Record ID No. |
2004 |
Author(s) |
Helber N., Requena N , 2008 |
Affiliation |
University of Karlsruhe, Institute of Applied Bioscience, Fungal Plant Interact Grp, Hertzstr 16, D-76187 Karlsruhe, GERMANY |
Title |
Expression of the fluorescence markers DsRed and GFP fused to a nuclear localization signal in the arbuscular mycorrhizal fungus Glomus intraradices |
Source. Vol.(no):Page |
New Phytologist.177(2):537-548p. |
Categories |
Arbuscular Mycorrhiza |
Subjects |
Biochemistry |
Sub-subjects |
Miscellaneous |
Organism |
n.a. |
Country |
GERMANY,Europe |
Abstracts |
Here, arbuscular mycorrhizal (AM) fungi were monitored in vivo introducing the
fluorescent reporters DsRed and GFP (green fluorescent protein) in Glomus intraradices using a
biolistic approach and Agrobacterium tumefaciens-mediated transformation. Both reporter genes
were fused to the nuclear localization signal of the Aspergillus nidulans transcription factor
StuA to target fluorescence to nuclei. Expression of DsRed was driven by two Glomus mosseae
promoters highly expressed during early symbiosis, GmPMA1 and GmFOX2, while expression of GFP was
driven by the A. Nidulans gpd promoter. All promoters worked in G. Intraradices as well as in A.
Nidulans. Red and green fluorescence was localized to nuclei of G. Intraradices spores and hyphae
3 d after bombardment. However, expression was transient. The efficiency of the
Agrobacterium-mediated transformation was very low. These results indicate that the biolistic
method allows the expression of foreign DNA into G. Intraradices with high frequency, but it is
insufficient to render stable transformants. DsRed vs GFP is a more appropriate living reporter
to be used in G. Intraradices because of the lower autofluorescence in the red channel but
targeted to the nucleus both marker genes can be visualized. This is the first report of
fluorescent marker expression in an AM fungus driven by arbuscular mycorrhizal promoters. |