Targeting OsNIP3;1 via CRISPR/Cas9: A strategy for minimizing arsenic accumulation and boosting rice resilience

• Published in: Journal of hazardous materials 2024-06, Vol.471, p.134325-134325, Article 134325
• Main Authors: Singh, Puja, Kumar, Amit, Singh, Twinkle, Anto, Sonik, Indoliya, Yuvraj, Tiwari, Poonam, Behera, Soumit Kumar, Chakrabarty, Debasis
• Format: Article
• Language: English
• Subjects: aquaporins; Aquaporins - genetics; Aquaporins - metabolism; Arsenic; Arsenic - metabolism; Arsenic - toxicity; carotenoids; Chlorophyll - metabolism; CRISPR-Cas Systems; CRISPR/Cas9; family; Gene Editing; Gene Knockout Techniques; genes; grain yield; human health; NIP; Oryza - genetics; Oryza - growth & development; Oryza - metabolism; photosynthesis; plant growth; Plant Proteins - genetics; Plant Proteins - metabolism; Plants, Genetically Modified - genetics; Rice; sap; Silicon; Silicon - metabolism; stress tolerance; toxicity; vigor; xylem; Xylem sap
• ISSN: 0304-3894; 1873-3336
• DOI: 10.1016/j.jhazmat.2024.134325

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. [Display omitted] •Study focuses on targeting various As transporters - OsLsi1, OsLsi2 and OsNIP3;1 by genome editing.•Knock-out (KO) plants showed reduced As content in various parts of rice including grain.•OsNIP3;1 edited lines (KO Osnip3;1) showed reduced arsenic content in xylem sap by 71-74%.•Enhanced physiological and agronomic traits were observed in Osnip3;1 under As stress.