Table S1

Primers used in this study"

Fig. S1

Nucleotide sequence alignment of SLY1 among different B. napus varieties A: Nucleotide sequence alignment of the A subgenome SLY1 among different varieties; B: Nucleotide sequence alignment of C subgenome SLY1 among different varieties. The black area represents exons, and the gray area represents introns."

Fig. 1

Comparison of protein sequences and construction of developmental tree for BnaSLY1 A: alignment of SLY1 proteins in Arabidopsis, B. rapa, B. oleracea, and B. napus (var. Westar); B: phylogenetic tree of BnaSLY1 proteins among different B. napus varieties. Numbers on branches are bootstrap values for 1000 replications; C: phylogenetic tree of SLY1 proteins among different species. The green part represents Group I, the blue part represents Group II, and the remaining colors represent Group III. CICLE: citrus; FCD: Ficus carica; EVM: marijuana; Cla97: watermelon; SORBI: sorghum; Zm: maize; Os: rice; Traes: wheat; HORVU: barley; AVESA: oats; Rchi: rosebush; Csa: cucumber; Vitvi: grapes; EUTSA: thellungiella; DCAR: carrot; CEY00: kiwi; SIN: sesame; HanXRQ: sunflower; LSAT: lettuce; FRAEX: white wax; OE9A: olive; AT: Arabidopsis; Bna: Brassica napus; Bra: Brassica rapa; Bo: Brassica oleracea."

Fig. 2

Expression patterns of BnaSLY1 homologs A: relative expression level of BnaSLY1 homologs in different tissues of Westar; B: heatmap of BnaSLY1 expression in different tissues of ZS11, represented by TPM values, data extracted from the BnIR database; C: subcellular localization of BnaSLY1 in Nicotona benthamiana. Scale bar, 25 μm."

Fig. 3

The target sites of BnaSLY1 and its mutations in several representative knockout lines A: gene structure of BnaSLY1 and the target sites; B: mutations at the targeted sites of BnaSLY1 in T1 lines. Green fonts represent the PAM sequence, red fonts represent mutations, and red dotted lines represent deletion of bases, respectively."

Table S2

Detection of target editing efficiency of CRISPR/Cas9 T0 generation transgenic seedlings"

Fig. 4

Phenotypes of the BnaSLY1 knockout mutants A: leaves of the control Westar and the mutants; B: seedlings of Westar and the mutants; C: flowering plants of Westar and the mutants; D: comparison of plant height between Westar and the mutants; E: comparison of flowering time between Westar and the mutants. Scale bar: 8 cm. ** indicates significant differences at the P < 0.01 probability level."

Fig. 5

BnaSLY1 impacts the cell morphology, chlorophyll, and wax synthesis in leaves A: paraffin sections of WT and the mutant leaves, scale bar: 100 μm; B: chloroplast ultrastructure of WT and the mutants, scale bar: 2 μm; C: comparison of leaf thickness between WT and the mutants; D: comparison of chlorophyll content between WT and the mutants. sm: spongy mesophyll; Cp: chloroplast; Tm: thylakoid membrane. * and ** indicate significant differences at the P < 0.05 and P < 0.01 levels, respectively; E: scanning electron microscopy of waxy on leaf epidermis; scale bar: 5 μm."

Fig. 6

Levels of endogenous GA in Westar and bnasly1 ** indicates significant differences at the P < 0.01 level."

Table S3

Summary of RNA-seq data for samples"

Fig. S2

Pearson correlation coefficients of transcriptome data CK: Westar; T: bnasly1."

Fig. S3

Volcano map of differentially expressed genes between Westar and bnasly1"

Fig. S4

GO classification of differentially expressed genes between Westar and bnasly1"

Fig. S5

KEGG Enrichment of differentially expressed genes between Westar and bnasly1"

Fig. 7

Differential expressed genes in the auxin signaling pathway a, b, and c indicate the log10(FPKM) of DEGs in the three biological replicates of Westar; d, e, and f indicate the log10(FPKM) of DEGs in the three biological replicates of bnasly1; g is the log2FC between Westar and bnasly1. AUX1/IAA: auxin/indole-3-acetic acid; TIR1: transport inhibitor response 1; ARF: auxin response factor; SAUR: small auxin upregulated RNA; GH3: gretchen hagen 3."

Table S4

DEGs in GA synthesis and signal transduction"

Fig. 8

Differential expressed genes in the wax synthesis pathway a, b, and c indicate the log10(FPKM) of DEGs in three biological replicates of Westar; d, e, and f indicate the log10(FPKM) of DEGs in three biological replicates of bnasly1; g is the log2FC between Westar and bnasly1. LACS: long-chain acyl-CoA synthetase; FAE: fatty acid elongase; CER3: CERIFERUM 3 (FAE complex subunit); CER26: CERIFERUM 26 (FAE complex subunit); WSD1: wax ester synthase/ diacylglycerol acyltransferase; FAR: fatty acyl-CoA reductase; MAH1: mid-chain alkane hydroxylase; CYP: cytochrome P450; GPAT: glycerol 3-phosphate acyltransferase; 2-MHG: 2-mono (10,16)-dihydroxyhexadecanoyl glycerol."

Table S5

DEGs related to flowering time"