W397 Laser Capture Microdissection and RNASeq Analysis of V. riparia Root Vascular Cylinders During Drought Stress

Date: Sunday, January 15, 2012
Time: 2:30 PM
Room: Pacific Salon 6-7 (2nd Floor)
Kimberely Vaughn , South Dakota State University, Brookings, SD
Vedbar Khadka , South Dakota State University, Brookings, SD
Kathy Mathiason , South Dakota State University, Brookings, SD
Dong He , South Dakota State University, Brookings, SD
Anne Fennell , South Dakota State University, Brookings, SD
Connecting cell-specific gene expression with tissue and plant responses to environmental changes is essential for furthering our functional genomic knowledge base.  Laser capture microdissection (LCM) allows harvest of tissue specific or pure cell populations to analyze transcriptomes driving specific cellular function.  We used LCM to isolate vascular cylinders from drought stressed and well watered roots of V. riparia, a species commonly used in grapevine rootstock and scion breeding.  RNASeq transcriptome analysis was performed on vascular cylinder and whole root tissues.  Sequence reads (100 bp) from vascular cylinder and whole root samples from drought stressed and well watered grapevines were filtered and aligned to the 12x Pinot Noir 40024 genomic sequence and then analyzed for FPKM and differential gene expression.   Analysis of the samples indicated that 14,123 and 20,717 genes were expressed in vascular cylinders and whole root respectively.  Analysis of ABA biosynthesis and metabolism transcripts indicated that 75% and 90% were found in the vascular cylinder and root respectively.  Of these ABA related transcripts, 33% and 12% were differentially expressed in the vascular cylinder and whole roots respectively. Tissue specific expression analyses like these will serve as a resource for grapevine genome functional annotation.