Flavanone 3-hydroxylase transcripts and flavonol accumulation are temporally coordinate in maize anthers

Flavanone 3-hydroxylase (F3H) activity is necessary for the production of both flavonols and anthocyanins. Flavonols are required for functional pollen in maize whereas anthocyanins are non-essential pigments. A cDNA for F3H was isolated from Zea mays using a heterologous sequence from Antirrhinum majus. Comparison of the deduced amino acid sequence of maize F3H with other F3H sequences confirmed that the protein is highly conserved among widely divergent plant species. The F3H gene is present in a single copy located at the tip of chromosome 2S. High levels of F3H gene expression were detected in pigmented husk and 26-day postpollination kernels; lower levels in 18-day postpollination kernels and in coleoptiles of germinating seedlings. Slot blot analysis showed that F3H transcript levels in young seedlings are increased by high fluence-rate white light treatment in the presence of the anthocyanin regulatory gene -r. HPLC analysis of extracts from developmentally staged anthers showed that flavonol accumulation begins at the uninucleate microspore stage, continues until maturity, and is not controlled by -r. When the expression pattern of several flavonoid biosynthetic genes was analyzed during microsporogenesis, only F3H transcript accumulation was coordinate with the appearance of flavonols in anthers.