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31.
A novel combination of optimization methods (Genetic Algorithm with Distance Geometry) has been developed and shown to find near-optimal solutions to a set of imposed structural constraints. With this modelling tool (GADGET), the fold-space of a variety of small zinc-binding proteins was investigated under the constraints required to form a zinc-binding site (or pair of sites). Analysis of the results concentrated on the ring-finger domain as the "classic" zinc-finger domains were too constrained to provide much topological variety, whilst the TFIIH domain (which has large unstructured loops) did not behave well. The intermediate ring-finger domain, however, was found to adopt a variety of different folds, many of which had near-optimal scores under the fitness function employed in GADGET (forming good secondary structures and zinc-coordination). Although the native fold was dominant amongst the solutions, the discovery of good alternate folds shows that even the eight residues constrained to form two zinc-binding sites was not sufficient to uniquely determine the native fold. Despite this, the fold-space of 48 theoretically possible folds was greatly reduced with just six topologies found in significant numbers.  相似文献   
32.
G2 pea exhibits an apical senescence delaying phenotype under short-day (SD) conditions; however, the structural basis for its apical development is still largely unknown. In the present study, the apical meristem of SD-grown G2 pea plants underwent a transition from vegetative to indeterminate inflorescence meristem, but the apical meristem of long-day (LD)-grown G2 pea plants would be further converted to determinate floral meristem. Both SD signal and GA3 treatment enhanced expression of the putative calcium transporter PPF1, and pea homologs of TFL1 (LF and DET), whereas LD signal suppressed their expression at 60 d post-flowering compared with those at 40 d post-flowering. Both PPF1 and LF expressed at the vegetative and reproductive phases in SD-grown apical buds, but floral initiation obviously increased the expression level of PPF1 compared with the unchanged expression level of LF from 40 to 60 d post-flowering. In addition, although the floral initiation significantly enhanced the expression levels of PPF1 and DET, DET was mainly expressed after floral initiation in SD-grown apical buds. Therefore, the main structural difference between LD- and SD-grown apical meristem in G2 pea lies in whether their apical indeterminate inflorescence medstem could be converted to the determinate structure.  相似文献   
33.
In the sequential response to plant growth regulators in young elongating tissue from peas and wheat the peak of sensitivity to 24-epi-brussinolide (1 μM) occurs after those of gibberellin and cytokinin and begins before that of auxin in isolated wheat ( Triticum vulgare L. ev. Egret) coleoptiles aged from 21-96 h. In dwarf pea ( Pisum sativum L. cv. Greenfeast) segments, the peak of sensitivity also lies between those of gibberellin and auxin, and it also occurs before sensitivity to auxin in sections from first leaves of wheat. All the leaf sections and all but the most mature coleoptiles and pea segments were sensitive to fusicocein (1 μM).  相似文献   
34.
Field-grown soybean plants (Glycine max (L.) Merr. cv. Evans) were treated with gibberellic acid (GA3; 10gl–1) and/or (2-chloroethyl)-trimethylammonium chloride (CCC; 0.8gl–1) in 1983 and 1984, and subsequent anthesis, pod set, seed size, seed number, and seed yield were determined at one node. The treatments were applied to five leaves in the center of each plant (typically leaves 7–11) and reproductive development at the node in the center of those leaves was monitored. Gibberellin A3 applied Early (about 3d before anthesis of the first flower at the monitored node) had no effect on the number of flowers produced, but decreased the fraction of flowers that set pods in both experimental years (by 32% in 1983 and 76% in 1984). Seed size was slightly decreased by the GA3 treatment in 1983 but not in 1984. The Middle GA3 treatment (applied about 3 days after the Early treatment) slightly decreased the number of pods set; and Late treatments (9 days after) had no effect. None of the monitored parameters were affected by CCC.The Early experiments were repeated with two additional genotypes, Lincoln and T210. Genotype T210 is a single-gene, dwarf mutant of Lincoln whose stem elongation and leaf expansion are insensitive to GA3. Gibberellin A3 affected the reproductive parameters in Lincoln very similarly to Evans but those in T210 were unaffected. This indicates that GA3 exerts its effect by increasing the mass of vegetative tissue and thus diverting assimilates away from the pods. However, since the mutation in T210 might affect a receptor that is in flowers as well as shoots, it is possible that GA3 exerted its effect on the normal genotypes directly on the developing pods, rather than indirectly by diverting photoassimilates.  相似文献   
35.
The gibberellin (GA) economy of young pea (Pisum sativum L.) fruits was investigated using a range of mutants with altered GA biosynthesis or deactivation. The synthesis mutation lh-2 substantially reduced the content of both GA4 and GA1 in young seeds. Among the other synthesis mutations, ls-1, le-1 and le-3, the largest reduction in seed GA1 content was only 1.7-fold (le-1), while GA4 was not reduced in these mutants, and in fact accumulated in some experiments (compared with the wild type). Mutation sln appeared to block the step GA20 to GA29 in young pods and seeds, but not as strongly as in older seeds. Mutations ls-1, le-1 and le-3 markedly reduced pod GA1 levels, but pod elongation was not affected. After feeds of [13C,3H]GA20 to leaves, the pods contained 13C,3H-labelled GA20, GA1, GA29 and GA81, and the seeds, [13C,3H]GA20 and [13C,3H]GA29. These findings are discussed in relation to recent suggestions regarding the role and origin of GA1 in pea fruits. Received: 6 June 1997 / Accepted: 15 July 1997  相似文献   
36.
The native gibberellin A4 (GA4), in radioactive form ([1,2-3H]GA4, 1.06 Ci/mmol), was fed to carrot somatic cell cultures (suspension and immobilized cell systems) and its metabolism over a 48 hr period was investigated. It was found that the [3H]GA4 was metabolized to at least two GAs, [3H]GA1 and [3H]GA8, six GA glucosyl conjugates, [3H]GA1-0(3)-glucoside, [3H]GA1-0(13)-glucoside, [3H]GA1-glucosyl ester, [3H]GA4-glucoside, [3H]GA4-glucosyl ester, a [3H]GA8 glucosyl conjugate(s) and a previously unknown [3H]GA1 glucosyl conjugate ([3H]GA1-0(3,13)-diglucoside-like compound). The GA1-diglucoside-like compound was found only in extracts of cells and was present in significant amounts (33 % of total extractable radioactivity). All other metabolites were present in both cells and medium. For extracts of the medium, no differences between the suspension and immobilized cultures existed in types of [3H]GA4 metabolites although quantitative differences were apparent.  相似文献   
37.
Angiosperm seeds integrate various environmental signals, such as water availability and light conditions, to make a proper decision to germinate. Once the optimal conditions are sensed, gibberellin (GA) is synthesized, triggering germination. Among environmental signals, light conditions are perceived by phytochromes. However, it is not well understood how phytochromes regulate GA biosynthesis. Here we investigated whether phytochromes regulate GA biosynthesis through PIL5, a phytochrome-interacting bHLH protein, in Arabidopsis. We found that pil5 seed germination was inhibited by paclobutrazol, the ga1 mutation was epistatic to the pil5 mutation, and the inhibitory effect of PIL5 overexpression on seed germination could be rescued by exogenous GA, collectively indicating that PIL5 regulates seed germination negatively through GA. Expression analysis revealed that PIL5 repressed the expression of GA biosynthetic genes (GA3ox1 and GA3ox2), and activated the expression of a GA catabolic gene (GA2ox) in both PHYA- and PHYB-dependent germination assays. Consistent with these gene-expression patterns, the amount of bioactive GA was higher in the pil5 mutant and lower in the PIL5 overexpression line. Lastly, we showed that red and far-red light signals trigger PIL5 protein degradation through the 26S proteasome, thus releasing the inhibition of bioactive GA biosynthesis by PIL5. Taken together, our data indicate that phytochromes promote seed germination by degrading PIL5, which leads to increased GA biosynthesis and decreased GA degradation.  相似文献   
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The mechanisms of secretory transport through the Golgi apparatus remain an issue of debate. The precise functional importance of calcium ions (Ca2+) for intra-Golgi transport has also been poorly studied. Here, using different approaches to measure free Ca2+ concentrations in the cell cytosol ([Ca2+]cyt) and inside the lumen of the Golgi apparatus ([Ca2+]GA), we have revealed transient increases in [Ca2+]cyt during the late phase of intra-Golgi transport that are concomitant with a decline in the maximal [Ca2+]GA restoration ability. Thus, this redistribution of Ca2+ from the Golgi apparatus into the cytosol during the movement of cargo through the Golgi apparatus appears to have a role in intra-Golgi transport, and mainly in the late Ca2+-dependent phase of SNARE-regulated fusion of Golgi compartments.  相似文献   
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