The Role of CRISPR/Cas9 for Genetic Advancement of Soybean: A Review

Muhammad Naveed *

Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan.

Jazib Javed

Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan.

Usama Waheed

Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan.

Maham Sajid

Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan.

Mehrab Ijaz

Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan.

Areeba Sehar

Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan.

Mazhar Attique

Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan.

. Mahnoor

Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan.

Salman Javed

Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan.

*Author to whom correspondence should be addressed.


Soybean is a crucial legume crop that is mainly grown for extracting oil and protein content Which can be used as a food source for human beings as well as livestock. We can also use the protein obtained from soybean for the extraction of biofuel. There is a dire need to increase genetic research on soybean for improvement and enhanced production. One big reason for genetic research on soybean is to make its resilience to the change in the climate. In modern days CRISPR/Cas9 has evolved as an emerging technique that allows us to manipulate the gene of selected traits in most crops including soybean. Advanced tools of biotechnology are widely utilized for the enhancement of crop production, improving quality and yield, introducing disease and insect resistance, and being environmentally friendly. This review gives a glimpse of how the mechanism of CRISPR/Cas9 performs its functions and a brief discussion of CRISPR/Cas9 which has increased the scope of study in the genetic advancement of soybean. It also illustrates some phenomena in which we can use CRISPR/Cas9 for the betterment of soybean.

Keywords: CRISPR/Cas9, genetic improvement, soybean, gene editing

How to Cite

Naveed, M., Javed, J., Waheed, U., Sajid, M., Ijaz, M., Sehar, A., Attique, M., Mahnoor, ., & Javed, S. (2022). The Role of CRISPR/Cas9 for Genetic Advancement of Soybean: A Review. Asian Journal of Biotechnology and Genetic Engineering, 5(2), 226–239. Retrieved from


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McCue P, Shetty K. Health benefits of soy isoflavonoids and strategies for enhancement: a review. Critical Reviews in Food Science and Nutrition. 2004;44(5): 361-367.

Singh G, Dukariya G, Kumar A. Distribution, importance and diseases of soybean and common bean: A review. Biotechnol. J. Int. 2020;24(6):86-98.

Pagano MC, Miransari M. The importance of soybean production worldwide, in Abiotic and biotic stresses in soybean production. Elsevier. 2016;1-26.

Hasanuzzaman M, et al. Soybean production and environmental stresses, in Environmental stresses in soybean production. Elsevier. 2016;61-102.

Rajput M, et al. RNA interference and CRISPR/Cas gene editing for crop improvement: A paradigm shift towards sustainable agriculture. Plants. 2021; 10(9):1914.

Wu X, Kriz AJ, Sharp PA. Target specificity of the CRISPR-Cas9 system. Quantitative Biology. 2014;2(2):59-70.

Asmamaw M, Zawdie B. Mechanism and applications of CRISPR/Cas-9-mediated genome editing. Biologics: Targets & Therapy. 2021;15:353.

Arora L, Narula A. Gene editing and crop improvement using CRISPR-Cas9 system. Frontiers in Plant Science. 2017;8: 1932.

Gupta D, et al. CRISPR-Cas9 system: A new-fangled dawn in gene editing. Life Sciences. 2019;232:116636.

Mahfouz MM, Piatek A, Stewart Jr CN. Genome engineering via TALENs and CRISPR/Cas9 systems: challenges and perspectives. Plant Biotechnology Journal. 2014;12(8):1006-1014.

Ran F, et al. Genome engineering using the CRISPR-Cas9 system. Nature Protocols. 2013;8(11):2281-2308.

Cai Y, et al. CRISPR/Cas9-mediated genome editing in soybean hairy roots. PLoS One. 2015;10(8):e0136064.

Bao, A, et al. CRISPR/Cas9-mediated targeted mutagenesis of GmSPL9 genes alters plant architecture in soybean. BMC Plant Biology. 2019;19(1):1-12.

Sun, Z, et al. Genetic improvement of the shoot architecture and yield in soya bean plants via the manipulation of GmmiR156b. Plant Biotechnology Journal. 2019;17(1): 50-62.

Li, M, et al. Identification of traits contributing to high and stable yields in different soybean varieties across three Chinese latitudes. Frontiers in Plant Science. 2020;10:1642.

Yang X, et al. Overexpression of GmGAMYB accelerates the transition to flowering and increases plant height in soybean. Frontiers in Plant Science. 2021;12:667242.

Hwang S, Lee TG. Integration of lodging resistance QTL in soybean. Scientific reports, 2019;9(1):1-11.

Cheng Q, et al. CRISPR/Cas9-mediated targeted mutagenesis of GmLHY genes alters plant height and internode length in soybean. BMC Plant Biology. 2019. 19(1):1-11.

Zhang, M, et al,. Progress in soybean functional genomics over the past decade. Plant Biotechnology Journal. 2022; 20(2):256.

Lu S, et al. Current overview on the genetic basis of key genes involved in soybean domestication. Abiotech. 2022:1-14.

Kara SR, Choudhuryb S, Chakrabortyc A. CRISPR/Cas9 for soybean improvement: A review.

Herman EM, et al. Genetic modification removes an immunodominant allergen from soybean. Plant Physiology. 2003; 132(1):36-43.

Sedivy EJ, Wu F, Hanzawa Y. Soybean domestication: the origin, genetic architecture and molecular bases. New Phytologist. 2017;214(2):539-553.

Chouard P. Vernalization and its relations to dormancy. Annual Review of Plant Physiology. 1960;11(1):191-238.

Buzzell R. Inheritance of a soybean flowering response to fluorescent-daylength conditions. Canadian Journal of Genetics and Cytology. 1971;13(4):703-707.

McBlain B, Bernard R. A new gene affecting the time of flowering and maturity in soybeans. Journal of Heredity. 1987;78(3):160-162.

Ray JD, et al. Genetic control of a long‐juvenile trait in soybean. Crop Science. 1995; 35(4):1001-1006.

Bonato ER, Vello NA. E6, a dominant gene conditioning early flowering and maturity in soybeans. Genetics and Molecular Biology. 1999; 22:229-232.

Cober ER, Voldeng HD. A new soybean maturity and photoperiod‐sensitivity locus linked to E1 and T. Crop Science. 2001;41(3):698-701.

Cober ER, Morrison MJ. Regulation of seed yield and agronomic characters by photoperiod sensitivity and growth habit genes in soybean. Theoretical and Applied Genetics. 2010; 120(5):1005-1012.

Kong F, et al. A new dominant gene E9 conditions early flowering and maturity in soybean. Crop Science. 2014;54(6):2529-2535.

Lu, S, et al. Natural variation at the soybean J locus improves adaptation to the tropics and enhances yield. Nature Genetics. 2017;49(5):773-779.

Yue Y, et al. A single nucleotide deletion in J encoding GmELF3 confers long juvenility and is associated with adaption of tropic soybean. Molecular Plant. 2017;10(4):656-658.

Wang F, et al. A new dominant locus, E11, controls early flowering time and maturity in soybean. Molecular Breeding. 2019;39(5):1-13.

Xia Z, et al. Positional cloning and characterization reveal the molecular basis for soybean maturity locus E1 that regulates photoperiodic flowering. Proceedings of the National Academy of Sciences. 2012;109(32):E2155- E2164.

Watanabe S, et al. A map-based cloning strategy employing a residual heterozygous line reveals that the GIGANTEA gene is involved in soybean maturity and flowering. Genetics. 2011; 188(2):395-407.

Liu, B, et al. Genetic redundancy in soybean photoresponses associated with duplication of the phytochrome A gene. Genetics. 2008;180(2):995-1007.

Watanabe S, et al. Map-based cloning of the gene associated with the soybean maturity locus E3. Genetics. 2009;182(4): 1251-1262.

Kong, F, et al. Two coordinately regulated homologs of FLOWERING LOCUS T are involved in the control of photoperiodic flowering in soybean. Plant Physiology. 2010;154(3):1220-1231.

Cai Y, et al. CRISPR/Cas9‐mediated targeted mutagenesis of GmFT2a delays flowering time in soya bean. Plant Biotechnology Journal. 2018;16(1):176-185.

Jacobs TB, et al. Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology. 2015; 15(1):1-10.

Li Z, et al. Cas9-guide RNA directed genome editing in soybean. Plant Physiology. 2015. 169(2):960-970.

Kanazashi Y, et al. Simultaneous site-directed mutagenesis of duplicated loci in soybean using a single guide RNA. Plant Cell Reports. 2018;37(3):553-563.

Do PT, et al. Demonstration of highly efficient dual gRNA CRISPR/Cas9 editing of the homeologous GmFAD2–1A and GmFAD2–1B genes to yield a high oleic, low linoleic and α-linolenic acid phenotype in soybean. BMC Plant Biology. 2019;19(1):1-14.

Li, Z, et al. Multiplex CRISPR/Cas9-mediated knockout of soybean LNK2 advances flowering time. The Crop Journal. 2021;9(4):767-776.

Nan H, et al. GmFT2a and GmFT5a redundantly and differentially regulate flowering through interaction with and upregulation of the bZIP transcription factor GmFDL19 in soybean. PloS one. 2014;9(5):e97669.

Cai Y, et al. Mutagenesis of GmFT2a and GmFT5a mediated by CRISPR/Cas9 contributes to expanding the regional adaptability of soybean. Plant Biotechnology Journal. 2020;18(1):298-309.

Subedi U, et al. The potential of genome editing for improving seed oil content and fatty acid composition in oilseed crops. Lipids. 2020;55(5):495-512.

Bortesi L, Fischer R. The CRISPR/Cas9 system for plant genome editing and beyond. Biotechnology Advances. 2015; 33(1):41-52.

Villanueva-Mejia, D. and J.C. Alvarez, Genetic improvement of oilseed crops using modern biotechnology. Adv. Seed Biol, 2017:295-317.

Al Amin, N, et al. CRISPR-Cas9 mediated targeted disruption of FAD2–2 microsomal omega-6 desaturases in soybean (Glycine max. L). BMC Biotechnology. 2019;19(1):1-10.

Ma X, et al. A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Molecular Plant. 2015;8(8): 1274-1284.

Mikami M, Toki S, Endo M. Comparison of CRISPR/Cas9 expression constructs for efficient targeted mutagenesis in rice. Plant Molecular Biology. 2015;88(6):561-572.

Singer SD, et al. Genetic engineering of lipid biosynthesis in seeds, in Biotechnology of crucifers. Springer. 2013;111-149.

Singer SD, Weselake RJ, Rahman H. Development and characterization of low α-linolenic acid Brassica oleracea lines bearing a novel mutation in a ‘class a’FATTY ACID DESATURASE 3gene. BMC Genetics. 2014;15(1):1-11.

Roesler K, et al. An improved variant of soybean type 1 diacylglycerol acyltransferase increases the oil content and decreases the soluble carbohydrate content of soybeans. Plant Physiology. 2016;171(2):878-893.

Chen G, et al. High‐performance variants of plant diacylglycerol acyltransferase 1 generated by directed evolution provide insights into structure-function. The Plant Journal. 2017;92(2):167-177.

Rodríguez-Leal D, et al. Engineering quantitative trait variation for crop improvement by genome editing. Cell. 2017;171(2):470-480. e8.

Wolter F, Schindele P, Puchta H. Plant breeding at the speed of light: the power of CRISPR/Cas to generate directed genetic diversity at multiple sites. BMC Plant Biology. 2019; 19(1):1-8.

Fan Y, et al. The soybean Rfg1 gene restricts nodulation by Sinorhizobium fredii USDA193. Frontiers in Plant Science. 2017;8:1548.

Herridge DF, Peoples MB, Boddey RM. Global inputs of biological nitrogen fixation in agricultural systems. Plant and Soil. 2008;311(1):1-18.

Wang L, et al. Use of CRISPR/Cas9 for symbiotic nitrogen fixation research in legumes. Progress in Molecular Biology and Translational Science. 2017;149:187-213.

Bai M, et al. Generation of a multiplex mutagenesis population via pooled CRISPR‐Cas9 in soya bean. Plant Biotechnology Journal. 2020;18(3):721-731.

Ren B, et al. Rhizobial tRNA-derived small RNAs are signal molecules regulating plant nodulation. Science. 2019;365(6456):919-922.

Li M, et al. GmNAC06, a NAC domain transcription factor enhances salt stress tolerance in soybean. Plant Molecular Biology. 2021;105(3):333-345.

Wei Y, et al. Quantitative response of soybean development and yield to drought stress during different growth stages in the Huaibei Plain, China. Agronomy. 2018;8(7):97.

Xiong R, et al. Root system architecture, physiological and transcriptional traits of soybean (Glycine max L.) in response to water deficit: A review. Physiologia Plantarum. 2021; 172(2):405-418.

Imran M, et al. Exogenous melatonin induces drought stress tolerance by promoting plant growth and antioxidant defense system of soybean plants. AoB Plants. 2021;13(4):plab026.

Sharma M, et al. Proteomics unravel the regulating role of salicylic acid in soybean under yield limiting drought stress. Plant Physiology and Biochemistry. 2018;130:529-541.

Mahajan S, Tuteja N. Cold, salinity, and drought stresses an overview. Archives of Biochemistry and Biophysics. 2005;444(2):139-158.

Dowling DK, Simmons LW. Reactive oxygen species as universal constraints in life-history evolution. Proceedings of the Royal Society B: Biological Sciences. 2009;276(1663):1737-1745.

Nakagawa A, et al. Drought stress during soybean seed filling affects storage compounds through regulation of lipid and protein metabolism. Acta Physiologiae Plantarum. 2018;40(6):1-8.

Egli D, Bruening W. Water stress, photosynthesis, seed sucrose levels and seed growth in soybean. The Journal of Agricultural Science. 2004;142(1):1-8.

Seo JS, et al. Expression of the Arabidopsis AtMYB44 gene confers drought/salt-stress tolerance in transgenic soybean. Molecular Breeding. 2012; 29(3):601-608.

Raza G, Singh MB, Bhalla PL. Somatic embryogenesis and plant regeneration from commercial soybean cultivars. Plants. 2019;9(1):38.

Yuan L, et al. GmLCLs negatively regulates ABA perception and signalling genes in soybean leaf dehydration response. Plant, Cell & Environment. 2021; 44(2):412-424.

Yang C, et al. GmNAC8 acts as a positive regulator in soybean drought stress. Plant Science. 2020;293:110442.

Jamil A, et al. Gene expression profiling of plants under salt stress. Critical Reviews in Plant Sciences. 2011;30(5):435-458.

Hamayun M, et al. Exogenous gibberellic acid reprograms soybean to higher growth and salt stress tolerance. Journal of Agricultural and Food Chemistry. 2010; 58(12):7226-7232.

Rasheed A, et al. Molecular tools and their applications in developing salt-tolerant soybean (Glycine max L.) cultivars. Bioengineering. 2022;9(10):495.

Sun T, et al. A golgi-localized sodium/hydrogen exchanger positively regulates salt tolerance by maintaining higher K+/Na+ ratio in soybean. Frontiers in Plant Science. 2021;12:638340.

Li X, et al. CRISPR/Cas9 technique for temperature, drought, and salinity stress responses. Current Issues in Molecular Biology. 2022;44(6):2664-2682.

Song JH, et al. Mutation of GmIPK1 gene using CRISPR/Cas9 reduced phytic acid content in soybean seeds. International Journal of Molecular Sciences. 2022; 23(18):10583.

Baek D, Chun HJ, Kim MC. Genome editing provides a valuable biological toolkit for soybean improvement. Plant Biotechnology Reports. 2022;1-12.

Sun Y, et al. Engineering herbicide-resistant rice plants through CRISPR/Cas9-mediated homologous recombination of acetolactate synthase. Molecular Plant. 2016;9(4):628-631.

Chen Y, et al. CRISPR/Cas9-mediated base-editing system efficiently generates gain-of-function mutations in Arabidopsis. Sci China Life Sci. 2017;60(5):520-523.

Tian S, et al. Engineering herbicide-resistant watermelon variety through CRISPR/Cas9-mediated base-editing. Plant Cell Reports. 2018;37(9):1353- 1356.

Wu J, et al. Engineering herbicide‐resistant oilseed rape by CRISPR/Cas9‐mediated cytosine base‐editing. Plant Biotechnology Journal. 2020;18(9):1857.

Ali Z, et al. Efficient virus-mediated genome editing in plants using the CRISPR/Cas9 system. Molecular Plant. 2015;8(8):1288-1291.

Bandara AY, et al. Dissecting the economic impact of soybean diseases in the United States over two decades. PloS one. 2020;15(4):e0231141.

Pompili V, et al. Reduced fire blight susceptibility in apple cultivars using a high‐efficiency CRISPR/Cas9‐FLP/FRT‐based gene editing system. Plant Biotechnology Journal. 2020; 18(3):845-858.

Pyott DE, Sheehan E, Molnar A. Engineering of CRISPR/Cas9‐mediated potyvirus resistance in transgene‐free Arabidopsis plants. Molecular Plant Pathology. 2016;17(8):1276-1288.

Ramakrishna W, et al. Structural analysis of the maize Rp1 complex reveals numerous sites and unexpected mechanisms of local rearrangement. The Plant Cell. 2002;14(12):3213-3223.

Wang, F, et al. Enhanced rice blast resistance by CRISPR/Cas9-targeted mutagenesis of the ERF transcription factor gene OsERF922. PloS one. 2016; 11(4):e0154027.

Zhang P, et al. Multiplex CRISPR/Cas9‐mediated metabolic engineering increases soya bean isoflavone content and resistance to soya bean mosaic virus. Plant Biotechnology Journal. 2020;18(6):1384-1395.

Nagy, E.D, et al, Novel disease resistance gene paralogs created by CRISPR/Cas9 in soy. Plant Cell Reports. 2021;40(6):1047-1058.

Liu JZ, Fang Y, Pang H. The current status of the soybean-soybean mosaic virus (SMV) pathosystem. Frontiers in Microbiology. 2016;7:1906.

Luo Y, et al. Development of a Csy4-processed guide RNA delivery system with soybean-infecting virus ALSV for genome editing. BMC Plant Biology. 2021;21(1):1-12.

Smith SM, Pryor AJ, Hulbert SH. Allelic and haplotypic diversity at the rp1 rust resistance locus of maize. Genetics. 2004;167(4):1939-1947.

Pivonia S, Yang X. Assessment of the potential year-round establishment of soybean rust throughout the world. Plant Disease. 2004;88(5):523-529.

Gao H, et al. Two classes of highly similar coiled coil-nucleotide binding-leucine rich repeat genes isolated from the Rps1-k locus encode Phytophthora resistance in soybean. Molecular Plant-microbe interactIons. 2005;18(10):1035-1045.

Chen X, et al. Generation of male-sterile soybean lines with the CRISPR/Cas9 system. The Crop Journal. 2021;9(6):1270-1277.

Jianing G, et al. CRISPR/Cas9 applications for improvement of soybeans, current scenarios, and future perspectives. Notulae Botanicae Horti Agrobotanici Cluj-Napoca. 2022;50(2):12678-12678.

Schmutz J, et al. Genome sequence of the palaeopolyploid soybean. Nature. 2010; 463(7278):178-183.

Fister AS, et al. Transient expression of CRISPR/Cas9 machinery targeting TcNPR3 enhances defense response in Theobroma cacao. Frontiers in Plant Science. 2018;9:268.

Holubová K, et al. Modification of barley plant productivity through regulation of cytokinin content by reverse-genetics approaches. Frontiers in Plant Science. 2018;9:1676.

Chen L, et al. Improvement of soybean Agrobacterium-mediated transformation efficiency by adding glutamine and asparagine into the culture media. International Journal of Molecular Sciences. 2018;19(10):3039.

Svitashev S, et al. Genome editing in maize directed by CRISPR–Cas9 ribonucleoprotein complexes. Nature Communications. 2016;7(1):1-7.

Foster AJ, et al. CRISPR-Cas9 ribonucleoprotein-mediated co-editing and counterselection in the rice blast fungus. Scientific Reports. 2018;8(1):1-12.

Lowe K, et al. Morphogenic regulators Baby boom and Wuschel improve monocot transformation. The Plant Cell. 2016;28(9):1998-2015.

Campbell BW, et al. Functional analysis and development of a CRISPR/Cas9 allelic series for a CPR5 ortholog necessary for proper growth of soybean trichomes. Scientific Reports. 2019; 9(1):1-11.

Bao A, et al. Genome editing technology and application in soybean improvement. Oil Crop Science. 2020;5(1):31-40.

He R, et al. Expanding the range of CRISPR/Cas9-directed genome editing in soybean. aBIOTECH. 2022;3(2):89-98.

Biden S, Smyth SJ, Hudson D. The economic and environmental cost of delayed GM crop adoption: The case of Australia's GM canola moratorium. GM Crops & Food. 2018;9(1):13-20.

Xu H, et al. Progresses, challenges, and prospects of genome editing in soybean (Glycine max). Frontiers in Plant Science. 2020;11:571138.

Curtin SJ, et al. Crispr/cas9 and talen s generate heritable mutations for genes involved in small rna processing of glycine max and medicago truncatula. Plant Biotechnology Journal. 2018. 16(6):1125-1137.

Waltz E. With a free pass, CRISPR-edited plants reach market in record time. Nature Biotechnology. 2018;36(1):6-8.