Key result
Synthetic HLVd circular RNA provides a highly stable and non-infectious diagnostic control for cannabis plants.
Why the study?
Hop latent viroid threatens Cannabis sativa cultivation, creating a need for reliable non-infectious positive controls for diagnostics.
The synthetic HLVd circRNA provides a safe, stable positive control for diagnostics and viroid research in Cannabis sativa.
Aim. Hop latent viroid (HLVd) poses a major threat to Cannabis sativa cultivation, with up to 90% infection rates and 70% THC loss.Reliable non-infectious positive controls are needed for diagnostics.Methods.We synthesized noninfectious HLVd circular RNA using a group I self-splicing ribozyme system.The product was characterized by RNase R digestion, RT-PCR, and sequencing to confirm circularization and the ribozyme-derived joining sequence.Results.Structural analysis showed the synthetic RNA retained high stability (G = -90.4kcal/mol) compared to native HLVd (G = -93.8kcal/mol), with introduced mutations preventing replication.The molecule remained stable from -20 to 37 C for 10 days and performed consistently as a PCR positive control.The infection assays in cannabis confirmed its non-infectious nature.Conclusion.The synthetic HLVd circRNA provides a safe, stable positive control for diagnostics and viroid research, addressing a key gap in current detection methods.
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Yan et al. (2025) studied Hop latent viroid (HLVd) infection (n=10). Synthetic non-infectious HLVd circular RNA vs. Mock inoculation with distilled sterilized water was evaluated on Infectivity and stability of synthesized HLVd circRNA. Synthetic HLVd circular RNA retained high stability (ΔG = –90.4 kcal/mol) and demonstrated no infectivity in cannabis plants, providing a safe positive control for diagnostics.
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