Cannabis Seeds Authentication by Chloroplast and Nuclear DNA Analysis Coupled with High-Resolution Melting Method for...

Area of Science:

• Genetics
• Plant Science
• Forensic Science

Background:

• Cannabis seeds have historical medicinal and nutritional uses, with rising demand for medical applications.
• An unregulated seed market and high product value lead to potential adulteration and quality control issues.
• Accurate identification of cannabis strains is crucial for regulated markets, medical use, and combating illegal trafficking.

Purpose of the Study:

• To develop and validate a rapid, cost-effective method for detecting contamination in commercial cannabis seeds.
• To implement DNA barcoding with high-resolution melting analysis for cannabis seed quality control.
• To differentiate between pure and adulterated cannabis seed batches.

Main Methods:

• DNA barcoding was performed on commercial and medical cannabis seeds using internal transcribed spacer (ITS) and rbcL markers.
• High-resolution melting (HRM) analysis was coupled with real-time PCR to analyze DNA sequence variations.
• Humulus lupulus L. was used as an outgroup for comparative genetic analysis.

Main Results:

• Homogeneous seed packages showed consistent melting temperatures, indicating single genotypes.
• Packages with contaminating seeds exhibited average melting temperature differences of 0.2°C.
• The developed method successfully identified variations indicative of seed contamination.

Conclusions:

• An effective, rapid, and low-cost method for screening and detecting contamination in cannabis seeds was established.
• The method utilizes nuclear ITS and chloroplast rbcL DNA regions for accurate analysis.
• This approach serves as a valuable quality control tool for cannabis seeds and other plant materials.