Gene expression in Jaltomata (species herrerae and procumbens) nectaries

RNA-Seq of nectary and leaf samples of Jaltomata herrerae, and Jaltomata procumbens Overall design: Jaltomata procumbens and Jaltomata herrerae plants were grown in Sun Gro LC8 soil in the MAES/MDA Plant Growth Facilities at the University of Minnesota with typical daytime highs of 25C and nighttime lows of 15C. Three types of RNA samples were separately prepared in triplicate from the floral nectaries of both species, including: 'pre-secretory' (~24-48 hours prior to anthesis/nectar secretion), 'secretory' (full anthesis, at ~3-6 hours after dawn), and 'post-secretory' (after petal abscission). RNA from fully expanded leaf tissue was also isolated in triplicate for both species. All tissues were manually dissected by hand with the RNA being immediately extracted by mechanical disruption with a microcentrifuge pestle and using an RNAqueous® RNA isolation kit (Thermo Fisher) with Plant RNA Isolation Aid (Thermo Fisher). 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 NovaSeq 6000 sequencing. Twenty-four TruSeq stranded mRNA libraries were created (triplicate samples for nectaries at three timepoints each, as well as leaf tissue, for each species) and sequenced via 150 bp, paired-end runs on the NovaSeq 6000 using Rapid chemistry. All libraries were pooled and sequenced. All expected barcodes were detected and mean depth for the libraries was = 20 M reads. The average quality scores were all above Q30.