Functional analysis of the DAT gene promoter using transient Catharanthus roseus and stable Nicotiana tabacum transformation systems
The Catharanthus roseus DAT gene encodes the enzyme acetyl-CoA:deacetylvindoline-4- O- acetyltransferase involved in the last step of the indole alkaloid pathway leading to vindoline. This gene is characterized by specific cell type expression in idioblasts and laticifers. To understand the specific...
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Published in | Plant cell reports Vol. 30; no. 7; pp. 1173 - 1182 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Berlin/Heidelberg
Springer-Verlag
01.07.2011
Springer Springer Nature B.V |
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Abstract | The
Catharanthus roseus
DAT
gene encodes the enzyme acetyl-CoA:deacetylvindoline-4-
O-
acetyltransferase involved in the last step of the indole alkaloid pathway leading to vindoline. This gene is characterized by specific cell type expression in idioblasts and laticifers. To understand the specific transcriptional regulation mechanism(s) of
DAT
, several
DAT
promoter
GUS
constructs were cloned into pCAMBIA1305.1. Agroinfiltration of different explant types of
C. roseus
resulted in organ-specific accumulation of GUS, albeit at various levels. Heterologous accumulation of GUS in transgenic tobacco revealed both general and non-specific expression with the exception of a stomata-specific expression when 2.3 kb of the
DAT
promoter was coupled with a portion of the
DAT
ORF. These results suggest that in addition to the 2.3 kb upstream of the
DAT
transcriptional start site, additional
cis
-acting elements may be responsible for the specific spatial expression of
DAT
in vivo. Furthermore, hairy roots transformed with
DAT
promoter
GUS
constructs demonstrated
GUS
expression in root tissues (visualized through GUS enzyme activity), even though
DAT
is repressed in non-transformed roots. |
---|---|
AbstractList | The Catharanthus roseus DAT gene encodes the enzyme acetyl-CoA:deacetylvindoline-4-O-acetyltransferase involved in the last step of the indole alkaloid pathway leading to vindoline. This gene is characterized by specific cell type expression in idioblasts and laticifers. To understand the specific transcriptional regulation mechanism(s) of DAT, several DAT promoter GUS constructs were cloned into pCAMBIA1305.1. Agroinfiltration of different explant types of C. roseus resulted in organ-specific accumulation of GUS, albeit at various levels. Heterologous accumulation of GUS in transgenic tobacco revealed both general and non-specific expression with the exception of a stomata-specific expression when 2.3 kb of the DAT promoter was coupled with a portion of the DAT ORF. These results suggest that in addition to the 2.3 kb upstream of the DAT transcriptional start site, additional cis-acting elements may be responsible for the specific spatial expression of DAT in vivo. Furthermore, hairy roots transformed with DAT promoter GUS constructs demonstrated GUS expression in root tissues (visualized through GUS enzyme activity), even though DAT is repressed in non-transformed roots.[PUBLICATION ABSTRACT] The Catharanthus roseus DAT gene encodes the enzyme acetyl-CoA:deacetylvindoline-4-O-acetyltransferase involved in the last step of the indole alkaloid pathway leading to vindoline. This gene is characterized by specific cell type expression in idioblasts and laticifers. To understand the specific transcriptional regulation mechanism(s) of DAT, several DAT promoter GUS constructs were cloned into pCAMBIA1305.1. Agroinfiltration of different explant types of C. roseus resulted in organ-specific accumulation of GUS, albeit at various levels. Heterologous accumulation of GUS in transgenic tobacco revealed both general and non-specific expression with the exception of a stomata-specific expression when 2.3 kb of the DAT promoter was coupled with a portion of the DAT ORF. These results suggest that in addition to the 2.3 kb upstream of the DAT transcriptional start site, additional cis-acting elements may be responsible for the specific spatial expression of DAT in vivo. Furthermore, hairy roots transformed with DAT promoter GUS constructs demonstrated GUS expression in root tissues (visualized through GUS enzyme activity), even though DAT is repressed in non-transformed roots. The Catharanthus roseus DAT gene encodes the enzyme acetyl-CoA:deacetylvindoline-4-O-acetyltransferase involved in the last step of the indole alkaloid pathway leading to vindoline. This gene is characterized by specific cell type expression in idioblasts and laticifers. To understand the specific transcriptional regulation mechanism(s) of DAT, several DAT promoter GUS constructs were cloned into pCAMBIA1305.1. Agroinfiltration of different explant types of C. roseus resulted in organ-specific accumulation of GUS, albeit at various levels. Heterologous accumulation of GUS in transgenic tobacco revealed both general and non-specific expression with the exception of a stomata-specific expression when 2.3kb of the DAT promoter was coupled with a portion of the DAT ORF. These results suggest that in addition to the 2.3kb upstream of the DAT transcriptional start site, additional cis-acting elements may be responsible for the specific spatial expression of DAT in vivo. Furthermore, hairy roots transformed with DAT promoter GUS constructs demonstrated GUS expression in root tissues (visualized through GUS enzyme activity), even though DAT is repressed in non-transformed roots. The Catharanthus roseus DAT gene encodes the enzyme acetyl-CoA:deacetylvindoline-4- O- acetyltransferase involved in the last step of the indole alkaloid pathway leading to vindoline. This gene is characterized by specific cell type expression in idioblasts and laticifers. To understand the specific transcriptional regulation mechanism(s) of DAT , several DAT promoter GUS constructs were cloned into pCAMBIA1305.1. Agroinfiltration of different explant types of C. roseus resulted in organ-specific accumulation of GUS, albeit at various levels. Heterologous accumulation of GUS in transgenic tobacco revealed both general and non-specific expression with the exception of a stomata-specific expression when 2.3 kb of the DAT promoter was coupled with a portion of the DAT ORF. These results suggest that in addition to the 2.3 kb upstream of the DAT transcriptional start site, additional cis -acting elements may be responsible for the specific spatial expression of DAT in vivo. Furthermore, hairy roots transformed with DAT promoter GUS constructs demonstrated GUS expression in root tissues (visualized through GUS enzyme activity), even though DAT is repressed in non-transformed roots. The Catharanthus roseus DAT gene encodes the enzyme acetyl-CoA:deacetylvindoline-4-O-acetyltransferase involved in the last step of the indole alkaloid pathway leading to vindoline. This gene is characterized by specific cell type expression in idioblasts and laticifers. To understand the specific transcriptional regulation mechanism(s) of DAT, several DAT promoter GUS constructs were cloned into pCAMBIA1305.1. Agroinfiltration of different explant types of C. roseus resulted in organ-specific accumulation of GUS, albeit at various levels. Heterologous accumulation of GUS in transgenic tobacco revealed both general and non-specific expression with the exception of a stomata-specific expression when 2.3 kb of the DAT promoter was coupled with a portion of the DAT ORF. These results suggest that in addition to the 2.3 kb upstream of the DAT transcriptional start site, additional cis-acting elements may be responsible for the specific spatial expression of DAT in vivo. Furthermore, hairy roots transformed with DAT promoter GUS constructs demonstrated GUS expression in root tissues (visualized through GUS enzyme activity), even though DAT is repressed in non-transformed roots. |
Author | Bernards, Mark A. Makhzoum, Abdullah St. Pierre, Benoit Petit-Paly, Geneviève |
Author_xml | – sequence: 1 givenname: Abdullah surname: Makhzoum fullname: Makhzoum, Abdullah email: abmakhzoum@hotmail.com organization: Department of Biology and The Biotron, The University of Western Ontario – sequence: 2 givenname: Geneviève surname: Petit-Paly fullname: Petit-Paly, Geneviève organization: Biomolécules et Biotechnologies Végétales, Universite Francois-Rabelais – sequence: 3 givenname: Benoit surname: St. Pierre fullname: St. Pierre, Benoit organization: Biomolécules et Biotechnologies Végétales, Universite Francois-Rabelais – sequence: 4 givenname: Mark A. surname: Bernards fullname: Bernards, Mark A. organization: Department of Biology and The Biotron, The University of Western Ontario |
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Keywords | Vindoline Acetyl-CoA:deacetylvindoline-4 Specific cell type Indole alkaloids Promoter analysis acetyltransferase Acyltransferases Vegetals Enzyme Transcription promoter Transferases Indole Functional analysis Catharanthus roseus Nicotiana tabacum Transients Apocynaceae Alkaloid Dicotyledones Angiospermae Acetyl-CoA:deacetylvindoline-4-O-acetyltransferase Spermatophyta Solanaceae Catharanthus |
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SSID | ssj0014376 |
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Snippet | The
Catharanthus roseus
DAT
gene encodes the enzyme acetyl-CoA:deacetylvindoline-4-
O-
acetyltransferase involved in the last step of the indole alkaloid... The Catharanthus roseus DAT gene encodes the enzyme acetyl-CoA:deacetylvindoline-4-O-acetyltransferase involved in the last step of the indole alkaloid pathway... |
SourceID | proquest crossref pubmed pascalfrancis springer |
SourceType | Aggregation Database Index Database Publisher |
StartPage | 1173 |
SubjectTerms | Acetyltransferases - genetics Acetyltransferases - metabolism Base Sequence Biological and medical sciences Biomedical and Life Sciences Biotechnology Catharanthus - genetics Catharanthus - metabolism Catharanthus roseus Cell Biology Cloning, Molecular Electroporation Enzymatic activity Fundamental and applied biological sciences. Psychology Gene Expression Regulation, Plant Genes, Plant Genes, Reporter Genetic Vectors Life Sciences Molecular Sequence Data Nicotiana - genetics Nicotiana - metabolism Nicotiana tabacum Open Reading Frames Original Paper Plant Biochemistry Plant Leaves - genetics Plant Leaves - metabolism Plant Roots - enzymology Plant Roots - genetics Plant Sciences Plant tissues Plants, Genetically Modified - genetics Plants, Genetically Modified - metabolism Promoter Regions, Genetic Roots Seedlings - genetics Seedlings - metabolism Stomata Transcription Initiation Site Transfection Transformation, Genetic |
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Title | Functional analysis of the DAT gene promoter using transient Catharanthus roseus and stable Nicotiana tabacum transformation systems |
URI | https://link.springer.com/article/10.1007/s00299-011-1025-y https://www.ncbi.nlm.nih.gov/pubmed/21308469 https://www.proquest.com/docview/871559792/abstract/ https://search.proquest.com/docview/871967089 https://search.proquest.com/docview/904476498 |
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