利用转录组测序与关联作图联合构建尾叶桉响应青枯病菌侵染过程的遗传调控网络

Investigation of stress resistance mechanism is the basis of breeding for tree disease resistance. Our project aims to reveal the genetic regulatory networks (GRNs) response to plants biotic stress. By using the Eucalyptus-Eucalyptus bacterial wilt (Eucalyptus urophylla and Ralstonia solanacearum) interaction system, single molecule real-time sequencing and strand-specific RNA Sequencing (ssRNA-seq) technology, we analyze the changes of Eucalyptus transcriptome, including the genes (transcription factors, TFs), miRNAs, and the alternative splicing of genes. TF-target, miRNA-target, and miRNA-TF-target regulatory network in response to biotic stress will be investigated based on the gene regulation website and degradome profiling. Based on the genome-wide sequencing and deep sequencing, SNP-based association mapping was used to validate deeply analyze the constructed GRNs. In the course of the investigation, single molecule real-time sequencing, ssRNA-seq and SNP-based association mapping will be firstly combined to study the function of GRNs in Eucalyptus response to biotic stress. This study will provide new insights into the regulation mechanisms of GRNs in Eucalyptus response to biotic stress, and provide candidate genes to genetic engineering breeding with the goals of improving the tree disease resistance.

揭示林木响应生物胁迫机制是林木抗逆育种的基础。本项目围绕遗传调控网络在生物胁迫中的响应机制这一研究目标，以尾叶桉与青枯病菌互作体系为材料，利用单分子实时测序技术研究基因可变剪接在林木响应生物胁迫中的作用；利用链特异性转录组测序技术筛选抗病相关基因(转录因子)和miRNA；利用转录因子靶基因预测网站及降解组数据初步构建转录因子-靶基因、miRNA-靶基因及miRNA-转录因子-靶基因遗传调控网络；结合本课题组前期完成的尾叶桉全基因组测序及重测序工作，利用SNP关联作图对构建的遗传调控网络进行深度解析与验证。本项目将在林木响应逆境胁迫中首次整合单分子实时测序技术，链特异性转录组分析及SNP关联作图技术对遗传调控网络在响应生物胁迫中的作用进行研究，为林木抗病分子机理的研究提供新思路，同时为林木抗病基因工程育种提供重要的基因资源。