Extended Data Fig. 6: Functional characterization of PsCIK2/3.
a, Flowering stage phenotype of Caméor (Fa/Fa, left) and JI0814 (fa/fa, right). b, Longitudinal section of 40-day-old stems from Caméor and JI0814, stained with safranin and fast green; the red crosses mark the region of the shoot apical meristem (SAM) in both genotypes. c, Transverse section of the apical meristem of 14-day-old paraffin-embedded stems, from Caméor (left) and JI0814 (right), stained with toluidine blue. d, Quantification of the number of vascular bundles in the longitudinal section of 40-day-old stems of Caméor and JI0814. e, Cross-sectional area of the apical meristem in 14-day-old stems of Caméor and JI0814. Three biological duplicates were used. ** represents a significant level at P < 0.01 using a Student’s t-test in (d) and (e). f, Gene expression level from RNA-seq data for key genes involved in the CLV3-WUS signalling pathway measured across different organs and developmental stages (three biological replicates for each sample) in Caméor. The normalization and statistical approach is the same as described in Extended Data Fig. 2h. g, qRT_PCR analysis of gene expression for key genes involved in the CLV3-WUS signalling pathway, comparing the apical bud and stem between the wild-type line (Caméor) and the fasciated line (JI0814). Three biological replicates (n = 3) were used for each sample. The samples were obtained 14 days post budding. Data are presented as mean ± SEM, and statistical significance was determined using a two-sided t-test. The calculation used H3 as the reference gene, with P_value <= 0.001 marked as “***”, P_value <= 0.01 marked as “**”, P_value <= 0.05 marked as “*”, P_value >0.05 marked as “ns”. h, In situ hybridization of PsCIK2/3 in the apical bud compared between the wild-type line (JI2716) and the fasciated line (JI0814). i, Subcellular localization of PsCIK2/3 in Nicotiana benthamiana, showing co-localization with the cell membrane. j, Yeast two-hybrid assay showing interaction of PsCIK2/3 with PsCLV1 and PsCLV2.
