ABSTRACT
Objective::To determine the chloroplast genomes of Liriope spicata var. prolifera, Ophiopogon japonicus in Sichuan and Zhejiang, analyze their sequence characteristics and complete the screening of specific DNA barcodes. Method::The chloroplast genomes of L. spicata var. prolifera, O. japonicus in Sichuan and Zhejiang were sequenced, spliced and annotated through high-throughtput sequencing technology, and the structural characteristics and phylogenetic relationships of chloroplast genomes were analyzed by bioinformatics. Result::The total length of chloroplast genome of L. spicata var. prolifera was 155 998 bp, the total content of guanine and cytosine (GC) was 37.7%, and 85 protein-coding genes, 37 transfer RNA (tRNA) genes and 8 ribosomal RNA (rRNA) genes were successfully annotated, a total of 274 simple sequence repeats (SSRs) were detected, the number of codons encoding leucine was the most, while the number of codons encoding tryptophan was the least. The total length of chloroplast genome of O. japonicus in Sichuan province was 156 078 bp, the total content of GC was 37.8%, and 85 protein-coding genes, 37 tRNA genes and 8 rRNA genes were successfully annotated, a total of 265 SSRs were detected, the number of codons encoding leucine was the most, while the number of codons encoding tryptophan was the least. The total length of chloroplast genome of O. japonicus in Zhejiang province was 156 207 bp, the total content of GC was 37.7%, and 85 protein-coding genes, 37 tRNA genes and 8 rRNA genes were successfully annotated, a total of 274 SSRs were detected with the highest number of codons encoding leucine and the lowest number of codons encoding tryptophan. Conclusion::The phylogenetic trees show that compared with O. japonicus in Sichuan province, L. spicata var. prolifera is more closely related to O. japonicus in Zhejiang province. The variation of non-coding regions of L. spicata var. prolifera, O. japonicus in Zhejiang and Sichuan is greater than that in the coding region. The entire chloroplast genome can be used as a super barcode for identifying species of Ophiopogon and Liriope.
ABSTRACT
Herba Anoectochili is a commonly used medicinal material. However, its adulteration is a serious concern. Due to the similar morphological characteristics of Herba Anoectochili and its adulterants, traditional identification techniques often fail to distinguish between them accurately, which is not conducive to the circulation management and safety of the medicinal materials. To improve the distinction between Herba Anoectochili and its adulterants accurately, this study identified 41 Herba Anoectochili and its adulterant samples based on the ITS2 sequence. Sequence characteristics, Basic Local Alignment Search Tool (BLAST) application, genetic distance, construction of phylogenetic tree, secondary structure prediction, and other methods showed the ITS2 sequence to accurately identify Herba Anoectochili from its adulterants. Furthermore, in this study, we designed a specific primer, based on the ITS2 sequence, and established a real-time PCR detection system for the rapid, sensitive, and specific identification of the original plant of Herba Anoectochili. Compared to DNA barcoding technology, this method has shorter detection time, stronger specificity, and higher sensitivity, which lays the foundation for the rapid identification of Herba Anoectochili.