Short Communication: Analysis of simple sequence repeats variations for the assessment of genotypic diversity in Centella asiatica (L.) Urb.

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ANSHARY MARUZY
RATNA SUSANDARINI

Abstract

Abstract. Maruzy A, Susandarini R. 2024. Short Communication: Analysis of simple sequence repeats variations for the assessment of genotypic diversity in Centella asiatica (L.) Urb. Biodiversitas 25: 734-740. Centella asiatica (L.) Urb. is a perennial plant widely known as herbal ingredient in traditional and modern medicine, as well as in the cosmetics industry. This herbaceous species grows in various types of habitat, and is known for its morphological variations among populations, indicating the existence of genotypic variation. In this study, the assessment of genotypic diversity using simple sequence repeats (SSR) was carried out on C. asiatica accessions from 32 populations collected from eight mountains in Central Java. Representative individuals from four populations grow at different altitudes were sampled in each mountain. Amplification of genomic DNA using the SSR primer mCaCIR002 produced a single amplicon of 180 bp. The results of SSR analysis showed that there is high genotypic diversity in C. asiatica which is characterized by variations in the number of microsatellite repeats of (CT)10 to (CT)19. Variations in the number of repeats are mainly caused by insertions and deletions in DNA sequences at the positions 102-145. Haplotype analysis also showed high genetic diversity within and between populations of C. asiatica, indicated by the haplotype diversity of 1.00 in all populations. The haplotype network analysis indicated that there was no gene flow between the four populations at each mountain where C. asiatica populations grow. The results of this study provide evidence for the role of SSR as a suitable molecular marker for studying genotypic diversity at the intraspecific level, and confirming the morphological variability in this species.

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References
Alqahtani A, Cho JL, Wong KH, Li KM, Razmovski-Naumovski V, Li GQ. 2017. Differentiation of three Centella species in Australia as inferred from morphological characteristics, ISSR molecular fingerprinting and phytochemical composition. Front Plant Sci 8, article 1980: 1-10. https://doi.org/10.3389/fpls.2017.01980
Amiteye S. 2021. Basic concepts and methodologies of DNA marker systems in plant molecular breeding. Heliyon 7 (10) article e08093: 1-20. https://doi.org/10.1016/j.heliyon.2021.e08093
Chachai N, Pensuriya B, Pinsuntiae T, Pratubkong P, Mungngam J, Nitmee P, Kaewsri P, Wongsatchanan S, Jindajia R, Triboun P, Sreesaeng J. 2021. Variability of morphological and agronomical characteristics of Centella asiatica in Thailand. Trends Sci 18 (23): 502-511. DOI: 10.48048/tis.2021.502
Gray NE, Harris CJ, Quinn JF, Soumyanath A. 2016. Centella asiatica modulates antioxidant and mitochondrial pathways and improves cognitive function in mice. J Ethnopharmacol 180: 78–86. https://doi.org/10.1016/j.jep.2016.01.013
Garrido-Cardenas JA, Mesa-Valle C, Manzano-Agugliaro F. 2018. Trends in plant research using molecular markers. Planta 247 (3): 543–557. https://doi.org/10.1007/s00425-017-2829-y
Hapsari L, Lestari DA, Probojati RT. 2020. Haplotype network analysis of wild banana relatives Ensete glaucum, Musa acuminata and Musa balbisiana based on cpDNA rbcL sequences in ex-situ collection. Indian J Genet 80 (3): 1-7.
Jiao J, Jia X, Liu P, Zhang Q, Liu F, Ma C, Xi P, Liang Z. 2018. Species identification of polygonati rhizoma in China by both morphological and molecular marker. C R Biologies 341 (2): 102–110. https://doi.org/10.1016/j.crvi.2017.10.004
Kongjaimun A, Takahashi Y, Yoshioka Y, Tomooka N, Mongkol R, Somta P. 2023. Molecular analysis of genetic diversity and structure of the lablab (Lablab purpureus (L.) Sweet) gene pool reveals two independent routes of domestication. Plants 12 (57): 1-18. https://doi.org/10.3390/plants12010057
Lee SJ, Shin YE, Kim YH, Lee SW. 2017. Molecular markers based on chloroplast and nuclear ribosomal DNA regions which distinguish Korean-specific ecotypes of the medicinal plant Cudrania tricuspidata Bureau. J Plant Biotechnol 44 (3): 235–242. https://doi.org/10.5010/JPB.2017.44.3.235
Li X, Wang J, Qiu Y, Wang H, Wang P, Zhang X, Li X. 2021. SSR-sequencing reveals the inter-and intraspecific genetic variation and phylogenetic relationships among an extensive collection of radish (Raphanus) germplasm resources. Biology 10 (12), article 1250: 1-17. https://doi.org/10.3390/biology10121250
Mariska E, Sitorus TD, Rachman JA. 2015. Effect of Centella asiatica leaves on gastric ulcer in rats. Althea Med J 2 (1): 114–118. DOI: 10.15850/amj.v2n1.444
Mathavaraj S, Sabu KK. 2021. Genetic diversity and structure revealed by genomic microsatellite markers in Centella asiatica (L.) Urb., a plant with medicinal potential. Mol Biol Rep 48 (1): 7387–7396. DOI: 10.25081/cb.2021.v12.6971
Mehrotra S, Goyal V. 2014. Repetitive sequences in plant nuclear DNA: Types, distribution, evolution and function. Genom Proteom Bioinform 12 (4): 164–171. https://doi.org/10.1016/j.gpb.2014.07.003
Milner ML, McIntosh EJ, Crisp MD, Weston PH, Rossetto M. 2013. Microsatellite variation for phylogenetic, phylogeographic and population-genetic studies in Lomatia (Proteaceae). Aust Syst Bot 26 (3): 186–195. DOI: 10.1071/SB13002.
Mubarok H, Handayani NSN, Maryanto I, Arisuryanti T. 2023. Phylogenetic and genetic variation analysis of lesser short-nosed fruit bat Cynopterus brachyotis (Müller 1838) on Java Island, Indonesia, inferred from mitochondrial D-loop. J Genet Engineer Biotechnol 21 (1): 1–10. DOI: 10.1186/s43141-022-00460-y.
Nav SN, Ebrahimi SN, Sonboli A, Mirjalili MH. 2021. Variability, association and path analysis of centellosides and agro-morphological characteristics in Iranian Centella asiatica (L.) Urban ecotypes. South African J Bot 139: 254–266. DOI: 10.1016/j.sajb.2021.03.006.
Noni? M, Šija?i?-Nikoli? M. 2021. Genetic Diversity: Sources, Threats, and Conservation. In: Leal Filho W, Azul AM, Brandli L, Lange Salvia A, Wall T. (eds) Life on Land. Encyclopedia of the UN Sustainable Development Goals. Springer, Cham. https://doi.org/10.1007/978-3-319-95981-8_53
Paradis E. 2018. Analysis of haplotype networks: The randomized minimum spanning tree method. Methods Ecol Evol 9 (5): 1308–1317. DOI: 10.1111/2041-210X.12969.
Popoola JO, Bello OA, Olugbuyiro JA, Obembe OO. 2017. Simple sequence repeats (SSR) analysis of genetic intraspecific relationships of Moringa oleifera populations from Nigeria. Sci Int (Lahore) 29 (3): 645-657.
Prasad A, Dhawan SS, Mathur AK, Prakash O, Gupta MM, Verma RK, Lal RK, Mathur A. 2014. Morphological, chemical and molecular characterization of Centella asiatica germplasms for commercial cultivation in the Indo-Gangetic Plains. Nat Prod Commun 9 (6): 779-784.
Putman AI, Carbone I. 2014. Challenges in analysis and interpretation of microsatellite data for population genetic studies. Ecol Evol 4 (22): 4399–4428. DOI: 10.1002/ece3.1305.
Rohini MR, Sane A, Chaudhary R, Himabindu K. 2019. Molecular characterization and DNA fingerprinting of Centella asiatica using SSR markers. Int J Chem Stud 7 (1): 705–710.
Sahu J, Das Talukdar A, Devi K, Choudhury MD, Barooah M, Modi MK, Sen P. 2015. E-Microsatellite markers for Centella asiatica (Gotu Kola) genome: Validation and cross-transferability in Apiaceae family for plant omics research and development. Omics 19 (1): 52–65. DOI: 10.1089/omi.2014.0113.
Sakthipriya M, Vishnu SS, Sujith S, Kumar PR, Sabu KK. 2018. Analysis of genetic diversity of Centella asiatica using SSR markers. Int J Appl Sci Biotechnol 6 (2): 103–109. DOI: 10.3126/ijasbt.v6i2.19583
Šarhanová P, Pfanzelt S, Brandt R, Himmelbach A, Blattner FR. 2018. SSR-seq: Genotyping of microsatellites using next?generation sequencing reveals higher level of polymorphism as compared to traditional fragment size scoring. Ecol Evol. 2018;8:10817–10833. https://doi.org/10.1002/ece3.4533
Storz JF. 2002. Contrasting patterns of divergence in quantitative traits and neutral DNA markers: Analysis of clinal variation. Mol Ecol 11 (12): 2537–2551. DOI: 10.1046/j.1365-294X.2002.01636.x.
Susandarini R, Subandiyah S, Daryono BS, Rugayah. 2020. Microsatellite polymorphism for molecular characterization of pomelo (Citrus maxima) accessions from Indonesia. Biodiversitas 21 (6): 2390–2395. DOI: 10.13057/BIODIV/D210608
Teacher AGF, Griffiths DJ. 2011. HapStar: Automated haplotype network layout and visualization. Mol Ecol Resour 11 (1): 151–153. DOI: 10.1111/j.1755-0998.2010.02890.x.
Vieira MLC, Santini L, Diniz AL, Munhoz CF2016. Microsatellite markers: what they mean and why they are so useful. Genet Mol Biol 39 (3): 312-328. DOI: 10.1590/1678-4685-GMB-2016-0027.
Viruel J, Haguenauer A, Juin M, Mirleau F, Bouteiller D, Boudagher?Kharrat M, Ouahmane L, La Malfa S, Médail F, Sanguin H, Feliner, GN, Baume A. 2018. Advances in genotyping microsatellite markers through sequencing and consequences of scoring methods for Ceratonia siliqua (Leguminosae). Appl Plant Sci 6 (12), article e01201: 1-10. DOI: 10.1002/APS3.1201.
Wang LY, Li SS, Wang TY, He CY, Luo HM, Zhang JG, Zeng YF. 2021. Genomic SSR and EST-SSR markers for phylogenetic and pedigree reconstructions - A comparison in sea buckthorn. Plant Breed 140 (1): 167–183. DOI: 10.1111/PBR.12889
Wöhrmann T, Weising K, 2011. In silico mining for simple sequence repeat loci in a pineapple expressed sequence tag database and cross-species amplification of EST-SSR markers across Bromeliaceae. Theor Appl Genet 123 (4): 635–647. DOI: 10.1007/s00122-011-1613-9
Xu Y, Xing M, Song L, Yan J, Lu W and Zeng A. 2021. Genome-wide analysis of simple sequence repeats in cabbage (Brassica oleracea L.). Front Plant Sci 12: 726084. DOI: 10.3389/fpls.2021.726084.

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