When we used 35S（pBinGlyRed） to construct overexpressing Arabidopsis, we found several plants emitting red light in T1 generation by green light irradiation. This has never been found in our previous research. We want to find out why. Is this phenomenon common?
Your construct has 35s:dsRED, that is a strong constitutive (although not expressed in pollen grains) promoter. Yes, you would expect the entire plant to be dsRED in your strong expression transformants. What sorts of patterns did you observe previously and which promoter did you use?
Kourosh Vahdati
The vector we used had a red light gene in it to ensure that the positive seeds would glow red through the filter under green light. The gene we put into the vector doesn't emit red light.
The 35S:dsRED component is between the tDNA borders so that is integrated into your T1 plants, it will still be expressed beyond the seed stage. It's a nice marker for transformation, but as the promoter is not specific to the seed stage you will see expression other times. In essence you are overexpressing your target gene and dsRED in your transformants.
Amy Klocko
Thank you for your explanation. I have understood why the whole plant is red. But there is another problem. Only two plants in the same batch of materials are red. This phenomenon has never been observed before. We uploaded a new 35S vector map. We marked the positions of red light gene promoter and enzyme digestion sites. We have just harvested T1 seeds. The seed positive rate of these two plants was close to 100%. Such high infection efficiency is an exciting thing.
Ah, you are observing variability in the T1 plants (some are completely bright red, others are less intense). You may have multiple insertions and/or position effects influencing the expression levels.
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