Adulteration Identification Of Lonicera Japonica Based On DNA Molecular Marker

Lonicera japonica Thunb is widely used in traditional Chinese medicines for its heat-clearing, detoxifying, and anti-inflammatory properties, which comes from L. hypoglauca Miq., L. confusa(Sweet) DC., L. macranthoides Hand.-Mazz., and L. fulvotomentosa Hsu et S. C. Cheng, is another medicinal material described in the Chinese Pharmacopoeia.. In the current status, the source plants of Flos Lonicerae and many other species of Lonicera have been misused as Lonicera japonica Thunb in different areas of China. Previous studies on the identification of L. japonica and its related species have been focused mainly on traditional classification, chemical methods, and various genetic molecular markers, such as RFLP and the diagnostic PCR. But the morphological features and chemical components are often unreliable and inconsistent because of the influence of the environment, plant growth phase, harvesting time, testing condition, and various artificial factors.Based on EST-SSR and EST-SNP markers, which is the new generation of markers, we applied high resolution melting(HRM) and pyrosequencing in adulteration detection of Lonicera japonica, and established a rapid, accurate, high-throughput new detection system. The main findings are as follows:1?The Excel-based digital fingerprinting was established.The coefficient of genetic similarity of Lonicera japonica and Lonicera confusa was only 0.28.87 commercial samples were clustered using the SSR fingerprinting,and divided into 3 producing areas as expected The Lonicera japonica SSR fingerprinting was reliable and convenient,and could be used in germplasm identification.2?Based on the SNP, and combined with HRM analysis technology, we establish a rapid and high-throughput true or false test methods for honeysuckle. Aimming the significant variation SNP site, HRM can distinguish Lonicera japonica and Lonicerae flos. within 2 h.The assay is fast, low cost, convenient operation and the advantages of the closed tube detection to prevent pollution.3?Based on the SNP, and combined with pyrosequencing analysis technology, we establish a precisen and automation authenticity test methods for honeysuckle. Aimming the significant variation SNP site, pyrosequencing assay can calculate the content of Lonicera japonica and Lonicerae flos. within 3 h.