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  • 1
    Electronic Resource
    Electronic Resource
    Isolation and characterization of a cDNA encoding granule-bound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potato (1993)
    Springer
    Plant molecular biology 23 (1993), S. 947-962 
    Details
    ISSN: 1573-5028
    Keywords: antisense effect ; granule-bound starch synthase ; cassava ; cDNA ; heterologous gene ; potato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A tuber-specific cDNA library of cassava (Manihot esculenta Crantz) was constructed and a full-length cDNA for granule-bound starch synthase (GBSS, also known as waxy protein), the enzyme responsible for the synthesis of amylose in reserve starch, was cloned. Sequencing of the cloned cDNA showed that it has 74% identity with potato GBSS and 60–72% identity with GBSS from other plant species. The cDNA encodes a 608 amino acid protein of which 78 amino acids form a chloroplast/amyloplast transit peptide of 8.37 kDa. The mature protein has a predicted molecular mass of 58.61 kDa (530 amino acids). Comparison of the GBSS proteins of various plant species and glycogen synthase of bacteria showed extensive identity among the mature form of plant GBSS proteins, in which the monocots and dicots form two separate branches in the evolutionary tree. From analysis of the genomic DNA of allotetraploid cassava, it is shown that GBSS is a low-copy-number gene. GBSS transcript is synthesized in a number of different organs, but most abundantly in tubers. Potato plants were transformed with the cassava GBSS cDNA in antisense orientation fused between the potato GBSS promoter and the nopaline synthase terminator. The expression of the endogenous GBSS gene in these transgenic potato plants was partially or completely inhibited. Complete inhibition of GBSS activity by the cassava antisense gene resulted in absence of GBSS protein and amylose giving rise to almost complete amylose-free potato starch. This shows that also heterologous genes can be used to achieve antisense effects in other plant species.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021811
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Introduction of sense and antisense cDNA for branching enzyme in the amylose-free potato mutant leads to physico-chemical changes in the starch (1996)
    Springer
    Planta 198 (1996), S. 340-347 
    Details
    ISSN: 1432-2048
    Keywords: Antisense inhibition ; Branching enzyme (potato) ; Co-suppression (gene activity) ; Gene activity (branching enzyme) ; Solanum (branching enzyme) ; Starch physico chemical properties
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract One isoform of the branching enzyme (BE; EC 2.4.1.18) of potato (Solarium tuberosum L.) is known and catalyses the formation of α-1,6 bonds in a glucan chain, resulting in the branched starch component amylopectin. Constructs containing the antisense or sense-orientated distal 1.5-kb part of a cDNA for potato BE were used to transform the amylose-free (amf) mutant of potato, the starch of which stains red with iodine. The expression of the endogenous BE gene was inhibited either largely or fully as judged by the decrease or absence of the BE mRNA and protein. This resulted in a low percentage of starch granules with a small blue core and large red outer layer. There was no effect on the amylose content, degree of branching or λmax of the iodine-stained starch. However, when the physico-chemical properties of the different starch suspensions were assessed, differences were observed, which although small indicated that starch in the transformants was different from that of theamf mutant.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00620049
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Field evaluation of antisense RNA mediated inhibition of GBSS gene expression in potato (1991)
    Springer
    Euphytica 59 (1991), S. 83-91 
    Details
    ISSN: 1573-5060
    Keywords: Agrobacterium rhizogenes ; antisense RNA ; granule-bound starch synthase ; Solanum tuberosum ; starch composition ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00025364
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    Paper (German National Licenses)
    Fulltext
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