Mutations of Asn321 and Glu322 Improve Resistance of 4-Hydroxyphenylpyruvate Dioxygenase SpHPPDm to Topramezone
Mutations of Asn321 and Glu322 Improve Resistance of 4-Hydroxyphenylpyruvate Dioxygenase SpHPPDm to Topramezone
复制标题
Asn321和Glu322突变提高4-羟苯基丙酮酸双加氧酶SHPPDm对Topramezone的抗性
DOI：
10.1021/acs.jafc.2c02327
复制
发表时间：
2022
影响因子：
6.1
通讯作者：
Jian He
中科院分区：
文献类型：
--
作者：
Haiyan Wang;Peng Lei;Bin Liu;Jianchun Zhu;Qin He;Le Chen;Jian He
苯唑草酮是一种高效的4-羟苯丙酮酸双加氧酶（HPPD）抑制剂，是抗除草剂基因工程的理想靶标。本研究从已获得的抗苯吡唑酮HPPD突变体SpHPPDm的随机突变库中筛选到了两个抗苯吡唑酮突变体K-19（N321 Y）和K-63（Q166 R/E322 V），其抗苯吡唑酮能力分别提高了205.3%和58.5%。通过单位点突变分析，N321和E322位点被确定为苯唑草酮抗性增加的关键位点。通过N321和E322位点的饱和突变，进一步获得突变体KB-145（N321 Y/E322 K）。KB-145对苯唑草酮的抗性比突变体SpHPPDm提高了955.3%。总之，本研究发现了两个显著影响SpHPPDm苯唑草酮抗性的新位点，为HPPD除草剂抗性的分子机制提供了新的见解，获得的突变体在抗除草剂作物构建中具有巨大的应用潜力。
As a highly efficient 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor herbicide, topramezone is an ideal target for herbicide-resistant genetic engineering. In this study, two mutants, K-19 (N321Y) and K-63 (Q166R/E322V), with topramezone resistance increased by 205.3 and 58.5%, respectively, were screened from the random mutation library of SpHPPDm, a topramezone-resistant HPPD mutant that we previously obtained. Sites N321 and E322 were identified as key sites for increased topramezone resistance by single-site mutation analysis. A mutant KB-145 (N321Y/E322K) was further obtained by saturation mutation at sites N321 and E322. The topramezone resistance of KB-145 increased by 955.3% compared to mutant SpHPPDm. In conclusion, this study identifies two new sites that significantly affect the topramezone resistance of SpHPPDm, which provides new insights into the molecular mechanism of herbicide resistance of HPPD, and the acquired mutants have great application potential in the construction of herbicide-resistant crops.