Bensulfuron-methyl (BSM) is a widely used herbicide in rice cultivation, but the mechanisms underlying its efficient uptake in rice are poorly understood. In this study, a yeast library expressing 1385 rice transporters was employed to screen proteins sensitive to BSM. As a result, a cyclic nucleotide-gated channel (CNGC) protein, OsCNGC12, was identified. By inactivating the OsCNGC12 function via gene editing, we developed BSM-tolerant rice lines. Our results showed that the OsCNGC12 mutant rice not only reduced BSM uptake but also promoted Ca²⁺ influx in the roots, leading to enhanced non-target-site tolerance to BSM. OsCNGC12 is localized in the plasma membrane and is abundantly expressed in the root caps, root vascular bundles, stems, and leaves, indicating its vital role in the distribution of BSM in rice. The indispensability of His-236, Gln-240, and Arg-268 in BSM perception in yeast was demonstrated through targeted mutagenesis of OsCNGC12. Furthermore, we developed a BSM seed coating agent for OsCNGC12 mutant rice, providing a simplified and cost-effective means of weed control in direct-seeded rice. Inclusion, this study showed that disruption of OsCNGC12 confers non-target-site tolerance to BSM in rice and has the potential for developing BSM-resistant rice varieties.

中文翻译：

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除草剂苯磺隆通过阻断 OsCNGC12 通道中的钙离子通量来抑制水稻幼苗发育

苯嘧磺隆 (BSM) 是一种在水稻种植中广泛使用的除草剂，但人们对其在水稻中有效吸收的机制知之甚少。在本研究中，使用表达 1385 个水稻转运蛋白的酵母文库来筛选对 BSM 敏感的蛋白质。结果，鉴定出了一种环核苷酸门控通道（CNGC）蛋白OsCNGC12。通过基因编辑使OsCNGC12功能失活，我们开发了 BSM 耐受性水稻品系。我们的结果表明，OsCNGC12突变体水稻不仅减少了 BSM 的吸收，而且促进了根部Ca ^{2+的流入，从而增强了对 BSM 的非靶位点耐受性。}OsCNGC12 定位于质膜，在根冠、根维管束、茎和叶中大量表达，表明其在 BSM 在水稻中的分布中发挥着重要作用。通过 OsCNGC12 的定向诱变证明了 His-236、Gln-240 和 Arg-268 在酵母 BSM 感知中不可或缺。此外，我们还为OsCNGC12突变体水稻开发了 BSM 种衣剂，为直播水稻杂草控制提供了一种简化且经济高效的方法。总的来说，这项研究表明，OsCNGC12 的破坏赋予了水稻对 BSM 的非靶位点耐受性，并具有开发抗 BSM 水稻品种的潜力。