Single nucleotide polymorphism-based visual identification of Rhodiola crenulata using the loop-mediated isothermal...

Area of Science:

• Pharmacognosy
• Molecular Biology
• Traditional Chinese Medicine

Background:

• Rhodiola crenulata is a key ingredient in traditional Chinese medicine, used for fatigue and stress.
• Accurate identification of Rhodiola species is crucial due to market confusion and safety concerns.

Purpose of the Study:

• To develop a simple, rapid, specific, and sensitive method for identifying Rhodiola crenulata.
• To distinguish Rhodiola crenulata from closely related species and hybrids.

Main Methods:

• Utilized cleaved amplified polymorphic sequence (CAPS) RT-PCR to identify single nucleotide polymorphism (SNP) sites.
• Developed a loop-mediated isothermal amplification (LAMP) assay based on identified SNP sites.
• Optimized primers, reaction temperature, time, and incorporated color indicators (HNB, N-red) for visualization.

Main Results:

• Identified specific SNP sites differentiating Rhodiola crenulata from related species.
• Established a real-time fluorescence LAMP system with optimal parameters.
• Achieved a sensitivity of 1,000 copies of target DNA within 35 minutes at 62°C.

Conclusions:

• Developed a simple, rapid, specific, sensitive, and visible method for Rhodiola crenulata identification.
• The LAMP assay effectively distinguishes Rhodiola crenulata from other species and hybrids.
• This method holds significant potential for the pharmaceutical industry to ensure herbal medicine quality.