Highlights

• A new enzyme-assisted RNA extraction method (PARI) was developed to effectively isolate high-quality RNA from ginseng roots rich in polysaccharides.
• The study is the first to sequence and characterize tRNAs from ginseng roots, identifying 41 isoacceptors including new transcriptionally silent ones.
• Using LC-MS/MS, the researchers detected a novel m⁷G modification at position 18 of tRNAGly(GCC), suggesting a structural deviation in the D-loop.

Summary

The research article introduces a new method for extracting high-quality RNA from ginseng roots, which are rich in polysaccharides that typically hinder RNA isolation. This polysaccharase-aided RNA isolation (PARI) technique uses enzymes like α-amylase to digest the polysaccharides, allowing for cleaner and more intact RNA extraction compared to traditional CTAB or TRIzol methods.

Using the RNA obtained, the study successfully sequenced ginseng transfer RNAs (tRNAs) and identified 41 isoacceptors, including a purified chloroplastic tRNAGly(GCC). Further analysis by LC-MS/MS revealed four post-transcriptional modifications—m⁷G, D, T, and Ψ—with m7G uniquely found at position 18, possibly due to a structural alteration in the D-loop.

This work not only provides a robust extraction method for polysaccharide-rich plant tissues but also lays groundwork for studying tRNA structure and function in medicinal plants like ginseng.

T. Yan, K. Hu, F. Ren, and Z. Jiang, “LC-MS/MS Profiling of Post-Transcriptional Modifications in Ginseng tRNA Purified by a Polysaccharase-Aided Extraction Method,” Biomolecules, vol. 10, no. 4, p. 621, Apr. 2020, doi: 10.3390/biom10040621.