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An artificially evolved gene for herbicide-resistant rice breeding
Friday, 2024/08/23 | 08:34:29

 

Jin DongXin-He YuJiangqing DongGao-Hua WangXin-Long WangDa-Wei WangYao-Chao YanHan XiaoBao-Qin YeHong-Yan Lin, and Guang-Fu Yang

PNAS August 12, 2024; 121 (34) e2407285121

Significance

The discovery and engineering of herbicide-resistant genes represent a critical frontier in modern crop breeding. Here, we focused on unraveling the intricate mechanisms underlying herbicide resistance, particularly within the prevalent 4-hydroxyphenylpyruvate dioxygenase Inhibitor Sensitive 1-Like (HSL) protein family. The structures of HSLs complexed with different herbicides not only revealed the initial binding mode of substrate in HSLs, but also explained the catalytic selectivity of some HSL members. From these structures, we identified four residues critical for catalysis. Leveraging an artificial evolution approach, these residues were mutated which significantly enhanced the resistance to β-triketone herbicides in rice. Our findings not only shed light on the catalytic mechanism of HSLs against herbicides but also provide a promising avenue for developing herbicide-resistant crops.

Abstract

Discovering and engineering herbicide-resistant genes is a crucial challenge in crop breeding. This study focuses on the 4-hydroxyphenylpyruvate dioxygenase Inhibitor Sensitive 1-Like (HSL) protein, prevalent in higher plants and exhibiting weak catalytic activity against many β-triketone herbicides (β-THs). The crystal structures of maize HSL1A complexed with β-THs were elucidated, identifying four essential herbicide-binding residues and explaining the weak activity of HSL1A against the herbicides. Utilizing an artificial evolution approach, we developed a series of rice HSL1 mutants targeting the four residues. Then, these mutants were systematically evaluated, identifying the M10 variant as the most effective in modifying β-THs. The initial active conformation of substrate binding in HSL1 was also revealed from these mutants. Furthermore, overexpression of M10 in rice significantly enhanced resistance to β-THs, resulting in a notable 32-fold increase in resistance to methyl-benquitrione. In conclusion, the artificially evolved M10 gene shows great potential for the development of herbicide-resistant crops.

 

See https://www.pnas.org/doi/10.1073/pnas.2407285121

 

Figure 5:

Herbicide resistance of OsHSL1 transgenic rice plants. (A) The pCAMBIA 3300 vector for expression of OsHSL1 or mutants in rice plants. KanR and HygR, kanamycin, and hygromycin resistance genes. (B) Herbicide susceptibility of Zhonghua 11 (ZH11) to MBQ transformed with WT OsHSL1 or M10. The leaf and stem were sprayed with herbicide-containing water at 7 to 10 d after transplantation. Plant growth was checked after 21 d. (C) The dose–response curves of (B), the bleaching percentage of rice leaves were shown. (D) Herbicide susceptibility of ZH11 expressing OsHSL1-WT or M10 to MBQ. Different concentrations of herbicide were solubilized in water at 7 to 10 d after transplantation. Plant growth was checked after 21 d. (E) The dose–response curves of (D), the bleaching percentage of rice leaves were shown. (F) Herbicide susceptibility of ZH11 expressing OsHSL1-WT or M10 to MBQ. Different concentrations of herbicide were dissolved in the culturing media. Plant growth was checked after 7 d. (G) The dose–response curves of (F), the bleaching percentage of rice leaves were shown. Herbicide concentration was indicated. Ctrl, no herbicide was applied. Different concentrations of MBQ are indicated.

 

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