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  • 1
    Electronic Resource
    Electronic Resource
    Copenhagen : Munksgaard International Publishers
    Physiologia plantarum 110 (2000), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The relationships between the changes induced in abscisic acid (ABA) and proline contents were investigated in canola (Brassica napus L.) leaf discs (CLDs) subjected to sequential hyper- and hypo-osmotic treatments. Changes in ABA content were found to precede that of proline, suggesting that ABA could exert regulatory effects on both osmo-induced proline accumulation and its subsequent mobilization during recovery. When exogenously supplied in the light during recovery, ABA caused inhibition of the apparent rate of proline mobilization with an I50 of about 20 μM. Under dark conditions, the ABA effect was not observed. This is in accordance with the fact that this effect could be suppressed in the light while using photosynthesis inhibitors. The ABA effect was partly restored under dark conditions when the CLDs were supplied with NADPH at high concentration. Under both conditions, ABA exerted its effect when supplied together with cycloheximide, suggesting that it does not rely on protein synthesis. When turgid CLDs, which heavily absorbed exogenously supplied l-proline, were incubated in the light on the reference medium with ABA, proline mobilization was also restricted, provided ABA was supplied during the period of proline uptake. Surprisingly, in CLDs that had recovered their full turgor after application of successive hyper- and hypo-osmotic treatments, application of ABA led to a very high accumulation of proline. In contrast, in freshly cut turgid CLDs incubated in presence of ABA, the basic proline level was only weakly enhanced. Thus, the apparent inhibitory effect of ABA on proline mobilization could be mediated through both an activation of biosynthesis and an inhibition of catabolism of this amino acid via light-dependent processes that remain to be elucidated.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Some aspects of the function of abscisic acid (ABA) in the control of proline (PRO) metabolism in canola leaf discs (CLD) subjected in vitro to consecutive hyper-osmotic (stress) and hypo-osmotic (recovery) treatments have been investigated. PRO accumulation in response to stress conditions relies on both stimulation of its synthesis via enhancement of transcription of the gene and activity of Δ1-pyrroline-5-carboxylate synthetase (P5CS) and inhibition of its degradation via inactivation of PRO dehydrogenase (PDH). These changes were partly reversed under recovery conditions. Thus both PDH mRNA and PDH activity increased while P5CS mRNA decreased. Surprisingly P5CS activity remained high even after a 20-h period of rehydration. Exogenously supplied ABA at recovery inhibited net PRO consumption and this could be associated with downregulation of PDH gene expression and PDH activity. Under these conditions ABA hardly upregulated P5CS gene expression while P5CS activity first transiently decreased and then reached a value close to that found under stressing conditions. Experiments with CLD supplied with either methionine sulphoximine or gabaculine, brought preliminary evidence for a significant synthesis of PRO from glutamate during recovery that replenished the proline pool(s) and provoked a negative effect on the net rate of PRO consumption. Consequently, it is suggested that the availability of both PRO precursors and ABA could be determinant in the control of the amount of residual PRO present in CLD after the period of recovery. This level also seemed to depend on the amount of P5CS transcripts induced under stress conditions. However, the results obtained with turgid leaf discs treated with ABA indicate that the ABA status of the tissues, necessary for inducing the proline response, is not sufficient to determine their PRO content because it remained relatively low despite the stimulation of P5CS expression and P5CS activity.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-5028
    Keywords: ABA-responsive element ; maize ; tissue-specific factors ; rab genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The maize gene rab28 has been identified as ABA-inducible in embryos and vegetative tissues. It is also induced by water stress in young leaves. The proximal promoter region contains the conserved cis-acting element CCACGTGG (ABRE) reported for ABA induction in other plant genes. Transient expression assays in rice protoplasts indicate that a 134 bp fragment (-194 to -60 containing the ABRE) fused to a truncated cauliflower mosaic virus promoter (35S) is sufficient to confer ABA-responsiveness upon the GUS reporter gene. Gel retardation experiments indicate that nuclear proteins from tissues in which the rab28 gene is expressed can interact specifically with this 134 bp DNA fragment. Nuclear protein extracts from embryo and water-stressed leaves generate specific complexes of different electrophoretic mobility which are stable in the presence of detergent and high salt. However, by DMS footprinting the same guanine-specific contacts with the ABRE in both the embryo and leaf binding activities were detected. These results indicate that the rab28 promoter sequence CCACGTGG is a functional ABA-responsive element, and suggest that distinct regulatory factors with apparent similar affinity for the ABRE sequence may be involved in the hormone action during embryo development and in vegetative tissues subjected to osmotic stress.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-5028
    Keywords: cDNA family ; maize ; pollen-expressed genes ; polygalacturonase ; signal peptide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A full-length cDNA clone, named PG1, abundantly expressed in late stages of pollen development, has been isolated from a cDNA library using a differential screening method with cDNA probes representative of microspores at early or late developmental stages. The encoded 410 amino acid polypeptide has significant homology with various polygalacturonases (PG) described elsewhere. Two polypeptides, of 49 and 53 kDa respectively, have been identified in the active PG fraction, isolated from mature pollen by immuno-cross-reaction with tomato PG antibodies. According to their N-terminal sequence, they can be identified as being mature peptides encoded by the PG1 cDNA clone. We propose that these two proteins derive from a unique precursor through several post-translational events, including the excision of a 22 amino-terminal signal peptide and glycosylation. PG-encoding genes form a small genomic family. Sequence analysis of three PG cDNA clones shows that they are closely related. The divergence of nucleotides between these three cDNA clones is 1%. They encode the same product.
    Type of Medium: Electronic Resource
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