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Clematis  florida  is a popular ornamental vine species known for diverse colors and shapes of its flowers but not for scent. Here we investigated the composition and biosynthesis of floral scent in ‘Kaiser’, a fragrant cultivar of  C. florida  that has sterile flowers. Volatile profiling revealed that flowers of ‘Kaiser’ emit more than 20 compounds, with monoterpenes being most abundant. Among the three floral organs, namely sepals, transformed-petals, and ovaries, ovaries had the highest rates of total volatile emission. To determine the molecular mechanism underlying floral scent biosynthesis in ‘Kaiser’, we sequenced a flower transcriptome and searched the transcriptome for terpene synthase genes ( TPSs ), which are key genes for terpene biosynthesis. Among the  TPS  genes identified, three were putative intact full-length genes and were designated  CfTPS1 ,  CfTPS2 , and  CfTPS3 . Phylogenetic analysis placed  CfTPS1 ,  CfTPS2 , and  CfTPS3  to the  TPS-g ,  TPS-b , and  TPS-a  subfamily, respectively. Through in vitro enzyme assays with  Escherichia coli -expressed recombinant proteins, both  CfTPS1  and  CfTPS2  were demonstrated to catalyze the conversion of geranyl diphosphate to linalool, the most abundant constituent of  C. florida  floral scent. In addition,  CfTPS1  and  CfTPS2  produced the sesquiterpene nerolidol from ( E,E )-farnesyl diphosphate.  CfTPS3  showed sesquiterpene synthase activity and produced multiple products  in vitro . All three  CfTPS  genes showed higher levels of expression in sepals than those in transformed-petals and ovaries. Our results show that despite being sterile, the flowers of ‘Kaiser’ have normal mechanisms for floral scent biosynthesis that make the flowers fragrant.

Composition and Biosynthesis of Scent Compounds from Sterile Flowers of an Ornamental Plant Clematis florida cv. ‘Kaiser’