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  • 1
    ISSN: 1432-2048
    Keywords: Cucumis (cotyledon) ; Glyoxylate cycle ; Senescence ; Sucrose
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When cotyledons are detached from cucumber (Cucumis sativus L. cv. Masterpiece) plants 14 d after seed imbibition, accumulation of the glyoxylate-cycle enzymes malate synthase (MS) and isocitrate lyase (ICL) occurs rapidly in the dark but not in the light. This is primarily due to an increase in transcript abundance. It has been proposed that glyoxylate-cycle enzymes are synthesised in response to lipid degradation. However, MS and ICL synthesis in detached cucumber cotyledons begins before a decline in lipid can be detected. Furthermore, the activation of MS and ICL gene expression does not correlate with loss of chlorophyll, carotenoid, protein or RNA. These results are discussed in terms of the activation of genes encoding glyoxylate-cycle enzymes and their possible function in detached and senescing organs. Previous work (Graham et al, 1992, Plant Cell 3, 349–357) has suggested that sucrose or other carbohydrates can repress the synthesis of MS and ICL. Consistent with this proposal, incubation of detached cotyledons in the light leads to a fivefold increase in sucrose content. However, when MS and ICL synthesis occurs in the dark, no change in the amount of sucrose is detected. Incubation of cotyledons in the dark in the presence of 25 mM sucrose leads to a complete inhibition of the accumulation of MS and ICL. Similarly, cotyledon protoplasts incubated in darkness accumulate MS and ICL, but not if 25 mM sucrose is included in the medium. These results collectively support the view that MS and ICL synthesis responds to the metabolic activities of plant cells.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: Cucumber (Cucumis sativus L.) ; Malate synthase ; Glyoxylate cycle ; Gene transcription ; Metabolic regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The malate synthase gene (ms) promoter in cucumber (Cucumis sativus L.) was investigated with the aim of distinguishing DNA sequences mediating regulation of gene expression by sugar, and expression following seed germination. Promoter deletions were constructed and their ability to direct expression of theβ-glucuronidase (gus) reporter gene was investigated in transgenicNicotiana plumbaginifolia. Gene expression was assayed in germinating seeds and developing seedlings (the germination response) and in seedlings transferred from light into darkness with and without sucrose (the sugar response). As progressively more of the promoter was deleted from the 5′ end, first the sugar response and then the germination response was lost. Thus, distinct regions of the promoter are required for carbohydrate control and for regulation of gene expression in response to germination. Sequence comparisons of thems promoter with that of the isocitrate lyase gene (icl) of cucumber have previously identified four IMH (ICL-MS Homology) sequences. One such sequence, IMH2, is shown here to be implicated in the sugar response of thems gene. The 17 bp sequence, which when deleted from thems gene results in loss of the germination response, contains a 14 bp sequence which is similar to a sequence in theicl promoter, which we refer to as IMH5. Furthermore, this sequence has similarity withamdI9-like sequences in filamentous fungi, which conferfacB-mediated acetate inducibility on several genes, including those encoding ICL and MS.
    Type of Medium: Electronic Resource
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