Fig. 5: The NHLP1-RACK1 pathway is conserved in regulating tillering/branching in rice and Arabidopsis.

Representative phenotypes (a) and tiller number comparison (b) of ZH11 (negative control) and Osnhlp1a mutants. p = 0.196 (Osnhlp1a-1 vs. Osnhlp1a-2), <0.0001 for other comparisons. Scale bar, 10 cm in (a). Representative phenotypes c and tiller number comparison d of ZF802 (negative control) and CRISPR/Cas9-generated Osnhlp1a and Osnhlp1b mutants. p = 0.4539 (Osnhlp1a vs. Osnhlp1b), 0.0017 (Osnhlp1b vs. Osnhlp1aOsnhlp1b), <0.0001 for other comparisons. Scale bar, 10 cm in (c). e OsRACK1A interacts with OsNHLP1a and OsNHLP1b, respectively, in a split-luciferase complementation assay. Different combinations of OsNHLP1a-nLUC, OsNHLP1b-nLUC, and cLUC-OsRACK1A were co-expressed in N. benthamiana leaves, and luciferase activity was detected after 2 days of infiltration. f Co-immunoprecipitation assays showed that OsRACK1A interacts with OsNHLP1a and OsNHLP1b, respectively, in N. benthamiana. OsNHLP1a-MYC and OsNHLP1b proteins were immunoprecipitated (IP) by HA beads. The black triangle represents the target bands. Representative phenotypes g and branch numbers h of the wild-type Arabidopsis thaliana Col-0 (negative control) and Atnhlp1a mutants, assessed approximately 65 days post-germination. The quantification of branch numbers encompasses the sum of the main stem, primary stem branches, secondary rosette branches, and primary rosette leaf branches. p = 0.1343 (Atnhlp1a−6 vs. Atnhlp1a−8), <0.0001 for other comparisons. The scale bar in (g) is 5 cm. i, j AtNHLP1a interacts with AtRACK1A, AtRACK1B, and AtRACK1C, respectively, according to split-luciferase complementation assays and co-immunoprecipitation assays in N. benthamiana. Data in (b, d, and h) are presented as mean ± SD (T₂ generation). Different letters indicate significant differences between groups, as determined by one-way ANOVA with Tukey’s multiple comparisons range tests (p < 0.01). These experiments in (f, i) were repeated three times independently with similar results. Individual data points are plotted, with the sample size (n) for each group is listed at the bottom of the rectangular column. Source data are provided as a Source Data file.