Development of a Gene-Based Soybean-Origin Discrimination Method Using Allele-Specific Polymerase Chain Reaction  
[PERSON]  
1 Experiment & Research Institute, National Agricultural Products Quality Management Service, Gimcheon 39660, Republic of Korea; [EMAIL_ADDRESS] (K-C.J. ); [EMAIL_ADDRESS] (B-Y.K. ); [EMAIL_ADDRESS] (M-J.K.); [EMAIL_ADDRESS] (N-K.K. ); [EMAIL_ADDRESS] (H-S-J.) [PERSON]  
1 Experiment & Research Institute, National Agricultural Products Quality Management Service, Gimcheon 39660, Republic of Korea; [EMAIL_ADDRESS] (K-C.J. ); [EMAIL_ADDRESS] (B-Y.K. ); [EMAIL_ADDRESS] (M-J.K.); [EMAIL_ADDRESS] (N-K.K. ); [EMAIL_ADDRESS] (H-S-J.) [PERSON]  
1 Experiment & Research Institute, National Agricultural Products Quality Management Service, Gimcheon 39660, Republic of Korea; [EMAIL_ADDRESS] (K-C.J. ); [EMAIL_ADDRESS] (B-Y.K. ); [EMAIL_ADDRESS] (M-J.K.); [EMAIL_ADDRESS] (N-K.K. ); [EMAIL_ADDRESS] (H-S-J.) [PERSON]  
1 Experiment & Research Institute, National Agricultural Products Quality Management Service, Gimcheon 39660, Republic of Korea; [EMAIL_ADDRESS] (K-C.J. ); [EMAIL_ADDRESS] (B-Y.K. ); [EMAIL_ADDRESS] (M-J.K.); [EMAIL_ADDRESS] (N-K.K. ); [EMAIL_ADDRESS] (H-S-J.) [PERSON]  
3 National Institute of Crop Science, Rural Development Administration, Suwon 16429, Republic of Korea; [EMAIL_ADDRESS] (Y-H.K. ); [EMAIL_ADDRESS] (H-.M.P)  
Correspondence: [EMAIL_ADDRESS] (H-C.S. ); [EMAIL_ADDRESS] (T-JK. ); tjkm@chnu. that soybean curd ranked sixth and soybean ranked seventh among the 156 items violating the Act on Origin Labeling of Agricultural and Fishery Products [3]. The origin of soybean is currently discriminated based on the difference in inorganic compounds using energy dispersive X-ray fluorescence spectrometry (ED-XRF). Furthermore, the absorbance difference across organic compounds is determined using Fourier transform near-infrared spectroscopy (FT-NIRS) [4; 5; 6]. The conventional physicochemical method of analysis enables a markedly simple and rapid analysis that requires no pre-treatment besides the grinding of samples. However, a single grain of soybean cannot be analyzed. Additionally, the minimum sample mass required for analysis is 5-50 g. These limitations could pose difficulties in the origin discrimination of mixed samples. Extensive research has been conducted to distinguish growing plant varieties through morphological classification. Experts in origin determination have traditionally relied on distinguishing varieties based on seed shape, gloss, and color. However, a challenge arises as results tend to vary depending on the evaluator's level of expertise and experience. To address this issue, the field has turned to DNA molecular marker technology for breed identification [7]. The most frequently used method in soybean variety analysis is the simple sequence repeat (SSR), or microsatellite, approach used to establish barcode systems. However, this method requires expensive equipment such as a DNA sequencer or chip electrophoresis and a large number of markers, making it difficult to adopt as a general laboratory method [8; 9]. The second most used PCR method is the cleaved amplified polymorphic sequence (CAPS) with a single-nucleotide polymorphisms (SNPs) approach. This method is time-consuming owing to the involvement of restriction enzymes in addition to PCR [10]. Recently, insertion-deletion (InDel) markers have achieved high reproducibility and are recognized as efficient molecular markers for distinguishing cultivars based on codominance [11]. In particular, these markers are attracting attention from researchers because they are relatively simple compared to other molecular markers. Soybeans possess 20 chromosomes with a known genome euchromatic DNA size of 705 Mb. These chromosomes are categorized into sVB (sparse variation block), characterized by the absence of chromosomal mutation, and dVB (dense variation block), which occurs infrequently.