Figure 3. Phylogenetic tree of KASI proteins in land plants.
Protein sequences were aligned using MAFFT. Phylogenetic trees were generated by neighbor-joining implemented in MEGA5 (Tamura et al., 2011). Partial gap deletion (95%) was used together with the JTT substitution model. Bootstrap values were calculated using 500 replicates. DIS likely originated before the angiosperm divergence (red star).
DOI: http://dx.doi.org/10.7554/eLife.29107.016
Figure 3—source data 1. Accession numbers for protein sequences used in the phyologenic tree.
elife-29107-fig3-data1.xlsx (12.3KB, xlsx)
DOI: 10.7554/eLife.29107.017
Figure 3—figure supplement 1. Transcript accumulation of KASI and RAM2 genes.
(A) Transcript accumulation of DIS, DIS-LIKE, KASI and RAM2 in control (mock) and R. irregularis colonized (AM) roots and in different organs of L. japonicus assessed by qRT-PCR. Expression values were normalized to those of the constitutively expressed gene EF1α (DIS, DIS-LIKE, KASI) and Ubiquitin10 (RAM2). Black circles represent three biological replicates. Different letters indicate significant differences (ANOVA; posthoc Tukey; n = 15; p≤0.05, F4,14(KASI) = 1.191, F4,14(DIS) = 8.412, F4,14(DIS-LIKE) = 4.563; p≤0.001, F4,14 = 67.41 (RAM2)). AM plants were inoculated with R. irregularis. Control and AM plants were harvested 5 wpi. (B) Arbuscule phenotype in wild type and dis-like-5 mutant roots after 5 wpi with R. irregularis as indicated by acid ink staining. White arrow heads indicate arbuscules.
Figure 3—figure supplement 2. Shoot phenotypes of dis and ram2 mutants.
dis and ram2 mutants do not show growth differences in shoot growth as compared to Gifu wild-type. The image has been taken 17 weeks post planting (size bar, 5 cm).
Figure 3—figure supplement 3. Genomic comparison of the DIS locus in host and non-host species.
Synteny analysis of a ~ 200 kb sized region in the Lotus japonicus, Medicago truncatula (green), Populus trichocarpa (orange), Phaseolus vulgaris (pink), Solanum lycopersicum (blue) and Carica papaya (yellow) genomes containing the DIS locus. The genomic block is well conserved in these host species. By contrast, no DIS homolog was detected in the corresponding genomic block of Arabidopsis thaliana (red). The red rectangle indicates the DIS and DIS-LIKE locus, DIS is indicated in yellow. The sequences above Lotus correspond to the forward strand and those below Lotus to the reverse strand. The orange strip on the left side corresponds to a non-assembled region of the L. japonicus genome.