LMD and dual RNA-seq of the L. maculans - B. napus infection site

In this project, LMD was used to isolate 100 micron sections of tissue directly at the host pathogen interface of B. napus and L. maculans. B. napus near isogenic lines (one containing resistance gene Rlm2) were challenged with L. maculans (AvrLm2). We used dual RNA-seq at 1 and 3 days post inoculation to identify genes activated early in the defense response.

本研究中，研究人员采用激光显微切割（Laser Microdissection, LMD）技术，直接在甘蓝型油菜（Brassica napus, B. napus）与黑胫病菌（Leptosphaeria maculans, L. maculans）的宿主-病原菌互作界面处，分离得到100微米厚的组织切片。选取携带抗病基因Rlm2（resistance gene Rlm2）的甘蓝型油菜近等基因系（near isogenic lines, NILs），以携带无毒基因AvrLm2的黑胫病菌进行接种。本研究在接种后1天和3天两个时间点开展双转录组测序（dual RNA-seq），以鉴定宿主防御反应早期激活的基因。