Reprogramming of flagellin receptor responses with surrogate ligands

Characterization of the genome-wide transcriptional responses induced by flg22, Maya1 and Maya2 using RNA sequencing. Total RNA from leaves and roots of wildtype (WT) and fls2 mutant plants were obtained as follows. For leaves, 4-weeks-old Arabidopsis plants grown on soil were treated for 2 hours with DMSO, flg22, Maya1, or Maya2. Total RNA was isolated from pools of more than 15 leaves isolated from independent plants. For roots, 5-day-old seedlings grown vertically on plates were treated for 1 hour with flg22 or Maya2. After 1 hour of incubation, the root tips including the meristematic zone, elongation zone, and a small portion of the maturation zone were isolated. More than 30 independent roots were pooled to extract total RNA for each sample. All the experiments were repeated three times (n=3) except for flg22 treatment in roots (n=2). Details about libraries construction and sequencing strategy are appended below.

基于RNA测序（RNA sequencing）解析flg22、Maya1与Maya2诱导的全基因组转录应答特征。野生型（wildtype, WT）与fls2突变体植株的叶片和根系总RNA提取方法如下：针对叶片样本：将栽培于土壤中的4周龄拟南芥植株用二甲基亚砜（DMSO）、flg22、Maya1或Maya2处理2小时，从多株独立植株采集的超过15片叶片混合后提取总RNA。针对根系样本：将垂直培养于平板上的5日龄幼苗用flg22或Maya2处理1小时；处理1小时后，采集包含分生区、伸长区及少量成熟区的根尖组织，每个样本的总RNA由超过30株独立幼苗的根系混合提取得到。所有实验均设置3次生物学重复（n=3），仅根系flg22处理组设置2次生物学重复（n=2）。文库构建及测序策略的详细信息详见下文。