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Tissue culture specificity of the tobacco ASA2 promoter driving hpt as a selectable marker for soybean transformation selection
Tissue culture specificity of the tobacco ASA2 promoter driving hpt as a selectable marker for soybean transformation selection
- Source :
- Plant Cell Reports. 27:1705-1711
- Publication Year :
- 2008
- Publisher :
- Springer Science and Business Media LLC, 2008.
-
Abstract
- This study was carried out to determine if the tobacco anthranilate synthase ASA2 2.3 kb promoter drives tissue culture specific expression and if it is strong enough to drive hpt (hygromycin phosphotransferase) gene expression at a level sufficient to allow selection of transformed soybean embryogenic culture lines. A number of transformed cell lines were selected showing that the promoter was strong enough. Northern blot analysis of plant tissues did not detect hpt mRNA in the untransformed control or in the ASA2-hpt plants except in developing seeds while hpt mRNA was detected in all tissues of the CaMV35S-hpt positive control line plants. However, when the more sensitive RT-PCR assay was used all tissues of the ASA2-hpt plants except roots and mature seeds were found to contain detectable hpt mRNA. Embryogenic tissue cultures initiated from the ASA2-hpt plants contained hpt mRNA detectable by both northern and RT-PCR analysis and the cultures were hygromycin resistant. Friable callus initiated from leaves of ASA2-hpt plants did in some cases contain hpt mRNA that was only barely detectable by northern hybridization even though the callus was very hygromycin resistant. Thus the ASA2 promoter is strong enough to drive sufficient hpt expression in soybean embryogenic cultures for hygromycin selection and only very low levels of expression were found in most plant tissues with none in mature seeds.
- Subjects :
- Nicotiana tabacum
Plant Science
Biology
Tissue Culture Techniques
Tissue culture
Transformation, Genetic
Tobacco
Gene expression
Northern blot
Promoter Regions, Genetic
Selectable marker
DNA Primers
Plant Proteins
Genetics
Base Sequence
Reverse Transcriptase Polymerase Chain Reaction
fungi
food and beverages
Promoter
General Medicine
Biolistics
Blotting, Northern
biology.organism_classification
Molecular biology
Blotting, Southern
Phosphotransferases (Alcohol Group Acceptor)
Transformation (genetics)
Callus
Soybeans
Agronomy and Crop Science
Subjects
Details
- ISSN :
- 1432203X and 07217714
- Volume :
- 27
- Database :
- OpenAIRE
- Journal :
- Plant Cell Reports
- Accession number :
- edsair.doi.dedup.....1db2607e6c49e4d00ae1a7fe9a85977e
- Full Text :
- https://doi.org/10.1007/s00299-008-0589-7