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  • 1
    ISSN: 1432-2048
    Keywords: cDNA clone (sucrose synthase) ; Gene expression (sucrose synthase, seed development) ; Nucleotide sequence (sucrose synthase) ; Seed development ; Sucrose synthase (cloning, expression, regulation) ; Vicia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Copy-DNA clones encoding a single class of sucrose-synthase (SUCS; EC 2.4.1.13) subunit have been isolated and sequenced from a Vicia faba L. seed cotyledonary library. Southern analyses indicated the existence of only one gene. Transcript levels determined by Northern blot hybridisation steadily increased until the middle of development [25–35 days after flowering (DAF)] and declined thereafter. Sucrose levels approximately paralleled levels of SUCS mRNA. The activity of SUCS increased with decreasing fructose and glucose concentrations and peaked about 10 d later than mRNA levels. In-vitro culture experiments demonstrated that increasing the sucrose concentration leads to increased levels of SUCS mRNA. The SUCS mRNA was also synthesised in seed-coat tissue, but in lower amounts than in cotyledons and with a different developmental profile. The early peak level of SUCS mRNA (20 DAF) in seed coats coincided with the peak in the amount of sucrose and with a peak of transiently synthesised starch.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2048
    Keywords: ADP-glucose pyrophosphorylase (cDNA-cloning, sequencing) ; Gene expression (ADP-glucose pyrophospharylase, seed development) ; Metabolic regulation (ADP-glucose pyrophospharylase) ; Seed development ; Starch biosynthesis ; Vicia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Several cDNA clones encoding two different ADP-glucose pyrophosphorylase (AGPase, EC 2.7.7.27) polypeptides denoted VfAGPC and VfAGPP were isolated from a cotyledonary library of Vicia faba L. Both sequences are closely related to AGPase small-subunit sequences from other plants. Whereas mRNA levels of VfAGPP were equally high in developing cotyledons and leaves, the mRNA of VfAGPC was present in considerable amounts only in cotyledons. During development of cotyledons, both mRNAs accumulated until the beginning of the desiccation phase and disappeared afterwards. The increase of AGPase activity in cotyledons during the phase of storage-product synthesis was closely followed by the accumulation of starch. The AGPase activity in crude extracts of cotyledons was insensitive to 3-phosphoglycerate whereas the activity from leaves could be activated more than five-fold. Inorganic phosphate inhibited the enzyme from both tissues but was slightly more effective on the leaf enzyme. There was a correlation at the cellular level between the distribution of VfAGPP and VfAGPC mRNAs and the accumulation of starch, as studied by in-situ hybridisation and by histochemical staining in parallel tissue sections of developing seeds, respectively. During the early phase of seed development (12–15 days after fertilization) VfAGPase mRNA and accumulation of starch were detected transiently in the hypodermal, chlorenchymal and outer parenchymal cell layers of the seed coat but not in the embryo. At 25 days after fertilization both synthesis of VfAGPase mRNA and biosynthesis of starch had started in parenchyma cells of the inner adaxial zone of the cotyledons. During later stages, the expression of VfAGPase and synthesis of starch extended over most of the cotyledons but were absent from peripheral cells of the abaxial zone, provascular and procalyptral cells.
    Type of Medium: Electronic Resource
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