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


ABSTRACT:

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.

SUBMITTER: Ibrahim S 

PROVIDER: S-EPMC9039650 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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

Ibrahim Saira S   Saleem Bilal B   Rehman Nazia N   Zafar Syed Adeel SA   Naeem Muhammad Kashif MK   Khan Muhammad Ramzan MR  

Journal of advanced research 20210714


<h4>Introduction</h4>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. <i>Inositol pentakisphosphate 2- kinase 1</i> (<i>IPK1</i>) gene has been reported to be an important gene for PA biosynthesis.<h4>Objective</h4>A recent genome editing tool CRISPR/Cas9 has been successfully applied to develop biofortified rice by disrupting <i>IPK1</i> gene, however, it remained a challenge in w  ...[more]

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