journal article·2025-01-09Open access

Comparative Study of Phytolacca Species Through Morphological, Chloroplast Genome, and Phylogenetic Analysis

Sumin Jeong, Yeseul Kim, Heewon Ji, Jun-Ho Song, Inkyu Park

Applied Sciences· Volume 15· Issue 2

Abstract

Phytolacca acinosa Roxb. and P. americana L. are recognized as the primary sources of Phytolaccae Radix, which is traditionally utilized for various medicinal purposes. However, because of their potent toxicity, it is essential to distinguish these species. This study has aimed to clarify the classification of Phytolacca species based on their morphology and genetic differences. The chloroplast genome of P. acinosa was sequenced and comparative analyses were conducted to identify the regions of variation and nucleotide diversity among the species. The results revealed that P. acinosa shares more sequence similarity with other Phytolacca species than with P. americana. Additionally, the dN/dS analysis showed that the ndhH gene of P. americana had a value of 1.0871, indicating positive selection. The phylogenetic tree, supported by strong bootstrap values and posterior probabilities, confirmed that P. acinosa and P. americana formed distinct clusters. Furthermore, the estimated divergence time between P. acinosa and P. americana was approximately 15.07 million years ago, indicating that they diverged earlier than P. insularis, P. polyandra, P. japonica, and P. latbenia. These findings indicated that P. acinosa and P. americana are phylogenetically distinct species, highlighting the need for accurate species identification and taxonomic reassessment to ensure the safe use of these toxic medicinal plants.
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Research context

OpenAlex topics: Genomics and Phylogenetic Studies · Plant Pathogens and Resistance · Genetic diversity and population structure

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Department of Biology and Chemistry, Changwon National University, Changwon 51140, Republic of Korea

Department of Biology, Chungbuk National University, Cheongju 28644, Republic of Korea

Funding

‘Student-Initiated Creative Research Project’ at Changwon National University in 2024 · Global Learning and Academic research institution for Master’s and Ph.D. students · Postdocs (LAMP) Program of the National Research Foundation of Korea (NRF) · National Research Foundation of Korea (NRF)

References (100+)

Show source-provided references
1. Song, Y., Jiang, F., Shi, J., Wang, C., Xiang, N., and Zhu, S. (2022). Phylogenomics Reveals the Evolutionary History of Phytolacca (Phytolaccaceae). Front. Plant Sci., 13.10.3389/fpls.2022.844918 ↗
2. Hong, S.P. (2018). Phytolaccaceae. The Genera of Vascular Plants of Korea, Academy Publishing Co.
3. Korea National Arboretum (2020). Checklist of Vascular Plants in Korea, Korea National Arboretum.
4. A Taxonomic Review of Phytolacca insularis (Phytolaccaceae)10.11110/kjpt.2007.37.4.431 ↗
5. Ministry Food and Drug Safety (2019). The Korean Pharmacopoeia, Ministry Food and Drug Safety. [12th ed.].
6. Medicinal properties and anti-inflammatory components of Phytolacca (Shanglu)10.1016/j.dcmed.2021.09.001 ↗
7. Antitumor activity of Americanin A isolated from the seeds of Phytolacca americana by regulating the ATM/ATR signaling pathway and the Skp2–p27 axis in human colon cancer cells10.1021/acs.jnatprod.5b00743 ↗
8. Anti-ascites effect of total saponins of Phytolaccae Radix on mice with ascites and mechanism
9. Carotenoid and Phenolic Profiles and Antioxidant and Anticholinesterase Activities of Leaves and Berries of Phytolacca acinosa10.1021/acsfoodscitech.3c00568 ↗
10. Saleri, F.D., Chen, G., Li, X., and Guo, M. (2017). Comparative analysis of saponins from different Phytolaccaceae species and their antiproliferative activities. Molecules, 22.10.3390/molecules22071077 ↗
11. Two new saponins from Phytolacca americana10.1055/s-2006-962732 ↗
12. Phytolacacinoside A, a new triterpenoid saponin from Phytolacca acinosa Roxb10.1080/10286020902849266 ↗
13. Ma, X.P., Zhang, W.F., Yi, P., Lan, J.J., Xia, B., Jiang, S., Lou, H.Y., and Pan, W.D. (2017). Novel Flavones from the Root of Phytolacca acinosa Roxb. Chem. Biodivers., 14.10.1002/cbdv.201700361 ↗
14. Effects of Phytolacca acinosa polysaccharides I with different schedules on its antitumor efficiency in tumor bearing mice and production of IL-1, IL-2, IL-6, TNF, CSF activity in normal mice10.3109/08923979709007659 ↗
15. Flawonoids and free phenolic acids from Phytolacca americana L. leaves
16. Metabolic exploration of the developmental abnormalities and neurotoxicity of Esculentoside B, the main toxic factor in Phytolaccae radix10.1016/j.fct.2023.113777 ↗
17. Identification of phytolaccosides in biological samples from pokeweed intoxication patients using liquid chromatography-tandem mass spectrometry10.1016/j.jchromb.2020.122123 ↗
18. Phytolacca Radix Poisoning: A Case Report10.22246/jikm.2020.41.2.241 ↗
19. Daniell, H., Lin, C.-S., Yu, M., and Chang, W.-J. (2016). Chloroplast genomes: Diversity, evolution, and applications in genetic engineering. Genome Biol., 17.10.1186/s13059-016-1004-2 ↗
20. Comparative organization of chloroplast genomes10.1146/annurev.ge.19.120185.001545 ↗
21. The evolution of the plastid chromosome in land plants: Gene content, gene order, gene function10.1007/s11103-011-9762-4 ↗
22. Rates and patterns of chloroplast DNA evolution10.1073/pnas.91.15.6795 ↗
23. Relative rates of synonymous substitutions in the mitochondrial, chloroplast and nuclear genomes of seed plants10.1016/j.ympev.2008.09.009 ↗
24. Chloroplast DNA and biosystematics: The effects of intraspecific diversity and plastid transmission10.2307/1223218 ↗
25. Epifluorescent microscopic evidence for maternal inheritance of chloroplast DNA10.1038/298481a0 ↗
26. Conservation of chloroplast genome structure among vascular plants10.1007/BF00418529 ↗
27. Molecular basis of maternal inheritance10.1073/pnas.69.9.2410 ↗
28. Utility of 17 chloroplast genes for inferring the phylogeny of the basal angiosperms10.2307/2656749 ↗
29. Raubeson, L.A., and Jansen, R.K. (2005). Chloroplast genomes of plants. Plant Diversity and Evolution: Genotypic and Phenotypic Variation in Higher Plants, CABI Publishing.10.1079/9780851999043.0045 ↗
30. Li, E., Liu, K., Deng, R., Gao, Y., Liu, X., Dong, W., and Zhang, Z. (2023). Insights into the phylogeny and chloroplast genome evolution of Eriocaulon (Eriocaulaceae). BMC Plant Biol., 23.10.1186/s12870-023-04034-z ↗
31. Biogeography of the Pistia clade (Araceae): Based on chloroplast and mitochondrial DNA sequences and Bayesian divergence time inference10.1080/10635150490445904 ↗
32. Evolution of the angiosperms: Calibrating the family tree10.1098/rspb.2001.1782 ↗
33. Time tree of Rubiaceae: Phylogeny and dating the family, subfamilies, and tribes10.1086/599077 ↗
34. Dong, W., Xu, C., Liu, Y., Shi, J., Li, W., and Suo, Z. (2021). Chloroplast phylogenomics and divergence times of Lagerstroemia (Lythraceae). BMC Genom., 22.10.1186/s12864-021-07769-x ↗
35. Jiménez-Barron, O., García-Sandoval, R., Magallón, S., García-Mendoza, A., Nieto-Sotelo, J., Aguirre-Planter, E., and Eguiarte, L.E. (2020). Phylogeny, diversification rate, and divergence time of Agave sensu lato (Asparagaceae), a group of recent origin in the process of diversification. Front. Plant Sci., 11.10.3389/fpls.2020.536135 ↗
36. A codon-based model of nucleotide substitution for protein-coding DNA sequences
37. Plastid genomics of Nicotiana (Solanaceae): Insights into molecular evolution, positive selection and the origin of the maternal genome of Aztec tobacco (Nicotiana rustica)10.7717/peerj.9552 ↗
38. One-third of the plastid genes evolved under positive selection in PACMAD grasses10.1007/s00425-017-2781-x ↗
39. Xie, D.-F., Yu, Y., Deng, Y.-Q., Li, J., Liu, H.-Y., Zhou, S.-D., and He, X.-J. (2018). Comparative analysis of the chloroplast genomes of the Chinese endemic genus Urophysa and their contribution to chloroplast phylogeny and adaptive evolution. Int. J. Mol. Sci., 19.10.3390/ijms19071847 ↗
40. Yang, Y., Zhou, T., Duan, D., Yang, J., Feng, L., and Zhao, G. (2016). Comparative analysis of the complete chloroplast genomes of five Quercus species. Front. Plant Sci., 7.10.3389/fpls.2016.00959 ↗
41. Preponderance of synonymous changes as evidence for the neutral theory of molecular evolution10.1038/267275a0 ↗
42. Lawrie, D.S., Messer, P.W., Hershberg, R., and Petrov, D.A. (2013). Strong purifying selection at synonymous sites in D. melanogaster. PLoS Genet., 9.10.1371/journal.pgen.1003527 ↗
43. Jansen, R.K., and Ruhlman, T.A. (2012). Plastid genomes of seed plants. Genomics of Chloroplasts and Mitochondria, Springer.10.1007/978-94-007-2920-9_5 ↗
44. An update on chloroplast genomes10.1007/s00606-007-0608-0 ↗
45. The chloroplast genome: A review10.1007/s11738-020-03089-x ↗
46. Rates of nucleotide substitution vary greatly among plant mitochondrial, chloroplast, and nuclear DNAs10.1073/pnas.84.24.9054 ↗
47. Regularities of context-dependent codon bias in eukaryotic genes10.1093/nar/30.5.1192 ↗
48. Chloroplast DNA codon use: Evidence for selection at the psbA locus based on tRNA availability10.1007/BF00175504 ↗
49. Codon usage in plant genes10.1093/nar/17.2.477 ↗
51. Comparative analysis of codon usage patterns in chloroplast genomes of six Euphorbiaceae species10.7717/peerj.8251 ↗
52. Mutational dynamics and phylogenetic utility of noncoding chloroplast DNA10.1007/s00606-009-0210-8 ↗
53. The contribution of slippage-like processes to genome evolution10.1007/BF00173185 ↗
54. Raubeson, L.A., Peery, R., Chumley, T.W., Dziubek, C., Fourcade, H.M., Boore, J.L., and Jansen, R.K. (2007). Comparative chloroplast genomics: Analyses including new sequences from the angiosperms Nuphar advena and Ranunculus macranthus. BMC Genom., 8.10.1186/1471-2164-8-174 ↗
55. Losing DNA methylation at repetitive elements and breaking bad10.1186/s13072-021-00400-z ↗
56. Retrotransposons: The intrinsic genomic evolutionist10.1007/s13258-014-0250-5 ↗
57. Microsatellites in different eukaryotic genomes: Survey and analysis10.1101/gr.10.7.967 ↗
58. Replication slippage involves DNA polymerase pausing and dissociation10.1093/emboj/20.10.2587 ↗
59. Emanuelli, F., Lorenzi, S., Grzeskowiak, L., Catalano, V., Stefanini, M., Troggio, M., Myles, S., Martinez-Zapater, J.M., Zyprian, E., and Moreira, F.M. (2013). Genetic diversity and population structure assessed by SSR and SNP markers in a large germplasm collection of grape. BMC Plant Biol., 13.10.1186/1471-2229-13-39 ↗
60. Simple sequence repeats as advantageous mutators in evolution10.1016/j.tig.2006.03.005 ↗
61. Qin, Z., Wang, Y., Wang, Q., Li, A., Hou, F., and Zhang, L. (2015). Evolution analysis of simple sequence repeats in plant genome. PLoS ONE, 10.10.1371/journal.pone.0144108 ↗
62. Comparative analysis of tandem repeats from hundreds of species reveals unique insights into centromere evolution10.1186/gb-2013-14-1-r10 ↗
63. Conservation and functional element discovery in 20 angiosperm plant genomes10.1093/molbev/mst082 ↗
64. Chang, C.-S., Kim, H., and Chang, K.S. (2014). Provisional Checklist of Vascular Plants for the Korea Peninsula Flora (KPF). Des. Paju.
65. Ernst, M., Saslis-Lagoudakis, C.H., Grace, O.M., Nilsson, N., Simonsen, H.T., Horn, J.W., and Rønsted, N. (2016). Evolutionary prediction of medicinal properties in the genus Euphorbia L. Sci. Rep., 6.10.1038/srep30531 ↗
66. Ma, M., Liu, Y., Bai, C., and Yong, J.W.H. (2021). The significance of chloroplast NAD(P)H dehydrogenase complex and its dependent cyclic electron transport in photosynthesis. Front. Plant Sci., 12.10.3389/fpls.2021.661863 ↗
67. Chloroplast NDH: A different enzyme with a structure similar to that of respiratory NADH dehydrogenase10.1016/j.bbabio.2015.10.013 ↗
68. Subcellular distribution and chemical forms of cadmium in Phytolacca americana L10.1016/j.jhazmat.2010.10.122 ↗
69. Bird-dispersal of Phytolacca americana L. and the influence of fruit removal on subsequent fruit development
70. Enhanced ferredoxin-dependent cyclic electron flow around photosystem I and α-tocopherol quinone accumulation in water-stressed ndhB-inactivated tobacco mutants10.1007/s00425-005-1548-y ↗
71. Structure and biogenesis of the chloroplast NAD(P)H dehydrogenase complex10.1016/j.bbabio.2010.10.015 ↗
72. Chloroplastic NAD(P)H dehydrogenase in tobacco leaves functions in alleviation of oxidative damage caused by temperature stress10.1104/pp.105.070490 ↗
73. Grosjean, N., Le Jean, M., Berthelot, C., Chalot, M., Gross, E.M., and Blaudez, D. (2019). Accumulation and fractionation of rare earth elements are conserved traits in the Phytolacca genus. Sci. Rep., 9.10.1038/s41598-019-54238-3 ↗
74. Proposal for a standardized temporal scheme of biological classification for extant species10.1073/pnas.96.13.7358 ↗
75. Estimating divergence times in large molecular phylogenies10.1073/pnas.1213199109 ↗
76. Rates of DNA sequence evolution differ between taxonomic groups10.1126/science.3082006 ↗
77. A modified protocol for rapid DNA isolation from plant tissues using cetyltrimethylammonium bromide10.1038/nprot.2006.384 ↗
78. Trimmomatic: A flexible trimmer for Illumina sequence data10.1093/bioinformatics/btu170 ↗
79. Jin, J.-J., Yu, W.-B., Yang, J.-B., Song, Y., DePamphilis, C.W., Yi, T.-S., and Li, D.-Z. (2020). GetOrganelle: A fast and versatile toolkit for accurate de novo assembly of organelle genomes. Genome Biol., 21.10.1186/s13059-020-02154-5 ↗
80. SOAPdenovo2: An empirically improved memory-efficient short-read de novo assembler10.1186/2047-217X-1-18 ↗
81. GeSeq–versatile and accurate annotation of organelle genomes10.1093/nar/gkx391 ↗
82. Artemis: An integrated platform for visualization and analysis of high-throughput sequence-based experimental data10.1093/bioinformatics/btr703 ↗
83. tRNAscan-SE On-line: Integrating search and context for analysis of transfer RNA genes10.1093/nar/gkw413 ↗
84. OrganellarGenomeDRAW (OGDRAW) version 1.3. 1: Expanded toolkit for the graphical visualization of organellar genomes10.1093/nar/gkz238 ↗
85. Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data10.1093/bioinformatics/bts199 ↗
86. MEGA11: Molecular evolutionary genetics analysis version 1110.1093/molbev/msab120 ↗
87. Heatmapper: Web-enabled heat mapping for all10.1093/nar/gkw419 ↗
88. MISA-web: A web server for microsatellite prediction10.1093/bioinformatics/btx198 ↗
89. Tandem repeats finder: A program to analyze DNA sequences10.1093/nar/27.2.573 ↗
90. REPuter: The manifold applications of repeat analysis on a genomic scale10.1093/nar/29.22.4633 ↗
91. VISTA: Computational tools for comparative genomics10.1093/nar/gkh458 ↗
92. Glocal alignment: Finding rearrangements during alignment10.1093/bioinformatics/btg1005 ↗
93. DnaSP 6: DNA sequence polymorphism analysis of large data sets10.1093/molbev/msx248 ↗
94. PAML 4: Phylogenetic analysis by maximum likelihood10.1093/molbev/msm088 ↗
95. Package ‘ggplot2’: Create Elegant Data Visualisations Using the Grammar of Graphics
96. MAFFT multiple sequence alignment software version 7: Improvements in performance and usability10.1093/molbev/mst010 ↗
97. Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis10.1093/oxfordjournals.molbev.a026334 ↗
98. jModelTest 2: More models, new heuristics and high-performance computing10.1038/nmeth.2109 ↗
99. MRBAYES: Bayesian inference of phylogenetic trees10.1093/bioinformatics/17.8.754 ↗
100. Drummond, A.J., and Rambaut, A. (2007). BEAST: Bayesian evolutionary analysis by sampling trees. BMC Evol. Biol., 7.10.1186/1471-2148-7-214 ↗