Cloning and characterization of a tuberous root-specific promoter from cassava (Manihot esculenta Crantz)

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Abstract

In order to obtain a tuberous root-specific promoter to be used in the transformation of cassava, a 1,728 bp sequence containing the cassava granule-bound starch synthase (GBSSI) promoter was isolated. The sequence proved to contain light- and sugar-responsive cis elements. Part of this sequence (1,167 bp) was cloned into binary vectors to drive expression of the firefly luciferase gene. Cassava cultivar Adira 4 was transformed with this construct or a control construct in which the luciferase gene was cloned behind the 35S promoter. Luciferase activity was measured in leaves, stems, roots and tuberous roots. As expected, the 35S promoter induced luciferase activity in all organs at similar levels, whereas the GBSSI promoter showed very low expression in leaves, stems and roots, but very high expression in tuberous roots. These results show that the cassava GBSSI promoter is an excellent candidate to achieve tuberous root-specific expression in cassava.
LanguageEnglish
Pages1955-1965
JournalPlanta
Volume236
Issue number6
DOIs
Publication statusPublished - 2012

Fingerprint

Manihot
Manihot esculenta
cassava
Organism Cloning
molecular cloning
promoter regions
luciferase
Luciferases
Starch Synthase
Firefly Luciferases
starch synthase
stems
Genes
leaves
granules
genes
Light
sugars
cultivars
granule-bound starch synthase I

Keywords

  • bound-starch-synthase
  • storage roots
  • transformation
  • expression
  • gene
  • agrobacterium
  • elements
  • sequences
  • database
  • program

Cite this

@article{3311e84e8cb54ed5b69da566078e6df9,
title = "Cloning and characterization of a tuberous root-specific promoter from cassava (Manihot esculenta Crantz)",
abstract = "In order to obtain a tuberous root-specific promoter to be used in the transformation of cassava, a 1,728 bp sequence containing the cassava granule-bound starch synthase (GBSSI) promoter was isolated. The sequence proved to contain light- and sugar-responsive cis elements. Part of this sequence (1,167 bp) was cloned into binary vectors to drive expression of the firefly luciferase gene. Cassava cultivar Adira 4 was transformed with this construct or a control construct in which the luciferase gene was cloned behind the 35S promoter. Luciferase activity was measured in leaves, stems, roots and tuberous roots. As expected, the 35S promoter induced luciferase activity in all organs at similar levels, whereas the GBSSI promoter showed very low expression in leaves, stems and roots, but very high expression in tuberous roots. These results show that the cassava GBSSI promoter is an excellent candidate to achieve tuberous root-specific expression in cassava.",
keywords = "bound-starch-synthase, storage roots, transformation, expression, gene, agrobacterium, elements, sequences, database, program",
author = "{Koehorst-van Putten}, H.J.J. and A.M.A. Wolters and I.J. Pereira-Bertram and H. Berg and {van der Krol}, A.R. and R.G.F. Visser",
year = "2012",
doi = "10.1007/s00425-012-1796-6",
language = "English",
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pages = "1955--1965",
journal = "Planta",
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T1 - Cloning and characterization of a tuberous root-specific promoter from cassava (Manihot esculenta Crantz)

AU - Koehorst-van Putten, H.J.J.

AU - Wolters, A.M.A.

AU - Pereira-Bertram, I.J.

AU - Berg, H.

AU - van der Krol, A.R.

AU - Visser, R.G.F.

PY - 2012

Y1 - 2012

N2 - In order to obtain a tuberous root-specific promoter to be used in the transformation of cassava, a 1,728 bp sequence containing the cassava granule-bound starch synthase (GBSSI) promoter was isolated. The sequence proved to contain light- and sugar-responsive cis elements. Part of this sequence (1,167 bp) was cloned into binary vectors to drive expression of the firefly luciferase gene. Cassava cultivar Adira 4 was transformed with this construct or a control construct in which the luciferase gene was cloned behind the 35S promoter. Luciferase activity was measured in leaves, stems, roots and tuberous roots. As expected, the 35S promoter induced luciferase activity in all organs at similar levels, whereas the GBSSI promoter showed very low expression in leaves, stems and roots, but very high expression in tuberous roots. These results show that the cassava GBSSI promoter is an excellent candidate to achieve tuberous root-specific expression in cassava.

AB - In order to obtain a tuberous root-specific promoter to be used in the transformation of cassava, a 1,728 bp sequence containing the cassava granule-bound starch synthase (GBSSI) promoter was isolated. The sequence proved to contain light- and sugar-responsive cis elements. Part of this sequence (1,167 bp) was cloned into binary vectors to drive expression of the firefly luciferase gene. Cassava cultivar Adira 4 was transformed with this construct or a control construct in which the luciferase gene was cloned behind the 35S promoter. Luciferase activity was measured in leaves, stems, roots and tuberous roots. As expected, the 35S promoter induced luciferase activity in all organs at similar levels, whereas the GBSSI promoter showed very low expression in leaves, stems and roots, but very high expression in tuberous roots. These results show that the cassava GBSSI promoter is an excellent candidate to achieve tuberous root-specific expression in cassava.

KW - bound-starch-synthase

KW - storage roots

KW - transformation

KW - expression

KW - gene

KW - agrobacterium

KW - elements

KW - sequences

KW - database

KW - program

U2 - 10.1007/s00425-012-1796-6

DO - 10.1007/s00425-012-1796-6

M3 - Article

VL - 236

SP - 1955

EP - 1965

JO - Planta

T2 - Planta

JF - Planta

SN - 0032-0935

IS - 6

ER -