Ultrasound-assisted extraction (UAE), using petroleum ether as the solvent, was systematically applied to extract main macamides and macaenes from Maca hypocotyls. Extraction yield was related with four variables, including ratio of solution to solid, extraction temperature, extraction time, and extraction power. On the basis of response surface methodology (RSM), the optimal conditions were determined to be the ratio of solution to solid as 10:1 (mL/g), the extraction temperature of 40 °C, the extraction time of 30 min, and the extraction power of 200 W. Based on the optimal extraction method of UAE, the total contents of ten main macamides and two main macaenes of Maca cultivated in twenty different areas of Tibet were analyzed by HPLC and UHPLC-ESI-Q-TOF-MS/MS. This study indicated that UAE was able to effectively extract macamides alkaloids from Maca hypocotyls. Quantitative analysis showed that geographical origins, not ecotypes, played a more important role on the accumulation of active macamides in Maca.

Ophiocordyceps sinensis is one of the best-known traditional Chinese medicines with distribution confined to the Tibetan Plateau and its surrounding regions. Harvesting the fungus contributes greatly to the livelihood of local communities. The quality and price varies amongst different production regions, usually resulting in an intentional mix-up of its production locality during trading processes, which leads to a demand of developing a reliable way that can trace the geographical origin of this fungus. In the present study, a DNA barcoding-based method applying two universal DNA barcodes for identifying fungal and insect, respectively i.e. the nuclear ribosomal internal transcribed spacer (ITS) and the mitochondrial cytochrome oxidase I (COI), was evaluated and used for geographical origin authentication of O. sinensis. A total of 24 ITS and 78 COI haplotypes were recognised from 215 individuals collected from 75 different geographic localities (county level). Ninety-nine haplotypes were defined using the combination of ITS and COI, discriminating the 75 investigated production counties into 99 distinct regions. A “core” production region was recognised which covers areas of Nagqu and Qamdo in Xizang, Yushu and Guoluo in Qinghai, Gannan (Maqu and Xiahe) in Gansu and certain regions in Nyingch (Bomi and Zayü) and Lhasa (Damxung) in Xizang and Garzê (Sêrxü) in Sichuan Province. Haplotype analyses using the combined barcodes of ITS and COI showed an excellent performance in the geographical origin authentication of O. sinensis and the definition of “core” and “non-core” production region.

Phlomoides rotata (Benth. ex Hook.f.) Mathiesen is a perennial herb endemic to Qinghai-Tibet Plateau with important medicinal properties. Here, we sequenced and analyzed the complete chloroplast (cp) genome of P. rotata and reconstructed the phylogeny of P. rotata based on 24 cp genomes. The genome of P. rotata is 151,825 bp in length, including a large single-copy (LSC) region of 83,129 bp and a small single-copy (SSC) region of 17,398 bp. A total of 131 genes were identified, of which 86 are protein-coding genes, 37 are transfer RNA genes, and eight are ribosomal RNA genes. Phylogenetic analyses revealed that the species P. rotata is closely related to Phlomoides alpina with bootstrap support (BS) values of 100%. Overall, the genomic resources presented in this study will be beneficial for further studies on evolutionary patterns of P. rotata and its closely related species.

Codonopsis convolvulacea subsp. vinciflora is a commonly used and endangered herb in Tibetan medicine. The chloroplast genome was determined to be 187,634 bp in length and contained a large single-copy and a small single-copy region of 102,174 bp and 8552 bp, respectively. The genome is predicted to contain 140 genes, including 90 protein-coding genes, 42 tRNA genes, and 8 rRNA genes. The overall GC content of the genome is 38.72%. A phylogenetic tree reconstructed by 11 chloroplast genomes reveals that Codonopsis convolvulacea subsp. vinciflora is mostly related to C. tsinglingensis with bootstrap support values of 100%.