Plant Ecology and Evolution, Department of Ecology and Genetics, Evolutionary Biology Centre and the Linnean Centre for Plant Biology in Uppsala, Uppsala University, Norbyvägen 18D, SE-75236, Uppsala, Sweden.
Physiological Botany, Department of Organismal Biology, Linnean Centre for Plant Biology in Uppsala, Uppsala University, Ulls Väg 24E, SE-756 51, Uppsala, Sweden.
New Phytol. 2021 Oct;232(2):595-609. doi: 10.1111/nph.17653. Epub 2021 Aug 19.
Previous studies of plant circadian clock evolution have often relied on clock models and genes defined in Arabidopsis. These studies identified homologues with seemingly conserved function, as well as frequent gene loss. In the present study, we aimed to identify candidate clock genes in the liverwort Marchantia polymorpha using a more unbiased approach. To identify genes with circadian rhythm we sequenced the transcriptomes of gemmalings in a time series in constant light conditions. Subsequently, we performed functional studies using loss-of-function mutants and gene expression reporters. Among the genes displaying circadian rhythm, a homologue to the transcriptional co-repressor Arabidopsis DE-ETIOLATED1 showed high amplitude and morning phase. Because AtDET1 is arrhythmic and associated with the morning gene function of AtCCA1/LHY, that lack a homologue in liverworts, we functionally studied DET1 in M. polymorpha. We found that the circadian rhythm of MpDET1 expression is disrupted in loss-of-function mutants of core clock genes and putative evening-complex genes. MpDET1 knock-down in turn results in altered circadian rhythm of nyctinastic thallus movement and clock gene expression. We could not detect any effect of MpDET1 knock-down on circadian response to light, suggesting that MpDET1 has a yet unknown function in the M. polymorpha circadian clock.
先前关于植物生物钟进化的研究通常依赖于在拟南芥中定义的生物钟模型和基因。这些研究鉴定了具有看似保守功能的同源物,以及频繁的基因丢失。在本研究中,我们旨在使用更无偏倚的方法在苔类植物 Marchantia polymorpha 中鉴定候选生物钟基因。为了鉴定具有昼夜节律的基因,我们在恒定光照条件下的时间序列中对芽生体的转录组进行了测序。随后,我们使用功能丧失突变体和基因表达报告基因进行了功能研究。在所显示昼夜节律的基因中,与拟南芥 DE-ETIOLATED1 转录共抑制因子具有同源性的基因表现出高振幅和早晨相位。由于 AtDET1 是无节律的,并且与缺乏在苔类植物中同源物的 AtCCA1/LHY 的早晨基因功能相关,因此我们在 M. polymorpha 中对 DET1 进行了功能研究。我们发现,在核心生物钟基因和推定的 evening-complex 基因的功能丧失突变体中,MpDET1 表达的昼夜节律被打乱。反过来,MpDET1 的敲低导致昼夜节律的扭转型叶运动和时钟基因表达发生改变。我们没有检测到 MpDET1 敲低对光的昼夜节律反应有任何影响,这表明 MpDET1 在 M. polymorpha 生物钟中具有未知的功能。