RNAseq of Bignonia magnifica cambium and differentiating xylem: self-supporting phase
Secondary xylem-forming tissues (cambium and differentiating xylem) were collected in April 2018 by peeling the bark and gently scraping the exposed xylem of 30 cm samples collected along the stems. Tissues were immediately frozen in liquid nitrogen and stored at -80 C. Before tissue harvest, each stem segment had its base cut and stored in 50 % alcohol to determine the xylem phase, self-supporting or lianescent. Segments presenting dubious or transition anatomy, and the segment next to it, were discarded. Six pools of self-supporting and six pools of lianescent secondary xylem-forming tissues, composed of 8-19 segments each, were collected for sequencing and differential expression analysis. Total RNA was extracted after sample lyophilization using ReliaPrepTM RNA Tissue Miniprep System (Promega Corporation) following manufacturer protocol. Libraries were constructed using Illumina TruSeq Stranded mRNA Sample Prep LT Protocol (Illumina).