Abstract
Leonurus cardiaca is well known for its medicinal importance. In this investigation, genotypic characterization of this species from six eco-geographical regions of Iran was evaluated by four molecular techniques (AFLP, RAPD, ISSR and IRAP). A total of 899 polymorphic fragments were detected by used molecular markers (AFLP = 356, RAPD = 325, ISSR = 113 and IRAP = 105) with an overall average polymorphism of 81.24 %. Genetic variation calculated using Shannon’s Information index (I) and Nei’s gene diversity index (H) showed high genetic diversity in studied germplasm. Also, analysis of molecular variance showed high genetic variation among (55 %) and within populations (45 %). UPGMA dendrogram constructed from combined data of molecular markers distinguished studied populations in accordance with the results obtained by each marker which all individuals were clearly differentiated into two major clusters. The correlation coefficients were statistically significant for all marker systems with the highest correlation between similarity matrixes of RAPD and ISSR markers (r = 0.82). The present results have an important implication for L. cardiaca germplasm characterization, improvement, and conservation. Furthermore, the characterized individuals exhibited a great deal of molecular variation and they seem to have a rich gene pool for breeding programs.
Similar content being viewed by others
References
Archak S, Gaikwad AB, Gautam D, Rao EVVB, Swamy KRM, Karihaloo JL (2003) Comparative assessment of DNA fingerprinting techniques (RAPD, ISSR and AFLP) for genetic analysis of cashew (Anacardium occidentale L.) accessions of India. Genome 46:362–369
Belaj A, Satovic Z, Cipriani G, Baldoni L, Testolin R, Rallo L, Trujillo I (2003) Comparative study of the discriminating capacity of RAPD, AFLP and SSR markers and of their effectiveness in establishing genetic relationships in olive. Theor Appl Genet 107:736–744
Biswas MK, Xu Q, Deng XX (2010) Utility of RAPD, ISSR, IRAP and REMAP markers for the genetic analysis of Citrus spp. Sci Hortic 124:254–261
Budak H, Shearman RC, Parmaksiz I, Dweikat I (2004) Comparative analysis of seeded and vegetative biotype buffalo grasses based on phylogenetic relationship using ISSRs, SSRs, RAPDs, and SRAPs. Theor Appl Genet 109:280–288
Chen L, Zhao L, Bai Y, Hu R, Si J (2009) Genetic relationship analysis of different provenances of Leonurus japonicus by ISSR marker. Zhongguo Zhong Yao Za Zhi. 34(11):5–1343
Coart E, Vekemans X, Smulders MJM, Wagner I, Van Huylenbroeck J, Van Bockstaele E, Roldan-Ruiz I (2003) Genetic variation in the endangered wild apple (Malus sylvestris (L.) Mill.) in Belgium as revealed by AFLP and microsatellite markers. Consequences for conservation. Mol Ecol 12:845–857
Du XY, Zhang QL, Luo ZR (2009) Development of retrotransposon primers and their utilization for germplasm identification in Diospyros spp. (Ebenaceae). Tree Genet Genomes 5:235–245
Finkeldey R, Hattemer HH (2007) Tropical forest genetics. Springer, Berlin
Hamrick JL (1982) Plant population genetics and evolution. Am J Bot 69:1685–1693
Heywood JS (1991) Spatial analysis of genetic variation in plant populations. Annu Rev Ecol Syst 22:335–355
Hogbin PM, Peakall R (1999) Evaluation of the contribution of genetic research to the management of the endangered plant Zieria prostrata. Conserv Biol 13:514–522
Horvath A, Christmann H, Laigret F (2008) Genetic diversity and relationships among Prunus cerasifera (cherry plum) clones. Botany 86:1311–1318
Kalendar R, Grob T, Regina M, Suoniemi A, Schulman A (1999) IRAP and REMAP: two new retrotransposon-based DNA fingerprinting techniques. Theor Appl Genet 98:704–711
Khadivi-Khub A, Zamani Z, Bouzari N (2008) Evaluation of genetic diversity in some Iranian and foreign sweet cherry cultivars by using RAPD molecular markers and morphological traits. Hortic Environ Biotechnol 49:188–196
Landry BS, Li RQ, Cheung WY, Granger RL (1994) Phylogeny analysis of 25 apple rootstocks using RAPD markers and tactical gene tagging. Theor Appl Genet 89:847–852
Mantel N (1967) The detection of disease clustering and generalized regression approach. Cancer Res 27:209–220
Meyer W, Mitchell TG, Freedman EZ, Vilgays R (1993) Hybridization probes for conventional DNA fingerprinting used as single primers in the polymerase chain reaction to distinguish strains of Cryptococcus neoformans. J Clin Microbiol 31:2274–2280
Milbourne D, Meyer R, Bradshaw JE, Baird E, Bonar N, Provan J, Powell W, Waugh R (1997) Comparison of PCR-based marker systems for the analysis of genetic relationships in cultivated potato. Mol Breed 3:127–136
Milkowska-Leyck K, Filipek B, Strzelecka H, Warsaw P (2002) Pharmacological effects of lavandulifolioside from Leonurus cardiaca. J Ethnopharmacol 80(1):85–90
Mills S, Bone K (2000) Principles and practice of phytotherapy, vol 204. Churchill Livingstone, London, pp 233–245
Morteza-Semnani K, Saeedi M, Akbarzadeh M (2008) The essential oil composition of Leonurus cardiaca L. J Essent Oil Res 20:107–109
Mozafarian V (1996) Lexicon of Iranian plant names. Publishing contemporary vocabulary. Tehran Press, Tehran, pp 121–132 (In Farsi)
Naik PK, Alam MF, Singh H, Goyal V, Parida S, Kalia S, Mohapatra T (2010) Assessment of genetic diversity through RAPD, ISSR and AFLP markers in Podophyllum hexandrum: a medicinal herb from the Northwestern Himalayan region. Physiol Mol Biol Plants 16(2):135–148
Nei M (1973) Analysis of gene diversity in subdivided populations. Proc Natl Acad Sci USA 70:3321–3323
Nybom H (2004) Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants. Mol Ecol 13:1143–1155
Peakall R, Smouse PE (2006) GenAIEx 6: genetic analysis in Excel. Population genetic software for teaching and research. Mol Ecol Notes 6:288–295
Pejic I, Ajmone-Marsan P, Morgante M, Kozumplick V, Castiglioni P, Taramino G, Motto M (1998) Comparative analysis of genetic similarity among maize inbred lines detected by RFLPs, RAPDs, SSRs, and AFLPs. Theor Appl Genet 97:1248–1255
Pirttila AM, Hirsikorpi M, Kamarainen T, Jaakola L, Hohtola A (2001) DNA isolation methods for medicinal and aromatic plants. Plant Mol Biol Rep 19:273a–273f
Popescu ML, Dinu M, Toth O (2009) Contibutions to the pharmacognostical and phytobiological study on Leonurus cardiaca (Lamiaceae). Farmacial 57:4
Powell W, Morgante M, Andre C, Hanafey M, Vogel J, Tingey S, Rafalski A (1996) The comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis. Mol Breed 2:225–238
Prevost A, Wilkinson MJ (1999) A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor Appl Genet 98:107–112
Rahimmalek M, Ebrahim B, Arzani A, Etemadi N (2009) Assessment of genetic diversity among and within Achillea species using amplified fragment length polymorphism (AFLP). Biochem Syst Ecol 37:354–361
Reddy MP, Sarla N, Siddiq EA (2002) Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica 128:9–17
Rohlf FJ (2000) NTSYS-pc numerical taxonomy and multivariate analysis system. Version 2. 1. Exeter Software, Setauket, NY
Romesburg HC (1990) Cluster analysis for researchers. Krieger Publishing, Malabar, FL
Russell JR, Fuller JD, Macaulay M, Hatz BG, Jahoor A, Powell W, Waugh R (1997) Direct comparison of levels of genetic variation among barley accessions detected by RFLPs, AFLPs, SSRs and RAPDs. Theor Appl Genet 95:714–722
Russi L, Moretti C, Raggi L, Albertini E, Falistocco E (2009) Identifying commercially relevant Echinacea species by AFLP molecular markers. NRC Res Press 52:912–918
Sehgal D, Rajpal VR, Raina SN, Sasanuma T, Sasakuma T (2009) Assaying polymorphism at DNA level for genetic diversity diagnostics of the safflower (Carthamus tinctorius L.) world germplasm resources. Genetica 135:457–470
Shahi-Gharahlar A, Zamani Z, Fatahi R, Bouzari N (2011) Estimation of genetic diversity in some Iranian wild Prunus subgenus Cerasus accessions using inter-simple sequence repeat (ISSR) markers. Biochem Syst Ecol 39:826–833
Shimada T, Hayama H, Haji T, Yamaguchi M, Yoshida M (1999) Genetic diversity of plums characterized by random amplified polymorphic DNA (RAPD) analysis. Euphytica 109:143–147
Singh A, Chaudhury A, Srivastava PS, Lakshmikumaran M (2002) Comparison of AFLP and SAMPL markers for assessment of intra-population genetic variation in Azadirachta indica. Asian J Plant Sci 162:17–25
Smulders MJM, van der Schoot J, Geerts RHEM, Antonisse-de Jong AG, Korevaar H, van der Werf A, Vosman B (2000) Genetic diversity and the reintroduction of meadow species. Plant Biol 2:447–454
Sneller CH, Miles JW, Hoyt JM (1997) Agronomic performance of soybean plant introductions and their genetic similarity to elite lines. Crop Sci 37:1595–1600
Song Z, Li X, Wang H, Wang J (2010) Genetic diversity and population structure of Salvia miltiorrhiza Bge in China revealed by ISSR and SRAP. Genetica 138:241–249
Soorni A, Nazeri V, Fatahi R, Khadivi-Khub A (2013) Multivariate analysis of motherwort germplasm in Iran using morphological variables and essential oil content. Plant Syst Evol. doi:10.1007/s00606-013-0932-5
Techen N, Crockett S, Khan I, Scheffler B (2004) Authentication of medicinal plants using molecular biology techniques to compliment conventional methods. Curr Med Chem 11:1391–1401
Vos P, Hogers R, Bleeker M, Lee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M, Zabeau M (1995) AFLP: A new technique for finger printing. Nucleic Acids Res 23:4407–4414
Wen XP, Pang XM, Deng XX (2004) Characterization of genetic relationships of Rosa roxburghii Tratt and its relatives using morphological traits, RAPD and AFLP markers. J Hort Sci Biotechnol 79:189–196
Williams JGK, Kubelik AR, Livak KJ, Rafalsk JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acid Res 18:6531–6535
Yeh FC, Yang RC, Boyle T (1999) POPGENE. Microsoft Windows-based Freeware for Population Genetic Analysis. Release 1.31. University of Alberta, Edmonton
Yu Q, Shen X, Shen Y, Chen J, Shi C, Wang Z (2009) AFLP Analysis of genetic diversity of Leonurus japonicus germplasm resources. Zhongcaoyao 40:1296–1299
Yuan L, Zhang Q, Guo D, Luo Z (2012) Genetic differences among ‘Luotian-tianshi’ (Diospyros kaki Thunb.) genotypes native to China revealed by ISSR and IRAP markers. Sci Hortic 137:75–80
Zabeau MM (1995) AFLP: a new concept for DNA fingerprinting. Nucleic Acids Res 23:4407–4414
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Khadivi-Khub, A., Soorni, A. Comprehensive genetic discrimination of Leonurus cardiaca populations by AFLP, ISSR, RAPD and IRAP molecular markers. Mol Biol Rep 41, 4007–4016 (2014). https://doi.org/10.1007/s11033-014-3269-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-014-3269-4