochalasin B inhibits growth and affects the position of cell organelles. After treatment with ionophore A23 187, growth is slower and shorter and wider cells are formed. D2O causes a transient reversion of organelle distribution but premitotic nuclei are not dislocated. In some tip cells the reversion of polarity persists; they continue to grow with a new tip at their base. During centrifugation, colchicine has only a slight influence on the stability of organelle anchorage. The former polar organization of most cells is restored within a few hours after centrifugation, and the cells resume normal growth. In premitotic cells the nucleus and other organelles cannot be retransported, they often continue to grow with reversed polarity. Colchicine retards the redistribution of organelles generally and increases the number of cells that form a basal outgrowth. The interrelationship between the peripheral cytoplasm and the nucleus and the role of microtubules in maintaining and reestablishing cell polarity are discussed.Abbreviations DMSO
dimethylsulfoxide
- CB
cytochalasin B
Dedicated to Prof. Dr. A. Pirson on the occasion of his 70. birthday 相似文献
19.
The effects of cyclic 3′,5′-adenosine monophosphate (cAMP) on amaranthin synthesis in the dark, or in the presence of kinetin or light were investigated in isolated cotyledons of Amaranthus tricolor and A. caudatus. The results suggest that sites or modes of action of cAMP and kinetin are not separated and differ from those of light and that the nucleotide cannot be considered a messenger involved in amaranthin formation stimulated by kinetin or by light. 相似文献
20.
Takuma Tanada 《Physiologia plantarum》1983,57(1):42-46
In order for far-red radiation at 760 nm to delay dark closing of Albizzia julibrissin pinnules, red light must be given simultaneously with or just prior to it. Studies have been made to determine whether a phytohormone can replace this red light requirement. Abscisic acid, gibberellin, kinetin, and indole-3-acetic acid have been found to replace the red light. Indole-3-carboxylic acid and a cytokinin antagonist are ineffective. In this hormone and far-red interaction, all hormones are effective at μ M or lower concentrations. The hormones show no interaction with red light at 660 nm. Simultaneous irradiation at 550 nm negates the effect of hormone and far-red interaction in delaying leaflet closing. These results are additional evidence that an unidentified far-red absorbing pigment could be involved with phytochrome in some far-red-mediated processes. 相似文献