Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (3): 590-597.doi: 10.3864/j.issn.0578-1752.2012.03.023

• RESEARCH NOTES • Previous Articles     Next Articles

Cloning and Expression Analysis of Eugenol Synthase Gene (RhEGS1) in Cut Rose (Rosa hybrida)

 YAN  Hui-Jun, ZHANG  Hao, JIAN  Hong-Ying, WANG  Qi-Gang, QIU  Xian-Qin, ZHANG  Ting, TANG  Kai-Xue   

  1. 1.云南省农业科学院花卉研究所/云南省花卉育种重点实验室,昆明 650205
  • Received:2011-08-10 Online:2012-02-01 Published:2011-12-01

Abstract:  【Objective】 The objective of this study was to clone and identify eugenol synthase gene (RhEGS1) related to flower scent metabolism in rose. The results will provide a foundation for further studying the gene’s function and biochemical characters. 【Method】 The full-length cDNA sequence of RhEGS1 was amplified by degenerate primers based on EGS from other plants and RACE cloning technology. The expression patterns of RhEGS1 in different tissues and different developmental stages were analyzed by semi-quantitative RT-PCR. Furthermore, prokaryotic expression vector was successfully constructed by Gateway cloning technology. 【Result】 The full-length of RhEGS1 was 1 207 bp, containing a 927 bp ORF which encoded a 83.87% and 81.55% homology putative EGS protein with 309 amino acids. Phylogeny analysis showed that RhEGS1 shared with CbEGS2 in Clarkia breweri and PhEGS1 in Petunia hybrida, respectively. RhEGS1 had a highest transcript level in stamens at blooming stage, and lower levels in those of bud at senescence stage. The expression of predicted 35 kD recombinant protein was induced by isopropyl β-D-thiogalacto-pyranoside (IPTG) at a final concentration of 0.5 mmol•L-1 for 4 h at 37℃. 【Conclusion】 The RhEGS1 was isolated from rose stamen and shared the conserved domains with other EGS. It showed the highest transcript level in stamens at blooming stage, and demonstrated active expression in prokaryotic cells.

Key words: rose (Rosa hybrida), RhEGS1, RT-PCR, Gateway clone, prokaryotic expression

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