Construction of Yeast One-Hybird Library for Screening of Eugenol Synthase Gene Bait vectors in Rosa chinensis

Abstract

Roses are cultivated as ornamental plants in gardens, cut flowers and potted flowers, but also serviced in the perfume industry. Rose flowers product different volatile compounds including eugenol, being floral attractant for pollinators and used as an important composition of essential oil. Here, the eugenol synthase gene (RcEGS1) promoter was cloned from R. chinensis 'Old Blush'. The sequences of RcEGS1 promoter were 1776bp, contained the conserved elements, and predicted to be with several cis-regulatory elements involved in secondary metabolism of plants. Yeast one-hybrid library of R. chinensis 'Old Blush' at flower opening was constructed with the three-frame reading. The library capacity was more than 1.12×107CFU. The length of fragments inserted was 1 Kb to 3 Kb, which is fully meeting the demand of selecting the yeast library. The bait yeast vectors with cis-transcription elements MBS-box, G-box and one fragment bait yeast vectors were constructed, respectively. Furthermore, the transcript factors R2R3-type MYB and bHLH3 were screened using the yeast one-hybrid system. The results suggested the two TFs might play an important role in the pathway of volatile biosynthesis in roses.