캐비어에 대한 수요가 증가하면서 캐비어 종류의 진위 여부를 확인 할 수 있는 분석법 마련이 필요해졌다. 본 연 구에서는 캐비어 종류을 나누는 철갑상어 종 특이 KASP 마커를 개발하였고 모니터링을 통해 국내 유통중인 불법 캐비어 제품을 확인하고자 하였다. 본 연구에서는 16S ribosomal RNA gene, cytochrome b gene, cytochrome c oxidase subunit I gene, cytochrome c oxidase subunit II gene, NADH dehydrogenase subunit 5 gene에서 철갑상어 종 특이적인 SNP를 선정하고 이를 표적하는 KASP 마커 11종을 개발하였다. 개발한 KASP 마커를 이용하여 한국 의 온라인 마켓에서 유통중인 캐비어 10종을 모니터링 분 석한 결과 2개의 제품에서 제품에 표기된 철갑상어 종과 다른 것으로 확인하였으며 두 제품 모두 실제보다 더 비 싼 캐비어로 둔갑하여 판매한 것으로 확인되었다. 본 연 구를 통해 제작한 KASP 분석방법으로 유통되는 캐비어 종류 분류가 가능하였고 모니터링을 통해 국내 유통되고 있는 불법 캐비어 제품도 확인하였다. 이에 따라 본 연구 에서 개발한 SNP기반 KASP 분석법으로 불법 캐비어 제 품 유통 근절에 기여 할 수 있을 것으로 기대한다.
Cucurbita is one of the crops with high demand in the world. Securing breeding sources is crucial and fundamental to any plant breeding program. This study was conducted to investigate the characterization of phenotypic traits and phylogenetic classification of germplasm provided by the National Agrobiodiversity Center of RDA. Fourteen phenotypic traits were measured in 199 accessions of Cucurbita germplasm. Among these germplasm, 92 accessions of C. moschata, 34 accessions of C. maxima, and 73 accessions of C. pepo were classified using the KASP marker from the Seed Industry Center. It was confirmed that there were five classifications in C. moschata, five in C. maxima and four in the C. pepo. The results of this study provide fundamental data on Cucurbita germplasm and are expected to be useful in breeding programs.
Known for its effectiveness in weight loss and diabetes prevention, Gymnema sylvestre products can be found in the US, Japanese, and Indian markets. However, the recommended dosage or safety of these products has not yet been proven. Therefore, development of an analytical method for detecting the content of Gymnema sylvestre in food products is required. Accordingly, this study proposes an analysis method that can examine Gymnema sylvestre in food using LC-ESI-MS/MS and KASP (Kompetitive Allele-Specific PCR) markers. In LC-ESI-MS/MS, a simultaneous analysis method for gymnemic acid and deacylgymnemic acid was optimized using negative ionization mode, and its validation test was completed for solid and liquid samples. In addition, KASP markers were prepared by finding the specific SNP of G. sylvestre in ITS2 and matK through DNA barcodes. The two KASP markers returned positive FAM fluorescence result when combined with G. sylvestre, and this aspect was confirmed in raw G. sylvestre as well. The applicability of the method was tested on 21 different food and healthy functional products containing G. sylvestre purchased on the internet. As a result, although there was a difference in the ratios of gymnemic acid and deacylgymnemic acid in LC-ESI-MS/MS, the index component was detected in all 21 products samples. In the KASP analysis, 9 products returned positive FAM result, and the rest of the products were found to be containing G. sylvestre extract. This study is the first study to use the dual system of LC-ESI-MS/MS and KASP for the analysis of G. sylvestre. The study has confirmed that these two methods are applicable to the examine G. sylvestre content in food products.