Improving the Herbicide Resistance of Rice 4-Hydroxyphenylpyruvate Dioxygenase by DNA Shuffling Basis-Directed Evolution

文献类型： 外文期刊

第一作者： Chen, Le

作者： Chen, Le;Liu, Rui;Tan, Qing;Luo, Hongmei;Chen, Yuyu;Jin, Yaru;Zheng, Zhongbing;Zhang, Baolong;Chen, Le;Zhang, Baolong;Guo, Dongshu;Zhang, Baolong

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关键词： 4-hydroxyphenylpyruvatedioxygenase (HPPD); directedevolution; DNA shuffling; enzyme properties; herbicide resistance

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：6.1； 五年影响因子：6.3 ）

ISSN： 0021-8561

年卷期： 2023 年 71 卷 41 期

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收录情况： SCI

摘要： 4-Hydroxyphenylpyruvate dioxygenase (HPPD) is an ideal target for herbicide resistance genetic engineering. In this study, a mutant MFRR-2 with mesotrione resistance was screened from an Oryza sativa HPPD and mutant- Zea mays HPPD DNA shuffling library. The enzyme properties showed that although the stability of the mutant decreased in vitro, the enzyme activity of MFRR-2 at the optimum temperature of 25 degree celsius was still equivalent to that of OsHPPD. Under 50 mu M mesotrione treatment, MFRR-2 enzyme activity remained at approximately 90%, while the enzyme activity of OsHPPD decreased by approximately 50%. Surprisingly, Fe2+ was found to have an inhibitory effect on the enzyme activity. Then, the transgenic rice of the MFRR-2 gene showed approximately 1.5 times mesotrione resistance compared to OsHPPD transgenic rice. In conclusion, this study has conducted a beneficial exploration on the use of DNA shuffling for HPPD-directed evolution, and the mutant has potential application value for herbicide resistance genetic engineering.

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