CRISPR/Cas9 mediated disruption of Inositol Pentakisphosphate 2-Kinase 1 (TaIPK1) reduces phytic acid and improves iron and zinc accumulation in wheat grains

Journal of Advanced Research

Nazia Rehman¹  Saira Ibrahim²  Bilal Saleem²  Syed Adeel Zafar³  Muhammad Ramzan Khan³  Muhammad Kashif Naeem³ 

[1] National Institute for Genomic and Advanced Biotechnology, National Agricultural Research Centre, Park Road, Islamabad Pakistan;Genome Editing and Sequencing Lab, National Centre for Bioinformatics, Quaid-i-Azam University Islamabad, Pakistan;National Institute for Genomic and Advanced Biotechnology, National Agricultural Research Centre, Park Road, Islamabad Pakistan;

关键词: Biofortification;    CRISPR/Cas9;    Phytic acid;    Iron;    Zinc;   

DOI  :  

来源: DOAJ

【 摘 要 】

Introduction: Phytic acid (PA) is an important antinutrient agent present in cereal grains which reduces the bioavailability of iron and zinc in human body, causing malnutrition. Inositol pentakisphosphate 2- kinase 1 (IPK1) gene has been reported to be an important gene for PA biosynthesis. Objective: A recent genome editing tool CRISPR/Cas9 has been successfully applied to develop biofortified rice by disrupting IPK1 gene, however, it remained a challenge in wheat. The aim of this study was to biofortify wheat using CRISPR/Cas9. Methods: In this study, we isolated 3 TaIPK1 homeologs in wheat designated as TaIPK1.A, TaIPK1.B and TaIPK1.D and found that the expression abundance of TaIPK1.A was stronger in early stages of grain filling. Using CRISPR/Cas9, we have disrupted TaIPK1.A gene in cv. Borlaug-2016 with two guide RNAs targeting the 1st and 2nd exons. Results: We got several genome-edited lines in the T0 generation at frequencies of 12.7% and 10.8%. Sequencing analysis revealed deletion of 1–23 nucleotides and even an addition of 1 nucleotide in various lines. Analysis of the genome-edited lines revealed a significant decrease in the PA content and an increase in iron and zinc accumulation in grains compared with control plants. Conclusion: Our study demonstrates the potential application of CRISPR/Cas9 technique for the rapid generation of biofortified wheat cultivars.

【 授权许可】

Unknown