The CLAVATA receptor FASCIATED EAR2 responds to distinct CLE peptides by signaling through two downstream effectors
- Cold Spring Harbor Laboratory, United States
- Pusan National University, Republic of Korea
- DuPont Pioneer, Agricultural Biotechnology, United States
- University of Massachusetts Amherst, United States
- The University of Sydney, Australia
Figures
Figure 1
Both fea3 and fea2 mutants are resistant to ZmFCP1 peptide.
(A) Transactivation of ZmFCP1 in primordia using a pYABBY14:LhG4 driver led to a strong reduction in vegetative SAM size as compared to a non-transgenic control, but this effect was only partially …
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Figure 1—source data 1
CLE peptide treatments of fea2;fea3 a segregating population.
- https://doi.org/10.7554/eLife.35673.004
Figure 2 with 5 supplements
Zmcrn mutants develop fasciated ears.
(A) Schematic of the Zmcrn mutant alleles. The arrows indicate the position of the Mutator transposon insertion and premature stop codon mutation. (B) Cleared SAMs from wild type (WT) and Zmcrn …
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Figure 2—source data 1
SAM size of Zmcrnin a segregating population.
- https://doi.org/10.7554/eLife.35673.011
Figure 2—figure supplement 1
Phylogeny of CRN related proteins, and ZmCRN features of a pseudokinase.
(A) The phylogeny shows that the maize genome has one CRN ortholog, which we named ZmCRN (GRMZM2G032132). The top three hits chosen by BLASTP search of AtCRN against the NCBI database (www.ncbi.nlm.n…
Figure 2—figure supplement 2
Mapping of the fea*148 fasciated ear mutant.
(A) Bulked segregant analysis of fea*148 using Maize SNP50 chip data delineated a 5 Mb mapping interval surrounding ZmCRN on chromosome 3. Scanning electron microscopy images of wild type (B) and Zmc…
Figure 2—figure supplement 3
Zmcrn/Zmcrn-148 F1 plants develop fasciated ears.
Scanning electron microscopy image of a Zmcrn/Zmcrn-148 F1 ear shows an enlarged ear inflorescence meristem (C), compared with normal ears of Zmcrn-148/+ (A) or Zmcrn/+ (B), suggesting that these …
Figure 2—figure supplement 4
Expression of FEA2, CT2 and ZmCRN in different domains of the SAM.
Expression values were extracted from the maize shoot apex RNAseq Atlas from the Timmermans Lab (available at https://maizegdb.org/, ). The samples are from B73 and different domains are captured by …
Figure 2—figure supplement 5
The association of ZmCRN locus with kernel row number (KRN).
The dots show the association of SNPs in the ZmCRN locus with KRN evaluated in five environments, and their BLUP data. The 3’UTR region of ZmCRN locus is significantly associated KRN. BLUP = best …
Figure 3
ZmCRN acts in a common pathway with FEA2, but not with CT2.
(A) Cleared SAMs from wild type (WT), Zmcrn, fea2, and Zmcrn;fea2 double-mutant plants. SAMs from Zmcrn and fea2 plants were significantly wider than in wild type (double-headed arrows), but SAM …
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Figure 3—source data 1
SAM size of Zmcrn;fea2 in a segregating population.
- https://doi.org/10.7554/eLife.35673.013
Figure 4 with 3 supplements
FEA2 is present in two different complexes.
(A) ZmCRN-mCherry was localized at the plasma membrane following tobacco transient expression (top), and in subsequent plasmolysis (bottom). In transient expression followed by immunoprecipitation …
Figure 4—figure supplement 1
ZmCRN-mCherry co-localized with FEA2-YFP and CT2-YFP on the plasma membrane.
ZmCRN-mCherry co-localized with FEA2-YFP (A) and with CT2-YFP (B) on the plasma membrane following transient expression in N. benthamiana. Scale bars: 20 μm.
Figure 4—figure supplement 2
FEA2-NmVen210 interacts with ZmCRN-CmVen210 by BiFC.
YFP signal was detected when FEA2-NmVen210 was co-expressed with ZmCRN-CmVen210. No YFP signal was detected when FEA2-NmVen210 was co-expressed with CT2-CmVen210, suggesting the interaction is …
Figure 4—figure supplement 3
Treatment with ZmFCP1 or ZmCLE7 peptide didn’t affect the protein complex formation.
5 μm ZmFCP1 or ZmCLE7 were infiltrated into the transformed N. benthamiana leaves expressing the indicated proteins following the protocol in Somssich et al. (2015). The presence of ZmFCP1 or ZmCLE7 …
Figure 5 with 2 supplements
ct2 and Zmcrn show different sensitivity to ZmCLE7 and ZmFCP1 peptides.
Embryos of each genotype were cultured with control, scrambled peptide (sCLV3) or with ZmFCP1 or ZmCLE7. Wild type SAM growth (double-headed arrows) was strongly inhibited by all peptides except …
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Figure 5—source data 1
ZmFCP1 and ZmCLE7 double peptide treatments inct2,Zmcrnandfea2in a segregating population.
- https://doi.org/10.7554/eLife.35673.023
Figure 5—figure supplement 1
ZmCRN and CT2 function in a different pathway to FEA3.
(A) Cleared SAMs from wild-type (WT), Zmcrn, fea3, and double-mutant plants. SAMs from Zmcrn and fea3 plants were wider than those in wild type (double-headed arrows), and Zmcrn;fea3 double mutants …
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Figure 5—figure supplement 1—source data 1
SAM size of Zmcrn;fea3 in a segregating population.
- https://doi.org/10.7554/eLife.35673.020
Figure 5—figure supplement 2
Triarabinosylated ZmCLE7 peptide is more potent.
Germinating maize seedling roots or embryos were treated with non-modified or triarabinose modified ZmCLE7 peptide at the concentrations indicated. In each case, the modified peptide was …
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Figure 5—figure supplement 2—source data 1
ZmCLE7-Arabinosylated peptide embryo assay.
- https://doi.org/10.7554/eLife.35673.022
Figure 6 with 1 supplement
Hypothetical model for FEA2 signaling through two different pathways.
Two different peptides, ZmFCP1 and ZmCLE7, are proposed to bind to two separate FEA2 receptor complexes, and the two signals are differentially transmitted to downstream components; with the ZmCLE7 …
Figure 6—figure supplement 1
Clustering, phylogenetic tree, and functional annotations of CLV2/FEA2 genes and their close homologs from five species.
CLANS clustering of CLV2/FEA2 and closely related homologs representing pairwise similarity (A). The clusters containing CLV2/FEA2 and FEA3 are circled in blue and green, respectively. The CLV2/FEA2 …
Additional files
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Supplementary file 1
table 1. P-values of the association between ZmCRN SNPs with kernel row number in multiple environments.
- https://doi.org/10.7554/eLife.35673.026
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Supplementary file 2
table 2. List of Primers.
- https://doi.org/10.7554/eLife.35673.027
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Transparent reporting form
- https://doi.org/10.7554/eLife.35673.028
