The SEEL Motif and Members of the MYB-related REVEILLE Transcription Factor Family are Important for the Expression of LORELEI in the Synergid Cells of the Arabidopsis Female Gametophyte

Abstract

Synergid cells in the micropylar end of the female gametophyte are required for critical cell-cell signaling interactions between the pollen tube and the ovule that precede double fertilization and seed formation in flowering plants. LORELEI (LRE) encodes a GPI-anchored protein that is expressed primarily in the synergid cells, and together with FERONIA, a receptor-like kinase, it controls pollen tube reception by the receptive synergid cell. Still, how LRE expression is controlled in synergid cells remains poorly characterized. We identified candidate cis-regulatory elements enriched in LRE and other synergid cell-expressed genes. One of the candidate motifs (‘TAATATCT’) in the LRE promoter was an uncharacterized variant of the Evening Element motif that we named as the Short Evening Element-like (SEEL) motif. Deletion or point mutations in the SEEL motif of the LRE promoter resulted in decreased reporter expression in synergid cells, demonstrating that the SEEL motif is important for expression of LRE in synergid cells. Additionally, we found that LRE expression is decreased in the loss of function mutants of REVEILLE (RVE) transcription factors, which are clock genes known to bind the SEEL and other closely related motifs. We propose that RVE transcription factors regulate LRE expression in synergid cells by binding to the SEEL motif in the LRE promoter. Identification of a cis-regulatory element and transcription factors involved in the expression of LRE will serve as a foundation to characterize the gene regulatory networks in synergid cells and investigate the potential connection between circadian rhythm and fertilization.One sentence summaryA newly identified SEEL motif in the promoter of LORELEI and at least three members of the REVEILLE transcription factor family are important for LORELEI expression in synergid cells of the Arabidopsis female gametophyte.

Discovery of a cis-regulatory element SaeM involved in dynamic regulation of synergid-specific MYB98

“…The element has been characterized as the MYB98 recognition site itself , suggesting the regulation of MYB98 in a positive-feedback loop. Furthermore, a more recent study by Noble et al (2022) reported an 8-bp-long SEEL element, "TAATATCT" in the LORELEI (LRE) promoter, deletion or mutation of which led to reduced expression of the gene in SCs. The element is recognized by MYB-related REVEILLE (RVE) TFs.…”

“…Among 64 exclusively SC-expressed genes, about 66% harbored a SaeM-like element within their 1 kb promoter region. Among them were those that had been experimentally verified to be SC-specific, such as MYB98 (Kasahara et al, 2005;Punwani et al, 2007) LUREs (1.1, 1.2, 1.3, 1.4, 1.7, and 1.8) (Takeuchi and Higashiyama, 2012;Zhong et al, 2019), LORELEI (Noble et al, 2022), DD11 and DD18 (Punwani et al, 2007). (Figure 5A and Supplementary Table S2).…”