Yin Congcong, Zhao Qiaoling, Yue Aiqin, Du Weijun, Liu Dingbin, Zhao Jinzhong, Zhang Yongpo, Wang Min
College of Arts and Sciences, Shanxi Agricultural University, Taigu 030801, Shanxi, China.
College of Agronomy, Shanxi Agricultural University, Taigu 030801, Shanxi, China.
J Anal Methods Chem. 2020 Nov 5;2020:8813239. doi: 10.1155/2020/8813239. eCollection 2020.
Soybean saponin is one of the important secondary metabolites in seeds, which has various beneficial physiological functions to human health. gene is the key enzyme gene for synthesizing class A saponins. It is of great significance to realize the visual and rapid detection of class A saponins at the genetic level. The hybridization chain reaction (HCR) was employed to the visual detection of gene, which was implemented by changing the length of the target fragment to 92 bp and using the hairpin probes we designed to detect the and genes. The best condition of HCR reaction is hemin (1.2 M), Triton X-100 (0.002%), ABTS (3.8 M), and HO (1.5 mM). It was found that HCR has high specificity for gene and could be applied to the visual detection of different soybean cultivars containing Aa type, Ab type, and Aa/Ab type saponins, which could provide technical reference and theoretical basis for molecular breeding of soybean and development of functional soybean products.
大豆皂苷是种子中重要的次生代谢产物之一，对人体健康具有多种有益的生理功能。基因是合成 A 类皂苷的关键酶基因。在基因水平上实现对 A 类皂苷的可视化快速检测具有重要意义。采用杂交链式反应（HCR）对基因进行可视化检测，通过将目标片段长度改为 92 bp 并使用我们设计的发夹探针来检测和基因来实现。HCR 反应的最佳条件是血红素（1.2 M）、 Triton X - 100（0.002%）、ABTS（3.8 M）和 HO（1.5 mM）。发现 HCR 对基因具有高特异性，可应用于对含有 Aa 型、Ab 型和 Aa/Ab 型皂苷的不同大豆品种的可视化检测，可为大豆分子育种和功能性大豆产品开发提供技术参考和理论依据。
