Poster: Comparative Genomics

Abs # 919: DNA fingerprint analysis of 8 commercial basil varieties (Ocimum basilicum and sanctum).

Presenter:	Twigg, Paul G, twiggp@unk.edu
Authors	Twigg, Paul G (A)   Nelsen, Kerri  (A)
Affiliations:	(A): University of Nebraska-Kearney

Basil is a common culinary herb and garden ornamental. Its origins are in central Africa and Southeast Asia. It is a source of essential oils used in food, flavorings, and fragrances. Basil has more than 50 species. The most common basil used (sweet basil; Ocimum basilicum) has over 60 varieties resulting from its polymorphic character and ease of cross-pollination. This diversity can result in confusion amongst growers and researchers alike. In this study, we demonstrate an easy mechanism using PCR and RFLP analysis whereby basil varieties can be discriminated even at the intraspecific level. We examined 7 varieties of sweet basil (Ocimum basilicum) and 1 variety of holy basil (Ocimum sanctum). For each variety, total DNA was extracted using a quick procedure which we will outline. From this total DNA, a variable region of the chloroplast DNA (cpDNA) flanked by conserved regions was amplified using PCR for each variety. This cpDNA region spanned a single tRNA gene and its downstream flanking region from the 5' end of the trnL gene to the 5' end of the trnF gene. PCR products of approximately 1200 bp were obtained for each variety using previously published primers. These products were each subjected to RFLP analysis using 6 common restriction endonucleases. Each digested set of fragments produced by a single enzyme for all varieties were electrophoresed to produce a fingerprint. We will present our fingerprint data and emphasize the differences we observed. This project was partially supported by NIH Grant Number 1 P20 RR16469 from the BRIN Program of the National Center for Research Resources, by the UNK Council for Undergraduate Research, and by the UNK Department of Biology.