Methods for studying the evolution of plant reproductive structures: comparative gene expression techniques.

A major component of evolutionary developmental (evo-devo) genetics is the analysis of gene expression patterns in nonmodel species. This comparative approach can take many forms, including reverse-transcriptase polymerase chain reaction, Northern blot hybridization, and in situ hybridization. The choice of technique depends on several issues such as the availability of fresh tissue, as well as the expected expression level and pattern of the candidate gene in question. Although the protocols for these procedures are fairly standard, optimization is often required because of the specific characteristics of the species under analysis. This chapter describes several methods commonly used to determine gene expression patterns in angiosperms, particularly in floral tissues. Suggestions for adapting basic protocols for diverse taxa and troubleshooting are also extensively discussed.