Zhang, Y., et al. · 2026 · Nature Plants research
doi:10.1038/s41477-026-02334-4 PMID:42399668
| Gene ID | Name | Evidence / Role | Function (this paper) |
|---|---|---|---|
| Mp4g15890 | MpFER | experimental subject | The single Marchantia CrRLK1L receptor kinase, shown here to be the MpRALF1 receptor in spermatozoids. MpFER is enriched in antheridiophores (proMpFER:GUS) and the MpFER-Flag protein localizes to sperm flagella; CRISPR Mpfer knockouts and amiRNA Ri-MpFER lines had reduced sperm ROS and motility that MpRALF1 peptide could no longer rescue. MpFER binds MpRALF1 (pulldown with MBP-MpFERecd, luciferase complementation with MpFER-deltaC) preferentially via the peptide's N-terminal region and forms a complex with MpLLG (CoIP); the MpFER-G282A substitution did not affect MpRALF1 binding, indicating a binding mode distinct from angiosperm RALF23-FER-LLG2 and RALF4-ANX1/BUPS1-LLG3. Gene ID not printed in the paper: resolved from the paper's own MpFER-RT-F/R qPCR primers (Suppl. Table 2), which match Mp4g15890 exactly, in agreement with Mecchia et al. 2022. |
| Mp4g22100 | MpLRE2 | experimental subject | GPI-anchored LORELEI-like co-receptor, renamed in this paper from MpLRE2 (the MpLRE gene that retains the GPI-anchoring site). MpLLG is enriched in antheridiophores and its Flag/eGFP fusions accumulate in sperm flagella; CRISPR Mpllg and amiRNA Ri-MpLLG lines showed reduced sperm ROS and motility that exogenous MpRALF1 failed to rescue. MpLLG co-immunoprecipitates with MpFERecd, binds MpRALF1 preferentially through the peptide C-terminal 38-aa region, and the MpLLG-N100A and MpLLG-A126Y substitutions disturb that binding, defining an MpRALF1-MpFER-MpLLG module upstream of ROS production. Gene ID not printed in the paper: resolved from the paper's own MpLLG-RT-F/R qPCR primers (Suppl. Table 2), which match Mp4g22100 (= MpLRE2 of Mecchia et al. 2022) exactly. |
| Mp7g07270 | MpRALF3 paper
MpRALF1 nomenclature |
experimental subject | Sperm-derived RAPID ALKALINIZATION FACTOR peptide, the central subject of this study. Identified in a screen of the top 1,000 antheridia-expressed genes, MpRALF1 is specifically and highly expressed in mature antheridia (RT-qPCR, proMpRALF1:GUS) and the peptide accumulates in spermatozoid flagella. Three CRISPR/Cas9 frameshift knockout lines and amiRNA knockdown (Ri-MpRALF1) reduced male fertility (fewer sporophytes after crossing), lowered sperm ROS levels and impaired sperm kinematics (reduced curvilinear velocity, fewer rapidly progressive and more slow/non-progressive spermatozoids); 10 nM synthetic MpRALF1 peptide rescued both ROS and motility. MpRALF1 is perceived by the MpFER-MpLLG receptor complex (pulldown, CoIP, luciferase complementation), and its N-terminal 18-aa region binds MpFER whereas its C-terminal 38-aa region binds MpLLG. NAMING NOTE: gene ID stated in the paper; Mecchia et al. 2022 (Development, doi:10.1242/dev.200580) calls this same locus MpRALF3 and uses MpRALF1 for Mp1g27120. |
| Mp7g00270 | MpRBOHA | experimental subject | The second Marchantia NADPH oxidase (RBOH) gene, also highly expressed in antheridiophores and antheridia. amiRNA knockdown (Ri-MpRBOH2) reduced sperm ROS levels and produced the same class of motility defects as Ri-MpRBOH1, supporting RBOH-derived ROS as the effector of MpRALF1-MpFER-MpLLG signalling in spermatozoids. NAMING NOTE: no gene ID is printed in the paper; resolved from the paper's own MpRBOH2-RT-F/R primers (Suppl. Table 2), which match Mp7g00270 (= MpRBOHA / MpNOX2; this locus is MpRBOH1 in Yotsui et al. 2023). Link by ID, never by name. |
| Mp3g20340 | MpRBOHB | experimental subject | One of the two Marchantia NADPH oxidase (respiratory burst oxidase homologue) genes, both highly expressed in antheridiophores and antheridia. amiRNA knockdown (Ri-MpRBOH1) significantly reduced sperm ROS levels - more strongly than Ri-MpRBOH2 - and impaired sperm kinematics (lower curvilinear velocity, fewer rapidly progressive and more slow-progressive spermatozoids), phenocopied by the NADPH-oxidase inhibitor DPI; this places RBOH-derived ROS downstream of the MpRALF1-MpFER-MpLLG module in sperm motility control. NAMING NOTE: no gene ID is printed in the paper; the locus was resolved from the paper's own MpRBOH1-RT-F/R primers (Suppl. Table 2), which match Mp3g20340 (= MpRBOHB / MpNOX1; MpRBOH1 in Chu et al. 2023 but MpRBOH2 in Yotsui et al. 2023). Link by ID, never by name. |
| Mp3g23400 | MpEF1 | experimental tool | Housekeeping gene used to normalize all RT-qPCR transcript measurements. |
| Mp1g03930 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 71 aa; no predicted signal peptide; FPKM 87.6177/104.321 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp1g09940 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 104 aa; signal peptide predicted; FPKM 203.428/188.183 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp2g12030 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 64 aa; no predicted signal peptide; FPKM 201.81/168.769 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp2g18090 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 50 aa; no predicted signal peptide; FPKM 89.157/99.0344 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp3g00220 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 89 aa; no predicted signal peptide; FPKM 88.5917/97.2983 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp3g16180 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 74 aa; no predicted signal peptide; FPKM 143.565/143.739 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp3g21270 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 91 aa; no predicted signal peptide; FPKM 447.429/471.366 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp3g22390 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (annotated as arabinogalactan peptide; 77 aa; signal peptide predicted; FPKM 136.58/154.867 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp4g01930 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 70 aa; signal peptide predicted; FPKM 180.97/183.813 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp4g05540 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 45 aa; no predicted signal peptide; FPKM 153.845/165.333 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp4g05860 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 47 aa; no predicted signal peptide; FPKM 153.012/175.651 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp4g10350 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 64 aa; signal peptide predicted; FPKM 433.106/457.258 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp5g01750 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 84 aa; no predicted signal peptide; FPKM 191.441/95.6325 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp5g03630 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 83 aa; signal peptide predicted; FPKM 99.2707/117.315 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp6g10430 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (no functional annotation; 78 aa; no predicted signal peptide; FPKM 115.709/133.423 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp6g11390 | missing | experimental background | Listed among the 17 putative peptide-encoding genes retrieved by this study's screen of the top 1,000 antheridia-expressed genes (annotated as signalling peptide taximin 2; 78 aa; no predicted signal peptide; FPKM 89.0919/54.7961 in the two antheridia replicates). Not characterized further - MpRALF1 (Mp7g07270) was the candidate pursued. |
| Mp5g09600 | MpLRE1 | citation background | Cited as the other of the two MpLRE genes; because it lacks the typical GPI-anchoring site (reported by Mecchia et al. 2022) it was not pursued, and the study focused on MpLRE2/MpLLG instead. |
| Mp1g27120 | MpRALF1 paper
MpRALF3 nomenclature |
experimental background | One of the three Marchantia RALF genes profiled by RT-qPCR (Extended Data Fig. 1a); not antheridia-enriched, so the study focused on MpRALF1. Locus resolved from the paper's MpRALF3-RT-F/R primers (Suppl. Table 2), which match Mp1g27120 - the locus that Mecchia et al. 2022 calls MpRALF1. |
| Mp2g21670 | MpRALF2 | experimental background | One of the three Marchantia RALF genes whose transcript levels were measured by RT-qPCR across male and female tissues (Extended Data Fig. 1a); unlike MpRALF1 it is not antheridia-enriched. Locus resolved from the paper's MpRALF2-RT-F/R primers (Suppl. Table 2). |