Phylogenetic analysis and development of molecular markers for five medicinal Alpinia species based on complete plastome sequences

Abstract

Background Alpinia species are widely used as medicinal herbs. To understand the taxonomic classification and plastome evolution of the medicinal Alpinia species and correctly identify medicinal products derived from Alpinia species, we systematically analyzed the plastome sequences from five Alpinia species. Four of the Alpinia species: Alpinia galanga (L.) Willd., Alpinia hainanensis K.Schum., Alpinia officinarum Hance, and Alpinia oxyphylla Miq., are listed in the Chinese pharmacopeia. The other one, Alpinia nigra (Gaertn.) Burtt, is well known for its medicinal values. Results The four Alpinia species: A. galanga, A. nigra, A. officinarum, and A. oxyphylla, were sequenced using the Next-generation sequencing technology. The plastomes were assembled using Novoplasty and annotated using CPGAVAS2. The sizes of the four plastomes range from 160,590 bp for A. galanga to 164,294 bp for A. nigra, and display a conserved quadripartite structure. Each of the plastomes encodes a total of 111 unique genes, including 79 protein-coding, 28 tRNA, and four rRNA genes. In addition, 293–296 SSRs were detected in the four plastomes, of which the majority are mononucleotides Adenine/Thymine and are found in the noncoding regions. The long repeat analysis shows all types of repeats are contained in the plastomes, of which palindromic repeats occur most frequently. The comparative genomic analyses revealed that the pair of the inverted repeats were less divergent than the single-copy region. Analysis of sequence divergence on protein-coding genes showed that two genes (accD and ycf1) had undergone positive selection. Phylogenetic analysis based on coding sequence of 77 shared plastome genes resolves the molecular phylogeny of 20 species from Zingiberaceae. In particular, molecular phylogeny of four sequenced Alpinia species (A. galanga, A. nigra, A. officinarum, and A. oxyphylla) based on the plastome and nuclear sequences showed congruency. Furthermore, a comparison of the four newly sequenced Alpinia plastomes and one previously reported Alpinia plastomes (accession number: NC_048461) reveals 59 highly divergent intergenic spacer regions. We developed and validated two molecular markers Alpp and Alpr, based on two regions: petN-psbM and psaJ-rpl33, respectively. The discrimination success rate was 100 % in validation experiments. Conclusions The results from this study will be invaluable for ensuring the effective and safe uses of Alpinia medicinal products and for the exploration of novel Alpinia species to improve human health.

Complete Chloroplast Genome Analysis of Two Important Medicinal Alpinia Species: Alpinia galanga and Alpinia kwangsiensis

“…An NJ phylogenetic tree was constructed using high-quality sequences to show the ability of the five highly variable regions in species-level identification ( Supplementary Figure 1 ), using 50% as a cut-off value for the condensed tree. Yang et al (2021) recently published an article on the development of molecular markers for five medicinal Alpinia species based on complete plastome sequences and developed molecular markers based on two highly variable regions ( petN-psbM and psaJ-rpl33 ). We identified different highly variable sites from those of Yang et al (2021) possibly because we used different Alpinia species in our analyses.…”

“… Yang et al (2021) recently published an article on the development of molecular markers for five medicinal Alpinia species based on complete plastome sequences and developed molecular markers based on two highly variable regions ( petN-psbM and psaJ-rpl33 ). We identified different highly variable sites from those of Yang et al (2021) possibly because we used different Alpinia species in our analyses. And adding more species might be more accurate for finding and developing highly variable markers.…”

Characterization of Codonopsis pilosula subsp. tangshen plastome and comparative analysis of Codonopsis species

“…Several successful examples have been reported to develop taxon-specific markers [ 63 ]. In the present study, the universal makers have not been successful in discriminating the medicinal Codonopsis species.…”

Comparative and Phylogenetic Analysis of Complete Chloroplast Genomes in Leymus (Triticodae, Poaceae)

“…Interestingly, the highly variable regions of Leymus species were different from those previously reported for designing phylogenetic trees and species identification of Leymus , such as trnL-trnF , trnH-psbA , trnK-rps16 , ndhF , rps16-trnQ , trnF-ndhJ , and ndhF-rpl32 [ 8 , 11 , 13 , 14 , 24 , 51 , 52 , 53 ]. Thus, by analyzing the variable regions of the 16 Leymus cp genomes, we were able to identify some molecular markers, such as SSRs [ 45 , 54 ] which can be more effectively used in DNA barcoding and to determine phylogenetic relationships in future studies.…”