**5.2 Challenges and prospective for GE and related product development in soybean**

In the last four decades, popular transgenic technology has been used to introduce foreign genes into crops, such as soybean, for desired qualities, and it has proven to be a viable option to expanding genetic resources. The random incorporation of transgenes in the genome, on the other hand, has triggered public outrage and rigorous government restriction, drastically increasing the cost and time required to generate a new variety. Instead of going through repeated back crosses to transfer a natural mutation in a traditional breeding method, GE technology allows crop breeders to integrate a desired feature into an elite background in a precise and predictable manner. Traditional mutagenesis breeding introduces mutations that are indistinguishable from those induced by GE. The largest constraint for GE application in soybean, such as other crops [130], is a lack of GE candidate target genes due to poor foundational

research, as described above. Technical issues such as the inability to precisely mutate any target site, the lack of ways to deliver genome-editing reagents into soybean cells, the low efficiency of selecting desired events and regenerating intact plants with targeted mutation, and off-site targeting are among the remaining bottlenecks. Through the use of newly developed GE technologies and a soybean regeneration system, several attempts have been made to reduce the restrictions and enhance the efficiency of recovering GE events. Additional challenges for GE product development include transgenic GE events, intellectual property restrictions, and government control of GE. Before GE can play a significant role in soybean improvement, these challenges must be addressed.

### **6. Success stories of CRISPR/Cas9 mediated in soybean (***G. max* **L.)**

The CRISPR-Cas9 method has been used to successfully mutate the genes GmFT2a, FAD2-2, and GmSPL9 in soybean modifying flowering time, seed oil profile, and plant architecture, respectively. This success implies that employing the CRISPR-Cas9 technology to improve soybean agronomical qualities is possible.

Targeted mutants of E1 gene controlling soybean flowering were generated. Two new types of mutations were discovered: 11 bp and 40 bp deletions in the E1 coding area, respectively, and frameshift mutations that resulted in premature translation termination codons and shortened E1 proteins, causing early blooming under long day conditions. Furthermore, by predicting and analyzing the probable off-target areas of E1 targets, no off-target effects were found. The shortened E1 protein disinhibited GmFT2a/5a, and boosting GmFT2a/5a gene expression led in evident early flowering in two new mutants with significantly decreased E1 gene expression [131].

### **7. Conclusions**

Thanks to the CRISPR/Cas9-based gene-editing method, researchers may now change crop-specific traits more accurately and effectively. The CRISPR/Cas9 system has become the most frequently used and versatile technology in crop breeding and functional genomics. Its unrivaled ability to manipulate genes contributed in the development of a number of crop varieties with beneficial agronomic traits. Most crop-improvement gene-editing research, on the other hand, is still in the early stages of uncovering genomic function and regulatory pathways. Gene-edited crops are still a long way from becoming commercially available. In addition, gene-editing approaches have yet to meet all of the requirements for changing plant genomes. Because several quality-related variables are governed by many QTLs and altering individual genes may not generate significant phenotypic change, further development will be critical for the use of CRISPR/Cas in plants. It might be possible to create a CRISPR/Cas-mediated chromosomal rearrangement technology that works well. Furthermore, delivering CRISPR cargoes remains a significant challenge. As a result, it would be advantageous to design new carrier materials. Aside from that, public concerns and the government's rigorous regulatory policies on gene-editing technologies are another roadblock to plant breeding progress. Despite the remaining hurdles, gene-editing technology is expected to become more frequently used in the future and will undoubtedly play a significant part in crop quality enhancement like in soybean.

*Role of CRISPR/Cas9 in Soybean (*Glycine max *L.) Quality Improvement DOI: http://dx.doi.org/10.5772/intechopen.102812*
