Quantitative determination of mosaic GFP gene expression in tobacco

A specific form of gene silencing that was observed visually as a mosaic distribution of fluorescent and non-fluorescent cells apparently dispersed at random within tissues was found in a few green fluorescent protein (GFP)-transformed tobacco lines. To characterize this event quantitatively, we stu...

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Bibliographic Details
Published in:Plant cell reports 2004-07, Vol.22 (12), p.939-944
Main Authors: Bastar, M.T, Luthar, Z, Skof, S, Bohanec, B
Format: Article
Language:English
Subjects:
Base Sequence
DNA Primers
Flow Cytometry
Gene expression
gene expression regulation
gene silencing
Genes, Reporter
Genetics
green fluorescent protein
Green Fluorescent Proteins
Luminescent Proteins - genetics
Luminescent Proteins - metabolism
molecular sequence data
Mosaicism - genetics
Nicotiana - cytology
Nicotiana - genetics
Nicotiana - metabolism
Nicotiana tabacum
nucleotide sequences
Plants, Genetically Modified - cytology
Plants, Genetically Modified - genetics
Polymerase Chain Reaction
Proteins
quantitative analysis
Recombinant Proteins - metabolism
reporter genes
tobacco
transgenes
transgenic plants
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Summary:A specific form of gene silencing that was observed visually as a mosaic distribution of fluorescent and non-fluorescent cells apparently dispersed at random within tissues was found in a few green fluorescent protein (GFP)-transformed tobacco lines. To characterize this event quantitatively, we studied flow cytometric measurements in GFP-expressing and -silenced cells in T1 and T2 progeny of four selected plants. The proportion of silenced cells varied considerably among the T1 lines but with notable genotype differences. Mosaic expression was inherited into the T2 generation in which the majority of progenies tested exhibited a level of silencing similar to that of their T1 parental plants. However, in some T2 progenies segregation, evident as a decrease or increase in the proportion of fluorescent cells, was observed. We discuss several factors, such as copy number, promoter activity or polyploidy, that may be the possible causes of the gene silencing, but none sufficiently explain the appearance of the mosaic distribution.
ISSN:0721-7714
1432-203X
DOI:10.1007/s00299-004-0782-2