A spontaneous dominant-negative mutation within a 35S::AtMYB90 transgene inhibits flower pigment production in tobacco.
BACKGROUND
In part due to the ease of visual detection of phenotypic changes, anthocyanin pigment production has long been the target of genetic and molecular research in plants. Specific members of the large family of plant myb transcription factors have been found to play critical roles in regulating expression of anthocyanin biosynthetic genes and these genes continue to serve as important tools in dissecting the molecular mechanisms of plant gene regulation.


RESULTS
A spontaneous mutation within the coding region of an Arabidopsis 35S::AtMYB90 transgene converted the activator of plant-wide anthocyanin production to a dominant-negative allele (PG-1) that inhibits normal pigment production within tobacco petals. Sequence analysis identified a single base change that created a premature nonsense codon, truncating the encoded myb protein. The resulting mutant protein lacks 78 amino acids from the wild type C-terminus and was confirmed as the source of the white-flower phenotype. A putative tobacco homolog of AtMYB90 (NtAN2) was isolated and found to be expressed in flower petals but not leaves of all tobacco plants tested. Using transgenic tobacco constitutively expressing the NtAN2 gene confirmed the NtAN2 protein as the likely target of PG-1-based inhibition of tobacco pigment production.


CONCLUSIONS
Messenger RNA and anthocyanin analysis of PG-1Sh transgenic lines (and PG-1Sh x purple 35S::NtAN2 seedlings) support a model in which the mutant myb transgene product acts as a competitive inhibitor of the native tobacco NtAN2 protein. This finding is important to researchers in the field of plant transcription factor analysis, representing a potential outcome for experiments analyzing in vivo protein function in test transgenic systems that over-express or mutate plant transcription factors.
Velten
Jeff
J
Plant Stress and Water Conservation Laboratory, United States Department of Agriculture - Agricultural Research Service, Lubbock, Texas, United States of America. VeltenLab@gmail.com
Cakir
Cahid
C
Cazzonelli
Christopher I
CI
D000483:Alleles; D000872:Anthocyanins / Q000235:genetics; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000378:metabolism; D001483:Base Sequence; D035264:Flowers / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D005799:Genes, Dominant; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D010858:Pigmentation; D030821:Plants, Genetically Modified; D012689:Sequence Homology, Nucleic Acid; D014026:Nicotiana / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism; D019076:Transgenes
D000872:Anthocyanins; D029681:Arabidopsis Proteins; C500589:Myb protein, Arabidopsis; D014157:Transcription Factors