Molecular Analysis of a Vanilla Hybrid Cultivated in Costa Rica

This chapter presents DNA sequence data confirming the hybrid origin of a Vanilla cultivated in Costa Rica. The plant used for this analysis was from a plantation in Costa Rica and was considered to be a hybrid plant. Phylogenetic analysis of the psaB DNA sequences revealed the close relationship of the sequence from the Costa Rican hybrid plant with that of V. planifolia and V. tahitensis. The PCR products were directly cloned into the pGEM‐T Easy vector and plasmids from individual transformed colonies were sequenced. The PCR products were gel purified and sequenced using the amplification primers and one additional primer. The internal transcribed spacer (ITS) region of Vanilla planifolia, Vanilla pompona, and the presumed hybrid, was PCR amplified, cloned into a bacterial plasmid, and sequences of 6 to 28 clones per plant sample were obtained as described in the chapter.