title: Overexpression of a synthetic insect-plant geranyl pyrophosphate synthase gene in Camelina sativa alters plant growth and terpene biosynthesis.
abstract: A novel plastidial homodimeric insect-plant geranyl pyrophosphate synthase gene is synthesized from three different cDNA origins. Its overexpression in Camelina sativa effectively alters plant development and terpenoid metabolism. Geranyl pyrophosphate synthase (GPPS) converts one isopentenyl pyrophosphate and dimethylallyl pyrophosphate to GPP. Here, we report a synthetic insect-plant GPPS gene and effects of its overexpression on plant growth and terpenoid metabolism of Camelina sativa. We synthesized a 1353-bp cDNA, namely PTP-MpGPPS. This synthetic cDNA was composed of a 1086-bp cDNA fragment encoding a small GPPS isomer of the aphid Myzus persicae (Mp), 240-bp Arabidopsis thaliana cDNA fragment encoding a plastidial transit peptide (PTP), and a 27-bp short cDNA fragment encoding a human influenza hemagglutinin tag peptide. Structural modeling showed that the deduced protein was a homodimeric prenyltransferase. Confocal microscopy analysis demonstrated that the PTP-MpGPPS fused with green florescent protein was localized in the plastids. The synthetic PTP-MpGPPS cDNA driven by 2 × 35S promoters was introduced into Camelina (Camelina sativa) by Agrobacterium-mediated transformation and its overexpression in transgenic plants were demonstrated by western blot. T2 and T3 progeny of transgenic plants developed larger leaves, grew more and longer internodes, and flowered earlier than wild-type plants. Metabolic analysis showed that the levels of beta-amyrin and campesterol were higher in tissues of transgenic plants than in those of wild-type plants. Fast isoprene sensor analysis demonstrated that transgenic Camelina plants emitted significantly less isoprene than wild-type plants. In addition, transcriptional analyses revealed that the expression levels of gibberellic acid and brassinosteroids-responsive genes were higher in transgenic plants than in wild-type plants. Taken together, these data demonstrated that this novel synthetic insect-plant GPPS cDNA was effective to improve growth traits and alter terpenoid metabolism of Camelina.
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, 27695, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Xi
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, 27695, USA.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Rossi
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, 27695, USA.
authors.forename: Xiuli
authors.initials: X
authors.lastname: Lin
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, 27695, USA. dxie@ncsu.edu.
authors.forename: De-Yu
authors.initials: DY
authors.lastname: Xie
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000235:genetics; D015153:Blotting, Western; D060406:Brassinosteroids / Q000494:pharmacology; D028244:Camellia / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004122:Dimethylallyltranstransferase / Q000235:genetics / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000235:genetics*; D005875:Gibberellins / Q000494:pharmacology; D006801:Humans; D007313:Insecta / Q000235:genetics; D008025:Ligases / Q000235:genetics* / Q000378:metabolism; D018613:Microscopy, Confocal; D010937:Plant Growth Regulators / Q000494:pharmacology; D030821:Plants, Genetically Modified; D018087:Plastids / Q000201:enzymology; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013729:Terpenes / Q000378:metabolism*
chemical_list: D060406:Brassinosteroids; D005875:Gibberellins; D010937:Plant Growth Regulators; D011106:Polyisoprenyl Phosphates; D011993:Recombinant Fusion Proteins; D013729:Terpenes; C015234:geranyl pyrophosphate; C007842:gibberellic acid; D004122:Dimethylallyltranstransferase; D008025:Ligases
keywords: Camelina sativa; Geranyl pyrophosphate synthase; Isoprene; Metabolic engineering; Prenyltransferase; Synthetic biology; Terpene