Open Access

ARTICLE

Construction and Functional Analysis of CRISPR/Cas9 Vector of FAD2 Gene Family in Soybean

Zhifeng Xiao, Yukun Jin, Qi Zhang, Abraham Lamboro, Baozhu Dong, Zhuoyu Yang, Piwu Wang^*

College of Agronomy, Jilin Agricultural University, Changchun, 130118, China

* Corresponding Author: Piwu Wang. Email:

(This article belongs to the Special Issue: Plant Physiology for Crop Production and Sustainable Agriculture)

Phyton-International Journal of Experimental Botany 2022, 91(2), 349-361. https://doi.org/10.32604/phyton.2022.017451

Received 11 May 2021; Accepted 18 June 2021; Issue published 26 September 2021

Abstract

Soybean oleic acid content is one of the important indexes to evaluate the quality of soybean oil. In the synthesis pathway of soybean fatty acids, the FAD2 gene family is the key gene that regulates the production of linoleic acid from soybean oleic acid. In this study, CRISPR/Cas9 gene editing technology was used to regulate FAD2 gene expression. Firstly, the CRISPR/Cas9 single knockout vectors GmFAD2-1B and GmFAD2-2C and double knockout vectors GmFAD2-2A-3 were constructed. Then, the three vectors were transferred into the recipient soybean variety Jinong 38 by Agrobacterium-mediated cotyledon node transformation, and the mutant plants were obtained. Functional analysis and comparison of the mutant plants of the T2 and T3 generations were carried out. The results showed that there was no significant difference in agronomic traits between the CRISPR/Cas9 single and double knockout vectors and the untransformed CRISPR/Cas9 receptor varieties. The oleic acid content of the plants that knocked out the CRISPR/Cas9 double gene vector was significantly higher than that of the single gene vector.

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