Transcriptome profiling, and cloning and characterization of the main monoterpene synthases of Coriandrum sativum L.

Abstract

Terpenoids are a large and diverse class of specialized metabolites that are essential for the growth and development of plants, and have tremendous industrial applications. The mericarps of Coriandrum sativum L. (coriander) produce an essential oil (EO) rich in monoterpenes, volatile C10 terpenoids. To investigate EO metabolism, the transcriptome of coriander mericarps, at three developmental stages (early, mid, late) was sequenced via Illumina technology and a transcript library was produced. To validate the usability of the transcriptome sequences, two terpene synthase candidate genes, CsγTRPS and CsLINS, encoding 558 and 562 amino acid proteins were expressed in bacteria, and the recombinant proteins purified by Ni-NTA affinity chromatography. The 65.16 (CsγTRPS) and 65.91 (CsLINS)kDa recombinant proteins catalyzed the conversion of geranyl diphosphate, the precursor to monoterpenes, to γ-terpinene and (S)-linalool, respectively, with apparent Vmax and Km values of 2.24±0.16 (CsγTRPS); 19.63±1.05 (CsLINS)pkat/mg and 66.25±13 (CsγTRPS); 2.5±0.6 (CsLINS)μM, respectively. Together, CsγTRPS and CsLINS account for the majority of EO constituents in coriander mericarps. Investigation of the coriander transcriptome, and knowledge gained from these experiments will facilitate future studies concerning essential and fatty acid oil production in coriander. They also enable efforts to improve the coriander oils through metabolic engineering or plant breeding.

Functional characterization of a new terpene synthase from Plectranthus amboinicus

“…Plants in the Lamiaceae family are known to produce a large variety of terpenoids, and this diversity could be due to expression of multiple terpene synthases and formation of multiple products by individual terpene synthases as previously reported [12][13][14][15]. Terpene synthase genes have been previously isolated and characterized from several important Lamiaceae members including Thymus caespititius [13,14], T. vulgaris [16], Coriandrum sativum L. [15], Origanum vulgarae [12] and Salvia officinalis [17]; however the terpene synthases responsible for terpenoids production in P. amboinicus have not been functionally characterized. Thus, the main objectives of this present study were to isolate and clone the full-length transcript of P. amboinicus monoterpene synthase, and to functionally express and characterize the recombinant terpene synthase in both in vivo and in vitro systems.…”

Expression, crystallization and structure elucidation of γ-terpinene synthase fromThymus vulgaris

“…To date, TPSs have been described as multifunctional enzymes, with single amino-acid substitutions being sufficient to result in an altered metabolic profile (Kö llner et al, 2004), and a number of TPSs and other terpene-modifying enzymes have been described (Keszei et al, 2008). Recently, TPS genes have been identified and characterized from several plant species from the Lamiaceae family (summarized in Lima et al, 2013): TPS from Lavandula (Muñ oz- Bertomeu et al, 2008;Lane et al, 2010), Mentha (Turner & Croteau, 2004), Origanum (Crocoll et al, 2010;Lukas et al, 2010), Perilla (Ito & Honda, 2007), Coriandrum (Galata et al, 2014) and Salvia (Kampranis et al, 2007;Schmiderer et al, 2010). Within the genus Thymus, only TPS from T. caespititius (including several isoforms) has been described previously (Lima et al, 2010(Lima et al, , 2011(Lima et al, , 2013Mendes et al, 2014).…”

Transcriptome-based investigation of cirrus development and identifying microsatellite markers in rattan (Daemonorops jenkinsiana)

“…Although the previous studies13141516171819 showed the analysis results based on two biological replicates were acceptable and relatively reliable at some aspects and extents, we had still conducted two investigations of data quality and then approved that data were useful and practical. Our analyses included the calculation of sequence randomness distribution (see Supplementary Fig.…”