Department of Plant and Soil Sciences, University of Kentucky, Lexington, Kentucky 40546-0312, USA.
Plant Physiol. 2022 Mar 4;188(3):1617-1631. doi: 10.1093/plphys/kiab553.
AGAMOUS-Like 18 (AGL18) is a MADS domain transcription factor (TF) that is structurally related to AGL15. Here we show that, like AGL15, AGL18 can promote somatic embryogenesis (SE) when ectopically expressed in Arabidopsis (Arabidopsis thaliana). Based on loss-of-function mutants, AGL15 and AGL18 have redundant functions in developmental processes such as SE. To understand the nature of this redundancy, we undertook a number of studies to look at the interaction between these factors. We studied the genome-wide direct targets of AGL18 to characterize its roles at the molecular level using chromatin immunoprecipitation (ChIP)-SEQ combined with RNA-SEQ. The results demonstrated that AGL18 binds to thousands of sites in the genome. Comparison of ChIP-SEQ data for AGL15 and AGL18 revealed substantial numbers of genes bound by both AGL15 and AGL18, but there were also differences. Gene ontology analysis revealed that target genes were enriched for seed, embryo, and reproductive development as well as hormone and stress responses. The results also demonstrated that AGL15 and AGL18 interact in a complex regulatory loop, where AGL15 inhibited transcript accumulation of AGL18, while AGL18 increased AGL15 transcript accumulation. Co-immunoprecipitation revealed an interaction between AGL18 and AGL15 in somatic embryo tissue. The binding and expression analyses revealed a complex crosstalk and interactions among embryo TFs and their target genes. In addition, our study also revealed that phosphorylation of AGL18 and AGL15 was crucial for the promotion of SE.
AGAMOUS-Like 18 (AGL18) 是一种与 AGL15 结构相关的 MADS 结构域转录因子 (TF)。在这里,我们表明,与 AGL15 一样,AGL18 在拟南芥中异位表达时可以促进体细胞胚胎发生 (SE)。基于功能丧失突变体,AGL15 和 AGL18 在 SE 等发育过程中具有冗余功能。为了了解这种冗余的本质,我们进行了多项研究来研究这些因子之间的相互作用。我们研究了 AGL18 的全基因组直接靶标,使用染色质免疫沉淀 (ChIP)-SEQ 结合 RNA-SEQ 来表征其在分子水平上的作用。结果表明,AGL18 结合到基因组中的数千个位点。AGL15 和 AGL18 的 ChIP-SEQ 数据比较表明,大量基因被 AGL15 和 AGL18 共同结合,但也存在差异。基因本体分析表明,靶基因富集了种子、胚胎和生殖发育以及激素和应激反应。结果还表明,AGL15 和 AGL18 相互作用形成一个复杂的调控环,其中 AGL15 抑制 AGL18 的转录积累,而 AGL18 增加 AGL15 的转录积累。共免疫沉淀显示 AGL18 和 AGL15 在体细胞胚胎组织中存在相互作用。结合表达分析揭示了胚胎 TF 及其靶基因之间的复杂串扰和相互作用。此外,我们的研究还表明,AGL18 和 AGL15 的磷酸化对 SE 的促进至关重要。