Abstract
Bioluminescence, which living organisms such as fireflies emit light, has been studied extensively for over half a century. This intriguing reaction, having its origins in nature where glowing insects can signal things such as attraction or defense, is now widely used in biotechnology with applications of bioluminescence and chemiluminescence. Luciferase, a key enzyme in this reaction, has been well characterized; however, the enzymes involved in the biosynthetic pathway of its substrate, luciferin, remains unsolved at present. To elucidate the luciferin metabolism, we performed a de novo transcriptome analysis using larvae of the firefly species, Luciola aquatilis. Here, a comparative analysis is performed with the model coleopteran insect Tribolium casteneum to elucidate the metabolic pathways in L. aquatilis. Based on a template luciferin biosynthetic pathway, combined with a range of protein and pathway databases, and various prediction tools for functional annotation, the candidate genes, enzymes, and biochemical reactions involved in luciferin metabolism are proposed for L. aquatilis. The candidate gene expression is validated in the adult L. aquatilis using reverse transcription PCR (RT-PCR). This study provides useful information on the bio-production of luciferin in the firefly and will benefit to future applications of the valuable firefly bioluminescence system.
Highlights
The firefly is a bioluminescent beetle belonging to the Order Coleoptera, Family Lampyridae
Over 100 genera and 2,000 species of fireflies have been reported around the world both in temperate and tropical areas (McDermott, 1964; McDermott, 1966; Branham, 2010). Of these 100 genera, Photinus and Photuris from North America (Lewis & Cratsley, 2008; Faust, De Cock & Lewis, 2012; Stansbury & Moczek, 2014; Martin et al, 2015; Sander & Hall, 2015) and Pyrocoelia (Fu et al, 2006a) and Luciola (Tsutomu, Hiroki & Eiichi, 1989; Fu et al, 2006b; Oba et al, 2006; Ohtsuki et al, 2008; Oba & Kainuma, 2009; How to cite this article Vongsangnak et al (2016), Transcriptome analysis reveals candidate genes involved in luciferin metabolism in Luciola aquatilis (Coleoptera: Lampyridae)
Bioluminescence is catalyzed by a luciferase enzyme in the presence of O2, ATP, and Mg2+ (Deluca, 1976; Baldwin, 1996) in a two-step reaction; D-luciferin is adenylated by ATP at the luciferase active site and converted into luciferyl-adenosine monophosphate
Summary
The firefly is a bioluminescent beetle belonging to the Order Coleoptera, Family Lampyridae. Bioluminescence is catalyzed by a luciferase enzyme in the presence of O2, ATP, and Mg2+ (Deluca, 1976; Baldwin, 1996) in a two-step reaction; D-luciferin is adenylated by ATP at the luciferase active site and converted into luciferyl-adenosine monophosphate (luciferyl-AMP). This luciferyl-AMP is oxidized and converted into excited state oxyluciferin. This excited state oxyluciferin later returns to its ground state by the emission of a visible photon, thereby generating visible light (Fraga, 2008; Naumov et al, 2009; Inouye, 2010; Pinto da Silva, Santos & Esteves da Silva, 2012)
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