Structure characterization and potential role of soybean phospholipases A multigene family in response to multiple abiotic stress uncovered by CRISPR/Cas9 technology

Abstract

Phospholipases A (PLAs) in plants play a crucial role in growth and development, as well as in response to biotic/abiotic stresses. To-date, there has not been any well documented, systematic analysis and comprehensive study of PLAs multigene family in soybean. Therefore, we conducted genome-wide characterization of PLAs in soybean (GmPLAs). Moreover, two paralog genes, GmpPLA-IIε &GmpPLA-IIζ, were knock-out by CRISPR/Cas9 (Clustered regular interspaced short palindromic repeat/CRISPR-associated protein9) technology to study their modulating role in soybean response to multiple abiotic stresses. A total 112 GmPLAs were identified in soybean genome comprising 78 PLA1, 29 patatin-like PLAs and 5 secretory sPLA2. Tandem and segmental duplication events accounted for expansion of PLAs in soybean genome. Some of the mutant lines exhibited superior performance under flooding and drought conditions. Also, the knock-out of the two paralog genes individually or simultaneously disturbed the root's response to a phosphorus-deficient environment relative to the wild type. All the mutants performed better under iron deficient condition compared to the wild type. Our study suggests that PLAs in soybean have diverse physiological and regulatory roles in abiotic stress response and tolerance. The mutants would be useful genetic materials to unravel molecular mechanism underlying soybean response to multiple abiotic stress conditions.