Tree Genetics and Molecular Breeding 2024, Vol.14, No.1, 32-42 http://genbreedpublisher.com/index.php/tgmb 34 (3) Versatility: Multiple genes can be targeted simultaneously, enabling complex trait modifications (An et al., 2020). (4) Minimal Growth Penalty: CRISPR/Cas9-mediated knockouts of lignin biosynthetic genes have shown reduced lignin content without significant growth penalties, making it a promising tool for biofuel production (Jang et al., 2021; Vries et al., 2021). Figure 1 The general phenylpropanoid and monolignol-specific pathways, with the changes in phenolic metabolism indicated for cse1 cse2 xylem and/or bark extracts compared with WT (Adopted from Vries et al., 2021) Image caption: Red and blue names indicate metabolites that are up or down, respectively, in cse1 cse2 xylem and/or bark extracts as compared to WT. Characterized metabolites uniquely detected in bark or xylem profiles are indicated with a (b) or (x), respectively. Co-occurring metabolites are indicated by (x, b). Metabolites that are framed in a box belong to the same class. The flow through the pathway towards the conventional monolignols is in black in WT, whereas (putative) alternative flows are in grey. Successive arrows show two or more metabolic steps. Solid arrows show enzymatic conversions that are validated by experimental evidence, whereas dashed arrows show suggested conversions (Saleme et al., 2017; Van Acker et al., 2017). *The catalytic activity of CSE1 and CSE2 is suggested in planta and proven in vitro in this work. Each metabolite is indicated with a unique number, corresponding to the metabolites mentioned in the manuscript. CAD, CINNAMYL-ALCOHOL DEHYDROGENASE; CCoAOMT, CAFFEOYL-CoA O-METHYLTRANSFERASE; CCR, CINNAMOYL-CoA REDUCTASE; CHS, CHALCONE SYNTHASE; COMT, CAFFEIC ACID O-METHYLTRANSFERASE; F5H, FERULATE 5-HYDROXYLASE; PAL, PHENYLALANINE AMMONIA-LYASE (Adopted from Vries et al., 2021)
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