Maternal Lineage Structure and Chloroplast Diversity of South Sumatran Rice, Indonesia, Revealed by the trnH–psbA Intergenic Spacer
Abstract
Despite the importance of comprehensive genetic characterization for the conservation and utilization of local rice germplasm in breeding programs, the maternal genetic diversity of South Sumatran local rice remains largely unexplored. In this study, we evaluated the genetic diversity, maternal lineage, and phylogenetic relationships of South Sumatran rice accessions using the chloroplast trnH–psbA intergenic spacer, a highly variable chloroplast marker widely used for assessing genetic diversity and phylogenetic relationships in rice and other plant species. A total of 18 South Sumatran rice accessions, along with reference sequences, were analyzed through sequence alignment, haplotype network construction, genetic distance clustering, and phylogenetic tree reconstruction using the Neighbor-Joining (NJ) and UPGMA methods. The trnH–psbA region showed a uniform sequence length of 415 bp and low nucleotide diversity (π = 0.096%), resulting in the identification of three haplotypes. Phylogenetic and genetic distance analyses consistently identified two accessions, Bone 1 and Pegagan 2, as genetically distinct, highlighting their potential value as candidates for future conservation and breeding initiatives. These findings provide a molecular basis for the preservation and strategic utilization of South Sumatran rice germplasm and emphasize the importance of conserving genetically divergent local accessions. Future studies integrating chloroplast and nuclear markers will enable a more comprehensive assessment of genetic diversity for rice breeding and conservation.
Keywords
Full Text:
PDFReferences
Adriansyah, F., Hasmeda, M., Suwignyo, R. A., Halimi, E. S., Fatimah, Wibisono, I., & Sarimana, U. (2022). Selection of the Sub1 locus for submergence-tolerant introgression in a backcrossing of South Sumatra rice based on SSR markers. Sains Malaysiana, 51(3), 695–706. https://doi.org/10.17576/jsm-2022-5103-05
Aygören Uluer, D. (2025). Is ITS2 more successful in angiosperm DNA barcoding than the entire ITS, ITS1, matK and trnH-psbA regions? A review. Anales Del Jardin Botanico de Madrid, 82(1), 1–15. https://doi.org/10.3989/ajbm.624
Basith, A., Arumingtyas, E. L., & Widodo, W. (2021). Genetic variation analysis of four local varieties of Indonesian black rice (Oryza sativa L.) based on partially rbcL cpDNA gene sequence. The Journal of Experimental Life Sciences, 11(1), 1–5. https://doi.org/10.21776/ub.jels.2021.011.01.01
Bodenhofer, U., Bonatesta, E., Horejsˇ-Kainrath, C., & Hochreiter, S. (2015). msa: an R package for multiple sequence alignment. Bioinformatics, 31(24), 3997–3999. https://doi.org/10.1093/bioinformatics/btv494
Brock, J. R., Mandáková, T., McKain, M., Lysak, M. A., & Olsen, K. M. (2022). Chloroplast phylogenomics in Camelina (Brassicaceae) reveals multiple origins of polyploid species and the maternal lineage of C. sativa. Horticulture Research, 9(uhab050), 1–13. https://doi.org/10.1093/hr/uhab050
Dong, W., Liu, J., Yu, J., Wang, L., & Zhou, S. (2012). Highly variable chloroplast markers for evaluating plant phylogeny at low taxonomic levels and for DNA barcoding. PloS One, 7(4), 1–9. https://doi.org/10.1371/journal.pone.0035071
Feng, Q., Song, W. C., Zhang, Y. J., & Shi, C. (2021). The complete chloroplast genome sequence of Oryza sativa temperate Japonica. Mitochondrial DNA Part B, 6(3), 927–928. https://doi.org/10.1080/23802359.2021.1888331
Fu, Y. B. (2021). Characterizing chloroplast genomes and inferring maternal divergence of the Triticum–Aegilops complex. Scientific Reports, 11(15363), 1–15. https://doi.org/10.1038/s41598-021-94649-9
Gholave, A. R., Pawar, K. D., Yadav, S. R., Bapat, V. A., & Jadhav, J. P. (2017). Reconstruction of molecular phylogeny of closely related Amorphophallus species of India using plastid DNA marker and fingerprinting approaches. Physiology and Molecular Biology of Plants, 23(1), 155–167. https://doi.org/10.1007/s12298-016-0400-0
Hall, T. (2011). BioEdit: An important software for molecular biology. GERF Bulletin of Biosciences, 2(1), 60–61.
Hanum, L., Windusari, Y., Setiawan, A., Muharni, Adriansyah, F., & Mubarok, A. A. (2018). Comparison of CTAB method and wizard genomic DNA purification system kit from promega on DNA isolation of local varieties of rice of South Sumatera. Science & Technology Indonesia, 3, 26–29. https://doi.org/10.26554/sti.2018.3.1.26-29
Ho, V. T., Nguyen, T. H., Nguyen, L. B. N., Nguyen, T. T. N., & Nguyen, M. P. (2023). Comparison of rbcL and trnH-psbA DNA barcodes in diverse Camellia species collection in Vietnam. Sabrao Journal of Breeding and Genetics, 55(3), 877–885. https://doi.org/10.54910/sabrao2023.55.3.22
Imanbayeva, A., Duisenova, N., Orazov, A., Sagyndykova, M., Belozerov, I., & Tuyakova, A. (2024). Study of the floristic, morphological, and genetic (atpF–atpH, Internal Transcribed Spacer (ITS), matK, psbK–psbI, rbcL, and trnH–psbA) differences in Crataegus ambigua populations in Mangistau (Kazakhstan). Plants, 13(12), 1–22. https://doi.org/10.3390/plants13121591
Kardos, M., Armstrong, E. E., Fitzpatrick, S. W., Hauser, S., Hedrick, P. W., Miller, J. M., Tallmon, D. A., & Chris Funk, W. (2021). The crucial role of genome-wide genetic variation in conservation. Proceedings of the National Academy of Sciences of the United States of America, 118(48), 1–10. https://doi.org/10.1073/pnas.2104642118
Kassem, M. A. (2025). Comparative analysis of chloroplast genomes across 20 plant species reveals evolutionary patterns in gene content, codon usage, and genome structure. International Journal of Plant Biology, 16(3), 1–16. https://doi.org/10.3390/ijpb16030105
Kumar, S., Stecher, G., Suleski, M., Sanderford, M., Sharma, S., & Tamura, K. (2024). MEGA12: Molecular evolutionary genetic analysis version 12 for adaptive and green computing. Molecular Biology and Evolution, 41(12), 1–9. https://doi.org/10.1093/molbev/msae263
Lee, J. H., Kim, K. J., Kim, B. Y., & Kim, Y. D. (2022). Molecular systematics of Poaceae based on eight chloroplast markers, emphasizing the phylogenetic positions of Korean taxa. Korean Journal of Plant Taxonomy, 52(3), 127–143. https://doi.org/10.11110/kjpt.2022.52.3.127
Leigh, J. W., & Bryant, D. (2015). POPART: Full-feature software for haplotype network construction. Methods in Ecology and Evolution, 6(9), 1110–1116. https://doi.org/10.1111/2041-210X.12410
Li, R., Wang, B., Xiao, S., Chen, L., Yin, F., Li, J., Jiang, C., Zhang, D., Zhong, Q., Zhang, Y., Xing, J., Cheng, Z., & Liu, L. (2025). Characterization of the complete chloroplast genome and comparative analysis of the phylogeny and codon usage bias of three Yunnan wild rice species. Frontiers in Plant Science, 16(1555104), 1–17. https://doi.org/10.3389/fpls.2025.1555104
Mursyidin, D. H., Nazari, Y. A., Badruzsaufari, & Masmitra, M. R. D. (2021). DNA barcoding of the tidal swamp rice (Oryza sativa) landraces from South Kalimantan, indonesia. Biodiversitas, 22(4), 1593–1599. https://doi.org/10.13057/biodiv/d220401
Pagès, H., Aboyoun, P., Gentleman, R., & Debroy, S. (2022). Biostrings: Efficient manipulation of biological strings. R Package, 1–32.
Paiman, Ardiyanta, Ansar, M., Effendy, I., & Sumbodo, B. T. (2020). Rice cultivation of superior variety in swamps to increase food security in Indonesia. Reviews in Agricultural Science, 8, 300–309. https://doi.org/10.7831/ras.8.0_300
Paradis, E. (2023). Plotting Haplotype Networks with pegas (pp. 1–16).
Paradis, E., Claude, J., & Strimmer, K. (2004). APE: Analyses of phylogenetics and evolution in R language. Bioinformatics, 20(2), 289–290. https://doi.org/10.1093/bioinformatics/btg412
Pattengale, N. D., Alipour, M., Bininda-emonds, O. R. P., Moret, B. M. E., & Stamatakis, A. (2010). How many bootstrap replicates are necessary ?. Journal of Computional Biology, 17(3), 337–354. https://doi.org/10.1089/cmb.2009.0179
Ridwan, M., & Suranto, S. (2025). The impact of the global food crisis and Indonesia’s strategies for achieving food security. International Journal of Environmental, Sustainability, and Social Science, 6(6), 1435–1447. https://doi.org/10.38142/ijesss.v6i6.1695
Rozas, J., Ferrer-Mata, A., Sanchez-DelBarrio, J. C., Guirao-Rico, S., Librado, P., Ramos-Onsins, S. E., & Sanchez-Gracia, A. (2017). DnaSP 6: DNA sequence polymorphism analysis of large data sets. Molecular Biology and Evolution, 34(12), 3299–3302. https://doi.org/10.1093/molbev/msx248
Sabar, M., Mustafa, S. E., Ijaz, M., Khan, R. A. R., Shahzadi, F., Saher, H., Javed, H. M., Zafar, S. A., Saleem, M. U., Siddique, S., & Sabir, A. M. (2024). Rice breeding for yield improvement through traditional and modern genetic tools. European Journal of Ecology, Biology and Agriculture, 1(1), 14–19. https://doi.org/10.59324/ejeba.2024.1(1).02
Saleem, A., Anwar, S., Nawaz, T., Fahad, S., Saud, S., Ur Rahman, T., Khan, M. N. R., & Nawaz, T. (2025). Securing a sustainable future: the climate change threat to agriculture, food security, and sustainable development goals. Journal of Umm Al-Qura University for Applied Sciences, 11(3), 595–611. https://doi.org/10.1007/s43994-024-00177-3
Samarina, L. S., Koninskaya, N. G., Shkhalakhova, R. M., Simonyan, T. A., & Kuzmina, D. O. (2025). DNA-barcoding for cultivar identification and intraspecific diversity analysis of agricultural crops. International Journal of Molecular Sciences, 26(14), 1–22. https://doi.org/10.3390/ijms26146808
Sánchez, M. L., Studer, B., & Kölliker, R. (2020). DNA barcode trnH ‑ psbA is a promising candidate for efficient identification of forage legumes and grasses. BMC Research Notes, 13(35), 1–6. https://doi.org/10.1186/s13104-020-4897-5
Sayed, H. A., Mostafa, S., Haggag, I. M., & Hassan, N. A. (2023). DNA barcoding of Prunus species collection conserved in the national gene bank of Egypt. Molecular Biotechnology, 65(3), 410–418. https://doi.org/10.1007/s12033-022-00530-z
Sha, L. N., Fan, X., Wang, X. L., Dong, Z. Z., Zeng, J., Zhang, H. Q., Kang, H. Y., Wang, Y., Liao, J. Q., & Zhou, Y. H. (2017). Genome origin, historical hybridization and genetic differentiation in Anthosachne australasica (Triticeae; Poaceae), inferred from chloroplast rbcL, trnH-psbA and nuclear Acc1 gene sequences. Annals of Botany, 119(1), 95–107. https://doi.org/10.1093/aob/mcw222
Smith, S., Nickson, T. E., & Challender, M. (2021). Germplasm exchange is critical to conservation of biodiversity and global food security. Agronomy Journal, 113(4), 2969–2979. https://doi.org/10.1002/agj2.20761
Song, Y., Chen, Y., Lv, J., Xu, J., Zhu, S., Li, M. F., & Chen, N. (2017). Development of chloroplast genomic resources for Oryza species discrimination. Frontiers in Plant Science, 8(1854), 1–10. https://doi.org/10.3389/fpls.2017.01854
Terryana, R. T., Lestari, P., Arsana, I. G. K. D., Nugroho, K., Mejaya, I. M. J., Sasmita, P., Sastro, Y., Mulya, K., Utami, D. W., & Mastur. (2022). Diversity and population structure of local rice varieties from Indonesia revealed by SSR markers. HAYATI Journal of Biosciences, 29(6), 749–761. https://doi.org/10.4308/hjb.29.6.749-761
Thomson, M. J., Polato, N. R., Prasetiyono, J., Trijatmiko, K. R., Silitonga, T. S., & McCouch, S. R. (2009). Genetic diversity of isolated populations of Indonesian landraces of rice (Oryza sativa L.) collected in East Kalimantan on the island of Borneo. Rice, 2(1), 80–92. https://doi.org/10.1007/s12284-009-9023-1
Tong, W., He, Q., Wang, X. Q., Yoon, M. Y., Ra, W. H., Li, F., Yu, J., Oo, W. H., Min, S. K., Choi, B. W., Heo, E. B., Yun, B. K., Kim, K. W., Kim, T. S., Lee, C. Y., & Park, Y. J. (2015). A chloroplast variation map generated using whole genome re-sequencing of Korean landrace rice reveals phylogenetic relationships among Oryza sativa subspecies. Biological Journal of the Linnean Society, 115(4), 940–952. https://doi.org/10.1111/bij.12564
Urtgam, S., Junmatong, C., Rattanachak, N., Wannathes, N., Sujipuli, K., Wattanachaiyingcharoen, W., Jongjitwimol, J., & Jongjitvimol, T. (2026). Genetic diversity of landrace rice varieties in lower northern Thailand using chloroplast DNA markers. International Journal of Agronomy, 2026(1), 1–19. https://doi.org/10.1155/ioa/3034611
Wickham, H. (2017). ggplot2 – Elegant graphics for data analysis (2nd edition). Journal of Statistical Software, 77, 3–5. https://doi.org/10.18637/jss.v077.b02
Zhang, J., Ning, Y., Li, J., Deng, Y., Wang, L. S., Mao, S., & Zhao, B. (2024). Comparative chloroplast genome analysis of Ardisia (Myrsinoideae, Primulaceae) in China and implications for phylogenetic relationships and adaptive evolution. BMC Plant Biology, 24(1198), 1–15. https://doi.org/10.1186/s12870-024-05892-x
Zhang, W., Sun, Y., Liu, J., Xu, C., Zou, X., Chen, X., Liu, Y., Wu, P., Yang, X., & Zhou, S. (2021). DNA barcoding of Oryza: conventional, specific, and super barcodes. Plant Molecular Biology, 105(3), 215–228. https://doi.org/10.1007/s11103-020-01054-3
Refbacks
- There are currently no refbacks.

























