alias: PN.D5
title: Transcriptomic comparisons of five Vitis vinifera cultivars under well-watered and water deficit conditions at harvest.
design: Vitis vinifera L. cultivars Cabernet Sauvignon, Merlot, Pinot Noir, Chardonnay, and Semillon were grown at the University of Nevada, Reno's Experimental Vineyards. Selected blocks within our experimental vineyard were exposed to a moderate water deficit treatment during the 2011 growing and harvest season. Water deficit was assessed through weekly mid-day stem water potentials on fully expanded, mature leaves using a pressure chamber. Drip irrigation was initiated when stem water potentials of the vines reached their target treatment levels.      Soluble sugars (°Brix) and titratable acidity (TA) measurements were measured weekly to assess maturity. A target Brix to TA ratio of 3.5 was used to determine the optimum maturity level for harvesting each cultivar, in September and October 2011. Biological replicates contained at least two cluster from non-adjacent vines. Berry clusters were collected between 11.00 h and 13.00 h. Berry kins were separated from pulp by squeezing and dabbing expelled pulp onto a lab wipe and flash frozen with liquid nitrogen. °Brix and TA were then measured from the combined pulp for each biological replicate.    Frozen skin tissue was ground thoroughly in a Retsch mixer mill (Retsch MM 301, Newtown, PA, USA). Total RNA was extracted with a modified CTAB extraction, followed by an on column DNase digestion treatment according manufactures instructions (Qiagen Mini Kit). RNA quality and quantity were assessed with RiboGreen and an Agilent 2100 Bioanalyzer and RNA LabChip assays according to manufactures instructions.    Library preparation and sequencing was performed by UCLA Neuroscience Genomics Core (UNGC). Sequencing libraries were produced from Total RNA using the TruSeq RNA library prep protocol (Illumina) according to manufactures instructions. TruSeq Version 3.0 sequencing chemistry (Illumina) and HiSeq TruSeq Flowcells were used for sequencing on an Illumina HiSeq 2000 according to manufactures instructions.    The goal of this research project was to profile transcriptional differences between well-watered and water deficit treated cultivars at harvest, following a season of applied water stress. Each cultivar and treatment was comprised of three biological replicates that were each sequenced.
identifiers.namespace: University of Nevada, Reno
identifiers.id: PN.D5