Comparison of gene expression between MOMT4 transgenic and wild type control poplar

Purpose: This experiment was designed to use RNA-seq approach to provide a priliminary examination whether or not overexpression MOMT4 in hybrid aspen disturbes the expression of lignin and other cell wall biosynthetic genes, followed by extensive qRT-PCR analysis. Our study showed that expression of MOMT4 resulted in a little disturbance on the gene expression involved in lignin biosynthesis. Overall design: The 1st to 15th internodes from the top of 2-month-old hybrid aspen (gray poplar, Populus tremula x Populus alba) stems of MOMT4-0 transgenic and control lines were harvested. After removing their barks, were immediately frozen in liquid nitrogen. About 100 mg stem samples pooled from 2~3 individual transgenic and control plants, respectively, were grounded to fine powder under liquid nitrogen. Total RNAs were extracted using Qiagen RNeasy Plant Mini Kit. The assessment of purity and concentration of each RNA sample, and the strand-specific RNA-Seq library construction were conducted by using the Polar Genomics (Ithaca, NY) service. Sequencing was done using the Illumina HiSeq2500 platform via 50-bp single-end reads of multiplexed RNA samples.