AUTHOR=Gao Longlong , Xu Wenjie , Xin Tianyi , Song Jingyuan TITLE=Application of third-generation sequencing to herbal genomics JOURNAL=Frontiers in Plant Science VOLUME=Volume 14 - 2023 YEAR=2023 URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2023.1124536 DOI=10.3389/fpls.2023.1124536 ISSN=1664-462X ABSTRACT=Traditional medicine has been used to protect human health for thousands of years. However, the genetic information of organism-derived species is still extremely rare, and the exploitation of natural resources is, therefore, limited. The application of third-generation sequencing (TGS) techniques has brought us a great opportunity to gather invaluable genetic information and promote the development of herbal genomics. In this review, we introduce two main TGS techniques, PacBio SMRT technology and Oxford Nanopore Technology, and compare the two techniques with Illumina, the predominant next-generation sequencing technique. In addition, we summarize nuclear and organelle genome assemblies of commonly used medicinal plants and choose several examples from genomics, transcriptomics, and molecular identification studies to dissect the specific processes and summarize the advantages and disadvantages of the two TGS techniques when applied to medicinal organisms. Finally, we describe how we expect that in the future, TGS techniques will be widely utilized to assemble telomere-to-telomere (T2T) genomes and the epigenomics research of medicinal plants.