germplasm name: seyval (seyve-villard 5-276) organism part: ud: 0000001 second order axillary bud cell type: UD:0000001 second order axillary bud development stage: UD:0000002 paradormancy age: 58 days after bud break growth media: peat/perlite/soil
Growth Description: Three to four shoots were trained vertically on spur pruned greenhouse grown vines. Vines were watered two times daily at 8:30 am and 3:00 pm. Environmental Conditions: Vines were grown in greenhouse at 44N latitude with 15h day using supplemental lighting (Na halide lamps) to maintain 600 micro mole m-2s-1.
Modified Pine Buffer RNA Extraction Procedure
Affy-Eukaryotic Target Preparation
Affy- Eukaryotic Target Hybridization
Affy- Array Scanning
Potted, spur-pruned two to six-year-old vines were removed from cold storage (Seyval on 3/19/2007; V. riparia on 3/26/2007) and grown under a LD (15 h) at 25/20 + 3C day/night temperatures (D/N). When vines reached 12-15 nodes (4/25/2007) they were randomized into two replicated treatment groups, long photoperiod (LD, 15h) or short photoperiod (SD,13h). Five days after this (4/30/2007) the SD (13h, 25/20C) photoperiod treatment was started and LD (15h 25/20C) treatment continued. SD treatment was imposed with automated photoperiod system (VRE Greenhouse Systems). At 1, 3, 7, 14, 21, 28 and 42 days of additional LD or SD, buds were harvested from nodes 3 to 12 from shoot base, immediately frozen in liquid nitrogen, and placed at -80C for future RNA, protein, and metabolite extraction. Three replications (5 vines/replication) were harvested between 05/07-06/07. After bud harvests, the pruned vines were returned to LD and monitored for bud endodormancy. The endodormant vines were identified after 28 days and moved to cold storage. The nondormant vines were allowed to grow again and induced into dormancy at a later date.
The MAS5.0 Normalization is performed using the justMAS function in the simpleaffy package of Bioconductor (www.bioconductor.org). The target value is 500 for all cases. The code was developed using R2.4.1, Bioconductor 1.9, and simpleaffy 2.8.0. The CDF files were downloaded from the Bioconductor website at http://www.bioconductor.org/packages/release/AffymetrixChip.html. The normalization scripts are available from the PLEXdb team by request.
Aug 04, 2009
Last update date
Aug 05, 2009
PLEXdb Curator, Sudhansu Dash
Iowa State University
Virtual Reality Application Center
1023 Crop Genomes Informatics Lab (Iowa State University)