Molecular Biology and Biotechnology Division, CSIR-National Botanical Research Institute, Lucknow 226001, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India.
Molecular Biology and Biotechnology Division, CSIR-National Botanical Research Institute, Lucknow 226001, India.
J Hazard Mater. 2024 Jun 5;471:134325. doi: 10.1016/j.jhazmat.2024.134325. Epub 2024 Apr 16.
Arsenic (As) contamination in rice poses a significant threat to human health due to its toxicity and widespread consumption. Identifying and manipulating key genes governing As accumulation in rice is crucial for reducing this threat. The large NIP gene family of aquaporins in rice presents a promising target due to functional redundancy, potentially allowing for gene manipulation without compromising plant growth. This study aimed to utilize genome editing to generate knock-out (KO) lines of genes of NIP family (OsLsi1, OsNIP3;1) and an anion transporter family (OsLsi2), in order to assess their impact on As accumulation and stress tolerance in rice. KO lines were created using CRISPR/Cas9 technology, and the As accumulation patterns, physiological performance, and grain yield were compared against wild-type (WT) under As-treated conditions. KO lines exhibited significantly reduced As accumulation in grain compared to WT. Notably, Osnip3;1 KO line displayed reduced As in xylem sap (71-74%) and grain (32-46%) upon treatment. Additionally, these lines demonstrated improved silicon (23%) uptake, photosynthetic pigment concentrations (Chl a: 77%; Chl b: 79%, Total Chl: 79% & Carotenoid: 49%) overall physiological and agronomical performance under As stress compared to WT. This study successfully utilized genome editing for the first time to identify OsNIP3;1 as a potential target for manipulating As accumulation in rice without compromising grain yield or plant vigor.
砷（As）污染在大米中造成了严重的健康威胁，这主要是由于其毒性和广泛的食用性。鉴定和操纵控制水稻砷积累的关键基因对于降低这种威胁至关重要。水稻中大量的水通道蛋白大型 NIP 基因家族由于功能冗余，为基因操作提供了一个很有前途的目标，这可能允许在不影响植物生长的情况下进行基因操作。本研究旨在利用基因组编辑技术生成 NIP 家族（OsLsi1、OsNIP3;1）和阴离子转运蛋白家族（OsLsi2）的基因敲除（KO）系，以评估它们对水稻砷积累和胁迫耐受性的影响。使用 CRISPR/Cas9 技术创建 KO 系，并在 As 处理条件下，将其与野生型（WT）相比，比较其砷积累模式、生理性能和籽粒产量。KO 系的籽粒砷积累量明显低于 WT。值得注意的是，Osnip3;1 KO 系在木质部汁液（71-74%）和籽粒（32-46%）中的砷含量显著降低。此外，与 WT 相比，这些系在 As 胁迫下，硅（23%）吸收、光合色素浓度（Chl a：77%；Chl b：79%，总 Chl：79%和类胡萝卜素：49%）以及整体生理和农艺性能均有所提高。本研究首次成功利用基因组编辑技术鉴定 OsNIP3;1 为一种操纵水稻砷积累的潜在靶标，而不会影响籽粒产量或植物活力。
