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A Cassava vein mosaic virus promoter cassette induces high and stable gene expression in clonally propagated transgenic cassava (Manihot esculenta Crantz)

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Title A Cassava vein mosaic virus promoter cassette induces high and stable gene expression in clonally propagated transgenic cassava (Manihot esculenta Crantz)
 
Creator Oyelakin, O.O.
Opabode, J.T.
Raji, A.A.
Ingelbrecht, I.L.
 
Subject cassava
african cassava mosaic virus
gene expression
root crops
 
Description The study described a T-DNA vectorwith a Cassava veinmosaic virus promoter cassette (pCsVMV) and a kanamycin
selectable marker gene driven by the 35S Cauliflower mosaic virus promoter with a view to stably express
transgenes over repeated cycles of clonal propagation. A ?-glucuronidase reporter gene under control of pCsVMV
(pCsVMV-GUS) was introduced into the cassava landrace ‘Tokunbo’ via Agrobacterium-mediated genetic transformation.
Transgenic tobacco plants (Nicotiana tabacumSR1) with the same gene construct were also produced.
In tobacco, the pCsVMV-GUS was highly expressed in all tissues tested such as leaf, stem, petiole, and roots. In
transgenic cassava, the pCsVMV-GUS gene was highly expressed in all tissues and most cell types of in vitro
plants including leaf, stem, petiole, and fibrous roots. The pCsVMV-GUS gene was also highly expressed in
these tissues as well as in tubers of greenhouse grown cassava. High and stable pCsVMV-GUS gene expression
wasmaintained over 3 cycles of ratooning under greenhouse conditions, thus showing the absence of undesired
gene silencing effects after repeated in vitro subculturing and vegetative propagation. Fromthe high constitutive
levels of GUS activity observed, the study concluded that the CsVMV promoter cassette was useful for high-level
expression in cassava over repeated cycles of clonal propagationThe study described a T-DNA vectorwith a Cassava veinmosaic virus promoter cassette (pCsVMV) and a kanamycin
selectable marker gene driven by the 35S Cauliflower mosaic virus promoter with a view to stably express
transgenes over repeated cycles of clonal propagation. A ?-glucuronidase reporter gene under control of pCsVMV
(pCsVMV-GUS) was introduced into the cassava landrace ‘Tokunbo’ via Agrobacterium-mediated genetic transformation.
Transgenic tobacco plants (Nicotiana tabacumSR1) with the same gene construct were also produced.
In tobacco, the pCsVMV-GUS was highly expressed in all tissues tested such as leaf, stem, petiole, and roots. In
transgenic cassava, the pCsVMV-GUS gene was highly expressed in all tissues and most cell types of in vitro
plants including leaf, stem, petiole, and fibrous roots. The pCsVMV-GUS gene was also highly expressed in
these tissues as well as in tubers of greenhouse grown cassava. High and stable pCsVMV-GUS gene expression
wasmaintained over 3 cycles of ratooning under greenhouse conditions, thus showing the absence of undesired
gene silencing effects after repeated in vitro subculturing and vegetative propagation. Fromthe high constitutive
levels of GUS activity observed, the study concluded that the CsVMV promoter cassette was useful for high-level
expression in cassava over repeated cycles of clonal propagation
 
Date 2015-03
2016-05-25T11:59:26Z
2016-05-25T11:59:26Z
 
Type Journal Article
 
Identifier Oyelakin, O.O., Opabode, J.T., Raji, A.A. & Ingelbrecht, I.L. (2015). A Cassava vein mosaic virus promoter cassette induces high and stable gene expression in clonally propagated transgenic cassava (Manihot esculenta Crantz). South African Journal of Botany, 97, 184-190.
0254-6299
https://hdl.handle.net/10568/74435
https://doi.org/10.1016/j.sajb.2014.11.011
 
Language en
 
Rights CC-BY-4.0
Open Access
 
Format p. 184-190
 
Publisher Elsevier BV
 
Source South African Journal of Botany