Key Laboratory of Plant Molecular Physiology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Plant Cell Physiol. 2018 Apr 1;59(4):744-755. doi: 10.1093/pcp/pcx168.
Auxin-induced callus formation represents an important cell reprogramming process during in vitro regeneration of plants, in which the pericycle or pericycle-like cells within plant organs are reprogrammed into the pluripotent cell mass termed callus that is generally required for subsequent regeneration of root or shoot. However, the molecular events behind cell reprogramming during auxin-induced callus formation are largely elusive. We previously identified that auxin-induced LATERAL ORGAN BOUNDARIES DOMAIN (LBD) transcription factors act as the master regulators to trigger auxin-induced callus formation. Here, by ChIP-seq (chromatin immunoprecipitation-based sequencing) and RNA sequencing approaches, we identified the potential LBD29 target genes at the genome-wide level and outlined the molecular events of LBD-triggered cell reprogramming during callus formation. We showed that LBD29 preferentially bound to the G-box (CACGTG) and TGGGC[C/T] motifs and potentially targeted >350 genes, among which the genes related to methylation, reactive oxygen species (ROS) metabolism, cell wall hydrolysis and lipid metabolism were rapidly activated, while most of the light-responsive genes were suppressed by LBD29. Further examination of a few representative genes validated that they were targeted by LBD29 and participated in the regulation of cell reprogramming during callus formation. Our data not only outline a framework of the early molecular events behind auxin-induced cell reprogramming of callus formation, but also provide a valuable resource for identification of genes that regulate cell fate switch during in vitro regeneration of plants.
生长素诱导的愈伤组织形成代表了植物体外再生过程中的一个重要的细胞重编程过程,在此过程中,植物器官内的中柱或中柱样细胞被重编程为多能细胞团,称为愈伤组织,这通常是随后再生根或芽所必需的。然而,生长素诱导的愈伤组织形成过程中细胞重编程背后的分子事件在很大程度上还不清楚。我们之前已经确定,生长素诱导的侧生器官边界域(LBD)转录因子作为主调控因子,触发生长素诱导的愈伤组织形成。在这里,我们通过 ChIP-seq(基于染色质免疫沉淀的测序)和 RNA 测序方法,在全基因组水平上鉴定了潜在的 LBD29 靶基因,并概述了 LBD 触发的细胞重编程在愈伤组织形成过程中的分子事件。我们表明,LBD29 优先结合 G 框(CACGTG)和 TGGGC[C/T]基序,并可能靶向 >350 个基因,其中与甲基化、活性氧(ROS)代谢、细胞壁水解和脂质代谢相关的基因被迅速激活,而大多数光响应基因则被 LBD29 抑制。对几个有代表性的基因的进一步研究证实,它们是由 LBD29 靶向的,并参与了愈伤组织形成过程中的细胞重编程调控。我们的数据不仅概述了生长素诱导的愈伤组织形成过程中细胞重编程的早期分子事件的框架,而且为鉴定植物体外再生过程中调节细胞命运转变的基因提供了有价值的资源。