Identification and application of herbicide‐resistant 4‐hydroxyphenylpyruvate dioxygenase (HPPD) alleles via directed evolution

作者：Bing Zhao, Yao Wang, Miaoyi Zhou, Xinxiang Liu, Hanshuai Li, Zi Shi, Yuhui Guo, Ting Li, Fengling Yang, Ronghuan Wang, Jiuran Zhao, Ya Liu · 发表于：Plant Biotechnology Journal · 年份：2025 · DOI：10.1111/pbi.70160 · 被引用次数：3 · 研究领域：Pharmacogenetics and Drug Metabolism、Pesticide and Herbicide Environmental Studies、Microbial Metabolic Engineering and Bioproduction

Herbicide development has played an important role in improving weed-control efficiency in crop fields, reducing agricultural production costs and increasing crop yields. HPPD inhibitors, a novel class of bleaching herbicides, are widely used for weed control in staple crops such as corn and rice. These herbicides inhibit the conversion of hydroxyphenylpyruvate to homogentisic acid (HGA) catalysed by HPPD (Figure S1). Currently, HPPD is a herbicide target with relatively few reported resistance cases. However, known HPPD proteins generally show low herbicide tolerance, which limits their practical application. To solve this, researchers have therefore introduced specific alterations in the HPPD amino acid sequence to obtain HPPD mutants with significantly improved herbicide tolerance (Dreesen et al., 2018; Hawkes et al., 2019). In recent years, advances in directed evolution techniques, such as MAGE (Wang et al., 2009), PACE (Esvelt et al., 2011) and OrthoRep (Ravikumar et al., 2018), have enabled the evolution of nearly any protein of interest (Table S1). In 2021, Yi et al. designed a targeted artificial DNA replisome (TADR) capable of error-prone replication of one strand of a target plasmid in cells (Yi et al., 2021). The system consists of a bacteriophage PhiX174 cisA protein, a bacterial Rep helicase and an error-prone DNA polymerase. The TADR can increase the mutation rate of the target plasmid by up to 2.3 × 105 fold. In this study, two HPPD genes with significant diff...