Transcriptome profiling, and cloning and characterization of the main monoterpene synthases of Coriandrum sativum L
A transcript library for coriander mericarp was produced. The main monoterpene synthase genes, CsγTRPS and CsLINS were cloned, expressed in bacteria, and functionally characterized. •Coriandrum sativum transcriptome was sequenced and a transcript library produced.•Isoprenoid biosynthetic transcript...
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Veröffentlicht in: | Phytochemistry (Oxford) 2014-06, Vol.102, p.64-73 |
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Zusammenfassung: | A transcript library for coriander mericarp was produced. The main monoterpene synthase genes, CsγTRPS and CsLINS were cloned, expressed in bacteria, and functionally characterized.
•Coriandrum sativum transcriptome was sequenced and a transcript library produced.•Isoprenoid biosynthetic transcript abundance was analyzed.•Main C. sativum monoterpene synthases were cloned and functionally characterized.•Linalool synthase only produces the (S) stereoisomer of linalool.•γ-Terpinene synthase yields γ-terpinene as its major product.
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. |
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ISSN: | 0031-9422 1873-3700 |
DOI: | 10.1016/j.phytochem.2014.02.016 |