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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 100 (1997), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We are investigating the molecular basis of low temperature responses in apples, by identifying and characterising fruit genes which show altered expression in response to cool-storage, Two independent cold-regulated clones (pAMTI and pAMT2) were isolated from a cDNA library derived from cool-stored apple (Malus domestics Borkh. cv. Granny Smith) fruit. These clones share only 27% amino acid identity with each other, but both show high similarity to plant metallothionein (MT)-like proteins. The polypeptide encoded by pAMTI shares similarity with type 2 MT-like sequences, while that encoded by pAMT2 is similar to others which share a different distribution of cysteine residues. We suggest, these form a ‘type 3’ group of MT-like clones. Genomic Southern analysis confirmed that there is a family of MT-like genes in apple. There are differing patterns of pAMTI and pAMT2 expression during apple fruit development, amt 1 RNA was abundant in flowers and during the early stages of development, and decreased as the fruit approached maturity, while amt2 RNA was barely detectable in flowers and young fruit and accumulated with fruit development. In ripe fruit. amt 1. expression was up-regulated, while amt2 expression was down-regulated. In leaves, both genes showed increased expression with leaf age. In Granny Smith, Cox's Orange Pippin and Braeburn apple cultivars. both genes were up-regulated in cool-stored fruit. In Granny Smith contical tissue, amt RNA levels were elevated within the first 45 min at both 0.5°C and 4°C, but not at 12.5°C. The different patterns of amt1 and amt2 expression during fruit development and in different tissues suggest that the respective genes have distinct controlling elements and may be functionally different. The in vivo roles of the encoded polypeptides, particularly in relation to chilling tolerance or acclimation, are as yet unknown.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5028
    Keywords: ACC-oxidase ; apple ; fruit ripening ; polygalacturonase ; promoter ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Levels of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and polygalacturonase (PG) mRNAs were characterized during ripening of Royal Gala, Braeburn and Granny Smith apples. Both ACC-oxidase and PG mRNAs were up-regulated in ripening fruit of all three cultivars. Expression in Royal Gala was detected earlier than in Braeburn and Granny Smith, relative to internal ethylene concentration. Genomic clones corresponding to the ACC-oxidase and PG mRNAs expressed in ripe apple fruit were isolated and ca. 2 kb of each promoter was sequenced. The start point of transcription in each gene was mapped by primer extension, and sequences homologous to elements in other ethylene-responsive or PG promoters were identified. The fruit specificity of the apple ACC-oxidase and PG promoters was investigated in transgenic tomato plants using a nested set of promoter fragments fused to the β-glucuronidase (gusA) reporter gene. For the ACC-oxidase gene, 450 bp of 5′ promoter sequence was sufficient to drive GUS expression, although this expression was not specific to ripening fruit. Larger fragments of 1966 and 1159 bp showed both fruit and ripening specificity. For the PG gene, promoter fragments of 1460 and 532 bp conferred ripening-specific expression in transgenic tomato fruit. However GUS expression was down-regulated by 2356 bp of promoter, suggesting the presence of a negative regulatory element between positions -1460 and -2356.
    Type of Medium: Electronic Resource
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