A coordinated pathway for the synthesis of a novel blood-red pigment in a Mauritian nectar.

RNA-Seq of a nectary sample of Nesocodon mauritianus Nesocodon mauritianus was curated and maintained by the College of Biological Sciences Conservatory at the University of Minnesota, St. Paul, Minnesota, U.S.A. Plants were grown in the Cloud Forest + Maritime Climate Room with the temperature maintained between 18 and 24 ˚C and 90-100% relative humidity. RNA was isolated from a single nectary that was manually dissected from a flower 24 hours after anthesis (secretory nectary). The RNA was immediately extracted by mechanical disruption with a microcentrifuge pestle and using an RNAqueous® RNA isolation kit (Ambion, Austin, TX) with Plant RNA Isolation Aid (Ambion, Austin, TX). Agarose gel electrophoresis and UV spectrophotometry were used to assess RNA quality for all samples prior to submission to the University of Minnesota Genomics Center for mRNA isolation, barcoded library creation and Illumina HiSeq 2500 sequencing. A single TruSeq RNA v2 library was created and sequenced via 125 bp, paired-end runs on the HiSeq 2500 using Rapid chemistry on a single lane.