Plastid phylogenomics and species discrimination in the “Chinese” clade of Roscoea (Zingiberaceae)

doi:10.1016/j.pld.2023.03.012

Plant Diversity ›› 2023, Vol. 45 ›› Issue (05): 523-534.DOI: 10.1016/j.pld.2023.03.012

Plastid phylogenomics and species discrimination in the “Chinese” clade of Roscoea (Zingiberaceae)

Hai-Su Hu^a,b, Jiu-Yang Mao^b, Xue Wang^b, Yu-Ze Liang^b, Bei Jiang^b,c, De-Quan Zhang^a,b,c

a. School of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621010, Sichuan, China;
b. College of Pharmacy, Dali University, Dali 671000, Yunnan, China;
c. Yunnan Key Laboratory of Screening and Research on Anti-pathogenic Plant Resources from Western Yunnan (Cultivation), Dali 671000, Yunnan, China

收稿日期:2022-11-21 修回日期:2023-03-15 出版日期:2023-09-25 发布日期:2023-11-04
通讯作者: De-Quan Zhang,E-mail:zhangdeq2008@126.com
基金资助:
This study was supported by National Natural Science Foundation of China (32060091 & 31660081), Reserve Talents Project for Young and Middle-Aged Academic and Technical Leaders of Yunnan Province (202105AC160063).

Plastid phylogenomics and species discrimination in the “Chinese” clade of Roscoea (Zingiberaceae)

Hai-Su Hu^a,b, Jiu-Yang Mao^b, Xue Wang^b, Yu-Ze Liang^b, Bei Jiang^b,c, De-Quan Zhang^a,b,c

a. School of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621010, Sichuan, China;
b. College of Pharmacy, Dali University, Dali 671000, Yunnan, China;
c. Yunnan Key Laboratory of Screening and Research on Anti-pathogenic Plant Resources from Western Yunnan (Cultivation), Dali 671000, Yunnan, China

Received:2022-11-21 Revised:2023-03-15 Online:2023-09-25 Published:2023-11-04
Contact: De-Quan Zhang,E-mail:zhangdeq2008@126.com
Supported by:
This study was supported by National Natural Science Foundation of China (32060091 & 31660081), Reserve Talents Project for Young and Middle-Aged Academic and Technical Leaders of Yunnan Province (202105AC160063).

摘要/Abstract

摘要： Roscoea is an alpine or subalpine genus from the pan-tropical family Zingiberaceae, which consists of two disjunct groups in geography, namely the “Chinese” clade and the “Himalayan” clade. Despite extensive research on the genus, Roscoea species remain poorly defined and relationships between these species are not well resolved. In this study, we used plastid genomes of nine species and one variety to resolve phylogenetic relationships within the “Chinese” clade of Roscoea and as DNA super barcodes for species discrimination. We found that Roscoea plastid genomes ranged in length from 163,063 to 163,796 bp, and encoded 113 genes, including 79 protein-coding genes, 30 tRNA genes, four rRNA genes. In addition, expansion and contraction of the IR regions showed obvious infraspecific conservatism and interspecific differentiation. Plastid phylogenomics revealed that species belonging to the “Chinese” clade of Roscoea can be divided into four distinct subclades. Furthermore, our analysis supported the independence of R. cautleoides var. pubescens, the recovery of Roscoea pubescens Z.Y. Zhu, and a close relationship between R. humeana and R. cautloides. When we used the plastid genome as a super barcode, we found that it possessed strong discriminatory power (90%) with high support values. Intergenic regions provided similar resolution, which was much better than that of protein-coding regions, hypervariable regions, and DNA universal barcodes. However, plastid genomes could not completely resolve Roscoea phylogeny or definitively discriminate species. These limitations are likely related to the complex history of Roscoea speciation, poorly defined species within the genus, and the maternal inheritance of plastid genomes.

关键词: Medicinal plant, Chloroplast genome, Molecular phylogeny, DNA barcoding, DNA sequencing, Species identification

Abstract: Roscoea is an alpine or subalpine genus from the pan-tropical family Zingiberaceae, which consists of two disjunct groups in geography, namely the “Chinese” clade and the “Himalayan” clade. Despite extensive research on the genus, Roscoea species remain poorly defined and relationships between these species are not well resolved. In this study, we used plastid genomes of nine species and one variety to resolve phylogenetic relationships within the “Chinese” clade of Roscoea and as DNA super barcodes for species discrimination. We found that Roscoea plastid genomes ranged in length from 163,063 to 163,796 bp, and encoded 113 genes, including 79 protein-coding genes, 30 tRNA genes, four rRNA genes. In addition, expansion and contraction of the IR regions showed obvious infraspecific conservatism and interspecific differentiation. Plastid phylogenomics revealed that species belonging to the “Chinese” clade of Roscoea can be divided into four distinct subclades. Furthermore, our analysis supported the independence of R. cautleoides var. pubescens, the recovery of Roscoea pubescens Z.Y. Zhu, and a close relationship between R. humeana and R. cautloides. When we used the plastid genome as a super barcode, we found that it possessed strong discriminatory power (90%) with high support values. Intergenic regions provided similar resolution, which was much better than that of protein-coding regions, hypervariable regions, and DNA universal barcodes. However, plastid genomes could not completely resolve Roscoea phylogeny or definitively discriminate species. These limitations are likely related to the complex history of Roscoea speciation, poorly defined species within the genus, and the maternal inheritance of plastid genomes.

Key words: Medicinal plant, Chloroplast genome, Molecular phylogeny, DNA barcoding, DNA sequencing, Species identification

Hai-Su Hu, Jiu-Yang Mao, Xue Wang, Yu-Ze Liang, Bei Jiang, De-Quan Zhang. Plastid phylogenomics and species discrimination in the “Chinese” clade of Roscoea (Zingiberaceae)[J]. Plant Diversity, 2023, 45(05): 523-534.

参考文献

[1] Amiryousefi, A., Hyvonen, J., Poczai, P., 2018. IRscope: An online program to visualize the junction sites of chloroplast genomes. Bioinformatics 34, 3030-3031.
[2] Bai, H.R., Oyebanji, O., Zhang, R., et al., 2021. Plastid phylogenomic insights into the evolution of subfamily Dialioideae (Leguminosae). Plant Divers. 43, 27-34.
[3] Bengtsson-Palme, J., Ryberg, M., Hartmann, M., et al., 2013. Improved software detection and extraction of ITS1 and ITS2 from ribosomal ITS sequences of fungi and other eukaryotes for analysis of environmental sequencing data. Methods Ecol. Evol. 4, 914-919.
[4] Bolger, A.M., Marc, L., Bjoern, U., 2014. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30, 2114-2120.
[5] Chen, Q., Wu, X.B., Zhang, D.Q., 2020. Comparison of the abilities of universal, super, and specific DNA barcodes to discriminate among the original species of Fritillariae cirrhosae bulbus and its adulterants. PLoS One 15, e0229181.
[6] Chen, Q., Hu, H.S., Zhang, D.Q., 2022. DNA barcoding and phylogenomic analysis of the genus Fritillaria in China based on complete chloroplast genomes. Front. Plant Sci. 13, 764255.
[7] Chen, S.F., Zhou, Y.Q., Chen, Y.R., et al., 2018. Fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics 34, i884-i890.
[8] Cowley, E.J., 2007. The genus Roscoea. Kew, UK: The Royal Botanic Gardens.
[9] Darriba, D., Taboada, G.L., Doallo, R., et al., 2012. jModelTest 2: more models, new heuristics and high-performance computing. Nat. Methods 9, 772.
[10] Du, G.H., Zhang, Z.Q., Li, Q.J., 2012. Morphological and molecular evidence for natural hybridization in sympatric population of Roscoea humeana and R. cautleoides (Zingiberaceae). J. Plant Res. 125, 595-603.
[11] Doyle, J.J., 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 19, 11-15.
[12] Escobari, B., Borsch, T., Quedensley, T.S., et al., 2021. Plastid phylogenomics of the Gynoxoid group (Senecioneae, Asteraceae) highlights the importance of motif-based sequence alignment amid low genetic distances. Am. J. Bot. 108, 2235-2256.
[13] Fu, C.N., Wu, C.S., Ye, L.J., et al., 2019. Prevalence of isomeric plastomes and effectiveness of plastome super-barcodes in yews (Taxus) worldwide. Sci. Rep. 9, 2773.
[14] Fu, C.N., Mo, Z.Q., Yang, J.B., et al., 2022. Testing genome skimming for species discrimination in large taxonomically difficult genus Rhododendron. Mol. Ecol. Resour. 22, 404-414.
[15] Ji, Y.H., Liu, C.K., Yang, Z.Y., et al., 2019. Testing and using complete plastomes and ribosomal DNA sequences as the next generation DNA barcodes in Panax (Araliaceae). Mol. Ecol. Resour. 19, 1333-1345.
[16] Jin, J.J., Yu, W.B., Yang, J.B., et al., 2020. GetOrganelle: a fast and versatile toolkit for accurate de novo assembly of organelle genomes. Genome Biol. 21, 241.
[17] Katoh, K., Standley, D.M., 2013. MAFFT multiple sequence alignment software version 7: Improvements in performance. Mol. Biol. Evol. 30, 772-780.
[18] Kearse, M., Moir, R., Wilson, A., et al., 2012. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28, 1647-1649.
[19] Kleinwee, I., Larridon, I., Shah, T., et al., 2022. Plastid phylogenomics of the Sansevieria clade of Dracaena (Asparagaceae) resolves a recent radiation. Mol. Phylogenet. Evol. 169, 107404.
[20] Kreuzer, M., Howard, C., Adhikari, B., et al., 2019. Phylogenomic approaches to DNA barcoding of herbal medicines: developing clade-specific diagnostic characters for Berberis. Front. Plant Sci. 10, 586.
[21] Kumar, S., Stecher, G., Tamura, K., 2016. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol. 33, 1870-1874.
[22] Li, D.B., Ou, X.K., Zhao, J.L., et al., 2020a. An ecological barrier between the Himalayas and the Hengduan Mountains maintains the disjunct distribution of Roscoea. J. Biogeogr. 47, 326-341.
[23] Li, S., Liu, S.L., Pei, S.Y., et al., 2020b. Genetic diversity and population structure of Camellia huana (Theaceae), a limestone species with narrow geographic range, based on chloroplast DNA sequence and microsatellite markers. Plant Divers. 42, 343-350.
[24] Li, X.W., Yang, Y., Henry, R.J., et al., 2015. Plant DNA barcoding: from gene to genome. Biol. Rev. 90, 157-166.
[25] Li, Z.Z., Gichira, A.W., Muchuku, J.K., et al., 2021. Plastid phylogenomics and biogeography of the genus Monochoria (Pontederiaceae). J. Syst. Evol., 59, 1027-1039.
[26] Liang, H., Zhang, Y., Deng, J.B., et al., 2020. The complete chloroplast genome sequences of 14 Curcuma species: insights into genome evolution and phylogenetic relationships within Zingiberales. Front. Genet. 11, 802.
[27] Liang, H., Chen, J., 2021. Comparison and phylogenetic analyses of nine complete chloroplast genomes of Zingibereae. Forests 12, 710.
[28] Luo, M.H., Wan, H.L., Lin, H.H., 2008. Species and distribution of Roscoea in China and the medicinal use. Chin. Wild Plant Resour. 27, 35-37+41.
[29] Misra, A., Srivastava, S., Verma, S., et al., 2015. Nutritional evaluation, antioxidant studies and quantification of poly phenolics in Roscoeae purpurea tubers. BMC Res. Notes 8, 324.
[30] Mohammad-Panah, N., Shabanian, N., Khadivi, A., et al., 2017. Genetic structure of gall oak (Quercus infectoria) characterized by nuclear and chloroplast SSR markers. Tree Genet. Genomes 13, 70.
[31] Murat, C., Riccioni, C., Belfiori, B., et al., 2011. Distribution and localization of microsatellites in the Perigord black truffle genome and identification of new molecular markers. Fungal Genet. Biol. 48, 592-601.
[32] Ngamriabsakul, C., Newman, M.F., Cronk, Q.C.B., et al., 2000. Phylogeny and disjunction in Roscoea (Zingiberaceae). Edinburgh J. Bot. 57, 39-61.
[33] Nock, C.J., Waters, D., Edwards, M.A., et al., 2015. Chloroplast genome sequences from total DNA for plant identification. Plant Biotechnol. J. 9, 328-333.
[34] Powell, W., Morgante, M., McDevitt, R., et al., 1995. Polymorphic simple sequence repeat regions in chloroplast genomes, application to the population genetics of pines. Proc. Natl. Acad. Sci. U.S.A. 92, 7759-7763.
[35] Rawat, S., Jugran, A.K., Bhatt, I.D., et al., 2018. Influence of the growth phenophases on the phenolic composition and anti-oxidant properties of Roscoea procera Wall. in western Himalaya. J. Food Sci. Technol. 55, 578-585.
[36] Ronquist, F., Teslenko, M., Van, D.M.P., et al., 2012. MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Syst. Biol. 61, 539-542.
[37] Rozas, J., Ferrer-Mata, A., Sanchez-DelBarrio, J.C., et al., 2017. DnaSP 6: DNA sequence polymorphism analysis of large datasets. Mol. Biol. Evol. 34, 3299-3302.
[38] Sahu, M.S., Mali, P.Y., Waikar, S.B., et al., 2010. Evaluation of immunomodulatory potential of ethanolic extract of Roscoea procera rhizomes in mice. J. Pharm. Bioallied Sci. 4, 346-349.
[39] Shahzadi, I., Abdullah., Mehmood, F., et al., 2019. Chloroplast genome sequences of Artemisia maritima and Artemisia absinthium: comparative analyses, mutational hotspots in genus Artemisia and phylogeny in family Asteraceae. Genomics 112, 1454-1463.
[40] Shaw, J., Lichey, E.B., Schilling, E.E., et al., 2007. Comparison of whole chloroplast genome sequences to choose noncoding regions for phylogenetic studies in angiosperms: the tortoise and the hare III. Am. J. Bot. 94, 275-288.
[41] Small, R.L., Ryburn, J.A., Cronn, R.C., et al., 1998. The tortoise and the hare: Choosing between noncoding plastome and nuclear ADH sequences for phylogeny reconstruction in a recently diverged plant group. Am. J. Bot. 85, 1301-1315.
[42] Srivastava, S., Ankita, M., Kumar, D., et al., 2015. Reversed-phase high-performance liquid chromatography-ultraviolet photodiode array detector validated simultaneous quantification of six bioactive phenolic acids in Roscoea purpurea tubers and their in vitro cytotoxic potential against various cell lines. Pharmacogn. Mag. 11, 488-495.
[43] Stamatakis, A., 2014. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30, 1312-1313.
[44] Tang, H.Q., Tang, L., Shao, S.C., et al., 2021. Chloroplast genomic diversity in Bulbophyllum section Macrocaulia (Orchidaceae, Epidendroideae, Malaxideae): insights into species divergence and adaptive evolution. Plant Divers. 43, 350-361.
[45] Thiel, T., Michalek, W., Varsheny, R., et al., 2003. Exploiting EST databases for the development and characterization of gene-derived SSR-markers in barley (Hordeum vulgare L.). Theor. Appl. Genet. 106, 411-422.
[46] Wei, R., Yang, J., He, L.J., et al., 2021. Plastid phylogenomics provides novel insights into the infrafamilial relationship of Polypodiaceae. Cladistics 37, 717-727.
[47] Wiche, S., Schneeweiss, G.M., dePamphilis, C.W., 2011. The evolution of the plastid chromosome in land plants: gene content, gene order, gene function. Plant Mol. Biol. 76, 273-297.
[48] Wick, R.R., Schultz, M.B., Zobel, J., et al., 2015. Bandage: interactive visualization of de novo genome assemblies. Bioinformatics 31, 3350-3352.
[49] Wu, D.L., Larsen, K., 2000. Roscoea Sm. In: Wu, Z.Y., Raven, P.H. (Eds.), Flora of China, Vol. 24. Science Press, Beijing; Missouri Botanical Garden Press, St. Louis, pp. 362-366.
[50] Wu, Q., Jiang, M., Chen, H.M., et al., 2020. Comparative analysis of three complete chloroplast genomes of Inula genus with phylogenetic analysis of 49 plants from Carduoideae. Acta Pharm. Sin. 55, 1042-1049.
[51] Xu, Y.L., Shen, H.H, Du, X.Y., et al., 2022. Plastome characteristics and species identification of Chinese medicinal wintergreens (Gaultheria, Ericaceae). Plant Divers. 44, 519-529.
[52] Yang, B.B., Li, L.D., Liu, J.Q., et al., 2021. Plastome and phylogenetic relationship of the woody buckwheat Fagopyrum tibeticum in the Qinghai-Tibet Plateau. Plant Divers. 43, 198-205.
[53] Yang, J.B., Li, D.Z., Li, H.T., 2014. Highly effective sequencing whole chloroplast genomes of angiosperms by nine novel universal primer pairs. Mol. Ecol. Resour. 14, 1024-1031.
[54] Yang, J.P., Zhu, Z.L., Fan, Y.J., et al., 2020. Comparative plastomic analysis of three Bulbophyllum medicinal plants and its utility for species identification. Acta Pharm. Sin. 55, 2736-2745.
[55] Zhang, D.Q., Duan, L.Z., Zhou, N., 2014. Application of DNA barcoding in Roscoea (Zingiberaceae) and a primary discussion on taxonomic status of Roscoea cautleoides var. pubescens. Biochem. Syst. Ecol. 52, 14-19.
[56] Zhang, D.Q., Yang, Y.S., Zhou, N., 2015. The identify of Roscoea cauteoides var. pubescens (Z.Y. Zhu) T.L. Wu based on molecular and morphological evidences. Phytotaxa 205, 259-267.
[57] Zhang, L., Huang, Y.W., Huang, J.L., et al., 2022a. DNA barcoding of Cymbidium by genome skimming: Call for next-generation nuclear barcodes. Mol. Ecol. Resour. 00, 1-16.
[58] Zhang, Y.M., Han, L.J., Yang, C.W., 2022b. Comparative chloroplast genome analysis of medicinally important Veratrum (Melanthiaceae) in China: Insights into genomic characterization and phylogenetic relationships. Plant Divers. 44, 70-82.
[59] Zhao, J.L., Xia, Y.M., Cannon, C.H., et al., 2016a. Evolutionary diversification of alpine ginger reflects the early uplift of the Himalayan-Tibetan Plateau and rapid extrusion of Indochina. Gondwana Res. 32, 232-241.
[60] Zhao, J.L., Gugger, P.F., Xia, Y.M., et al., 2016b. Ecological divergence of two closely related Roscoea species associated with late Quaternary climate change. J. Biogeogr. 43, 1990-2001.
[61] Zhao, J.L., Zhong, J.S., Fan, Y.L., et al., 2017. A preliminary species-level phylogeny of the alpine ginger Roscoea: implications to speciation. J. Syst. Evol. 55, 215-224.
[62] Zhao, J.L., Paudel, B.R., Yu, X.Q., et al., 2021. Speciation along the elevation gradient: divergence of Roscoea species within the south slope of the Himalayas. Mol. Phylogenet. Evol. 164, 107292.
[63] Zheng, D.S., Zhang, J.Y., Guo, Q.S., 2013. CpDNA non-coding sequence analysis of Pinellia ternata and its related species. Chin. Tradit. Herb. Drugs 44, 881-886.

Plastid phylogenomics and species discrimination in the “Chinese” clade of Roscoea (Zingiberaceae)

Plastid phylogenomics and species discrimination in the “Chinese” clade of Roscoea (Zingiberaceae)

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