Isolation and characterisation of cDNAs encoding the large and small subunits of ADP-glucose pyrophosphorylase from cassava (Manihot esculenta Crantz) View Full Text


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Article Info

DATE

2001-06

AUTHORS

Tichafa R.I. Munyikwa, Jan Kreuze, Martin Fregene, Luc Suurs, Evert Jacobsen, Richard G.F. Visser

ABSTRACT

Screening of a tuber specific cassava cDNA library resulted in the isolation of full length cDNA clones with homology to the genes encoding the small and large subunits of ADP glucose pyrophosphoryalse. Sequence analysis revealed that AGPase B the clone with homology to the small subunit shared 54% homology at amino acid level with the AGPase S clone that is more closely related to the large subunit. Segregation analysis of a cross between the cassava cultivars TMS 30572 and CM 2177-2 revealed that AGPase S is a single copy gene that is localised on the female derived linkage group E of the cassava genetic map. AGPase B is a low copy gene of which one member is localised on the female derived linkage group P. The two genes are expressed in all cassava tissues but AGPase B exhibits a higher steady state mRNA level than AGPase S and is highly expressed in leaf and tuber tissue. The AGPase enzyme activity was much higher in young cassava leaves as compared to older leaves and tubers. Cassava AGPase was activated by 3-PGA and inhibited by up to 90% in the presence of inorganic phosphate (Pi). The tuber enzyme was relatively unaffected by 3PGA but was highly inhibited by Pi. Transformation of potato (Solanum tuberosum) plants with an antisense AGPase B construct resulted in 10 out of 134 antisense AGPase B plants having on average 3.5 times more tubers than the control non transgenic plants. Analysis of these transgenic plants revealed they had greatly reduced levels of AGPase B mRNA, 1.5 to 3 times less starch, and five times higher levels of soluble sugars, sucrose, glucose and fructose, to those found in control plants. More... »

PAGES

71-83

References to SciGraph publications

  • 1997-08. A molecular genetic map of cassava (Manihot esculenta Crantz) in THEORETICAL AND APPLIED GENETICS
  • 1992-05. Comparison of proteins of ADP-glucose pyrophosphorylase from diverse sources in JOURNAL OF MOLECULAR EVOLUTION
  • 1991-11. Comparison of the primary sequences of two potato tuber ADP-glucose pyrophosphorylase subunits in PLANT MOLECULAR BIOLOGY
  • 1992-12. Cloning, partial sequencing and expression of a cDNA coding for branching enzyme in cassava in PLANT MOLECULAR BIOLOGY
  • 1996-08. Synthesis of fructans in tubers of transgenic starch-deficient potato plants does not result in an increased allocation of carbohydrates in PLANTA
  • 1995-11. Factors affecting the inhibition by antisense RNA of granule-bound starch synthase gene expression in potato in MOLECULAR GENETICS AND GENOMICS
  • 1997-03. Molecular cloning and characterization of two novel isoforms of the small subunit of ADPglucose pyrophosphorylase from sweet potato in MOLECULAR GENETICS AND GENOMICS
  • 1995-09. Molecular cloning and characterization of novel isoforms of potato ADP-glucose pyrophosphorylase in MOLECULAR GENETICS AND GENOMICS
  • 1993-12. Molecular characterization of multiple cDNA clones for ADP-glucose pyrophosphorylase from Arabidopsis thaliana in PLANT MOLECULAR BIOLOGY
  • 1999-08. The influence of alterations in ADP-glucose pyrophosphorylase activities on starch structure and composition in potato tubers in PLANTA
  • 1990-10. One of two different ADP-glucose pyrophosphorylase genes from potato responds strongly to elevated levels of sucrose in MOLECULAR GENETICS AND GENOMICS
  • 1993-12. Isolation and characterization of a cDNA encoding granule-bound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potato in PLANT MOLECULAR BIOLOGY
  • 1995-01. Cell-type specific, coordinate expression of two ADP-glucose pyrophosphorylase genes in relation to starch biosynthesis during seed development of Vicia faba L. in PLANTA
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    DOI

    http://dx.doi.org/10.1023/a:1017551520240

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