Abstract
Clustered Regularly Interspaced Short Palindromic Repeat/CRISPR-associated protein 9 (CRISPR/Cas9) system has been regularly applied for genome editing and gene function identification in wild soybean (Glycine max) cultivars. However, till date no studies have demonstrated successful mutagenesis in wild soybean (Glycine soja) which is the ancestor of Glycine max and rich in stress tolerance genes. In the current study, we report the successful creation of mutations in the loci encoding plasma membrane Na+/H+ antiporter (SOS1) and nonselective cation channels (NSCC) in wild soybean hairy roots using the CRISPR/Cas9 system. Two genes, GsSOS1 and GsNSCC, were mutagenized with frequencies of 28.5% and 39.9%, respectively. Biallelic mutations in GsSOS1 were detected in transgenic hairy roots. GsSOS1 mutants exhibited altered Na+/K+ ratios in the roots under both control and salt-treated conditions. However, no significant effects of GsNSCC mutation on Na+/K+ ratios were observed. RNA-Seq analysis revealed that both GsSOS1 and GsNSCC mutation altered the transcription profiles in mutant roots. Many differentially expressed gene sets that are associated with various cellular functions were identified. Our results demonstrated that CRISPR/Cas9 systems as powerful tools for wild soybean genome editing and would significantly advance the gene mining and functional identification in wild soybean.
Highlights
It is well known that soybean (Glycine max) is one of the most important crops of the world (Lam et al, 2010; Smil, 2000)
Based upon above-mentioned information and other reports in Glycine max, we investigated for the existence of studies wherein successful transformation and mutagenesis in the wild soybean (Glycine soja) using root hair was reported
The PCR-RE assay showed that gene mutations were induced (Figure1) and mutagenesis efficiencies for GsSOS1 and GsNSCC were 28.5% and 39.9%, respectively (Table 1)
Summary
It is well known that soybean (Glycine max) is one of the most important crops of the world (Lam et al, 2010; Smil, 2000). Wild soybean (Glycine soja) is the known ancestor of Glycine max, possessing much greater adaptability to a variety of environmental stresses and is believed to be rich in stress tolerance genes. It has, been suggested as a potential source of germplasm to improve the agronomic traits of cultivated soybean (Ge et al, 2011; Wen et al, 2009). The hairy soybean root system was used to detect the target gene editing efficiency with different sgRNA promoters (Sun et al, 2015; Du et al, 2016) or different target loci (Jacobs et al, 2015) and of endogenous and exogenous genes (Cai., 2015), which significantly improved the optimization of CRISPR/Cas system in soybean functional genomic research. Overexpression of GmMYB118 in soybean hairy roots, significantly improved the plants’ drought and salt tolerance when compared with jas.ccsenet.org
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
