I want to express GFP in the endosperm using an endosperm specific promotor and I was wondering whether the endosperm is transformed as well when performing the floral dip or just the egg cell?
I have not seen any report suggesting endosperm in seed of Arabidopsis is transformed using floral dip method.
In theory, any cell from the dipped plant that comes into contact with the bacteria could be transformed by the Agro. That being said, the usual methods used to screen for Arabidopsis transformants are based on the seedling being resistant to a selective agent (kanamycin or similar). You could look for GFP fluorescence in the seeds of your dipped plants. However, it may be more practical to screen for T1s and analyze the endosperm of the T2 seeds. This approach will also give you far more material to work with than just screening T1 seeds.
http://link.springer.com/article/10.1023%2FA%3A1024272023966
The Endosperm and the Embryo of Arabidopsis thaliana are Independently Transformed through Infiltration by Agrobacterium tumefaciens
Principally, chances is there that, at least in a few transformants, both the endosperm and zygote get the transgene; the MMC can be transformed by Agro. It can be checked under microscope, comparing with non-dipped plants. However, practically one need to go to next generation for detailed analysis, where you should see the expression in endosperm anayway (if the promoter is active in endosperm).
I am a bit confused regarding your objective. If I am right, you are aiming at raising a transgenic that express your gene of interest in an endosperm specific manner? If so, go ahead with raising transgenic through defined floral dip method without thinking which tissue is getting transformed during the very process as such, cause ultimately you will screen the positive plants obtained through transformation of egg (otherwise you will not get any transgenic). And, in the positive transgenic plants each and every cell regardless of the tissue will have your gene downstream to endosperm promoter. If your promoter works as expected, you will definitely get the expression of the gene specifically in endosperm of your positive transgenic plant.
In short, you don't require to transform the specific tissue where you want your gene to express. Your transgenic will contain your promoter-gene cassette in its every cell but gene will be expressed based on promoter activity.
Feel free to ask for further clarification.
Thank you for your answers!
I'll transform the plants as you suggested and screen the T2 seeds.
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