Fig. 3

Functional annotation of genes responsible for CG metabolism. A Schematic representation of the CG biosynthesis pathway, in which the key enzymes are highlighted in red. B Manhattan plot displaying the GWAS results for linamarin content. The lower panel shows the location of five CG biosynthesis genes and three HNLs in a 140-kb region. C HPLC chromatograms of the products of the reactions of UGT85K4 and UGT85K5, respectively, with UDP-glucose and acetone cyanohydrin. D Expression correlation of CG metabolism genes in SR of 22 cultivated cassava accessions. E Reduced transcript expression of UGT85K5 in UGT85K5-silenced cassava plants (CsCMV-UGT85K5) compared with the control (CsCMV-NC) in leaf. Bars indicate mean ± SD, n = 3. F Reduced levels of linamarin in UGT85K5-silenced cassava plants (CsCMV-UGT85K5) compared with the control (CsCMV-NC) in leaf. Bars indicate mean ± SD, n = 3. G–I Boxplot displaying the relative contents of linamarin based on the causative SNPs 12:35990792 (for CYP79D1), 12:35945868 (for CYP71E7b), and 12:35953769 (for UGT85K5), respectively. J The relative content of linamarin based on the allelic combination of causative SNPs 12:35990792 (for CYP79D1), 12:35945868 (for CYP71E7b), and 12:35953769 (for UGT85K5). Different letters indicate significant differences at P < 0.05 based on Duncan’s multiple range tests