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. 2017 Sep 21;6:e25012. doi: 10.7554/eLife.25012

Figure 1. brush contains a point mutation in CNGC.IVA1 (BRUSH).

(A) Schematic of the intron-exon structure of BRUSH highlighting the brush mutation in the first exon (red asterisk). (B) Phylogenetic tree of BRUSH (red node end) in relation to Arabidopsis thaliana CNGC proteins. BRUSH is orthologous to the Group IVA members AtCNGC19 and AtCNGC20. (C) Overview of the BRUSH protein domain structure highlighting the conserved Group IVA domain (CD, green), transmembrane domains (T, light blue), putative filter region (F, orange), and the predicted cyclic nucleotide-binding domain (CNBD, purple). Shown below is the CD sequence in brush (G134E mutation, asterisk) relative to BRUSH, AtCNGC19, and AtCNGC20 and the Group IVA CNGC consensus (Yuen and Christopher, 2013). Numbers at the branch points in (B) indicate the percentage bootstrap values (100 iterations) for the inferred tree. Scale bar in (B) indicates the number of amino acid substitutions per site.

Figure 1—source data 1. Figure 1B source data.
elife-25012-fig1-data1.xlsx (57.9KB, xlsx)
DOI: 10.7554/eLife.25012.007

Figure 1.

Figure 1—figure supplement 1. Map-based cloning of the brush mutation on chromosome 2.

Figure 1—figure supplement 1.

Overview of contigs CM0435 and CM0312 (Lotus japonicus genome v2.5) on the short arm of chromosome 2 surrounding the brush target region. The relative location of SSR and SNP markers used for mapping are indicated on the contigs. The brush mutation was previously linked to TM0312 by rough mapping. The red asterisk indicates the position of the causative mutation responsible for the brush phenotype. SSR markers were amplified with fluorescent primers and analysed on an ABI capillary sequencer. SNPs were genotyped by sequencing and/or KASP (LGC).

Figure 1—figure supplement 2. Syntenic chromosomal locations of CNGC.IVA genes in selected plant species.

Figure 1—figure supplement 2.

A cluster of 5 CNGC.IVA genes are present on chromosome 2 of L. japonicus. CNGC.IVA2 contains a large transposon (Tn) insertion. The CNGC.IVA cluster is bordered by genes encoding for β-amylase (red) and glutamate decarboxylase (blue). The Glycine max (soybean) and Medicago truncatula genomes both contain a cluster of CNGC.IVAs (Charpentier et al., 2016) associated with the same anchor genes. In contrast, two non-legume plant species of the Rosaceae family (Fragaria vesca and Prunus persica) contain a single CNGC.IVA copy in association with the glutamate decarboxylase anchor. Chromosome 3 of Arabidopsis thaliana contains 2 CNGC.IVA copies. The chromosomal depictions are simplified to highlight the syntenic loci. The slash between CNGCIVAc and CNGCIVAf on Medicago chromosome 6 represents a gap of 480 kb.

Figure 1—figure supplement 3. Protein sequence comparison of Group IVA CNGCs from Lotus japonicus.

Figure 1—figure supplement 3.

Protein sequence alignment of BRUSH (CNGC.IVA1), CNGC.IVA3, CNGC.IVA4, and CNGC.IVA5. Residues identical to BRUSH are shown as dots and gaps are shown as dashes. Shown is the Group IVA CNGC conserved domain (CD, green), the position of the brush mutation (red asterisk), predicted transmembrane domains (TM, light blue), putative selectivity filter (F, orange) in the pore region, and the cyclic nucleotide-binding domain (CNBD, purple). CNGC.IVA3 (81%), CNGC.IVA4 (65%), and CNGC.IVA5 (72%) show high-sequence identity to BRUSH.