MADS-box genes are a large family of transcription factors found in yeast, animals, and plants. In plants, MADS-box genes control an array of developmental processes ranging from flower and fruit development to root development. MADS-box genes have been best characterized for their role in the ABC model of flower development. Of the three classes of MADS-box genes in the ABC model, the B class genes have been best characterized. B class genes have been isolated from a diverse array of angiosperms, and the phylogenetic history and patterns of gene expression of this class of MADS-box genes are becoming clear. We have isolated B class MADS-box genes from the ANITA grade of basal angiosperms. Here we present a detailed protocol for isolating RNA from basal angiosperms. We further estimate the phylogenetic position, using both maximum parsimony and maximum likelihood methods, of the B class genes isolated from Amborella, Nuphar, and Illicium. Lastly, we present preliminary gene expression data for AP3 and PI genes isolated from Nuphar.

Key words: Basal Angiosperms, MADS-Box, Phylogeny, RT-PCR