Plant Molecular Biology

, Volume 56, Issue 4, pp 643–659 | Cite as

Identification and characterization of a novel cassava (Manihot esculenta Crantz) clone with high free sugar content and novel starch

  • Luiz Joaquim castelo branco carvalho
  • Claudia Regina Batista de souza
  • Julio César mattos De cascardo
  • Carlos Bloch Junior
  • Laura Campos


This study reports the identification of a new class of cassava (Manihot esculenta Crantz) with a storage root showing unusual free sugar accumulation and novel starch. Twenty-seven clones high in free sugar were identified under cultivation in primitive rural community areas in the Amazon. Iodine test and glucose oxidase–peroxidase reagent strips were used, in the field, for identification of starch and glucose, respectively. Five out of these 27 clones of cassava were cultivated at EMBRAPA Genetic Resources and Biotechnology and used for biochemical characterization, starch synthesis enzyme activities and gene expression analysis. Carbohydrates were fractioned into free sugar, polymerized water-soluble and -insoluble α-polyglucan. Clones of series CAS36 accumulate over 100 times more free sugar (mainly glucose) than commercial varieties. Monosaccharide composition analysis revealed one clone with distinct water-soluble sugars not present in the commercial cultivar. Structure analysis of the water-soluble and -insoluble α-polyglucan revealed the presence of a glycogen-like starch in clone CAS36.1. This clone indicated disruption in the starch synthesis pathway for enzyme activities and protein blot analyses in ADPG-pyrophosphorylase and branching enzyme, and their corresponding protein. Gene expression analysis indicated the lack of transcript for the gene coding for branching enzyme, but not for the gene coding for the ADPG-pyrophosphorylase small subunit. In addition, the pattern of distribution of sugar and starch content showed to be related to tissue age in the storage root.


cassava free sugar novel starch branching enzyme 


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  1. Allem, A.C. 1994The origin of Manihot esculenta Crantz (Euphorbiaceae)Genet. Res. Crop Evol.41133150CrossRefGoogle Scholar
  2. Ball, S., Guan, H.P., James, M., Myers, P., Mouille, G., Buleons, A., Colonna, P., Preiss, J. 1996From glycogen to amylopectin: a model for the biogenesis of the plant starch granuleCell86349352CrossRefPubMedGoogle Scholar
  3. Ballicora, M.A., Fu, Y., Nesbitt, N.M., Preiss, J. 1998ADP-glucose pyrophosphorylase from potato tubers Site-directed mutagenesis studies of the regulatory sites. Plant Physiol.118265274CrossRefPubMedGoogle Scholar
  4. Bogracheva, T.Y., Morris, V. J., Ring, S.G., Hedley, C.L. 1998The granular structure of C-type pea starch and its role in gelatinizationBiopolymers45323332CrossRefGoogle Scholar
  5. Cabral, G.B., Carvalho, L.J.C.B. and Schaal, B.A. 2000. The formation of storage roots in cassava. In: L.J.C.B. Carvalho, A.M. Thro and A.D. Vilarinhos (Eds.) Cassava Biotechnology, Proceedings of the IV International Scientific Meeting -- CBN (3--7 November 1998, Salvador, BA, Brazil), EMBRAPA-Genetic Resources and Biotechnology Brasilia-DF, Brazil, pp. 345--356.Google Scholar
  6. Carvalho, L.J.C.B. 1990. Maize acid invertases: purification, characterization, localization and function. Ph.D. Thesis, Cornell University, Ithaca, NY.Google Scholar
  7. Carvalho, L.J.C.B., Cabral, G.B., Campos, L. 2000Raiz de reserva de mandioca: um sistema biológico de múltiplas utilidade EMBRAPA-Recursos Genéticos e Biotecnologia. Serie Documentos 44. 16 pp. BrasiliaDF, BrazilGoogle Scholar
  8. Craig, J., Lloyd, J.R., Tomlinson, K., Barber, L., Edwards, A., Wang, T.L., Martin, C., Hedley, C.L., Smith, A.M. 1998Mutations in the gene encoding starch synthase II profoundly alter amylopectin structure in pea embryosPlant Cell10413426CrossRefPubMedGoogle Scholar
  9. da Cunha, A.G. 1978Dicionário Histórico das Palavras Portuguesas de Origem Tupi Companhia MelhoramentosUniversidade de Brasília, Brasília Brasil357Google Scholar
  10. Daiuto, E.R. 2000Biosintese de amido durante o crescimento secundario da raiz de reserva da mandioca M.Sc. DissertationUniversidade Estadual de São PauloBotucatu-SP, BrazilGoogle Scholar
  11. de Albuquerque, M. 1969A mandioca na AmazôniaSUDAMBelém-PA, Brasil 277Google Scholar
  12. de Souza, CRB. 2001Isolamento e caracterização molecular de genes e proteínas de raiz de reserva de mandioca (Manihot esculenta Crantz) Ph.D. DissertationUniversidade de BrasiliaBrazilGoogle Scholar
  13. Edwards, A., Fulton, D.C., Hylton, C.M., Jobling, S.A., Gidley, M., Rossner, U., Martin, C., Smith, A.M. 1999A combined reduction in activity of starch synthase II and III of potato has novel effects on the starch of tubersPlant J.17251261Google Scholar
  14. Fox, J.D., Robyt, J.R. 1991Miniaturization of three carbohydrate analyses using a microsample plate readerAnal. Biochem.1959396PubMedGoogle Scholar
  15. Fukuda, M.G.W., Costa, I.R.S, Vilarinhos, A.D., de Oliveira, R.P. 1996Banco de germoplasma de mandioca: Manejo, Conservação e CaracterizaçãoDocumentos CNPMF No. 68. p103Cruz das Almas BA, BrasilGoogle Scholar
  16. Hawker, J.S., Ozbun, J.L., Ozaki, H., Greenberg, E., Preiss, J. 1974Interaction of spinach leaf adenosine diphosphate glucose α-1,4 glucan-α-4-glucosyl transferase in synthesis of branched α-glucanArch. Biochem. Biophys.160530551PubMedGoogle Scholar
  17. Hizukuri, S. 1985Relationship between the distribution of the chain length of amylopectin and the crystalline structure of starch granulesCarbohydr. Res.141295306Google Scholar
  18. Hizukuri, S. 1986Polymodal distribution of chain lengths of amylopectins, and its significanceCarbohydr. Res.147342347Google Scholar
  19. Lisboa, C. 1631. História Animal e Árvores do Maranhão. Page 176.Google Scholar
  20. Macdonald, F.D., Preiss, J. 1983Solubilization of the starch-granule-bound starch synthase of normal maize kernelPlant Physiol.73175178Google Scholar
  21. McNeil, M., Darvill, A.G., Aman, P., Franzen, L.E., Albersheim, P. 1982Structural analysis of complex carbohydrate using high-performance liquid chromatography, gas chromatography, and mass spectrometryMeth. Enzymol.83345PubMedGoogle Scholar
  22. Munyikwa, T.R.I. 1997Isolation and characterization of starch biosynthesis genes from cassava (Manihot esculenta Crantz)Ph.D. DissertationWageningen University Wageningen, NetherlandsGoogle Scholar
  23. Munyikwa, T.R.I., Langeveld, S., Jacobsen, E., Visser, R.G.F. 1997Cassava starch biosynthesis: new avenues for modifying starch quantity and qualityEuphytica966575Google Scholar
  24. Nakamura, Y., Umemoto, T., Takahata, Y., Komae, K., Amano, E., Satoh, H. 1996Changes in structure of starch and enzyme activities affected by sugary mutations in developing rice endosperm Possible role of starch debranching enzyme (R-enzyme) in amylopectin biosynthesis. Plant Physiol.97491498Google Scholar
  25. O’Hair, S.K. 1989Cassava root starch content and distribution varies with tissue ageHortScience24505506Google Scholar
  26. Olsen, K.B.A., Schaal,  1999Evidence on the origin of cassava: phylogeography of Manihot esculentaProc Natl. Acad. Sci. USA9655865591PubMedGoogle Scholar
  27. Rateaver, B. 1951Anatomy and regeneration in the stem and root of Manihot utilíssima Pohl Ph.D. DissertationUniversity of MichiganAnn Arbor, MIGoogle Scholar
  28. Reiter, W.D., Chapple, C.C.S., Somerville, C.R. 1993Altered growth and cell walls in a fucose-deficient mutant of ArabidopsisScience26110321035Google Scholar
  29. Reiter, W.D., Chapple, C.C.S., Somerville, C.R. 1999Mutants of Arabidopsis thaliana with altered cell wall polysaccharide compositionPlant J.12335345Google Scholar
  30. Salehuzzaman, S.N.I.M., Jacobsen, E., Visser, G.F. 1992Cloning partial sequencing and expression of a cDNA coding for branching enzyme in cassavaPlant Mol. Biol.20809819PubMedGoogle Scholar
  31. Salehuzzaman, S.N.I.M., Jacobsen, E., Visser, R.G.F. 1993Isolation and characterization of cDNA encoding granule-bound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potatoPlant Mol. Biol.20947962Google Scholar
  32. Sarmento, S.B.S. 1997Caracterização da fécula de mandioca (Manihot esculenta C.) no período de colheita de cultivares de uso industrial Ph.D. DissertationUniversidade de Sao PauloPiracicaba, SP, BrazilGoogle Scholar
  33. Singletary, G.W., Banisadr, R., Keeling, P. 1997Influence of gene dosage on carbohydrate synthesis and enzyme activities in endosperm of starch-deficient mutant of maizePlant Physiol.113291304Google Scholar
  34. Sivad, M.N., Preiss, J. 1998Starch: basic science and biotechnologyAdv. Food Nutr. Res.411167PubMedGoogle Scholar
  35. Sriroth, K., Santisopasri, V., Petchalanuwat, C., Kurotjanawong, K., Piyachomkwan, K., Oates, C.G. 1999Cassava starch granule structure-function properties: influence of time and conditions at harvest on four cultivars of cassava starchCarbohydr. Polym.38161170Google Scholar
  36. Travaços, S. 1596. Declaração do Brasil XLVII (Brazilian Declaration XLVII), p. 35.Google Scholar
  37. Waigh, T.A., Perry, P., Riekel, C., Gidley, M.J., Donald, A.M. 1998Chiral side-chain liquid-crystalline polymeric properties of starchMacromolecules2279807984Google Scholar
  38. Zakhia, N., Whaetley, C., O’Brien, G.O. and Dufour, D. 1995. Screening of CIAT cassava germplasm diversity: the relationship between cyanogenic potential and cassava starch functional properties. In: The Cassava Biotechnology Network. Proceedings of the 2nd International Scientific Meeting (Bogor, Indonesia, 22--23 August 1994), Centro Internacional de Agricultura Tropical, Cali, Colombia, pp. 823--830.Google Scholar
  39. Zeeman, S.C., Umemoto, T., Lue, W.L., Yeung, P.A., Martin, C., Smith, A.M., Chen, J. 1998A mutant of Arabidopsis␣lacking a chloroplastic isoamylase accumulates both␣starch␣and phytoglycogenPlant Cell1016991711PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Luiz Joaquim castelo branco carvalho
    • 1
  • Claudia Regina Batista de souza
    • 1
    • 3
  • Julio César mattos De cascardo
    • 2
  • Carlos Bloch Junior
    • 1
  • Laura Campos
    • 1
    • 4
  1. 1.EMBRAPA Genetic Resources and BiotechnologyBrasiliaBrazil
  2. 2.Departamento de Biologia CelularUniversidade Estadual de Santa CruzIlhéusBrazil
  3. 3.Departamento de GenéticaUniversidade Federal do ParaBelémBrazil
  4. 4.Conselho Nacional, Desenvolvimento Cientifico & Tecnologico (CNPq)Brazil

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