Estrogen-inducible GFP expression patterns in rice (Oryza sativa L.)
Ayako Okuzaki1, Ken-ichi Konagaya, Yoshihiko Nanasato
1Division of Plant Sciences, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan.
Plant Cell Reports
|December 9, 2010
Summary
We studied the estrogen-inducible XVE system in rice, finding optimal 17-β-estradiol concentrations for green fluorescent protein (GFP) expression in calli and seedlings. This provides key insights for controlling gene expression in monocots.
Area of Science:
- Plant Molecular Biology
- Genetics and Genomics
- Biotechnology
Background:
- The XVE system is a chemical-inducible gene expression tool effective in dicots like Arabidopsis.
- Its application in monocots, such as rice (Oryza sativa L.), for functional genomics and marker-free transgenics is less understood.
- Specific correlations between inducer concentration and gene expression levels in monocots remain largely uncharacterized.
Purpose of the Study:
- To investigate the dose-response and time-course patterns of estrogen-inducible green fluorescent protein (GFP) expression in transgenic rice.
- To establish optimal conditions for the XVE system in rice, a key monocot model plant.
- To compare GFP expression levels between the XVE-inducible system and a constitutive promoter (35S) in different rice tissues.
Main Methods:
- Generation of transgenic rice lines harboring the estrogen-inducible GFP expression vector (pLex:GFP).
- Treatment of rice calli and seedlings with varying concentrations of 17-β-estradiol (the inducer).
- Analysis of GFP expression patterns using fluorescence microscopy over different time courses and inducer concentrations.
Main Results:
- GFP expression was detected in rice calli within 2 days at >5 μM 17-β-estradiol, with peak expression sustained for 8 days at 25 μM.
- In seedlings, leaf GFP signals from the pLex:GFP system were weaker than those from the p35S:GFP system.
- However, root GFP signals were comparable between pLex:GFP and p35S:GFP transgenic lines at >10 μM 17-β-estradiol.
Conclusions:
- The study provides the first detailed characterization of the XVE system's dose-response and time-course in rice.
- Optimal 17-β-estradiol concentrations and treatment durations for inducing gene expression in rice calli and seedlings were identified.
- These findings offer valuable guidance for utilizing the XVE system effectively in rice and potentially other monocot species for precise gene expression control.
Related Concept Videos
02:11Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...
02:50Transgenic Plants
Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...