Molecular basis of the biodiversity of the essential oil production in Savia sp.
2007
Ninga, E.
The genes encoding the enzymes responsible for the production of essential oils in Salvia fruticosa have been shown to exhibit significant variation due to their geographical position. The objective of this M.Sc. was to investigate the natural diversity of the key biosynthetic enzymes (terpene synthases) and to obtain a detailed understanding of the molecular and structural basis of this variation. The C-terminal domain (the domain responsible for the catalytic specificity) of genes that encode cineole synthase from Salvia fruticosa was amplified using PCR. To investigate biodiversity, three different sources were used. One was the cDNA constructed from plant growing in MAICh`s green houses and the other two were first strand cDNA made out of RNA extracted from plants, belonging to a different population on the Eastern part of Crete, that has shown to exhibit a different monoterpene profile. Comparisons were made between these genes and the same region of other monoterpene synthases found in nature. Site directed mutagenesis was employed to introduce different mutations in SfCinS1 and SpSabS1. These explored the plasticity of different residues in these sites. Furthermore, we attempted to convert the highly specific SpSabS1 to the promiscuous SoSabS1 by introducing two different amino acid substitutions, and confirmed that residues further away from the active site can play a significant role in the catalytic specificity of terpene synthases.
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