Cadmium (Cd) contamination in rice poses significant health risks to consumers. This study aimed to reduce Cd accumulation in the elite Vietnamese rice variety TBR225 (TBR225) by editing the Natural Resistance-Associated Macrophage Protein 5 (OsNRAMP5) gene using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR- associated 9 (Cas9) technology. We successfully generated OsNRAMP5 knockout mutants through Agrobacterium-mediated transformation. Sequencing analysis revealed various mutation types, including deletions, insertions, and substitutions, all resulting in premature stop codons or frameshift mutations. Three homozygous, transgene-free mutant lines were selected for further analysis. These lines exhibited significantly reduced Cd accumulation in roots (78.4-84.5%), shoots (72.3-83.8%), and grains (50.5-66.0%) compared to wild-type plants when exposed to different Cd concentrations. Importantly, the OsNRAMP5 mutations did not adversely affect major agronomic traits, including growth time, plant height, tiller number, grain yield, or amylose content. Additionally, the accumulation of essential micronutrients such as Fe and Zn remained unaffected in the mutant lines. Our results demonstrate the potential of CRISPR/Cas9-mediated OsNRAMP5 editing as an effective strategy for developing low-Cd accumulating rice varieties without compromising agronomic performance or nutritional quality. This approach could significantly contribute to enhancing food safety in regions facing Cd contamination challenges.

Citations

Citations to this article as recorded by  

• Genome editing in rice: toward climate-resilient, nutrient-rich yields
  Xin Huang, Wenshu He, Ludovic Bassie, Paul Christou, Teresa Capell
  Trends in Plant Science.2026; 31(7): 925.     CrossRef
• CRISPR/Cas9 Genome Editing in Rice (Oryza sativa L.) for Multi-Stress Resilience and Prospects for Agricultural Sustainability
  Abdul Malik, Nur Ardiyana Rejab, Muhamad Afiq Aziz
  Journal of Crop Health.2026;[Epub]     CrossRef
• Engineering Abiotic Stress Tolerance in Plants Using CRISPR Cas: From Molecular Targets to Physiological Resilience
  Komal Sharma, Upma Bhatt, Vineet Soni
  Journal of Soil Science and Plant Nutrition.2026; 26(2): 4593.     CrossRef
• Exploring Flooding Challenges, Causes, and Mitigation Strategies in Rice
  Abdoul-Razak Oumarou Mahamane, Junior S. Kamara, Moise Hubert Byiringiro, Alpha Sow, Khaled F. M. Salem
  International Journal of Genomics.2026;[Epub]     CrossRef
• Liebig Review: Essential and Beneficial Elements in the Regulation of Cadmium Uptake and Tolerance in Crop Plants
  Maria Manzoor, Karl Hermann Mühling
  Journal of Plant Nutrition and Soil Science.2026;[Epub]     CrossRef
• CRISPR/Cas9 Mediated Genome Editing for Enhancing Abiotic Stress Tolerance in Rice: An Omics Guided Perspective
  Mahavir Joshi, Pari Panwar, Smile Sharma, Bharat Sagar, Sukhminderjit Kaur, Manikant Tripathi
  Molecular Biotechnology.2026;[Epub]     CrossRef
• Harnessing breeding and biotechnological innovations for global food security under climate change
  Obaid Ur Rehman, Feifei Zhu, Xinjuan Hu, Xiangru Xu, Muhammad Uzair, Jiangya Qian, Sajid Fiaz, Shuhao Huo
  Functional & Integrative Genomics.2025;[Epub]     CrossRef