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1.
We tested two hypotheses for the mechanism by which xyloglucan–pectin covalent bonds are formed in Arabidopsis cell cultures. Hypothesis 1 proposed hetero-transglycosylation, with xyloglucan as donor substrate and a rhamnogalacturonan-I
(RG-I) side-chain as acceptor. We looked for enzyme activities that catalyse this reaction using α-(1→5)-l-[3H]arabino- or β-(1→4)-d-[3H]galacto-oligosaccharides as model acceptor substrates. The 3H-oligosaccharides were supplied (with or without added xyloglucans) to living Arabidopsis cell-cultures, permeabilised cells, cell-free extracts, or four authentic XTHs. No hetero-transglycosylation occurred. Therefore,
we cannot support hypothesis 1. Hypothesis 2 proposed that some xyloglucan is manufactured de novo as a side-chain on RG-I.
To test this, we pulse-labelled Arabidopsis cell-cultures with [3H]arabinose and monitored the radiolabelling of anionic (pectin-bonded) xyloglucan, which was resolved from free xyloglucan
by ion-exchange chromatography. [3H]Xyloglucan–pectin complexes were detectable <4 min after [3H]arabinose feeding, which is shorter than the transit-time for polysaccharide secretion, indicating that xyloglucan–pectin
bonds were formed intra-protoplasmically. Thereafter, the proportion of the wall-bound [3H]xyloglucan that was anionic remained almost constant at ∼50% for ≥6 days, showing that the xyloglucan–pectin bond was stable
in vivo. Some [3H]xyloglucan was rapidly sloughed into the medium instead of becoming wall-bound. Only ∼30% of the sloughed [3H]xyloglucan was anionic, indicating that bonding to pectin promoted the integration of xyloglucan into the wall. We conclude
that ∼50% of xyloglucan in cultured Arabidopsis cells is synthesised on a pectic primer, then secreted into the apoplast, where the xyloglucan–pectin bonds are stable and
the pectic moiety aids wall-assembly. 相似文献
2.
Infiltrating detached maize (Zeamays L.) leaves with L-galactono-1,4-lactone (L-GAL) resulted in a 4-fold increase in the content of leaf ascorbate. Upon exposure to high irradiance (1000 μmol photons m−2 s−1) at 5 °C, L-GAL leaves de-epoxidized the xanthophyll-cycle pigments faster than the control leaves; the maximal ratio of de-epoxidized
xanthophyll-cycle pigments to the whole xanthophyll-cycle pool was the same in both leaf types. The elevated ascorbate content,
together with the faster violaxanthin de-epoxidation, did not affect the degree of photoinhibition and the kinetics of the
recovery from photoinhibition, assayed by monitoring the maximum quantum efficiency of photosystem II primary photochemistry
(Fv/Fm). Under the experimental conditions, the thermal energy dissipation seems to be zeaxanthin-independent since, in contrast
to the de-epoxidation, the decrease in the efficiency of excitation-energy capture by open photosystem II reaction centers (Fv′/Fm′) during the high-irradiance treatment at low temperature showed the same kinetic in both leaf types. This was also observed
for the recovery of the maximal fluorescence after stress. Furthermore, the elevated ascorbate content did not diminish the
degradation of pigments or α-tocopherol when leaves were exposed for up to 24 h to high irradiance at low temperature. Moreover,
a higher content of ascorbate appeared to increase the requirement for reduced glutathione.
Received: 20 May 1999 / Accepted: 29 October 1999 相似文献
3.
Identification and Partial Characterization of the Pectin
Methyltransferase “Homogalacturonan-Methyltransferase” from
Membranes of Tobacco Cell Suspensions 下载免费PDF全文
A membrane preparation from tobacco (Nicotiana tabacum L.) cells contains at least one enzyme that is capable of transferring the methyl group from S-adenosyl-methionine (SAM) to the C6 carboxyl of homogalacturonan present in the membranes. This enzyme is named homogalacturonan-methyltransferase (HGA-MT) to distinguish it from methyltransferases that catalyze methyletherification of the pectic polysaccharides rhamnogalacturonan I or rhamnogalacturonan II. A trichloroacetic acid precipitation assay was used to measure HGA-MT activity, because published procedures to recover pectic polysaccharides via ethanol or chloroform:methanol precipitation lead to high and variable background radioactivity in the product pellet. Attempts to reduce the incorporation of the 14C-methyl group from SAM into pectin by the addition of the alternative methyl donor 5-methyltetrahydrofolate were unsuccessful, supporting the role of SAM as the authentic methyl donor for HGA-MT. The pH optimum for HGA-MT in membranes was 7.8, the apparent Michaelis constant for SAM was 38 μm, and the maximum initial velocity was 0.81 pkat mg−1 protein. At least 59% of the radiolabeled product was judged to be methylesterified homogalacturonan, based on the release of radioactivity from the product after a mild base treatment and via enzymatic hydrolysis by a purified pectin methylesterase. The released radioactivity eluted with a retention time identical to that of methanol upon fractionation over an organic acid column. Cleavage of the radiolabeled product by endopolygalacturonase into fragments that migrated as small oligomers of HGA during thin-layer chromatography, and the fact that HGA-MT activity in the membranes is stimulated by uridine 5′-diphosphate galacturonic acid, a substrate for HGA synthesis, confirms that the bulk of the product recovered from tobacco membranes incubated with SAM is methylesterified HGA. 相似文献
4.
Summary. The main objective of the present study was to evaluate the in vivo and in vitro effect of Arg on serum nucleotide hydrolysis. The action of Nω-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase, on the effects produced by Arg was also examined.
Sixty-day-old rats were treated with a single or a triple (with an interval of 1 h between each injection) intraperitoneal
injection of saline (group I), Arg (0.8 g/kg) (group II), L-NAME (2.0 mg/kg or 20 mg/kg) (group III) or Arg (0.8 g/kg) plus
L-NAME (2.0 mg/kg or 20 mg/kg) (group IV) and were killed 1 h later. The present results show that a triple Arg administration
decreased ATP, ADP and AMP hydrolysis. Simultaneous injection of L-NAME (20 mg/kg) prevented such effects. Arg in vitro did not alter nucleotide hydrolysis. It is suggested that in vivo Arg administration reduces nucleotide hydrolysis in rat serum, probably through nitric oxide or/and peroxynitrite formation.
Both are first authors. 相似文献
5.
The biosynthesis of galactan was investigated using microsomal membranes isolated from suspension-cultured cells of potato
(Solanum tuberosum L. var. AZY). Incubation of the microsomal membranes in the presence of UDP-[14C]galactose resulted in a radioactive product insoluble in 70% methanol. The product released only [14C]galactose upon acid hydrolysis. Treatment of the product with Aspergillus niger endo-1,4-β-galactanase released 65–70% of the radioactivity to a 70%-methanol-soluble fraction. To a minor extent, [14C]galactose was also incorporated into proteins, however these galactoproteins were not a substrate for Aspergillus niger endo-1,4-β-galactanase. Thus, the majority of the 14C-labelled product was 1,4-β-galactan. Compounds released by the endo-1,4-β-galactanase treatment were mainly [14C]galactose and [14C]galactobiose, indicating that the synthesized 1,4-β-galactan was longer than a trimer. In vitro synthesis of 1,4-β-galactan
was most active with 6-d-old cells, which are in the middle of the linear growth phase. The optimal synthesis occurred at
pH 6.0 in the presence of 7.5 mM Mn2+. Aspergillus aculeatus rhamnogalacturonase A digested at least 50% of the labelled product to smaller fragments of approx. 14 kDa, suggesting that
the synthesized [14C]galactan was attached to the endogenous rhamnogalacturonan I. When rhamnogalacturonase A digests of the labelled product
were subsequently treated with endo-1,4-β-galactanase, radioactivity was not only found as [14C]galactose or [14C]galactobiose but also as larger fragments. The larger fragments were likely the [14C]galactose or [14C]galactobiose still attached to the rhamnogalacturonan backbone since treatment with β-galactosidase together with endo-1,4-β-galactanase
digested all radioactivity to the fraction eluting as [14C]galactose. The data indicate that the majority of the [14C]galactan was attached directly to the rhamnose residues in rhamnogalacturonan I. Thus, isolated microsomal membranes contain
enzyme activities to both initiate and elongate 1,4-β-galactan sidechains in the endogenous pectic rhamnogalacturonan I.
Received: 24 June 1999 / Accepted: 30 August 1999 相似文献
6.
The tryptophan auxotroph mutant trp3-1 of Arabidopsis thaliana (L.) Heynh., despite having reduced levels of l-tryptophan, accumulates the tryptophan-derived glucosinolate, glucobrassicin and, thus, does not appear to be tryptophan-limited.
However, due to the block in tryptophan synthase, the mutant hyperaccumulates the precursor indole-3-glycerophosphate (up
to 10 mg per g FW). Instability of indole-3-glycerophosphate leads to release of indole-3-acetic acid (IAA) from this metabolite
during standard workup of samples for determination of conjugated IAA. The apparent increase in “conjugated IAA” in trp3-1 mutant plants can be traced back entirely to indole-3-glycerophosphate degradation. Thus, the levels of neither free IAA
nor conjugated IAA increase detectably in the trp3-1 mutant compared to wild-type plants. Precursor-feeding experiments to shoots of sterile-grown wild-type plants using [2H]5-l-tryptophan have shown incorporation of label from this precursor into indole-3-acetonitrile and indole-3-acetic acid with
very little isotope dilution. It is concluded that Arabidopsis thaliana shoots synthesize IAA from l-tryptophan and that the non-tryptophan pathway is probably an artifact.
Received: 1 March 2000 / Accepted: 10 April 2000 相似文献
7.
Atomic force microscopy (AFM) enables the topographical structure of cells and biological materials to be resolved under
natural (physiological) conditions, without fixation and dehydration artefacts associated with imaging methods in vacuo. It
also provides a means of measuring interaction forces and the mechanical properties of biomaterials. In the present study,
AFM has been applied for the first time to the study of the mechanical properties of a natural adhesive produced by a green
plant cell. Swimming spores of the green alga Enteromorpha linza (L.) J. Ag. (7–10 μm) secrete an adhesive glycoprotein which provides firm anchorage to the substratum. Imaging of the adhesive in its
hydrated state revealed a swollen gel-like pad, approximately 1 μm thick, surrounding the spore body. Force measurements revealed
that freshly released adhesive has an adhesion strength of 173 ± 1.7 mN m−1 (mean ± SE; n=90) with a maximum value for a single adhesion force curve of 458 mN m−1. The adhesive had a compressibility (equivalent to Young's modulus) of 0.54 × 106 ± 0.05 × 106 N m−2 (mean ± SE; n=30). Within minutes of release the adhesive underwent a progressive `curing' process with a 65% reduction in mean adhesive
strength within an hour of settlement, which was also reflected in a reduction in the average length of the adhesive polymer
strands (polymer extension) and a 10-fold increase in Young's modulus. Measurements on the spore surface itself revealed considerably
lower adhesion-strength values but higher polymer-extension values than the adhesive pad, which may reflect the deposition
of different polymers on this surface as a new cell wall is formed. The study demonstrates the value of AFM to the imaging
of plant cells in the absence of fixation and dehydration artefacts and to the characterisation of the mechanical properties
of plant glycoproteins that have potential utility as adhesives.
Received: 22 February 2000 / Accepted: 20 April 2000 相似文献
8.
Light-induced expression of the Gsa gene encoding the heme and chlorophyll biosynthetic enzyme glutamate 1-semialdehyde aminotransferase in Chlamydomonas reinhardtii was previously shown to involve Ca2+ and calmodulin (CaM) (C. lm et al. 1996, Plant Cell 8: 2245–2253). To further analyze the signal transduction pathway for
light-induced Gsa expression, the effects of several pharmacological agents were examined. Treatment of light-dark synchronized cells with
the heterotrimeric G-protein agonist Mas-7 caused partial induction of Gsa in the dark. The phospholipase C inhibitor U73122 inhibited light induction of Gsa. Exposure of cells to light caused a sustained 3-fold increase in cellular d-inositol 1,4,5-trisphosphate (InsP3) concentration. KN-93, a specific inhibitor of Ca2+/CaM-dependent protein kinase II, inhibited light induction of Gsa. In contrast, cyclosporin A, a specific inhibitor of the Ca2+/CaM-dependent phosphoprotein phosphatase calcineurin, did not affect light induction of Gsa. These results, together with the earlier results, suggest the involvement of a canonical signal transduction pathway for
light-regulated Gsa expression that involves a heterotrimeric G-protein activation, phospholipase C-catalyzed InsP3 formation, InsP3-dependent Ca2+ release, and activation of a downstream signaling pathway through a Ca2+/CaM-dependent protein kinase.
Received: 21 October 1999 / Accepted: 3 December 1999 相似文献
9.
Fast track to the trichome: induction of N-acyl nornicotines precedes nicotine induction in Nicotiana repanda 总被引:3,自引:0,他引:3
Nicotiana repanda Wildenow ex Lehmann acylates nornicotine in its trichomes to produce N-acyl-nornicotine (NacNN) alkaloids which are dramatically
more toxic than nicotine is to the nicotine-adapted herbivore, Manduca sexta. These NacNNs, like nicotine, were induced by methyl jasmonate (MeJA) and wounding, but the 2-fold increase in NacNN pools
was much faster (within 6 h) than the MeJA-induced increase in nornicotine pools (24 h to 4 d), its parent substrate. When
15NO3
− pulse-chase experiments with intact and induced plants were used to follow the incorporation of 15N into alkaloids in different plant parts over the plant's lifetime, it was found that the root nicotine pool was most rapidly
labeled, followed by the shoot nornicotine and NacNN pools. After 3 d, 3.12% of 15N acquired was in nicotine (0.93%), nornicotine (0.32%) and NacNNs (1.73%) while only 0.14% was in anabasine. Once NacNNs
are externalized to the leaf surface, they are not readily re-distributed within the plant and are lost with senescing leaves.
The wound- and MeJA-induced N-acylation of nornicotine is independent of induced changes in nornicotine pools and the rapidity
of the response suggests its importance in defense against herbivores.
Received: 3 July 1999 / Accepted: 17 September 1999 相似文献
10.
Somatic embryogenesis induced by the simple application of abscisic acid to carrot (Daucus carota L.) seedlings in culture 总被引:3,自引:0,他引:3
Seedlings of carrot (Daucus carota L. cv. Red Cored Chantenay) formed somatic embryos when cultured on medium containing abscisic acid (ABA) as the sole source
of growth regulator. The number of embryos per number of seedlings changed depending on the concentration of ABA added to
the medium, with a maximum embryo number at 1 × 10−4 M ABA. Seedling age was critical for response to exogenous ABA; no seedling with a hypocotyl longer than 3.0 cm was able to
form an embryo. Removal of shoot apices from seedlings completely inhibited the embryogenesis induced by application of exogenous
ABA, suggesting that the action of ABA requires some substance(s) that is translocated basipetally from shoot apices through
hypocotyls. Histologically, somatic embryos shared common epidermal cells and differentiated not through the formation of
embryogenic cell clumps, but directly from epidermal cells. These morphological traits are distinct from those of embryogenesis
via formation of embryogenic cell clumps, which has been found in embryogenic carrot cultures established using 2,4-dichlorophenoxyacetic
acid or other auxins. These results suggest that ABA acts as a signal substance in stress-induced carrot seedling somatic
embryogenesis.
Received: 22 April 2000 / Accepted: 8 June 2000 相似文献
11.
To test the hypothesis that the contribution of phosphoribulokinase (PRK) to the control of photosynthesis changes depending
on the light environment of the plant, the response of transgenic tobacco (Nicotiana tabacum L.) transformed with antisense PRK constructs to irradiance was determined. In plants grown under low irradiance (330 μmol m−2 s−1) steady-state photosynthesis was limited in plants with decreased PRK activity upon exposure to higher irradiance, with a
control coefficient of PRK for CO2 assimilation of 0.25 at and above 800 μmol m−2 s−1. The flux control coefficient of PRK for steady-state CO2 assimilation was zero, however, at all irradiances in plant material grown at 800 μmol m−2 s−1 and in plants grown in a glasshouse during mid-summer (alternating shade and sun 300–1600 μmol m−2 s−1). To explain these differences between plants grown under low and high irradiances, Calvin cycle enzyme activities and metabolite
content were determined. Activities of PRK and other non-equilibrium Calvin cycle enzymes fructose-1,6-bisphosphatase, sedoheptulose-1,7-bisphosphatase
and ribulose-1,5-bisphosphate carboxylase-oxygenase were twofold higher in plants grown at 800 μmol m−2 s−1 or in the glasshouse than in plants grown at 330 μmol m−2 s−1. Activities of equilibrium enzymes transketolase, aldolase, ribulose-5-phosphate epimerase and isomerase were very similar
under all growth irradiances. The flux control coefficient of 0.25 in plants grown at 330 μmol m−2 s−1 can be explained because low ribulose-5-phosphate content in combination with low PRK activity limits the synthesis of ribulose-1,5-bisphosphate.
This limitation is overcome in high-light-grown plants because of the large relative increase in activities of sedoheptulose-1,7-bisphosphatase
and fructose-1,6-bisphosphatase under these conditions, which facilitates the synthesis of larger amounts of ribulose-5-phosphate.
This potential limitation will have maintained evolutionary selection pressure for high concentrations of PRK within the chloroplast.
Received: 15 November 1999 / Accepted: 27 January 2000 相似文献
12.
Ermel FF Follet-Gueye ML Cibert C Vian B Morvan C Catesson AM Goldberg R 《Planta》2000,210(5):732-740
The development of pectin structural features during the differentiation of cambial derivatives was investigated in aspen
(Populus tremula L. × P. tremuloides Michx.) using biochemical and immunocytochemical methods. Comparisons were also made between active and resting tissues.
Active tissues, in particular cambial cells and phloem derivatives, were characterized by a high pectin content. Use of antibodies
raised against arabinan side chains of rhamnogalacturonan 1 (LM6), as well as biochemical analysis, revealed an obvious decrease
from the cortex to the differentiating xylem. Galactan side chains, detected with LM5 antibodies, were present mainly in the
cambial zone and enlarging xylem cells. In contrast, they were totally absent from sieve-tube cell walls. Image analysis of
LM5 immunogold labelling in the cambial zone showed a clustered distribution of galactan epitopes in the radial walls, a distribution
which might result from the association of two different periodic processes, namely the exocytosis of galactan and wall expansion.
Cessation of cambial activity was characterized by cell wall thickening accompanied by a sharp decrease in the relative amount
of pectin and a lowering of the degree of methylesterification. The data provide evidence that the walls of phloem and xylem
cells differ in their pectin composition even at a very early stage of commitment. These differences offer useful tools for
identifying the initial cells among their immediate neighbours.
Received: 12 June 1999 / Accepted: 20 October 1999 相似文献
13.
A microsomal preparation from suspension-cultured potato stem cells (Solanum tuberosum L. cv. AZY) was incubated with [14C]acetyl-CoA resulting in a precipitable radiolabeled product. Analysis of the product revealed that it consisted mostly of
acetylated proteins and cell wall polysaccharides, including xyloglucan, homogalacturonan and rhamnogalacturonan I. Thus,
acetyl-CoA is a donor-substrate for the O-acetylation of wall polysaccharides. A rhamnogalacturonan acetylesterase was used to develop an assay to measure and characterize
rhamnogalacturonan O-acetyl transferase activity in the microsomal preparation. Using this assay, it was shown that the transferase activity was
highest during the linear growth phase of the cells, had a pH-optimum at pH 7.0, a temperature optimum at 30 °C, an apparent
K
m of 35 μM and an apparent V
max of 0.9 pkat per mg protein. Further analysis of the radiolabeled acetylated product revealed that it had a molecular mass
>500 kDa.
Received: 3 July 1999; Accepted: 27 September 1999 相似文献
14.
In order to determine why the activated methyl cycle is up-regulated in plants undergoing defence responses to fungal pathogens
we have monitored the utilisation of methyl groups derived from methionine in cell-suspension cultures of alfalfa (Medicago sativa L.) treated for various times with fungal elicitor, by carrying out a parallel labelling study with [35S]methionine and [methyl-3H]methionine. The distribution of the two radiolabels among the medium, soluble cellular components and cell wall was then
determined. In the absence of elicitor the utilisation of the two radiolabels was similar. However, in the presence of the
elicitor the total incorporation of radioactivity from [methyl-3H]methionine into metabolites was far greater than from [35S]methionine, indicating that the methyl label had been utilised in methylation reactions. Elicitor treatment resulted in
up to a sixfold increase in the use of 3H-methyl groups in the methylation of hydrophobic metabolites. In the period 0–24 h after elicitor treatment, increased methylation
was directed largely into the synthesis of the isoflavonoid phytoalexin medicarpin and related metabolites. Newly synthesized
phytoalexins were exported into the medium, while a significant proportion of the medicarpin accumulating in the cell in the
early stages of elicitation was derived from the hydrolysis of its respective conjugate. Elicitor treatment also modified
the incorporation of 3H-methyl groups into the cell wall. Between 0 and 24 h after elicitor treatment the methylation of pectin in the cell wall
declined. After 24 h, pectin methylation recovered and was associated with an increase in the methylation of other wall-bound
polysaccharide components. Since no other major metabolic sink for the increased methylation was determined we conclude that
the increased activity of the activated methyl cycle during defence interactions in alfalfa is required to support phytoalexin
synthesis and cell wall modifications.
Received: 1 August 1996 / Accepted: 24 October 1996 相似文献
15.
Tove M. I. E. Christensen John E. Nielsen Jette D. Kreiberg Preben Rasmussen Jørn D. Mikkelsen 《Planta》1998,206(4):493-503
Pectin methyl esterase (PME) from orange (Citrus sinensis L.) fruit peels has been purified by ammonium sulphate precipitation, and ion-exchange and gel-filtration chromatography.
Characterization of the enzyme revealed a 36-kDa protein with an isoelectric point >9, a pH optimum at 7 and temperature optimum
at 50 °C. The substrate specificity and kinetic experiments showed that the affinity of PME for pectin was highly dependent
on the degree of esterification (DE) of the pectin, with K
m values of 0.7 mg ml-1 for pectin with a DE of 70% and 17 mg ml-1 for pectin with a DE of 25%. The sequences of the NH2-terminal end of digested peptides from the mature protein were obtained. A DNA fragment of 501 bp was cloned by polymerase
chain reaction amplification using degenerate primers and was further used for screening of a cDNA library. Two cDNA clones
were isolated encoding PMEs of 584 amino acids and 362 amino acids, respectively, including a putative signal peptide. The
deduced amino acid sequence showed full identity to the sequenced peptides. Polyclonal antibodies raised against orange peel
PME were used for immunohistochemistry. The main localization of PMEs was in the outer cell layers of the juice vesicles,
in the outer cell layers of the lamellae between the segments and in the inner cell layers of the albedo in the peel. In-situ
hybridization showed that the mRNA is very abundant in the fruit and was found in the same cell layers as the native enzyme.
A very intensive staining for PME mRNA was also seen in the core and in the flavedo close to the oil glands.
Received: 15 November 1997 / Accepted: 7 April 1998 相似文献
16.
When a brassinosteroid biosynthesis inhibitor, brassinazole (Brz), was applied at concentrations ranging from 0.1 to 2 μM,
Arabidopsis thaliana (L.) Heynh seedlings grown in the dark exhibited morphological features of light-grown plants, i.e. short hypocotyls, expanded
cotyledons, and true leaves, in a dose-dependent manner. Control (non Brz-treated) seedlings grown in the dark for 40 d did
not develop leaf primordia. However, treatment with the lowest concentration of Brz induced the development of leaf buds,
although it hardly induced any short hypocotyls, and treatment with the highest concentration of Brz induced both short hypocotyls
and leaves. Labeling experiments with the thymidine analogue 5-bromo-2′-deoxyuridine revealed that amplification of cell nuclei
and organellar nucleoids is activated in the shoot apical meristems of dark-grown Brz-treated seedlings. These results suggest
that Brz-treatment induces development of true leaves. Furthermore, condensation and scattering of plastid nucleoids, which
is known to occur during the differentiation of etioplasts into chloroplasts, was observed in the plastids of dark-grown Brz-treated
cotyledons. In addition, high levels of ribulose-1,5-bisphosphate carboxylase-oxygenase proteins accumulated in the plastids
of the cotyledons. Electron microscopy showed that the plastids were etioplasts with a prolamellar body and few thylakoid
membranes. These results suggest that Brz treatment in the dark induces the initial steps of plastid differentiation, which
occur prior to the development of thylakoid membranes. This is a novel presumed function of brassinosteroids. These cytological
changes seen in Brz-treated Arabidopsis were exactly the same as those seen in a brassinosteroid-biosynthesis-deficient mutant, det2, supporting the hypothesis that Brz has no side-effects except inhibiting brassinosteroid biosynthesis, and should prove
a useful tool in clarifying the role of brassinosteroids.
Received: 10 February 2000 / Accepted: 11 April 2000 相似文献
17.
Inoculation and nitrate alter phytohormone levels in soybean roots: differences between a supernodulating mutant and the wild type 总被引:9,自引:0,他引:9
The levels of different cytokinins, indole-3-acetic acid (IAA) and abscisic acid (ABA) in roots of Glycine max [L.] Merr. cv. Bragg and its supernodulating mutant nts382 were compared for the first time. Forty-eight hours after inoculation with Bradyrhizobium, quantitative and qualitative differences were found in the root's endogenous hormone status between cultivar Bragg and the
mutant nts382. The six quantified cytokinins, ranking similarly in each genotype, were present at higher concentrations (30–196% on average
for isopentenyl adenosine and dihydrozeatin riboside, respectively) in mutant roots. By contrast, the ABA content was 2-fold
higher in Bragg, while the basal levels of IAA [0.53 μmol (g DW)−1, on average] were similar in both genotypes. In 1 mM NO3
−-fed Bragg roots 48 h post-inoculation, IAA, ABA and the cytokinins isopentenyl adenine, and isopentenyl adenosine quantitatively
increased with respect to uninoculated controls. However, only the two cytokinins increased in the mutant. High NO3
− (8 mM) markedly reduced root auxin concentration, and neither genotypic differences nor the inoculation-induced increase
in auxin concentration in Bragg was observed under these conditions. Cytokinins and ABA, on the other hand, were little affected
by 8 mM NO3
−. Root IAA/cytokinin and ABA/cytokinin ratios were always higher in Bragg relative to the mutant, and responded to inoculation
(mainly in Bragg) and nitrate (both genotypes). The overall results are consistent with the auxin-burst-control hypothesis
for the explanation of autoregulation and supernodulation in soybean. However, they are still inconclusive with respect to
the inhibitory effect of NO3
−.
Received: 16 April 1999 / Accepted: 13 December 1999 相似文献
18.
A quantitative study of indole-3-acetic acid (IAA) turnover, and the contribution of tryptophan-dependent and tryptophan-independent
IAA-biosynthesis pathways, was carried out using protoplast preparations and shoot apices obtained from wild-type and transgenic,
IAA-overproducing tobacco (Nicotiana tabacum L.) plants, during a phase of growth when the level of endogenous IAA was stable. Based on the rate of disappearance of [13C6]IAA, the half-life of the IAA pool was calculated to be 1.1 h in wild-type protoplasts and 0.8 h in protoplasts from the
IAA-overproducing line, corresponding to metabolic rates of 59 and 160 pg IAA (μg Chl)−1 h−1, respectively. The rate of conversion of tryptophan to IAA was 15 pg IAA (μg Chl)−1 h−1 in wild-type protoplasts and 101 pg IAA (μg Chl)−1 h−1 in protoplasts from IAA-overproducing plants. In both instances, IAA was metabolised more rapidly than it was synthesised
from tryptophan. As the endogenous IAA pools were in a steady state, these findings indicate that IAA biosynthesis via the
tryptophan-independent pathway was 44 pg IAA (μg Chl)−1 h−1 and 59 pg IAA (μg Chl)−1 h−1, respectively, in the wild-type and transformed protoplast preparations. In a parallel study with apical shoot tissue, the
presumed site of IAA biosynthesis, the rate of tryptophan-dependent IAA biosynthesis exceeded the rate of metabolism of [13C6]IAA despite the steady state of the endogenous IAA pool. The most likely explanation for this anomaly is that, unlike the
protoplast system, injection of substrates into the apical tissues did not result in uniform distribution of label, and that
at least some of the [2H5]tryptophan was metabolised in compartments not normally active in IAA biosynthesis. This demonstrates the importance of using
experimental systems where labelling of the precursor pool can be strictly controlled.
Received: 18 January 2000 / Accepted 24 February 2000 相似文献
19.
Maize (Zea mays L.) cell cultures incorporated radioactivity from [14C]cinnamate into hydroxycinnamoyl-CoA derivatives and then into polysaccharide-bound feruloyl residues. Within 5–20 min, the
CoA pool had lost its 14C by turnover and little or no further incorporation into polysaccharides then occurred. The system was thus effectively a
pulse–chase experiment. Kinetics of radiolabelling of diferulates (also known as dehydrodiferulates) varied with culture age.
In young (1–3 d) cultures, polysaccharide-bound [14C]feruloyl- and [14C]diferuloyl residues were both detectable within 1 min of [14C]cinnamate feeding. Thus, feruloyl residues were dimerised <1 min after their attachment to polysaccharides. For at least
the first 2.3 h after [14C]cinnamate feeding, polysaccharide-bound [14C]diferuloyl residues remained almost constant at ≈7% of the total polysaccharide-bound [14C]ferulate derivatives. Since feruloyl residues are attached to polysaccharides <1 min after the biosynthesis of the latter,
and >10 min before secretion, the data show that extensive feruloyl coupling occurred intra-protoplasmically. Exogenous H2O2 (1 mM) caused little additional feruloyl coupling; therefore, wall-localised coupling may have been peroxidase-limited. In
older (e.g. 4 d) cultures, less intraprotoplasmic coupling occurred: during the first 2.5 h, polysaccharide-bound [14C]diferuloyl residues were a steady 1.4% of the total polysaccharide-bound [14C]ferulate derivatives. In contrast to the situation in younger cultures, exogenous H2O2 induced a rapid 4- to 6-fold increase in all coupling products, indicating that coupling in the walls was H2O2-limited. In both 2- and 4-d-old cultures, polysaccharide-bound 14C-trimers and larger coupling products exceeded [14C]diferulates 3- to 4-fold, but followed similar kinetics. Thus, although all known dimers of ferulate can now be individually
quantified, it appears to be trimers and larger products that make the major contribution to cross-linking of wall polysaccharides
in cultured maize cells. We argue that feruloyl arabinoxylans that are cross-linked before and after secretion are likely
to loosen and tighten the cell wall, respectively. The consequences for the control of cell expansion and for the response
of cell walls to an oxidative burst are discussed.
Received: 19 January 2000 / Accepted: 13 April 2000 相似文献
20.
Intracellular chloroplast photorelocation in the moss Physcomitrella patens is mediated by phytochrome as well as by a blue-light receptor 总被引:3,自引:0,他引:3
The light-induced intracellular relocation of chloroplasts was examined in red-light-grown protonemal cells of the moss Physcomitrella patens. When irradiated with polarized red or blue light, chloroplast distribution in the cell depended upon the direction of the
electrical vector (E-vector) in both light qualities. When the E-vector was parallel to the cross-wall (i.e. perpendicular
to the protonemal axis), chloroplasts accumulated along the cross-wall; however, no accumulation along the cross-wall was
observed when the E-vector was perpendicular to it (i.e. parallel to the protonemal axis). When a part of the cell was irradiated
with a microbeam of red or blue light, chloroplasts accumulated at or avoided the illumination point depending on the fluence
rate used. Red light of 0.1–18 W m−2 and blue light of 0.01–85.5 W m−2 induced an accumulation response (low-fluence-rate response; LFR), while an avoidance response (high-fluence-rate response;
HFR) was induced by red light of 60 W m−2 or higher and by blue light of 285 W m−2. The red-light-induced LFR and HFR were nullified by a simultaneous background irradiation of far-red light, whereas the
blue-light-induced LFR and HFR were not affected at all by this treatment. These results show, for the first time, that dichroic
phytochrome, as well as the dichroic blue-light receptor, is involved in the chloroplast relocation movement in these bryophyte
cells. Further, the phytochrome-mediated responses but not the blue-light responses were revealed to be lost when red-light-grown
cells were cultured under white light for 2 d.
Received: 7 September 1999 / Accepted: 15 October 1999 相似文献